{"text": "The epithelial-mesenchymal transition (EMT) is characterized by the loss of cell-cell adhesion and cell polarity in epithelial cells and the acquisition of motile and invasive properties. While essential for development, the EMT is one mechanism by which tumors can acquire the capability to undergo tissue invasion and metastasis. It is therefore important to identify novel therapies that can inhibit the EMT, but few assays for EMT inhibitors in high throughput screens (HTS) have developed. A change in fibroblast growth factor receptor 2 (FGFR2) splicing occurs during the EMT and using an innovative luciferase-based splicing reporter assay we previously carried out a genome-wide high throughput cDNA expression screen for regulators of this splicing switch. This screen identified the epithelial cell type specific splicing regulators ESRP1 and ESRP2 demonstrating the feasibility of cell-based splicing assays in high throughput, array-based screens. An extensive set of ESRP-regulated exons switch splicing during the EMT, indicating that global changes in alternative splicing occur during this process. A change in this splicing network is a thus a dynamic feature of the EMT and changes in splicing of ESRP-regulated targets can be used as a biomarker for the EMT. In this application we will develop more robust next generation splicing reporter assays using ESRP- regulated exons that undergo profound \"switch-like\" changes in splicing and configure them for HTS assays using the Molecular Libraries Production Centers Network (MLPCN). In Aim 1, we will adapt existing minigene reporters containing ESRP regulated exons and flanking intronic regulatory sequences for HTS in the context of our established luciferase-based reporter minigenes. The reporters will include exons whose inclusion is activated as well as those that undergo skipping during the EMT. Additional reporters will also be developed for use in counter-screens to prioritize HTS hits. In Aim 2, these screens will be configured for screening in 384 well format and pilot screens will be carried out using several small compound libraries as well as several previously described compounds that have been shown to function as general modulators of splicing. These compounds will be screened in mesenchymal cells for splicing changes indicative of the reverse process of mesenchymal to epithelial transition (MET) and in epithelial cells for inhibition or reversal of an inducible EMT. Successful completion of this pilot phase of this funding mechanism (PAR-10-182) in year one will enable us to submit these assays for the larger scale screening phase using the MLPCN library of compounds. Such screens hold great promise to yield novel small molecule regulators of splicing, including a subset that broadly promote epithelial-specific splicing pathways to inhibit or reverse the EMT and block cancer metastasis. Such compounds will potentially include those that affect signaling pathways or other upstream events that might potently activate broad transcriptional and post-transcriptional gene expression programs that inhibit the EMT. PUBLIC HEALTH RELEVANCE: The epithelial to mesenchymal transition (EMT) is the process by which cancer cells can escape from the primary site and metastasize to distant sites and is therefore a target for novel cancer therapies. We have identified regulators of alternative splicing that control an epithelial splicing network that is lost during the EMT, suggesting that a mesenchymal splicing program can promote the EMT and that these splicing changes serve as biomarkers for this process. The current application will use innovative splicing assays to carry out screens for novel compounds that inhibit this splicing transition and thereby identify lead compounds for drugs to prevent tumor metastasis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] The specific aim of this application is to help the UPR for repair, renovation, and modernization of its unique animal research resource (ARC) at the CS facility of the CPRC. The CPRC is an unrivaled national and international research resource for comparative studies in the biomedical and behavioral sciences that has received NCRR funding through a P40 award or the equivalent for the past three decades. The proposed renovations will improve the care of research animals and will provide much needed infrastructure support for the ongoing and planned biomedical and behavioral research program at CS. The free-ranging population of rhesus monkeys (Macaca mulatta) on the island of CS provides scientists with an unparalleled opportunity for performing biomedical and behavioral research projects utilizing primates residing in a semi-natural habitat. This population has the most extensive computerized demographic and genetics database available to researchers anywhere in the world. The population management program for CS has been designed to optimize the health and well-being of the monkeys, to enhance the value of the colony for research. In addition, the goal is to provide healthy animals to the scientific community for biomedical research, including AIDS and SlV pathogenesis and vaccine development as well as the support of National defense programs. Many of the existing structures on CS island were constructed in the 1930s and 1940s and are in dire need of renovation. This application seeks funds to renovate the dock to the island and funds to repair the roads, drinking water purification system and the three feeding and trapping corrals on CS. Renovations and modernization of these facilities are essential in order to prevent the continued deterioration of the infrastructure on CS, to maintain and enhance animal health and well being, to retain AAALAC accreditation, to comply with federal regulations and guidelines, and to continue to promote the research program and resources of the UPR. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "EXCEED THE SPACE PROVIDED. The proposed research is a continued study of mobile introns in bacteria. The mobile group I and group II introns are both self-splicing, but their RNA-splicing and DNA-mobility pathways are distinct. In each case, the intron typically transfers to an allelic intron-less site, in a homingprocess initiated by intron-encoded endonucleases. For group I intron homing, recombination events are strictly DNA-based whereas for group II intron homing, also termed retrohoming, RNA is involved at levels of both the intron template and the cleavage enzyme for mobility. Transposition to ectopic sites also occurs at low frequency, and is the process responsible for intron dissemination in nature. During the current funding period we showed the following: Homing of the group I td intron is regulated by the very endonuclease that promotes the process, by acting as an autorepressor. Also, retrohoming of group II introns is dependent on host polymerases in a way that suggests that intron movement is related to the cell's DNA damage response. Furthermore, group II intron retrotransposition occurs into single- or double-stranded DNAs in a host-dependent fashion. Additionally, transposed group II introns retain their ability to move, reflecting the high fidelity of the group II intron- encoded reverse transcriptase. Finally, in a study of a small regulatory RNA, DsrA, included in a non-intron- related specific ami, we showed not only that DsrA undergoes conformational changes to effect global regulation, but also that DsrA promotes acid tolerance in pathogenic bacteria, thereby enhancing virulence. In the upcoming funding period we will follow up on these findings using genetic studies in bacteria, as well as biochemical and structural approaches. Further, we will address exciting questions relating to intron evolution and the relationship of group II nitrons to spliceosomeal introns in yeast, all as described in the original application. Our overall goal of studying DNA- and RNA-based rearrangements of model introns remains unchanged. In addition to the innate mechanistic importance of these studies, the invasiveness of group II introns and their similarities to human retrotransposons and spliceosomal introns have great evolutionary, biotechnological and medical significance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A number of recent studies suggest that cognitive processes rely on spatial and temporal patterns of activity in extended neural networks. Optical imaging techniques can provide information on a microscopic level about the individual and collective behavior of cells involved in these processes. We propose to develop an advanced image probe and digital acquisition system designed for high performance functional neural imaging using intrinsic light scattering signals. The first goal of the project is to improve the image probe design for high sensitivity to small and rapid changes in light scattering. Two methods of reflectance mode illumination will be explored for fluorescence and polarized light measurements. The system will incorporate an electronically tunable filter to illuminate tissue with specific wavelengths for spectroscopic measurements, and an intensified detector for dynamic fluorescence measurements. The second goal is to implement hardware and software improvements to the data acquisition system. Application performance requirements and the need to integrate of a number of data modalities require that we develop custom data acquisition hardware; the proposed system will control CCD camera circuitry, and capture, process and archive 2000 frames per second, with 256 channels of concurrent electrophysiological data. We will integrate this hardware into inexpensive, multiple processor. Pentium computer systems, designed for distributing tasks across multiple processors. The third goal for the project is to apply the device to investigate the nature and origin of light scattering changes associated with neural activation. Our preliminary studies in the hippocampus and medulla have demonstrated several different optical changes associated with neural activation, including fast light scattering changes concurrent with neural swelling and electrical transmission, and slower changes in light absorbance associated with hemodynamic coupling to metabolic demand. We will examine the spectral nature of the optical signals, characterize the timing of at least four components that we have identified, and employ physical and physiological manipulations to identify and characterize underlying mechanisms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Complex developmental processes that define organ morphology and cellular differentiation appear to be governed by only several signal transduction pathways, suggesting the likelihood of extensive interactions among those pathways. However, the mechanisms that coordinate the activity of different signaling cascades involved in this process remain ill defined. In this proposal, I wish to tackle this important issu by focusing on the interplay between two prominent signals that control development; the transcription coactivator Yes- associated protein (YAP), the prime target of the Hippo kinase cascade, and Sonic hedgehog (Shh) signaling. The main goals of this proposal are to 1) dissect the function of YAP in spinal cord development, and 2) define how the Shh and Hippo signaling pathways act in concert to control development in the vertebrate spinal cord. The overall hypothesis of this proposal is that YAP serves as a novel downstream effector of Shh during the control of NPC proliferation and specification of ventral cell fates in the CNS. Specifically, I postulate that the YAP-dependent signaling communicates with the Shh signaling pathway at multiple levels, providing an effective means of coordinating these two transduction cascades during neural development. I will test this hypothesis using an ensemble of molecular methods, chick embryos and mutant mice. The specific aims of this proposal are as follows: 1) to determine the role of YAP in cell-type specification during neural tube development and 2) to define how the Shh and Hippo signaling pathways act in concert to control development in the vertebrate spinal cord. Besides defining the basic mechanisms by which YAP and Shh signaling coordinate key aspects of normal CNS development, this study should also uncover important principles applicable to other signaling networks that regulate organogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Directly taken from the application) Polycystic kidney disease (PKD) is characterized by proliferating epithelial cells, alterations in basement membrane composition, and aberrant localization of cell surface proteins. These cellular properties are consistent with a reversion of the renal epithelial cell to a more embryonic phenotype. Thus, genes regulating the differentiation and proliferation of the normal renal epithelium may be aberrantly affected in PKD and may directly contribute to the initiation and progression of the disease. This proposal will address the function of the transcription factor, Pax-2, in the development of normal and cystic renal epithelium. Pax-2 is required and sufficient for the conversion of the kidney mesenchyme to an epithelial phenotype by acting as a differentiation and proliferation stimulus. Pax-2 expression in the developing kidney is repressed as the normal epithelium matures, but is detected at higher levels in the cystic epithelium of experimental animals. The proposed experiments will determine the developmental expression pattern of Pax-2 in the cpk mouse, a well-characterized model for PKD. Furthermore, the cpk mutation will be crossed into the Krd mutant mouse that has one Pax-2 allele deleted. This genetic cross will test for modification of cpk by Pax-2 using gene dosage. If continued expression of Pax-2 in cpk mice is a proliferation stimulus, reducing the Pax-2 gene dosage may result in a slower progressing form of PKD. Additional experiments will address the regulation of the Pax-2 gene by identifying promoter elements required for tissue specific transcription. Finally, preliminary data indicates several cell adhesion molecules of the cadherin gene family may be regulated directly by Pax-2. The cadherin regulatory sequences will be cloned and tested for their ability to bind Pax-2 and mediate trans- activation. These experiments will elucidate basic molecular mechanisms of renal epithelial morphogenesis and may lead to a more complete understanding of the aberrant developmental events underlying PKD. Ultimately, understanding the regulation and function of transcription factors in renal diseases may lead to novel therapeutic interventions based on modulating gene expression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The object of the proposed investigation is to gain additional insight into the molecular events and regulatory implications in messenger-RNA metabolism in cultured mammalian cells. Human (HeLa S3) cells infected by and rat embryo cells transformed by adenovirus will be employed in these investigations. In the lytic infection the following aspects of viral RNA metabolism will be explored. Transcription of the genome will be examined by electron microscopy following dispersal of nuclear contents. Processing of apparent large molecular weight nuclear-precursor RNA will be examined by analysis of 5'-termini and by analysis of RNA synthesized in the presence of adenosine analogues. Fate of cytoplasmic viral RNA will be determined in enucleated cells (cytoplasts). Much of the proposed study will depend upon limited RNA structure analysis, especially if viral RNA sequences which remain in the nucleus can be isolated. In addition, information bearing on the mechanism of formation of virions and restriction of host-cell RNA synthesis should also be elucidated. The feasibility of extending these investigations to adeno 2-transformed rat embryo cells, which will depend on the incorporation of sufficient radioactivity, will be constantly evaluated. These investigations should not only bear on the cell biological implications of the biogenesis of messenger-RNA but also on the infection and transformation of cultured mammalian cells. BIBLIOGRAPHIC REFERENCES: Nuclear and cytoplasmic adenovirus RNA: Differences between 5'-termini of messenger and non-messenger transcripts. McGuire, P. M., Piatak, M. and L. D. Hodge. J. Mol. Biol. 101 379-396 (1976). Visualization of a unique adenovirus DNA-protein complex. O. L. Miller and L. D. Hodge. J. Cell Biol. 67 284a (1975).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Molecular Core Abstract: Dr. Harris will direct the Molecular Core and a staff of two full-time scientists. This essential Core will service the Program Project entitled \"Critical Interactions of APOBECSs: Molecular Approaches to Novel HiV Therapies\" by producing plasmid DNA constructs for APOBECS and Vif protein expression in E. coli and mammalian cells. We anticipate completing 10-20 constructs per week and providing a strong foundation for the program investigators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The consequences of alcohol abuse on the American public are profound, both in terms of individual well-being and impact on the family structure, as well as the enormous cost to society in terms of lost productivity and associated health care expenses. Despite increasing efforts, our understanding of the neurobiological mechanisms that underlie the effects of alcohol and the development of alcohol use disorders (AUD) remains incomplete. Epidemiological research has pointed to adolescence as a critical period in the development of alcohol disorders. The prefrontal cortex (PFC) is a brain region that is not yet mature at the onset of human adolescence and continues to develop during this period, during which some individuals may be highly susceptible to the effects of alcohol. The PFC mediates control over goal-directed behaviors and dysfunction of the PFC is thought to underlie compulsive drug-taking and relapse in substance abusers. Binge drinking is highly prevalent in adolescents, and episodes of high alcohol intake have been associated with decreased PFC activity and function (hypofrontality). Imaging studies suggest that hypofrontality persists in chronic alcohol abusers, and may therefore be a contributory factor in the development of AUDs and behavioral pathologies in adulthood. The underlying mechanisms of this PFC hypoactivity are unknown, and the development of robust animal models would therefore be useful in investigating the underlying changes in neuronal excitability. A better understanding of these changes would enable possible molecular and therapeutic interventions in order to prevent the development of alcoholism. One plausible mechanism for hypofrontality involves the depression of persistent activity, a mode of firing that can be observed in recordings from pyramidal neurons in the PFC of rodents. This type of activity is seen at more depolarized membrane potentials and is associated with performance in working memory tasks, and is dependent on the Ih current, which is mediated by a family of hyperpolarization-activated and cyclic nucleotide modulated (HCN) channels. We propose that chronic changes in persistent firing might result from prolonged alcohol exposure. To date there have been few detailed studies of excitability in the PFC after drinking and none in adolescent rodents. In three separate but integrated aims, we plan to test the overall hypothesis that the HCN1 channel that contributes to the Ih current in PFC PNs is important for the regulation of alcohol drinking, and specifically that (a) binge drinking of alcohol during adolescence inhibits persistent firing and excitability in the PFC via reduction of Ih and (b) reduction in HCN1 channel activity in layer 5 of PFC can mimic the effects of alcohol consumption during adolescence, while (c) activation or over-expression of HCN1 channels can restore persistent activity and normal levels of excitability in PFC of binge drinking adolescent animals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have constructed (and continue to develop) a laser-based facility for time-resolved fluorescence spectroscopy of biomolecules. This facility provides rapid collection and analysis of luminescence data related to macromolecular size, flexibility, folding and structural fluctuations. Our time-correlated laser fluorometer was used to study the folding and dynamics of several proteins. We focused most of our effort on DNA- binding proteins that control the transcription of DNA blueprints into the \"field copies\" (MRNA) used to build proteins in cells. Fluorescence was used to measure distances between proteins and the sections of DNA they control, to look at the wobbling of proteins in the complex, and to reveal internal changes in the bound protein. We studied \"oct-pou\", a bipartite factor able to bind two different classes of DNA control sites and accelerate transcription 100-fold. It was seen to \"read\" DNA by changing shape. We also began studies on HIV integrase and Heat Shock Factor. The former incorporates HIV DNA into human DNA, and the latter \"turns on\" a stress response by self-association. This year we also continued efforts to adapt our laser instruments to the imaging of tissues. In particular, we demonstrated an electronic laser scanning device that uses discoloration to find BB-sized objects almost an inch inside tissue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "G Proteins participate in the transduction of cellular signals by cycling between an active conformation (complexed with GTP) D and an inactive conformation (complexed with GDP). The ras proteins are guanine nucleotide binding proteins believed to participate in the transduction of proliferative signals, although their specific role is still unknown. Using a variety of experimental approaches, we showed that they are not regulatory elements of either of the two main signaling pathways in mammalian cells, namely the adenylate cyclase and phosphoinostide pathways; however, using Xenopus oocytes, we were able to show that they exert a regulatory role (even in the absence of protein synthesis) in cascades of phosphorylation associated with the induction of proliferation and cell division. The HIV nef gene product has also been proposed to act as a regulatory G protein based on reports attributing GTP binding and GTPase activity to these proteins. We expressed different cDNA clones of nef using the same bacterial and mammalian vectors used previously for ras. Unlike ras, nef from the different HIV isolates lacked GTP binding activity but showed autophosphorylation using either GTP or ATP as the phosphate donor. In addition, unlike ras, HIV nef did not exhibit oncogenic potential in focus-forming assays with NIH 3T3 cells, nor did it cause meiotic maturation of Xenopus oocytes. It therefore appears that the biological function of nef does not follow the pattern of G proteins; however, its phosphorylation, either by protein kinase C or through its autokinase activity, may be functionally significant in vivo.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "OVERALL DESCRIPTION: This application continues and amplifies the work already undergoing at the Center. The overall goal of the MU proposal is to foster interdisciplinary research that will enable the systematic evaluation of the safety and efficacy of botanicals. The PI's of the current proposal have already established collaborations, which has been shown a productive one. The proposed studies are aimed to understand the molecular mechanisms of phytochemicals and phytonutrients in human disease. The Center grant contains five projects, pilot studies, core units and career development. The concepts directing the research program include 1) the influence of phytonutrients in the progression of prostate carcinoma, this proposal examines the effect of ERKO mice will be studies in the TRAMP mouse model. 2) The effects phytoestrogens in the innate immunity of ER deficient mice, this proposal aims to determine whether phytoestrogens act to innate immunity through either the ER alpha or beta, or both, in SCID mice. 3) This project will determine whether treatment of CF cystic fibrosis with soy derived isoflavones, (primarily Genistein) acts through a non-ER-mediated pathway, on the mutated cystic fibrosis transmembrane conductance regulator (CFTR). 4) To identify and characterize botanicals with primary emphasis on those used in food supplements. 5) To examine alternative molecular mechanisms of phytoestrogens when considered as polyphenols in neurodegenerative disease. The research is facilitated by resources and several well established core facilities. The center Advisory Committee will provide guidance from distinguished representatives that will meet twice a year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (provided by investigator): Fluorogenic minor groove binder (MGB) probes containing an MGB-quencher at the 5'-end and a fluorophore at the 3'-end have been recently reported. These probes fluoresce on hybridization to the complementary targets. The 5'-MGB-quencher group prevents 5' - nuclease digestion by Taq polymerase during homogeneous amplification. The 5'-MGB-quencher-oligonucleotide-fluors can be used as probes in general nucleic acid and SNP detection assays. We propose to use a new software prediction program to develop MGB Eclipse probe assays to detect Clostridium botulinum, Bacillus anthracis and Yersinia pestis. The designed probes and primers will be optimized with use of three modified bases to improve probe performance of AT- and GC-rich sequences. In the Phase 1 effort, we will design and optimize the primers and probes and demonstrate function of the MGB Eclipse probe assays on a set of known target samples. The specificity of the probes will be evaluated against panels of closely related non-pathogenic organisms. In Phase II, we will expand the number of Category A organisms and improve throughput by multiplexing. In addition, commercial prototype MGB Eclipse(TM) probe assays will be developed for each of these organisms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lung cancer is the leading cause of cancer related mortality in both men and women and remains a major health issue. More than 162,000 individuals will die from lung cancer in the coming year, more than breast, prostate and colon cancer combined. The majority of lung cancer cases is attributable to tobacco smoking and in some cases other environmental risk factors. Although the relative risk of developing lung cancer declines dramatically in smokers who quit, former smokers remain at risk for the disease. Several recent studies show that greater than 50% of newly diagnosed lung cancers occur in former smokers. It is estimated that there are approximately equal numbers of smokers and former smokers in the United States. Since smoking cessation is a major public health initiative, former smokers will increasingly account for a higher percentage of lung cancer cases. Therefore, two high-risk population groups exist for lung cancer and improved disease management can be beneficial to both current and former smokers. Additionally, the prognosis for lung cancer patients is very poor. Resistance to chemotherapy used in lung cancer treatment remains a major problem and a better understanding of the mechanisms for resistance could lead to more effective therapies. The MAP3K8 gene is a mitogen activated protein (MAP) kinase kinase kinase expressed in a variety of cells and found to be oncogenic and constitutively activated when altered at the 3 end. However, mutation of the gene appears to be a rare event in humans, but altered MAP3K8 expression is associated with multiple tumor types. MAP3K8 possesses the unique characteristic of activating multiple cascades, including both proliferative and apoptotic signal transduction pathways such as the MEK-1 and SEK-1 pathways, respectively. In NIH3T3 transfection assays utilizing lung tumor DNA, our lab identified a 3 alteration of MAP3K8 similar to the previous reports. We first hypothesized that MAP3K8 might be a target for mutation since we were the first group to report an activating mutation in a primary human tumor. However, it has become clear that mutations are not a common event in tumorigenesis for this gene. Subsequently we showed varied levels of expression of the gene in lung tumor cell lines. This led us to investigate other downstream pathways that could explain the tumorigenic potential of MAP3K8. These included transcription factor array analysis and protein kinase array experiments. We were able to confirm other reports in the literature demonstrating upregulation of NF-kappaB and AP-1 as well as identify other important transcription factors not reported in the literature. These and other experiments, as well as published reports lead us to modify our hypothesis that increased expression of MAP3K8 occur in lung cancer and contribute to disease progression. We have recently shown that increased protein expression of MAP3K8 in lung tumor cell lines leads to changes in downstream signaling pathways and ultimately transcription of important genes in cell survival. To test the effects of MAP3K8 over expression on survival in the presence of a commonly used chemotherapeutic, paclitaxel, we stably transfected a normal tracheal epithelial cell line with MAP3K8. These data suggest MAP3K8 expression is altered in lung cancer cells lines and because of its role in the inflammatory response and cell survival, MAP3K8 over expression may be involved in tumor progression. Future experiments will demonstrate the importance of MAP3K8 in resistance to paclitaxel. We are also positioned to test the effect of MAP3K8 on tumorigenesis using the knockout mouse model and the skin two step carcinogenesis model.Lung cancer is the leading cause of cancer related mortality in both men and women and remains a major health issue. More than 162,000 individuals will die from lung cancer in the coming year, more than breast, prostate and colon cancer combined. The majority of lung cancer cases is attributable to tobacco smoking and in some cases other environmental risk factors. Although the relative risk of developing lung cancer declines dramatically in smokers who quit, former smokers remain at risk for the disease. Several recent studies show that greater than 50% of newly diagnosed lung cancers occur in former smokers. It is estimated that there are approximately equal numbers of smokers and former smokers in the United States. Since smoking cessation is a major public health initiative, former smokers will increasingly account for a higher percentage of lung cancer cases. Therefore, two high-risk population groups exist for lung cancer and improved disease management can be beneficial to both current and former smokers. Additionally, the prognosis for lung cancer patients is very poor. Resistance to chemotherapy used in lung cancer treatment remains a major problem and a better understanding of the mechanisms for resistance could lead to more effective therapies. The MAP3K8 gene is a mitogen activated protein (MAP) kinase kinase kinase expressed in a variety of cells and found to be oncogenic and constitutively activated when altered at the 3 end. However, mutation of the gene appears to be a rare event in humans, but altered MAP3K8 expression is associated with multiple tumor types. MAP3K8 possesses the unique characteristic of activating multiple cascades, including both proliferative and apoptotic signal transduction pathways such as the MEK-1 and SEK-1 pathways, respectively. In NIH3T3 transfection assays utilizing lung tumor DNA, our lab identified a 3 alteration of MAP3K8 similar to the previous reports. We first hypothesized that MAP3K8 might be a target for mutation since we were the first group to report an activating mutation in a primary human tumor. However, it has become clear that mutations are not a common event in tumorigenesis for this gene. Subsequently we showed varied levels of expression of the gene in lung tumor cell lines. This led us to investigate other downstream pathways that could explain the tumorigenic potential of MAP3K8. These included transcription factor array analysis and protein kinase array experiments. We were able to confirm other reports in the literature demonstrating upregulation of NF-kappaB and AP-1 as well as identify other important transcription factors not reported in the literature. These and other experiments, as well as published reports lead us to modify our hypothesis that increased expression of MAP3K8 occur in lung cancer and contribute to disease progression. We have recently shown that increased protein expression of MAP3K8 in lung tumor cell lines leads to changes in downstream signaling pathways and ultimately transcription of important genes in cell survival. To test the effects of MAP3K8 over expression on survival in the presence of a commonly used chemotherapeutic, paclitaxel, we stably transfected a normal tracheal epithelial cell line with MAP3K8. These data suggest MAP3K8 expression is altered in lung cancer cells lines and because of its role in the inflammatory response and cell survival, MAP3K8 over expression may be involved in tumor progression. Future experiments will demonstrate the importance of MAP3K8 in resistance to paclitaxel. We ar [summary truncated at 7800 characters]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have also completed the first characterization of the active site of NiSOD from Streptomyces seoulensis. This study reveals that Ni site is bound to thiolate donors, like other Ni redox enzymes but unlike any previously characterized SOD. Examination of oxidized and reduced samples reveal a coordination number change (5 -> 4) and a shift in the edge energy appropriate for a one-electron process. There is also evidence that the active site consists of two Ni centers, one of which is not redox active in analogy with the CuZnSOD, and therefore must be composed of two subunits in the tetrameric enzyme.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Given the ubiquitous nature of the peptide linkage in biological molecules, replacement of the amide bond with isosteres in potential drug candidates has been a continual goal of many laboratories. Successful replacements will provide improved stability, lipophilicity, and absorption. Many surrogates have been introduced already, yet the synthesis of many of these isosteres in a combinatorial way is difficult and requires several steps. Thus, the discovery of new peptide surrogates with easier syntheses is an important achievement that could open new opportunities for the study of amide-containing molecules and the development of inhibitors with novel physicochemical properties.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research is aimed at the elucidation of the biological function and genetic control of the polydisperse circular DNA complement of Bacillus megaterium. The specific objectives involve: (1) the characterization of the mode of production of these elements; (2) the determination of their physiological function in the cell, and (3) a test of the possibility that these elements reflect a novel regulatory mechanism for DNA replication and/or gene expression which may provide clues to an understanding of certain types of neoplasias in higher organisms. The research involves the use of electron microscopic analysis, including partial denaturation and heteroduplex techniques, and cleavage of specific circular DNA classes with site-specific restriction endonucleases in order to determine the relationships between these molecules. DNA-DNA hybridization will also be employed as will sedimentation velocity procedures for characterization of these elements. Specific inhibitors will be employed to determine the mode of production. Their biological properties will be assessed by attempts to isolate auxotrophic mutants as well as genetic variants defective in megacin production and other properties, and by attempts to transfer these elements to other bacteria by genetic transformation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In cancer, normal epigenetic silencing and cytosine methylation are disrupted. Dietary intake of methyl donors can directly affect epigenetic mechanisms through cytosine methylation. Our longterm goal is to understand how the risk of human disease, cancer in particular, is affected by epigenetics and diet. We use A10-vy (obese yellow) mice, a model that exhibits a highly variable phenotype of obesity, tumors, and type II diabetes;the expression of the syndrome is under epigenetic control. The epigenetic state of the A-vy allele can be inherited, indicating that the epigenetic marks determining the behavior of the allele are maintained in the germline. Supplementation of the maternal A-vy diet with methyl donors during gestation alters phenotypes in offspring. We hypothesize that continuous supplementation of the maternal diet with methyl donors will produce a cumulative increase in methylation of the A-vy allele, resulting in a multigenerational trend toward suppression of the obese yellow phenotype, and denser methylation of the allele. The changes may persist after supplementation is withdrawn. Our specific aims are: 1. Investigate the effects of continuous methyl donor supplementation on inheritance of the obese yellow phenotype in A-vy mice. Continuous feeding of methyl donors to A-vy mothers may produce changes in phenotype that increase with more generations. 2. Ask if the effects of methyl donor supplementation persist for generations when supplementation is withdrawn. Changes induced by methyl donors may be maintained in the germline, resulting in epigenetic \"memory\" that persists for one or more generations. 3. Investigate effects of methyl donor supplementation on CpG methylation of the A-vy allele We will use bisulphite allelic sequencing to obtain a detailed picture of the methylation status of the allele in mice bred for Aims 1 and 2.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The airway epithelium represents the first line of defense for the lungs against inhaled-pollutants and infectious agents. Injury to the airway epithelium occurs during mechanical ventilation and in inflammatory diseases such as asthma. Movement of air into and out of the lungs during respiration causes wide variations in the distension of airways, and the epithelial cells lining the airways are thus exposed to both circumferential wall strain depending upon changes in airway diameter and longitudinal elongation or compression depending upon the expansion of the lung volume. Although there has been much interest in the function of epithelium in response to injury and in the pathogenesis of asthma and other chronic obstructive diseases, there has not been an investigation of the role of mechanical strain on the function of airway epithelium. The central hypothesis of this proposal is that physiological levels of mechanical strain regulate the rate of wound closure and the synthesis of eicosanoids in airway epithelial cells by mechanisms involving reactive oxygen species. This hypothesis will be investigated by stretching both primary cultures of cat tracheal epithelial cells and human bronchial epithelial cells as well as lines of human bronchial epithelial cells grown on elastic membranes using a novel biaxial strain device. Preliminary results demonstrate that both cyclic mechanical elongation and compression delay wound closure by inhibiting cell spreading and migration. Furthermore, indomethacin inhibits and prostaglandin E2 (PGE2) enhances wound closure. Cyclic strain inhibits synthesis of PGE2, and other prostanoids by inactivating the enzyme cyclooxygenase (COX). Preliminary evidence also suggests that COX inactivation is oxidant-mediated. In Specific Aim 1, the mechanisms by which cyclic stretch and compression inhibit wound closure in epithelial monolayers will be determined. Measurements of wound width, cell area, and internuclear distances will indicate cell spreading and migration at the wound edge. In Specific Aim 2, the mechanisms by which cyclic strain regulates the metabolism of arachidonic acid to prostanoids will be determined. Western blots of COX-l and COX-2 protein expression and Northern blots of mRNA levels will be used to determine the adaptation response to cyclic strain. In Specific Aim 3, the role of epithelial oxidant/antioxidant balance in the stretch-induced regulation of wound closure and COX inhibition will be examined. Investigation of the mechanisms underlying epithelial repair and injury during cyclic strain may lead to better strategies for the management of patients during mechanical ventilation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a multi-center study to determine if Selegiline and/or Tocopheral will slow the progression of Alzheimer's disease. It is a double blind, multi-center, placebo controlled study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goals of this work include developing methods to image the stress response in the brains of experimental animals and humans, and to develop methods of manipulating the stress response in order to protect the brain against ischemia and other injuries. Experiments will determine whether local protein synthesis decreases in non-infarcted regions of cortex following focal ischemia in the rat and following global ischemia in the gerbil; and will determine whether the changes of protein synthesis correlate regionally and temporally with the induction of the HSP70 heat shock protein. It is proposed that the decreases in total protein synthesis could be used to indirectly image the stress response in non-infarcted human tissues. Studies of stress gene induction in ischemic human brain will be performed to show that the patterns of HSP70 and other stress gene induction are similar in human and rodent brain. The induction of the heme oxygenase-1 (HO-1) mRNA and protein will be examined in ischemic brains of normal mice, SOD transgenic and SOD knockout mice. Since HO-1 is induced by oxidative and ischemic stress, it is predicted that HO-1 will be induced to a greater degree in ischemic brains of SOD knockouts compared to SOD transgenics. Suppression of HO-1 induction with HO-1 antisense oligonucleotides is predicted to exacerbate ischemic injury more in SOD knockout compared to SOD transgenic mice. Since heme proteins induce the hemeosygenase HO-1 stress protein, it is proposed that prior treatment of animals with heme proteins will protect the brain against ischemic injury, Blockade of HO-1 induction may block protection produced by heme. Lastly, the cloning of the stress gene, methyl malonyl CoA mutase, will be completed and its induction following focal and global ischemia will be characterized, as well as the factors including lipid peroxidation that regulate its expression, Methods used will include rodent focal and global ischemia models, protein synthesis, cloning, DNA nick end-labeling, in situ hybridization and immunocytochemistry.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The functional nature of the projection from the frontal eye field to the brain stem has been studied in the rhesus monkey. Like the frontotectal projection, the frontopontine projection contains cells which discharge in association with eye movements or visual fixation, but not cells which have exclusive peripheral visual responses. The nature of the visual stimuli evoking smooth pursuit were was studied using open-loop visual methods. Superimposition of open- loop position and velocity errors during pursuit maintenance resulted in the generation of eye velocities that indicated that stimulus position as well as stimulus velocity is an important stimulus for the maintenance of smooth pursuit. The time course and dynamics of uniocular saccadic adaptation were studied in monkeys who were made to adapt to a weakened eye. At first the weakened eye had a hysteresis in orbital position, and an orbital-position-dependent saccadic inaccuracy. Both the hysteresis and the orbital position dependent effects were compensated for in a point by point manner with experience. The results suggest that the oculomotor system has a complicated and sensitive corrective mechanisms for the non-linearity of orbital mechanics. Any physical derangement causes maladjustment of this compensation, which can be adapted in time.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Streptococcus pneumoniae (the pneumococus) ranks among the five leading causes of infectious death worldwide. This single species accounts for a large proportion of respiratory tract (pneumonia, otitis media) and invasive (sepsis, meningitis) bacterial diseases. The development of efficacious conjugate vaccines for children has been based upon protection against the few capsule types that commonly cause disease. However, this strategy is now being eroded by the selection for and replacement by non-vaccine types. The initial step in the interaction of the pneumococcus with its host is colonization of the nasopharynx. Experience with conjugate vaccines has demonstrated that interrupting colonization results in herd immunity that amplifies prevention of disease in the population. To better understand the biology of colonization, during the prior funding period, we utilized a murine model to characterize bacterial and host factors that allow for pneumococcal persistence and its eventual clearance from the mucosal surface. These studies show that carriage, much like infection in normally sterile sites, induces acute inflammation. However, opsonophagocytic killing by this neutrophil-dominated response is not completely effective in clearing colonizing organisms. Complete clearance of carriage requires cellular immunity, and is mediated by the gradual Th17-dependent influx of tissue macrophages into the nasal lumen. The key to the success of the pneumococcus in colonization (and disease), therefore, is its ability to evade the initial inflammatory response it elicits. In specific aim#1, we will identify and characterize the complete set of pneumococcal genes and gene products contributing to evasion of opsonization and phagocytic killing by neutrophils in vitro and in vivo. The key to resolution of pneumococcal carriage appears to be recognition and uptake by macrophages. It remains unclear how the unique population of upper respiratory tract macrophages recognizes colonizing pneumococci. In specific aim#2, we will identify the macrophage receptor(s) (including scavenger receptors and C-type lectins) required for non-opsonic clearance during colonization. Finally, in specific aim#3, we will examine the role of capsule type in colonization and whether type-specific differences are explained by i) evasion of opsonophagocytic clearance by neutrophils and ii) recognition and uptake by upper respiratory tract macrophages. Together these studies will provide mechanistic insight into the three main features of pneumococcal carriage;why it is common, why it is transient, and why it varies greatly among isolates by capsule type. PUBLIC HEALTH RELEVANCE: Streptococcus pneumoniae colonizes the mucosal surface of the human nasopharynx and blocking this initial step in host interaction is the key to prevention of pneumococcal disease in the population. Our prior studies of bacterial and host factors affecting colonization show that pneumococci evade the neutrophil response it elicits, but is eventually cleared by cellular immunity leading to an influx of nasal macrophages. The proposed studies provide a mechanistic understanding of i) its ability to inhibit uptake and killing by neutrophils, ii) its recognition by macrophages, and iii) the role of its capsule type in these interactions with phagocytes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Experimental autoimmune thyroiditis in mice is a very useful model of autoimmune disease since it shows the strongest genetic control by gene(s) within the major histocompatibility complex so far described for such a disease. Preliminary experiments suggested that monoclonal anti-Ia antibodies given before immunization of mice with mouse thyroglobulin emulsified in complete Freund's adjuvant, completely prevented the development of thyroiditis. These experiments will be expanded, giving antibodies after the immunization, to observe a suppressive effect on the established disease. The minimum amount of monoclonal antibody needed for suppression and the effect of several antibodies directed against different Ia antigens will be investigated to show the particular Ia antigen(s) on antigen presenting cells involved in presenting thyroglobulin to T lymphocytes. In mice given monoclonal anti-Ia and immunized with thyroglobulin in Freund's adjuvant, several parameters will be monitored namely, the in vitro lymphocyte stimulation by thyroglobulin, the number of T and B lymphocytes (and the number of their subsets) in the spleen, the number of plaque forming cells in the spleen, the skin delayed-type hypersensitivity reaction after challenge with thyroglobulin, the titer of circulating thyroglobulin antibodies, the IgG concentration in serum, the number of cytotoxic T lymphocytes in lymph nodes, and the magnitude of thyroid infiltration with mononuclear cells. Adoptive transfer experiments will be performed to detect the possible induction of suppressor cells after treatment with monoclonal anti-Ia. The expression of Ig antigens in the thyroid of mice (on thyroid cells, antigen presenting cells and endothelial cells) during the development of EAT and the possible modulation of Ia antigens by anti-Ia antibody will be investigated by immunofluorescence and electron microscopy. The effect of various monoclonal anti-Ia antibodies on in vitro lymphocyte stimulation by thyroglobulin will also be investigated in mice immunized with thyroglobulin. The experiments just described will shed light on the role of class II major histocompatibility antigens in the development of autoimmune diseases and will help to better understand the genetic control of these diseases. The experiments will also suggest methods for specific treatment of human autoimmune diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application for a KO1 Career Development Award seeks five years of support for education and research activities that will prepare Dr. Justeen Hyde to examine the consequences of child abuse and neglect among adolescents in the foster care system in Los Angeles. Previous research has identified children in foster care with histories of abuse and neglect as a population at high risk for a number of developmental and behavioral problems. Researchers, child welfare advocates and journalists have demonstrated that child welfare programs often fail to address the complex needs of maltreated youth. Given the scarcity of published data on adolescents in foster care, the proposed research and education activities are designed to build a foundation of knowledge about the experience of foster care from multiple perspectives. Education and training activities include: coursework in child/adolescent development, public policy making, and quantitative research methods; participation in formal trainings offered to social workers and foster parents/guardians; and one-on-one mentorship provided by an interdiscplinary group of advisors at the University of Southern California, among others. These activities will prepare her to conduct an ethnographic study of young adolescents, ages 12 to 15, entering foster care for the first time. The specific aims of the proposed research are to: 1) document the histories of maltreatment among adolescents entering foster care for the first time, particularly the age at which maltreatment was first experienced and the types and frequency of abuse; 2) characterize maltreated adolescents' experiences entering and adapting to foster care; 3) identify mediating factors among maltreated adolescents that influence variations in behavioral, emotional and psychological adaptations to foster care placements over time; 4) explore, from multiple perspectives, the most pressing needs of adolescents entering foster care for the first time and appropriate strategies for addressing these needs. A total of 30 adolescents will be recruited from two types of placements and interviewed at three points in time over a 6-month period. A subsample will be recruited for in-depth case studies, which include interviews with case managers and guardians. This project will provide depth to current understandings of the consequences of abuse and neglect on adolescents in foster care. Study findings will be widely disseminated to a range of key stakeholders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To determine whether phlebotomy to an iron depleted state improves response of chronic hepatitis C infection to alpha-interferon therapy in patients who have previously failed such therapy. A secondary objective will be to correlate iron reduction therapy via phlebotomy with hepatic iron concentration. The primary efficacy endpoints are the response rate (based on loss of hepatitis C RNA in serum) at the end of a 6-12 month course of interferon, and the ability to sustain that response for 6 months after the end of therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application seeks support for Pediatric Oncology Group activities by Southwestern Medical School. Since 1981 our institution has grown to become the Group's third or fourth largest member, among 40 participating institutions, with regard to patients enrolled on therapeutic studies. Moreover, investigators from our center have risen to positions of leadership on major Group committees, including the Executive Committee, New Agents and Pharmacology Committee, and Lymphoid Diseases Committee (Relapsed ALL Subcommittee). Investigators from Southwestern Medical School also have served and are serving as study coordinators of key Pediatric Oncology Group protocols, including the SIMAL-3 study for relapsed patients with ALL, the Group's second largest study in patient accrual. We are now proposing a major commitment of resources from our center to continue Pediatric Oncology Group research studies. This grant proposal describes the personnel and facilities in our center and its affiliate institution, Cook Children's Hospital in Fort Worth, with which we aim to conduct Group activities. The past history of active participation in clinical cancer research within and separate from the Pediatric Oncology Group and our initial contributions to the Group during the past 3 years provide evidence of this commitment. Specifically, we aim to participate in Pediatric Oncology Group research activities by: 1) Actively enrolling as many patients as possible on Pediatric Oncology Group treatment and ancillary protocols, among the 130-140 new children with cancer seen each year at Southwestern Medical School and its affiliate institution; 2) collecting recording, and submitting all necessary data in a timely and organized fashion in order that our protocol entries continue to receive a high rate of evaluability; 3) making scientific contributions to the Pediatric Oncology Group by actively serving on key committees, as protocol coordinators, and as consultants to the Group's leaders in a variety of areas, particularly regarding new therapeutic approaches to ALL and development of Phase I and Phase II drug trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Amyloid b-peptide (Ab), the major molecular component of the cerebral amyloid plaques, appears to play a central role in the neuropathology of Alzheimer's disease (AD). Compounds that prevent the formation of Ab aggregates or that selectively destroy these aggregates are attractive candidates for the development of therapeutic reagents for prevention and treatment of Alzheimer's disease. Also, compounds that interfere with the interactions of amyloid precursor protein (APP) with other factors that are involved in directing it into pathological pathway as well as those that are capable to prevent or destroy intraneuronal accumulations of Ab, are of great interest for developing therapeutic molecules for Alzheimer's disease. In this project we are proposing the possible immunological intervention for prevention and treatment of AD applying phage display technology for the construction of the first anti-Ab single chain fragment variable (scFv) antibody library and the selection of individual phage clones expressing Ab-specific scFv antibody, capable of preventing the aggregation of Ab or dissolving the preexisting aggregates, scFv phage display library enriched in anti-Ab antibodies will be constructed using the first strand cDNA synthesized from mRNA of spleen and lymph node cells isolated from mice immunized with Ab. The constructed library as well as a human scFv library will be used in bioselection procedure to isolate Ab-specific scFvs expressed on phage. The amino acid sequences of antibody complementarity-determinig regions (CDR) will be prepared and used as \"mini-antibodies\". Evaluation of in vitro Ab aggregation and neurotoxicity in the presence of the selected compounds: scFv antibodies expressed on phage or in soluble form and \"mini-antibodies\" will be performed. The most promising molecules selected in this project will be proposed for further evaluation in animal models to all researchers interested in these studies. If successful, these molecules could be of interest for passive immunization in humans, may be after modification with substances that would increase their blood-brain barrier permeability. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In order to understand how a pathogenic change in a gene causes disease, it is necessary to recognize how pathogenic mutations could affect a protein structure-function, protein-protein interactions in protein networks and how these changes could be associated with clinical parameters describing the disease phenotype. We imply molecular modeling to build protein structure, simulate the effect of pathogenic missense changes, and provide a quantitative analysis of their impact on protein structure and stability. Here we use oculocutaneous albinism, autosomal dominant maculopathy, and X-linked retinoschisis (XLRS) as our disease models. 1. Oculocutaneous albinism (OCA) is a rare genetic disorder of melanin synthesis that results in hypopigmented hair, skin, and eyes. There are currently four types of OCA. For the first time, three full-length protein atomic structures, TYR (OCA1), TYRP1 (OCA3), and SLC45A2 (OCA4), were successfully modeled by homology and in silico analysis of missence changes from the NEI/NHGRI molecular diagnostic study has been performed. TYR is a type I trans-membrane monooxygenase. The 4-helix bundle is structurally conserved in different species to carry CuA and CuB ions essential for the catalytic reaction. The active site is formed by 6 His residues structurally coordinating the copper positions. Several missense changes from the collaborative NEI/NHGRI molecular diagnostic study were analyzed. S50L is found in the Cys-rich motif 1 of tyrosinase, whereas R298W is located within the Cys-rich motif 2. The mutations N364H, P384A, D394N, D437N and R403V disrupt the coordination of the copper ion center. A490D affects conformation of helix located in melanosomal membrane. TYRP1 is a type 1 membrane protein. This protein also has 2 Cys-rich motifs and an active site with 6 His residues coordinating 2 metal ions of unknown nature that could be either copper or zinc. A heterozygous missense mutation, A24T, was found in the border between a signal peptide and Cys-rich motif 1 of TYRP1. The structure of SLC45A2 is predicted as a multi-pass trans-membrane protein. The missense mutation L60R is predicted to be deleterious. To test our predictions on tyrosinase activity we engineered a construct of human C-terminal truncated tyrosinase, hTyrCtr. The expression of hTyrCtr in E. Coli was confirmed by Western blot analysis. Metal affinity chromatography shown poor binding to the column which suggests that C-terminal His-tag peptides have decreased binding capacity to the IMACS resin. In addition, L-Dopa enzymatic assay demonstrated that hTyrCtr is expressed as a non-active enzyme which might be due to either the loss of Cu2+ ions in a catalytic site or protein partial misfolding. In contrast, similar protein construct was implied for the protein expression in larvae. We shown that the hTyrCtr, and 2 mutant variants, R422Q and R422W, are active soluble proteins which catalyzes the rate-limiting conversions of tyrosine to DOPA and DOPA to DOPA-quinone. In perspective, a detailed understanding of protein structure and the mechanisms controlling tyrosine-modified tyrosinase interactions would allow to establish molecular chaperone screening for a future medical treatment of patients with the OCA-1B albinism. 2. Assembly of elastic fibers is critical for structural development as well as proper functioning of the extracellular matrix. Elastin and 10-nm fibrillin containing microfibrils form the major components of elastic fibers, which form integral part of extracellular matrices including Bruchs membrane. One of fibrillins, fibrillin-2 or FBN2, is a 2,912 amino acid polypeptide which consists of one amino-terminal trans-membrane domain, 4 epidermal growth factor-like (EGF) domains, 43 calcium-binding consensus sequences (Ca_EGF domains), and 9 transforming growth factor 1 binding protein-like (TB) domains. FBN-2 has 363 cysteine residues. The amino acid sequence of fibrillin-2 been used to generate a native and mutant variant structures for the Ca-EGF motifs 12-19 by homology modeling. Protein fold of Ca_EGF domain is maintained by 6 conserved cysteines which form 3 SS-bridges. In addition, negatively charged conserved residues are either involved in direct ligation to calcium or involved in stabilizing the calcium-binding site. Calcium ion improves the fold stability, help to fix a relative orientation of two neighbor Ca_EGF domains, and stabilize a spatial orientation of FBN-2. Disease-causing mutation E1144K introduces a positive charge into the negatively charged cavity and decreases the Ca-binding affinity. The interaction of K1144 and E1178 change a relative orientation of a neighbor domain. E1438K mutation is expected to have a similar structural effect. Both mutations are associated with a severe phenotype of disease and could change of microfiber packing and elasticity. The M1247T change is affecting the hydrophobic surface loop. The SS-bridge C1246-C1257 stabilizes the native fold of a protein by lowering entropy of the polypeptide chain and by condensing hydrophobic residues from the surface loop into local hydrophobic core using hydrophobic interactions. Thus, the mutation M1247T might affect the SS-bond stability and/or intermolecular interactions. Other mutations are mild changes. Mutations with severe phenotype are likely to cause a change in the fiber flexibility, packaging, and pathogenicity. This might cause the loss of elastic fibers, thickening and calcification of Bruchs membrane which are associated with dominant maculopathy and AMD pathophysiology. 3. Gene mutations that encode retinoschisin (RS1) cause X-linked retinoschisis (XLRS), a form of juvenile macular and retinal degeneration that affects males. Molecular modeling predicted an association between the type of structural RS1 alterations and the severity of full-field ERG phenotype in all but the oldest group of patients. This is now a second study (Hum Mol Genet 19:1302, 2010) that indicates a genotype-ERGphenotype correlation, and it was done with a totally separate and independent cohort. There was a significant association between the predicted severity of RS1 perturbation and both photopic and scotopic ERG b/a-ratios, but only for one age group (15-30 years). Severe RS1 missense changes were associated with a lower ERG b/a ratio than for mild and moderate missense changes, suggesting a quantitatively distinct ERG phenotype. Age-related differences in dark-adapted ERG parameters are consistent with those reported previously in the RS1 knockout mouse. 4. One of possible clinical implications in a human eye disease is using chaperones for the stabilization of native protein structure in mutant variants affected by genetic mutations. This stabilization could be performed in controllable fashion by using small heat shock proteins (sHSPs) with genetically engineered structure. Our recent study have indicated a role for changing of protein hydrophobicity in the thermal adaptation of alpha-crystallin A and suggested ways to produce sHSP variants with altered chaperone-like activity. In this work we use molecular modeling, computational biology, and side-directed mutagenesis to evaluate the effect of mutations and to establish a link between sHSPs hydrophobicity and physiological temperatures. sHSPs maintain cellular homeostasis by preventing stress and disease-induced protein aggregation. In addition, our work provided an evidence for an evolutionary mechanism that has adapted chaperone activity to different environmental temperatures though the alteration of hydrophobicity at crucial locations in the protein structure. This combination of experimental and computational design potentially could be used to create a new generation of artificial chaperones with a purpose to improve stability of mutant variants in inherited eye disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Illinois Prevention Research Center (IPRC) of the University of Illinois at Chicago has a primary mission to reduce health disparities through behavior change approaches by conducting health promotion and disease prevention research across the lifespan, translating research into practice and measuring the real-world effectiveness, dissemination and translation of health promotion and disease prevention programs and initiatives. The IPRC's activities concentrate in seven areas and aim specifically to: (1) Establish and maintain collaborative relationships with community and academic partners, including a Community Committee and CommunityAdvisory Board, university faculty, students, public health agencies and non-traditional partners to address data driven health disparities; (2) Engage the community and establish a research agenda to conduct participatory research on modification of health behaviors associated with increased risk for major chronic conditions such as obesity and diabetes; (3) Conduct community training, graduate student mentoring and continuing education opportunities in evidence-based public health research and practice; (4) Implement and evaluate innovative programs and interventions and communicate findings to community and professional audiences; (5) Develop a cadre of well trained researchers, practitioners, students and community members to enhance the capacity of communities, public health agencies and organizations, and prevention centers to conduct evidence-based participatory research; (6) Disseminate and translate intervention protocols and research findings to the public health community, local, state, and national policy makers, community-partners, and the research community; and (7) Identify contextual factors that may influence implementation of activities and achievement of outcomes. IPRC activities are guided by the CDC's Logic Model for the Prevention Research Center Program and informed by state-of-the-art conceptual frameworks for community-participatory research, development of intervention strategies at the individual, organizational, and environmental level, and diffusion of innovations. The evaluation of the IPRC focuses on our success in achieving the goals related to each of the seven overall aims.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mature B cell lymphomas are a diverse set of diseases which share the property of having failed to complete the differentiation process into plasma cells or undergo apoptosis. PRDI-BF1 and its murine homologue Blimp-1 are transcriptional repressors and act as a molecular switch to commit activated B cells to become non-dividing plasma cells or undergo apoptosis. In this role PRDI-BF1 has broad significance for normal humoral immune responses and in malignancies of mature B cells and plasma cells. We hypothesize that failure to induce PRDI-BF1 expression or function contributes to the persistence of lymphomas and as such therapies designed to rescue PRDI-BF1 function may be important in treating lymphoma. Artificial overexpression of PRDI-BF1 leads to apoptosis in multiple lymphoma lines. Furthermore the hypothesis is supported by our finding that treatment of lymphoma cells with chemotherapeutic agents such as proteasome or histone deacetylase inhibitors leads to an early induction of PRDI-BF1 expression. We have made the recent discovery that PRDI-BF1 directly recruits the histone methyltransferase G9a to mediate silencing of interferon-beta. This suggests chromatin remodeling is the means by which PRDI-BF1 drives terminal differentiation of B cells and kills lymphoma cells. The impact of chromatin remodeling driven by PRDI-BF1 in this process will be investigated. PRDI-BF1 is transcriptionally regulated by unknown mechanisms. This will be investigated and will provide the first detailed understanding of transcriptional regulation at this critical developmental time point and may suggest new avenues to induce PRDI-BF1. Finally, PRDI-BF1 function requires a highly conserved SET-like domain but its activity is unknown. We will identify the proteins interacting at this site and determine their role in PRDI-BF1 activity. Thus this proposal will shed new light on the events occurring during differentiation and may indicate new targets to affect the growth and persistence of lymphomas. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our objectives are to promote excellence in biomedical research through development and maintainance of high quality animal resources. To meet these objectives we propose three major projects within the animal resources program: 1. Renovate animal surgery facilities to provide a suitable environment for recovery surgery and meet DHHS guidelines. 2. Improve satellite animal housing facilities to meet DHHS guidelines. (No funds are requested for this project.) 3. Improve the central animal facilities to provide better utilization of space and meet the needs of the research faculty by upgrading animal housing and procedural areas. These improvements will benefit animal research projects totalling approximately $22 million, and will add to the substantial improvements the College has already made to upgrade animal resources, which have cost more than $5 million over the last two years.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the course of studying inflammatory muscle diseases (polymyositis, dermatomyositis, and related diseases), we have encountered patients with many other muscle diseases. We have studied patients with two genetic metabolic myopathies in detail: phosphofructokinase (PFK) deficiency, and acid maltase (acid alpha-glucosidase) deficiency (also known as Glycogen Deficiency Type II or GSD II or Pompe Syndrome). For the last several years, we have focused particular attention on GSD II because of its close resemblance to myositis. It is a recessively inherited lysosomal storage disease in which glycogen accumulates in the lysosomes, particularly those in skeletal muscle. When the enzyme, known also as GAA, is completely absent, affected infants are usually sick at birth and die in infancy of heart failure, rarely living longer than a year. Apparently the enzyme is needed in the heart only in infancy since affected individuals with even a small amount of effective enzyme survive without cardiac involvement. Survivors generally develop a progressive proximal myopathy with pulmonary failure secondary to diaphragmatic involvement in later years. The long-term aim of our studies is to prevent and to treat this devastating disease, particularly the adult variety, in which the level of enzyme is only slightly (less than two-fold) below the minimum necessary for a normal life. From quite early in the project, it was driven by the belief that GSDII, being a lysosomal storage disease, should be amenable to enzyme replacement and possibly gene therapy, and since there are well over 30 diseases of abnormal lysosomal storage, our results might have wider application. There are estimated to be several thousand cases of GSD II worldwide, and hundreds in the United States, of which many are fatal in infancy (Pompe Syndrome). Our guiding plan has been to do research directed towards therapy, but without trying to move into areas likely to be covered by pharmaceutical companies. We aimed to develop tools that would advance the development of therapy while at the same time learning new biology or developing techniques that might be applicable to other lysosomal or other enzyme deficiency diseases, especially those in muscle. We have followed lines of research designed to provide answers necessary for the development of optimal therapy for GSD II. Starting with the knockout mice, a human GAA expression vector was put in transgenically under the control of either a skeletal muscle or a liver specific promoter controllable by doxicycline [164]. These mice were designed to determine whether the liver, a much easier organ to target, could serve as a suitable depot organ for the synthesis and secretion of GAA that could be taken up by the heart and by skeletal muscle. We discovered that the liver was a superior site to the skeletal muscle for the repair of both heart and skeletal muscle. [164] The achievement of therapeutic levels with skeletal muscle transduction required the entire muscle mass to produce high levels of enzyme of which little found its way to the plasma, whereas liver, comprising <5% of body weight, secreted 100-fold more enzyme, all of which was in the active 110 kDa precursor form. In the past year, we have continued studies on the therapy of the mouse disease. A major step forward was the development of a mouse model that could be used for pre-clinical study of the effects of long term therapy of the disease by repeated injection of the recombinant human enzyme. The barrier to such study has been the immunological response of the mice to the foreign human protein. After a few doses, the animals die of what appears to be anaphylactic shock. In the experiments described above, we developed a strain of mice that produces a minute amount of GAA in the liver ? an amount below the level of biochemical detection, but the mRNA is detectable by PCR. The liver glycogen is very slightly reduced, but the glycogen in other tissues is unaffected so that the mice are phenotypically unchanged. These mice develop no detectable immune response and can receive weekly injections of rhGAA for many months at low or high dose and have no adverse immunological effects. These mice, immunologically speaking, resemble those human infants who have tiny levels of the enzyme and are thereby tolerant to injection of the recombinant protein. The work on this strain has been published, and the mice have been distributed to many investigators and are now used for pre-clinical studies [173]. Our own studies with endogenous expression of the GAA gene in the skeletal muscle {172] and with the intravenous injection of high doses of rhGAA over long periods have shown the remarkable and discouraging finding that unless the enzyme is turned on (controllable transgene) early or the injections are begun early, the stored glycogen in skeletal muscle is only incompletely removed, and clinical recovery of muscle strength is marginal. By contrast, the glycogen stored in cardiac muscle is cleared well. This finding accords with the small clinical experience so far in human infants, in which heart failure is reversed, but skeletal muscle strength has responded little or not at all. This failure of response has now become a major focus of our work. We have discovered that clearance tends to be very good in Type I skeletal fibers and very poor in Type II fibers. We are exploiting the different proportions of these two types of muscle in different fibers to study the possible reasons for this imbalance. The amount of the mannose-6-phosphate receptor is clearly lower in Type II fibers than in Type I fibers or heart muscle, and so are the levels of several other proteins in the lysosomal pathway. We are, with the collaboration of the NIAMS Light Imaging Unit, developing a technique to measure the pH of lysosomes in living, unfixed single muscle fibers in order to determine whether the pH in lysosomes with a large quantity of stored glycogen in Type II fibers is optimal for GAA. We are exploring the difference in gene expression between the two types of muscle fiber in Pompe mice of various ages to look for pathogenetic clues as well as to look for therapeutic avenues. In other studies, we have been working for the past year with the Rapoport lab in NIA to attempt to deliver therapeutic levels of GAA across the blood brain barrier to clear CNS neurons of the large quantities of stored glycogen seen in Pompe mice, and which it is presumed will accumulate in the brains of Pompe infants who survive infancy with the help of exogenous enzyme replacement.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Currently, the standard of care for cancer therapy has limited specificity to only tumor tissue, and can cause devastating side effects. Cancer vaccines utilize the immune system to generate a sustainable and specific anti-tumor response, providing a promising means to improve cancer therapy. However, delivery of vaccine antigen/peptide to dendritic cells (DCs) remains a limiting factor for persistent anti-tumor immune response. Thus, there is a profound need for designing delivery vehicles for vaccine peptides. Current nanocarriers mainly consist of liposomes and polymers, which are larger in size (>100nm in diameter) and tend to stay at injection sites while smaller nanoparticles drain into lymph nodes with subcutaneous injections. They also have known toxicities post degradation. Gold nanoparticles (AuNPs), on the other hand, have three advantages: potential lymph drainage due to their small size, simplistic synthesis/conjugation process and passively collection in antigen presenting cells. Although peptide conjugations on to AuNPs are not new, this proposal aim to be the first to utilize layering techniques onto AuNPs to deliver large does of vaccine peptides for enhanced anti-tumor effect. Thus, the goal of this proposal is to design an effective, simple, versatile gold-based nanovaccine (AuNV) platform for improved anti-tumor immune response. Three specific aims will be carried out to achieve this goal. In aim 1, class I peptides, which stimulate tumor-killing cytotoxic T cells, are conjugated onto polyethylene glycol (PEG) coated AuNPs. Peptides used in this proposal are from a model antigen (ovalbumin) and common melanoma antigens (gp100 and Trp-2). From preliminary characterizations of AuNVs, conjugation yield is very high (~90%) and the overall particle size remained sub-100nm. This will allow potential lymphatic drainage post subcutaneous injection. In vitro Immune response from AuNVs will be measured by interferon-? release, a marker for vaccine anti-tumor efficacy. In vivo efficacy of AuNVs will be assessed by splenocyte sensitization, tumor rejection and treatment assays. As part of aim 1, biodistribution of AuNPs from subcutaneous injections will be explored and be the first to look at distribution at the cellular level. In aim 2, class II peptdes, which stimulate helper T cells, will be incorporated in the AuNV design. In vivo enhancement of antitumor efficacies will be assessed with known class II peptides. However for unknown class II peptides situations, peptide pools will be used to incorporate both class I and class II epitopes. In aim 3, CpG, a known inflammatory stimulant used for cancer immunotherapy, on AuNPs will be combined with AuNVs for a complete vaccine regimen. Future applications of AuNVs are substantial. This design would dramatically lower the cost of immunotherapy compared cellular vaccines. They are easily synthesized, versatile and can potentially target later stage cancers by using inducing anti-tumor immunity over the whole body. PUBLIC HEALTH RELEVANCE: The American Cancer Society estimated over 1.5 million new cancer cases and over half a million deaths in 2010 with national cancer care expenditures estimated to be 104.1 billion dollars since 2006. Cancer vaccines are effective but are limited by vaccine delivery methods and enormous cost for cellular based methods. Gold-based nanovaccines (AuNV) designs are cost-effective, versatile, and can improve the delivery and efficacy of cutting edge cancer immunotherapies and hopefully improve the survival and quality of life for cancer patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "An understanding of the relationship between malarial parasites and their hosts will ultimately aid in developing vaccine strategies. Malarial parasites release heat-stable soluble antigens into the sera of human infected with Plasmodium falciparum. The immunologic role of these antigens is unclear. We recently described a heat-stable antigen (Py117) that circulates in the sera of mice during Plasmodium yoelii infection and an immunologically-related, crossreactive protein (Pf93) that circulates in the sera of humans with P. falciparum malaria. Preliminary data suggest that soluble Py117 and an Ag of merozoites share common epitopes. This has led us to hypothesize that Py117 may provide a protective mechanism for the parasite by \"tying-up antibodies before they reach important receptor molecules on the surface of merozoites. We also hypothesize that the secreted antigen is poorly immunogenic thereby increasing the probability of high levels of circulating antigen. These hypotheses will be tested using the P. yoelii-mouse model system. Experiments proposed include evaluating the antigenic relationship between the secreted and merozoite- associated forms of the antigen, determining if enhancing the immune response to Py117 by immunization or removal of immunoresponsiveness to Py117 by neonatal tolerance will alter the course of malaria infection, determining if the antibody response to Py117 is H-2 restricted and comparing the kinetics of antigenemia, anti-Py117 Ab production and possible formation of immune complexes in strains of mice that differ in susceptibility to malaria. Basic information gained from the mouse model should aid in our understanding of the human situation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Band 3, the major intrinsic glycoprotein constituent of the erythrocyte membrane is responsible for anion transport. Selective in situ proteolysis of this protein has been used to define domains of the protein, certain of which are glycosylated. It is the objective of this research to determine the sequence, linkage and anomeric configuration of the carbohydrate chains of band 3 and to assign them to regions of the peptide. Isolation of micromolar quantities of band 3 and its proteolytic fragments from single units of blood will be coupled wth carbohydrate structural techniques operating on the micromolar level. Since detergent solubilization of the extremely hydrophobic peptides of band 3 limits the effectiveness of exhaustive proteolysis, two new chemical methods will be used to depolymerize the peptide chain for oligosaccharide isolation. High pressure liquid chromatography will be used to fractionate long oligosaccharides while oligomers up to six residues will be gas chromatographed. The carbohydrate structures of oligosaccharides will be determined by methylation analysis, mass spectrometry of the oligosaccharides, high field proton nuclear magnetic resonance and by circular dichroism.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This program continues to study the behavioral effects of brain lesions in children and adults. The aim of our experiments is directed toward five topics: 1. Language and related nonverbal capacities considered as significant indices of left-hemisphere function. 2. Certain perceptual achievements, particularly the recognition of faces and objects in atypical views, as tentative indicators of right-hemisphere processes. 3. Personal and exptrapersonal orientation, as measured in several tasks, to be varied in such a way as to permit a better distinction of learned from relatively unlearned components in performance, and of predominantly parietal from predominantly frontal lesion effects in either hemisphere. 4. Memory, as assessed in tests involving relatively short-term as contrasted with longer-term memory, and varying conditions of retrieval. 5. Mood: Specifically, we shall continue our assessment of therapeutic outcome, neurologic status, and behavioral test performance after surgical interruption of the cingulate gyrus and bundle in selected cased of persistent pain or severe affective disorders. All except the last two of these questions will be investigated, in a manner as nearly comparable as we can accomplish, in children and adults with brain pathology. We realize that there are severe difficulties, both in establishing comparability of lesions across these age groups, and in achieving any certainty of the comparability of tasks; and it will not be possible to do equal justice to all the questions in the next few years, since so much depends on the rates at which appropriate clinical cases will appear. Nevertheless, by monitoring a properly prepared set of behavioral tasks and by keeping the five topics as signposts, we are more likely to make maximal use of the patients who might come our way.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Breast cancer among Filipina American women represents a major but largely neglected cancer disparity; -first, though not as highly visible as other Asian subgroups, the Filipino population in the US is large, second among Asians only to the Chinese. Second, Filipinas have higher rates of breast cancer incidence than most other Asian subgroups. Third, resources for and data regarding Filipina women with breast cancer are almost non-existent. In 2004, West Bay Filipino Multi-Service Center (West Bay) joined forces with the UCSF: comprehensive Cancer Center (UCSF-CCC) and the San Francisco General (SFGH) Breast Care Program to start the first Filipina breast cancer support group (Sinag Tala) in San Francisco. However, group attendance is uneven and the little research available on Filipino Americans suggests that a \"one size fits all\" approach to outreach would not be effective in this highly relational (collectivist-oriented) culture. The proposed study will elucidate the meanings of survivorship and breast cancer support in this community, and inform how to design more culturally appropriate outreach building upon existing community resources social networks) for the women who need them most. We will utilize multiple qualitative methods because each taps different types of data (participant observation, individual ethnographic interviews, and small group interviews) to achieve our specific aims to: 1) identify beliefs and values associated with breast cancer, survivorship, and support; and 2) create and pre-test culturally resonant outreach themes and channels based on core cultural values and existing community resources. The conceptual framework is drawn from theories of quality of life, social capital, and relational culture. The product of this study will be new, culturally appropriate outreach themes and outreach communication channels designed to encourage Filipina breast cancer patients and survivors to take part in support groups. Just as access to and participation in adequate and meaningful social support has been shown to improve the quality of life of White breast cancer patients, developing meaningful outreach themes and channels to link Filipina breast cancer survivors to support services is likely to improve the quality of life of Filipina breast cancer survivors and to address survivorship disparities in this community. Formative research into culturally relevant outreach for support services has implications across the stages of survivorship. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Prostanoids exert profound physiological effects on the urinary tract including nociception, induction of smooth muscle contractility, enhancement of cell proliferation, and blood flow regulation. The rate-limiting step in prostanoid synthesis is catalyzed by cyclooxygenase (COX). This enzyme exists in two isoforms: COX-1 (usually constitutive) and COX-2 (highly inducible). COX-2 is substantially induced with distension and stretch of urinary tissues. A new area of interest involving COX activity in urinary tract has been identified: urothelial cell stretch-induced COX-2 expression. In cell culture, urothelial stretch produces a 10-fold induction in COX-2 levels within 6 hours of applying stretch. Since COX-2 upregulation would lead to an increase in soluble prostanoids, this process represents a critical link between urothelial stretch and afferent pain perception, muscle contractility, and cell proliferation. While the specific model of this proposal involves urothelial cells cultured from the ureter, this novel concept has broad implications in the pathophysiology of the urinary tract. The goal of this project is to identify cellular pathways of stretch-induced COX-2 expression and thereby provide several potential targets for drug therapy. Based on titerature reports in other cells, and preliminary data, the hypothesis of this proposal is that stretch activation of PKC and p38 MAP kinase signaling cascades induces COX-2 in urothelial cells. The Specific Aims are: I. To characterize COX-2 induction in human urothelial cells subjected to mechanical stretch. This will be accomplished by determining the optimal duration and degree of stretch for COX-2 induction in human urothelial cells and determining if stretch-induced COX-2 expression is regulated transcriptionally and/or post-transcriptionally in human urothelial cells. II. To determine if stretch induction of COX-2 is mediated by increased intracellular calcium concentrations [Ca++] i and subsequent PKC and p38 MAP kinase activation. This will be accomplished by determining if stretch of urothelial cells increases [Ca++] i, and activates PKC and p38 MAP kinase, if inhibition of calcium, PKC, or p38 MAP kinase attenuate stretch-induction of COX-2, and if p38-dependent transcriptional factors are involved in stretch-induced COX-2 expression. The findings of this exploratory proposal have great potential to expand into several dynamic lines of research and may identify novel approaches to treating urinary tract disorders such as bladder outlet obstruction, ureteropelvic junction obstruction, and interstitial cystitis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The isolation of alpha-aminolevulinate synthetase (ALA-S) from livers of induced chick embryos is being continued; with purified enzyme, an antibody will be developed to study the induction mechanism and the transport of the enzyme from the microsome into the mitochondria where it is active. Chlorinated hydrocarbons both induce ALA-S synthesis and block the biosynthetic chain so that uroporphyrin accumulates. The mechanism of the block will be sought. Iron and metabolites of the chlorinated hydrocarborns may be involved. These studies may lead to our understanding of the environmental toxic effects of the chlorinated hydrocarbons including tetrachlorodibenzodioxin. In Friend leukemia cells dimethyl sulfoxide caused their differentiation into erythropoetic cells. The steps in the development of the enzymes of the heme biosynthetic chain will be examined to note whether there is a sequence in time of the developing enzymes that follows the sequence of enzymes of the heme biosynthetic chain. In animals, the synthesis of the first committed product of the heme biosynthetic chain is ALA. It is well establised that ALA is formed by an enzyme that uses glycine plus succinyl CoA as its substrates. In plants there is evidence that ALA may be formed from other substrates; the mechanism of ALA formation will be studied with appropriate labelled substrates and with metabolic inhibitors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To meet the dual threats of emerging infectious diseases and engineered biowarfare/bioterror agents, there is a pressing need for more efficient systems for vaccine development. TRIAD, or the Translational Immunology Research and Accelerated [Vaccine] Development program, based in the Biotechnology Program at the University of Rhode Island, has pioneered the development and application of an integrated \"gene to vaccine\" in silico, in vitro and in vivo vaccine design program to address this need. TRIAD has selected Category A pathogens F. tularensis, Category B agents Burkholderia pseudomallei and Burkholderia mallei, and emerging infectious diseases (HCV, H. pylori, tick borne diseases) as the focal point of this proposal. Using the TRIAD immunoinfomatics Toolkit, TRIAD investigators will pursue the development of second generation epitopebased immunome-derived vaccines for these pathogens, while addressing the failings of prior generations of epitope based vaccines. We will maximize payload quantity using validated immunoinformatics tools that permit selection of optimal T cell epitopes that are highly conserved and immunogenic. We will ensure payload quality by choosing epitopes that demonstrate antigenicity in human PBMC as well as protection in established murine models of disease/infection. We will select a combination of promiscuous Class II epitopes and Class I supertype epitopes will provide >99% coverage of human populations. We will avoid cross-reactive epitopes and explore the role of regulatory T cells in the context of improving vaccine design. Where appropriate, we will combine our epitope-driven vaccines with broad-spectrum anti-LPS vaccines. We will optimize payload, delivery, formulation, and adjuvanting by exploring a range of delivery options [Dendritic cells, DEC205, DNA, electroporation, mucosal delivery). The TRIAD project aims to develop vaccines demonstrating broad spectrum activity include crossprotective and multiple component vaccines, and delivery technologies that have the potential to be effective against multiple emerging and re-emerging infectious diseases. Our efforst to merge rational design with rcentadvances in vaccine deliverywill manifest in a coordinated toolkit and a cadre of informed users, who will be ready and able to apply the tools to discover new treatments for emerging infectious disease and biodefense.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose was to determine if whole body and skeletal muscle glutamine and leucine metabolism are altered in HIV-infected subjects. Six HIV-infected men with chronic stable opportunistic infections and 10% weight loss, 8 HIV-infected men and 1 woman without wasting and 6 HIV- negative age-and weight-matched men were studied. Constant intravenous infusions of stable isotopically labeled leucine and glutamine were used to assess plasma GLN and LEU rates of appearance by mass spectrometric measurements of stable isotope content. Fasting whole body protein breakdown and synthesis rates were increased above control in the asymptomatic HIV-infected subjects and further increased in symptomatic HIV-infected subjects. These findings suggest that the rate of muscle protein breakdown was increased while the rate of muscle protein synthesis was unchanged in symptomatic HIV-infected subjects. Since lymphocytes require GLN release may be increased to provide energy for proliferati ng lymphocytes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the proposed research is to understand the mechanisms of how genes are regulated in different tissues. This will be accomplished through the study of the human gene encoding debranching enzyme and molecular analysis of Type III glycogen storage disease. In this disease there are patients who lack debranching enzyme activity in both liver and muscle, and patients with enzyme deficiency confined to liver; yet the enzyme is a monomeric protein and appears to be identical in all tissues. The proposed research hypothesizes that debranching enzyme in liver and muscle is encoded by a single gene; its expression, however, is under separate genetic control. Patients with various clinical and enzymatic subgroups of Type III glycogen storage disease are being followed at Duke Medical Center; fibroblasts from these patients have been collected and stored. The debranching enzyme has been purified from porcine muscle and the antibody characterized. The cDNA coding for debranching enzyme from both human liver and muscle will be isolated by screening lambda gtll human cDNA libraries with antibody against porcine debranching enzyme. The debranching enzyme mRNA in these two tissues will be compared by Northern blot analysis, cDNA sequencing and/or sequencing of mRNA by primer extension. The chromosomal gene(s) coding for debrancher enzyme will be isolated, and the molecular structure and number of copies of the genes will be determined. Information about the number of debrancher genes and the sequences of the mRNA from liver and muscle will be used to design strategies to investigate the mechanism responsible for control of tissue-specific debrancher gene expression. This may involve sequencing of the 5' flanking region for two possible transcription initiation sites, or in the case of two different mRNA sequences encoded by a single gene, sequencing the regions of the gene that specify the mRNA unique to liver or muscle. The defective chromosomal debrancher genes from patients with disease confined to liver will also be cloned and the mutations characterized. Molecular classification of Type III glycogen storage disease will be performed at the protein, mRNA, and gene levels. These molecular findings will be correlated to the type and severity of clinical phenotypic expression. Information gained on molecular structure of glycogen debrancher gene and human mutants lacking debrancher activity may lead to identifying DNA regions important in the control of tissue-specific gene expression, and may provide a model for mechanisms responsible for clinical variability in other genetic disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The effect of experimental conditions related to the induction of atherosclerosis will be studied with an emphasis on changes in elastin fiber formation. The conditions which will be studied include hypertension, hyperlipidemia, and hyperinsulinemia. Hypertension will be induced by galactose intoxication, which in avians produces marked plasma hyperosmolarity. Hyperinsulinemia will be studied using both in vitro and in vivo systems. In addition, properties of proforms to tropoelastin will be studied after isolation and characterization in an attempt to provide information on the metabolism of elastin. One of the key features of the work will be the assessment and identification of events which lead to increases or decreases in net elastin synthesis. One aspect of the work will be the investigation of changes in elastin synthesis in older animals or through the lifecycle of animals exposed to conditions which are known to alter the metabolism of the vascular wall.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We plan to isolate and analyze zebra fish mutants with altered visual systems. Of particular interest initially will be mutants with altered organizations of retinal cell types, and specific enrichment procedures will be utilized to aid in the isolation of such mutants. The mutants will be analyzed by behavioral, anatomical, physiological and genetic methods. In order to increase the frequency of mutants in the populations subjected selection, we will expose fish to mutagens at appropriate stages in their life cycle. Since most mutations are recessive, we plan to generate homozygous fish from mutagenized eggs in order to enable us to recognize mutant phenotypes. Procedures for generating homozygous fish have already been developed in our laboratory.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The importance in the understanding of the basic mechanism of human plasminogen activation is underscored by the wide use of fibrinolysis in the therapy of some blood clotting and cardiovescular disorders. Staphylokinase is a plasminogen activator protein secreted by strains of Staphylococcus aureas. Limited data from ours and other laboratories suggest that this protein activates plasminogen by strong binding of the two proteins, similar to the mechanism of streptokinase. The lack of information on staphylokinase has been due to the difficulty to obtain sufficient amounts of pure protein. This project proposes (1) to obtain pure staphylokinase form cloned gene in E. coli, (2) to examine themechanism of human plasminogen activated by kinetic and binding studies, (3) to determine the functional groups involved in plasminogen binding and activation by differential chemical methods, and (4) to change the functional groups of staphylokinase using site-directed mutagenesis of cloned gene. These changes involve lysines, tyrosines, tryptophan, N-terminal regiona and C-terminal region. The changes on the N- and C-terminal regions will test the functional roles of these residues. The changes in Tyr and Trp will be done in conjunction with 19F-nuclear magnetic resonance studies of the role of these residues. The studies described above will permit a greater understanding of mode of plasminogen activation by staphylokinase in specific and of contact activation of serine protease zymogens in general.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Heart disease is the leading cause of morbidity and mortality in the Western World. As this is mostly related to disorders of the blood supply to the heart, it is very important to be able to assess the perfusion of the heart wall. Conventional methods for assessing myocardial perfusion have many limitations, including relatively low resolution and poor ability to provide quantitative data. Magnetic resonance imaging (MRI) of the \"first-pass\" kinetics of contrast enhancement by a bolus injection of contrast agent is a very promising method for noninvasive assessment of myocardial perfusion, with higher resolution than conventional radionuclide imaging methods. However, current approaches to MRI perfusion studies have only been \"semiquantitative\", due to several technical difficulties. We have been achieving very encouraging initial results in making absolute measurements of regional myocardial perfusion, using novel MRI methods that we have designed to overcome the limitations of conventional MRI approaches. In the proposed research, we will further develop our MRI methods, and the associated image analysis methods, for quantitatively measuring myocardial perfusion. We will perform a series of studies on normal subjects to establish the expected range of variability of the results. We will also perform a series of studies of patients with coronary artery disease, comparing the results of our MRI methods with those of conventional methods used to assess the blood supply to the heart, including radionuclide imaging, CT angiography and conventional cardiac catheterization (including invasive measures of flow reserve). The significance of the proposed work is its potential to achieve more accurate and higher resolution assessments of blood flow to the heart wall than current conventional methods can provide. This would be very useful for aiding treatment decisions and for following the results of therapy in patients with ischemic heart disease, a common and serious clinical condition. The quantitative MRI methods we propose to develop for the assessment of cardiac vascular disease may also be applicable to assessment of the perfusion of other important organs, such as the brain, which is subject to stroke, and in other important disease processes, such as cancer, where the disease evolution may be related to blood flow.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The work being carried on is quite varied in method and approach, but has one consistent aspect: the application of conditioning and training theories and methods to the empirical analysis of concepts important to theories of social behavior, motivation, psychopathology, and personality development. The major focus has been aversive control but now the emphasis is shifting to the aversive, motivational after-effects of terminating positively-reinforcing stimuli rather than the aversive, motivational effects of the onset of aversive stimuli. Specific research areas: (1) the interaction of Pavlovian conditioning and instrumental learning; how appetitive and aversive Pavlovian conditioning modify instrumental responses of either appetitive or aversive origin; (2) tests of an opponent-process theory of affect and motivation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is known that individuals with peripheral arterial disease are at increased risk for cardiovascular morbidity and mortality. In order to facilitate design of a trial to evaluate the effects of intervention on risk factors on morbidity and mortality in such patients a feasibility study (ADMIT) has been designed to evaluate the ability to recruit, treat and retain volunteers in anticipation of a large scale multi-center study focusing on risk factor reduction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Importance: Intimate partner violence (IPV) is a serious national problem noted in Healthy People 2010 and the CDC Injury Research Agenda. The Deaf community is an understudied linguistic, cultural, and disability minority group whose IPV risks and experiences are virtually unknown. Preliminary studies, most conducted by this research team, suggest that the deaf population experiences higher levels of IPV than the general population and more frequent and serious IPV injury consequences such as emergency room visits and suicidality. To be maximally effective, IPV intervention and prevention strategies must be responsive to the language, culture, and other characteristics of the target population. Moreover, if, as our clinical and research experience suggests, IPV perpetrator characteristics and manifestations of IPV affecting the Deaf community differ from general population norms in significant ways, then etiologic research identifying those differences must precede and inform the development of IPV prevention and intervention strategies targeting the deaf population, as well as guide further research on this important topic. Objectives: (1) To investigate and describe risk and protective factors associated with characteristics of IPV perpetrators who are involved in violent relationships where one or both partners are deaf and communicate via American Sign Language (ASL). (2) To investigate and describe risk and protective factors regarding the ways IPV behaviors manifest in relationships involving one or more deaf individuals who use ASL, especially behaviors associated with greater injury and mortality risk. (3) To compare and contrast these study findings with current knowledge regarding risk and protective factors associated with IPV perpetrators and perpetration behaviors in the general population. (4) To utilize study findings to make recommendations for future research, intervention, and prevention efforts aimed at preventing and/or reducing the injury consequences of IPV in the Deaf community. Study Design: This etiologic, cross-cultural study will be conducted by a multidisciplinary team of deaf and hearing researchers. A period of study team cross-training precedes the data-collection activities. The majority of study data accrues via 90 semi-structured interviews with three distinct participant groups (see below). We employ a mixed-methods study design based on a social ecological model of IPV origin, continuation, and severity. Rigorous qualitative and quantitative methods will be employed, the latter focused on three well-regarded IPV measures used in this study on an exploratory basis. The study goal is to discover Deaf community-specific IPV risk and protective factors by triangulating data from our three participant groups, and further assess these findings via presentation to three respondent verification focus groups consisting of Deaf community IPV perpetrators (2 groups), and victims/service providers (1 group). Setting: The 90 semi-structured interviews forming the core source of study data will involve geographically diverse/balanced samples from our three respondent categories (see below). Most of these interviews will involve deaf people communicating in ASL and will take place via videophone technology which allows point- to-point, distant audio-visual communication and, therefore, communication in ASL. Participants who are hearing or hard-of-hearing can choose a telephone interview. Two of the three, face-to-face respondent verification focus groups will take place in southern California, where collaborators run the country's only two programs serving deaf IPV perpetrators. The third will take place in Rochester, New York. Participants: Three respondent samples will participate in our semi-structured interviews: (1) individuals (professional and lay) who provide services to deaf IPV perpetrators or victims (N=30), (2) deaf IPV victims (N=20), and (3) individuals (regardless of hearing status) who have perpetrated violence in relationships where one or both partners is a deaf ASL-user (N=40). An additional 15 individuals will participate in the respondent verification focus groups that comprise the final phase of the study - 10 IPV perpetrators (2 groups of 5), as defined above, and one group of 5 deaf IPV victims and service providers. Interventions: This study does not involve interventions. Outcome Measures: We will employ a mixed-method data analysis approach, involving rigorous qualitative analysis of interview and focus group data and quantitative analysis of these data in relation to data accrued through the exploratory use of three well-regarded measures of IPV during the semi-structured interviews with deaf IPV victims and perpetrators (N=60 total). Also, an Evaluation Task Group will investigate and gauge the ongoing and final success of the project through a quarterly series of formative and summative evaluations. PUBLIC HEALTH RELEVANCE: RELEVANCE TO PUBLIC HEALTH STATEMENT Intimate partner violence (IPV) is a serious national problem identified by federal authorities and many professional societies. When IPV occurs in unique subpopulations, such as minority groups, etiologic research must guide the design of prevention and intervention strategies if they are to be maximally effective. The American Deaf population, which is a severely understudied language and disability minority group, experiences high rates of IPV and deserves quality etiologic research aimed at reducing this high-priority public health burden.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "With the increased use of prenatal screening by ultrasound, more fetuses are being diagnosed as potentially having congenital renal obstruction (2-5% of all pregnancies in the US). To date, the most common cause of renal failure in children is from renal damaged caused by urinary obstruction. Many controversies exist regarding detection, prognosis, and proper management of children with this condition. We hypothesize that congenital renal obstruction results in the release of specific proteins into the urine that reflect changes in protein secretion and shedding from renal tubular cells and other sources. We predict that these changes will vary with onset, duration and severity of the obstruction. In our previous work, we have demonstrated our ability to use state-of-the-art mass spectrometric and proteomic approaches to identify proteins in the urinary proteome in an animal model of postnatal maturation. With this experience, my objective is to identify new markers of obstructive renal damage that could be potential diagnostic or prognostic clinical tools. Our approach will be to use an animal model of neonatal renal obstruction and follow urine composition over time after initiation of the injury. We will study the urinary proteome during obstruction using advanced qualitative and quantitative proteomic methodologies in order to prioritize and identify candidate clinical markers. The discriminatory power of the candidate markers, and their potential for clinical translation, will be determined using directed quantitative proteomics in select human infant cohorts with and without severe renal obstruction. Overall, my long-term career objective is to become an independent pediatric urologic clinician-scientist with a strong commitment to translational research that focuses on biomarker discovery for renal injury. My immediate goals are to acquire a strong background and research experience in mass spectrometry, proteomics, and biomarker validation. During the early portion of my career, I plan to add to my foundation as a clinician investigator through lectures, strong mentorship by Dr. Michael Freeman and others, scientific collaborations, hands-on experience, and didactic coursework that focuses on proteomics, clinical trials, biomarkers, epidemiology, and bioinformatics. My research will be conducted at Children's Hospital Boston in the Urological Diseases Research Center and in the Children's Hospital Boston Proteomics Center. In summary, this project will provide the necessary foundation for a successful career as an independent investigator and will lead to the identification of novel biomarkers that may be used to inform clinical decisions in children affected with congenital renal obstruction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Pathology Core goal will continue to serve as a resource for the investigators interested in neurogenetics research. The resources include a tissue bank and providing neuropathological expertise in the analysis of murine and human normal and lesional tissue associated with Neurofibromatosis 2 (NF2), Schwannomatosis and Tuberous Sclerosis (TSC). To accomplish this goal the core will: 1) Coordinate the collection of frozen lesional tissues and cell culture from patients with inherited nervous system tumor syndromes and from corresponding solitary, sporadic tumors from the general population, as well as collect corresponding fresh and paraffin-embedded fixed tissues from normal controls (autopsy tissues). 2) Maintain a separate database of tissues, cell lines and, DNA for these projects, which can be linked to other databases (clinical database, mutation analysis database, RNA database). 3) Provide immunohistochemical expertise in the study of cellular and subcellular localization of merlin, tuberin, hamartin and their interacting proteins and analyze the expression of novel proteins (identified by CGH and microRNA studies), phosphorylated proteins and neurotransmitter receptors in murine models, in the normal human brain and in TSC or NF-associated lesions 4) Provide histological and immunohistochemical expertise in the classification and analysis of lesions associated with NF2, Schwannomatosis and TSC and their sporadic counterparts. 5) Provide histological and immunohistochemical expertise in the classification and analysis of newly generated lesions in Nf2 and Tsc murine models and evaluate the delivery and effects of experimental gene therapy in these lesions. The core will ensure optimal and uniform processing of all tissues and cell cultures to be used by the investigators of the various projects. In addition, the histological review of all tissues (murine and human) will ensure uniform and standardized classification, which are essential for meaningful comparison and interpretation of the data. The core will continue its work, in collaboration with the various projects, of elucidating the pathogenesis and altered protein expression in TSC-associated lesions in humans and in the Tsc murine models and in defining the molecular, clinical and histological characteristics of NF-associated lesions in humans and murine models. Finally, the delivery and effect of gene therapy on lesions in Tsc and Nf2 mouse models will be assessed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The pol gene of the human immunodeficiency virus (HIV) encodes three viral enzymes, reverse transcriptase, protease and integrase, which are critical for the replication of HIV. This project focused on the discovery of inhibitors of one of these viral enzymes, HIV reverse transcriptase (HIV RT). The broad objectives are to contribute to the chemistry and biochemistry of conceptually new nucleosides and nucleotides with useful therapeutic potential against the infectivity of HIV. These novel compounds, with stereochemically defined surrogate carbohydrate components, that are regioisomeric with the natural nucleosides and nucleotides, are referred to as isonucleosides and isonucleotides. A compelling rationale for this investigation comes from the discovery in this project of a potently anti-HIV active compound, (S,S)-isodideoxyadenosine [(S,S)-isoddA}. IsoddA 5'-triphosphate is a powerful inhibitor of HIV RT (KI 16 nM). This renewal proposal seeks to expand successful work of the current project to include cyclic monophosphate (MP) pro-drugs (cyclo Sal MPs) and hydrolytically stable phosphonyl derivatives in order to deliver the MPs or their isosteres directly inside the cell. In addition, new classes of isonucleosides with multiple supporting rationaleS for anti-HIV activity are proposed. The synthetic work will involve the development of approaches to the novel, chiral isodideoxynucleosides and their purification and complete characterization by magnetic resonance, mass spectral, and X-ray methods. Collaborative antiviral studies on the target compounds and pro-drug forms will be carried out against HIV-1 and HIV-2, including drug-resistant HIV isolates. Data on inhibition of the cytopathic effect of HIV, on inhibition of HIV RT, on inhibition of proviral DNA synthesis, on inhibition of the expression of HIV-1 p-24 Gag protein, on host cell cytotoxicicty including inhibition of cellular DNA polymerases alphaepsilon beta and gamma, and on therapeutic indexes will be determined and analyzed. Cellular combination drug studies, particularly those having the potential for synergistic inhibition of HIV infectivity, are also planned.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Background and Objective: [unreadable] Previously, all experiments examining protein protein interactions in living cells by FRET have been limited to the use of two fluorescent proteins.[unreadable] Here, we report the development of a novel FRET technology for studying complexes involving three proteins.[unreadable] The system utilizes CFP, YFP and mRFP (monomer red fluorescent protein). CFP->YFP FRET1, YFP->mRFP FRET2 and CFP->YFP->mRFP linked FRET signals are monitored separately by flow cytometry. The technology has been validated by constructing plasmids encoding several FRET-positive and -negative controls, including CFP-YFP-mRFP, CFP-T2TD-YFP-mRFP and CFP-YFP-T2TD-mRFP, whereas T2TD (TRAF2 TRAF domain) acts as FRET insulator because of its structure with a distance of 9 nm from the head to the end.[unreadable] Furthermore, this technology has been used to examine the trimer formation of TRAF2 in living cells.[unreadable] [unreadable] Results: [unreadable] 1. The validity of CFP->YFP->mRFP 3-way FRET: For the first time, we validated the practicality of using flow cytometry to determine CFP->YFP->mRFP linked FRET by directly visualizing 2-step-FRET signals and the quenching of CFP->YFP FRET1 signals.[unreadable] 2. The determination of three protein complexes: with the system, we examined the trimer formation of tumor-necrosis-factor-associated factor 2 in living Hela cells. TRAF2 was tagged separately by CFP, YFP and mRFP. Co-transfection of all three plasmids displayed the 2-step-chained FRET signals, but not in cells co-transfected with any two of them and blank fluorescent protein. [unreadable] [unreadable] [unreadable] Conclusions: we have developed a method to determine interactions among three proteins in living cells using flow cytometry and aCFP-YFP-mRFP FRET system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The free Ca2 ion concentration (Ca2 ion) of the cytosol, an important parameter in the regulation of carbohydrate metabolism, is itself regulated through the combined actions of the Ca2 ion transport systems of cellular membranes. Since these systems have been studied under unphysiological conditions, little is known of the details of cytosolic (Ca2 ion) regulation in an intracellular environment. This laboratory's approach has therefore been to measure directly the functional result of subcellular Ca2 ion transport under simulated in vivo conditions. An ion-selective electrode continuously monitors ambient (Ca2 ion) in a cytosol-like medium containing either isolated fractions of mitochondria and endoplasmic reticulum (microsomes) from rat liver or isolated rat liver cells pre-treated to make only their plasma membranes permeable to solutes. Having already documented joint steady-state regulation of ambient (Ca2 ion) by mitochondria plus microsomes at values approximating those of liver cell cytosol, this project will investigate 1) inter-organellar effects on Ca2 ion homeostasis through reciprocal alteration of internal Ca2 ion content and 2) modulatory effects on Ca2 ion homeostasis of a) phosphate, adenosine triphosphate and oxidized/reduced pyridine nucleotide, which reflect the status of carbohydrate metabolism, and b) glucagon, epinephrine and norepinephrine, which alter the rate of carbohydrate metabolism.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The study of tolerance to self-antigen in B-cells is of the utmost importance to understand how the immune system is regulated in health and disease. Unfortunately, the study of human B-cell tolerance has been hampered by the inability of investigators to identify an abundant population of autoreactive B-cells with homogenous antigenic reactivity and surface markers. We have identified such a population (termed VH4- 34 cells) and over the last three years we have validated the use of these cells for the study of human B-cell tolerance. [unreadable] [unreadable] VH4-34 cells recognize glycolipid autoantigens widely expressed in the surface of multiple cell types as well as microbial glycolipid antigens and probably play a homeostatic role in host defense by connecting the innate and adaptive immune systems. In healthy individuals, VH4-34 cells are prevented from inducing autoimmunity by positive selection into the marginal zone of the spleen and negative selection form the germinal centers. The latter mechanism prevents VH4-34 cells from expanding into the long-lived postgerminal center memory and plasma cell compartments thereby inhibiting the production of significant amounts of pathogenic autoantibodies. [unreadable] [unreadable] In this application we propose to study the phenotypic and functional properties of VH4-34 cells in healthy individuals and to compare them to VH4-34 cells obtained with patients with Systemic Lupus Erythematosus (SLE) in whom B-cell tolerance is defective. This will be accomplished by using multiparameter flow cytometry and immunocytochemistry. Furthermore, we will analyze the signaling processes responsible for the biological responses observed with VH4.34 cells and again determine how signaling differs in patients with SLE. Finally, we will ascertain what type of T-cells regulate the behavior of VH4-34 cells both in healthy subjects and SLE patients. [unreadable] [unreadable] The studies proposed herein should yield important insights into the regulation of autoimmunity and the pathogenesis of autoimmune diseases such as SLE. Furthermore, we believe that our results will improve our ability to treat patients with SLE and similar diseases. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "While most women are overweight or obese when they reach menopause, we know little about how obesity affects this transition and its impact on metabolic health. We have hypothesized that the enhanced production of estrogens in peripheral tissues of obese women protects adipose tissue, skeletal muscle, and bone from the menopause-related decline in function. We have merged well-characterized preclinical models of obesity (obesity-resistant/prone rats) and menopause surgical ovariectomy (OVX) to study how pre-existing obesity affects this transition. In the first aim, we examine obesity's effects on the loss of ovarian estrogen production and its impact on energy balance, fuel utilization, and metabolic health. The role of extragonadal estrogen production in obesity's effects will be dissected with the use of aromatase inhibitors. Diet-induced weight loss is common in women with weight control problems, but the body compensates with homeostatic adaptations that drive weight regain. The loss of ovarian function in weight reduced women may exacerbate this metabolic drive to gain weight and create a potentiated vulnerability to metabolic disease. Regular exercise and estrogen have many overlapping effects on energy balance and fuel utilization that may counter these consequences to energy homeostasis and disease. In the second aim, we examine how energy restricted weight loss from obesity affects the drive to gain weight and the decline in metabolic health after the loss of ovarian function. The impact of regular exercise, with actions that reflect an estrogen mimetic, will be examined in weight reduced subjects. These studies will provide insight into how one of the most common preconditions of menopause, obesity, affects the loss of ovarian function and its consequences to energy homeostasis and metabolic disease. Observations from these studies may provide evidence that certain populations of obese woman (treated with aromatase therapy, energy restricted) who may be particularly vulnerable to menopausal weight gain and metabolic disease unless they maintain a program of regular physical activity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "West Nile Virus (WNV) is one of the many neurotropic flaviviruses that is widely spread throughout the world, and continues to cause significant morbidity and mortality. In the US alone there were 24,000 cases of human WNV infection and 1000 fatalities since 2006. Both innate and adaptive immune components, including CDS, CD4 T cells and B cells contribute to WNV clearance and prevent infection of neurons. The adaptive immune components also form memory, which is the hall-mark for vaccination strategies. The generation of adaptive immunity is linked to innate activation, primarily via viral sensing by host pattern recognition receptors (PRRs), which in turn lead to ARC activation and generation of inflammatory mediators. One class of these early inflammatory mediators, type-l interferons (IFN-I) induce an anti-viral state in infected and neighboring cells. Many viruses, including WNV, developed potent IFN-I evasive strategies. Recent studies from our and other labs show that IFN-I can also exert profound influence on T cells responses, and generation of immune memory;but very little is known about the roles of specific pattern recognition receptors and the IFN-signaling in generating flavivirus-specific adaptive response. An understanding of this is critical for generating improved vaccines. We hypothesize that IFN-I signaling plays a critical role in generating WNV-specific adaptive immunity and that strategies to interfere with viral IFN-I evasive mechanisms should yield better vaccines. In Aim 1 we will define the role of dendritic cell signaling via RNA helicases and TLRs for generating WNV-specific CDS T cell responses. In Aim 2 we will examine the role of IFN-I signaling in the generation of WNV-specific CDS T cell responses. In Aim 3 we will assess the importance of the timing of IFN-I signals in programming T cell responses. Using the knowledge gained from these and the studies proposed in the other four projects of this U19, in Aim 4 we will modulate IFN-I signaling as means to enhance vaccination. Thus, this proposal will contribute to attainment of multi-project objectives of this U19.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] This is a 5-year training program renewal to support 5 postdoctoral trainees (Ph.D. and/or M.D.), 4 predoctoral students (Ph.D. or M.D./Ph.D) and 3 short-term minority (BS or in MS program) students per year for years 21-25 with an emphasis on a multidisciplinary approach to understanding hematopoietic cell regulation and disease mechanisms. The average training period for postdoctoral and predoctoral students will be 3 and 4 years, respectively. Twenty-three productive preceptors will provide extensive training in highly active and interactive research laboratories pursuing state of the art research on fundamental aspects of hematological, cellular, physiological, molecular, and biochemical aspects of cell growth and disease-related abnormalities. Preceptors are in 6 departments and 4 research centers housed in 4 buildings in close-proximity to each other. Strong track-records of peer-reviewed publications and extramural funding of preceptors are a major strength of the proposed training program, as are the training records of the preceptors. Opportunities for pursing multi-disciplinary research exist due to active, ongoing scientific collaborations among preceptors. A significant emphasis is to promote and foster the development of the next generation of biomedical researchers committed to scientific careers in academic medicine to become independent investigators. The approach for comprehensive training encompasses close and direct contact with seminars, laboratory meetings, and participation in national and international scientific meetings is already in place. The decision of a student to associate with a laboratory will be by mutual consent of trainee and preceptor. The scientiifc development of the trainee will be facilitiated by a research committee composed of preceptors with multi-disciplinary research interests. Major resources that are available include internationally recognized clinical and basic science research programs, an NCI-designated cancer center, an NIH-designated National Gene Vector Laboratory, an NIDDK Center of Excellence in Molecular Hematology, and state-of-the-art infrastructure, and frequent contacts with active investigators at IU and throughout the world [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Coronary heart disease is the leading cause of death in the United States. The elderly, those older than 65 years of age, are more likely to have coronary disease and this problem will become even more pervasive as the population ages. Stenting of coronary blockages has become a conventional technique in both the emergency setting of a heart attack as well as more routine therapy to reduce chest pain. In general, there are two types of coronary stents: drug-coated (or drug-eluting stents, DES) and non-drug- coated (or bare metal stents, BMS). Placement of either type of stent requires treatment with clot- preventing medications such as aspirin and clopidogrel or prasugrel to keep them open. The latter two are newer types of medications that work in conjunction with aspirin to reduce the function of cells that help form blood clots in the body (platelets), thereby preventing clot formation on stents (stent thrombosis). Currently, the American College of Cardiology / American Heart Association recommend continuation of these medications for at least one year following DES and for at least one month following BMS placement. Initial studies comparing DES to BMS showed that by reducing scar formation around stents, DES are less likely to close up and require repeat procedures. Recruitment of elderly in these studies was limited and subjects were generally healthier when compared to the community-dwelling elderly. Despite lack of such information, the elderly routinely receive DES during cardiac procedures, requiring the obligatory one-year of anti-platelet drugs. On average, the elderly are more likely to be taking multiple medications, have a lower body weight, and have worse kidney function than the relatively healthier population in randomized trials, thereby putting them at higher risk of drug-drug interactions and unwanted bleeding. Bleeding, which was originally considered just a nuisance in the early days of stenting, has more recently been recognized to increase mortality in patients with coronary heart disease. The balance of possible increased risk of repeat procedures with the use of BMS and potentially increased risk of bleeding with long-term use of dual antiplatelet therapy needs further investigation. The Dual Antiplatelet (DAPT) Study will randomize >20,000 patients to either 12 versus 30 months of dual antiplatelet drugs. This trial is unique in that it will study all-comers, there is no upper age limit, and will collect data on bleeding, interruptions of medications, and adverse cardiac events. We intend to study the DAPT Study elderly subset and compare differences in effectiveness and risk of dual antiplatelet therapy and stent types. Data on the rate of heart attacks, stroke, repeat procedures, and bleeding from a state-mandated registry which collects information on all stenting procedures in Massachusetts will also be analyzed for 3 and 4 year follow-up. Information obtained from this project will provide guidance to physicians caring for elderly patients toward stenting and medical treatment of coronary heart disease, particularly in regards to type and duration of antiplatelet drugs. PUBLIC HEALTH RELEVANCE: An important aspect of the management of coronary heart disease entails placement of drug coated and uncoated stents in the heart arteries to open up blockages. Medications required to prevent the stents from clotting can also increase the risk of bleeding and safety of these stents and medication in the elderly is not entirely clear based on current available data. The goal of our project is to study the safety and efficacy of these medications and stents in the elderly as part of a 20,000 patient randomized trial of these clot prevention medications and a statewide registry that manages information on all coronary stenting procedures done in Massachusetts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The early development of nerve-muscle synapses is characterized by three major events: (a) the accumulation of junctional acetylcholine receptors (AChRs), (b) the localization of synaptic acetylcholinesterase (AChE), and (c) the elimination of extrajunctional AChRs. This last event is mechanistically distinct from the accumulation of junctional AChRs and seems to result from the suppression of AChR synthesis in extrajunctional regions of muscle cells caused by muscle contraction. The final result is a muscle cell with a highly elaborated apparatus specialized for efficient synaptic transmission. Experiments in this proposal are designed to understand at the molecular level the way in which neurons and muscle cells communicate to establish this apparatus. The proposal is divided into three major sections. The first will make use of immunocytochemistry and antibody microinjection to demonstrate a functional association between the presence of a newly discovered muscle component (a 37 kilodalton nonmyofibrillar tropomyosin) and the ability to cluster AChRs. This molecule was first identified by its absence from vitally transformed muscle cells which are unable to cluster AChRs at all. The second section of this proposal describes similar techniques to probe other cytoskeletal elements involved in clustering and subsequent structural changes in the muscle cell. These studies are based on the observation from this laboratory that clustering causes a subset of organelles, including myonuclei and the Golgi apparatus, to assume a constant sub-cluster localization. The final section is a study of changes in the levels of AChRs and AChE caused by the increase in muscle cell Ca2+ which occurs during contraction. Also, experiments are designed to test the hypothesis that regional differences in the amount of Ca2+ released during contraction or in the levels of particular Ca2+-binding proteins underly the ability of these cells to specify where particular macromolecules are synthesized. Ca2+ concentration will be measured using the Ca2+-sensitive fluorescent dye fura-2 and optical image processing. Ca2+-binding proteins will be investigated using biochemical and immunological techniques. These experiments should add considerably to our knowledge of how neurons influence properties of their target cells and may contribute to an understanding of a variety of developmental and neurological disorders. In addition, certain results may provide information useful in comprehending cell transformation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this protocol are to compare the combination of 10-Ethyl-10- Deaza-Aminopterin (10-EdAM), mitomycin and vinblastin (EMV) with the combination of mitomycin and vinblastine (MV) in the treatment of patients with advanced Non-Small Cell Lung Cancer and to demonstrate that the two regimens are acceptable in terms of toxicity and to compare their relative toxicity. This is an open-label, multicenter, randomized trial.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The E4 allele of the Apolipoprotein E (ApoE) gene is the strongest genetic risk factor for the onset of sporadic Alzheimer's disease (AD) identified to date. The roles by which ApoE influences amyloid-beta (A2) metabolism and non-A2-mediated mechanisms in AD pathogenesis, however, remain to be fully clarified. The literature, and our preliminary data, suggest that early alterations in peripheral lipids (sphingolipids, fatty acids, cholesterol and cholesterol esters) reflect brain functioning and pathology, and may interact with ApoE genotype in the development of AD pathogenesis, warranting human studies. Clinical and epidemiological studies of short duration, while important, will not contribute to our understanding of the earliest phases of AD pathogenesis because Alzheimer's pathology (A2 plaques and neurofibrillary tangles) begins decades before the emergence of symptoms and substantial neurodegeneration. Identifying factors years before the onset of AD that may modify the effects of ApoE4 in initiating and promoting AD pathology and the subsequent emergence of symptoms will uniquely contribute to the development of prevention strategies. Longitudinal studies of cognitively normal individuals with serial measures of Alzheimer's pathology in the living brain, as proposed here in the unique cohort of the Baltimore Longitudinal Study of Aging (BLSA), initiated in 1958, are necessary to understand the relationship between ApoE4 genotype, perturbations in peripheral lipids, their interaction, and later development of AD clinical symptoms and brain alterations. The BLSA is one of few human studies that could provide the unprecedented opportunity to systematically examine this relationship over a long follow-up. BLSA participants, cognitively normal at their first visit in the study (mean age: 63.4), have a mean follow-up of 14.3 years (SD = 6.5) and a maximum follow-up of 38.9 years. In the proposed study we will measure plasma lipid levels (sphingolipids, fatty acids, cholesterol and cholesterol esters) at three early visit in the BLSA study, roughly 5 years apart, for those aged 55 and over (n=1095), and at the last visit, as well as during the neuroimaging sub-study. The specific aims include examining the proposed peripheral lipids, changes in these lipids over a long follow-up, and their interaction with ApoE to predict: 1) decline in tests of memory; 2) incident MCI, all-cause dementia, and AD; 3) change in serial MRI measures of brain atrophy and white matter lesion burden over 10 years; and 4) amyloid-beta deposition on 11C-PIB PET scans. The lipids will be assayed using an already-developed targeted and quantitative lipidomic approach. PUBLIC HEALTH RELEVANCE: Many potent risk factors for Alzheimer's disease (AD), including hypertension and high cholesterol, are most detrimental in mid-life, presumably at the emergence of AD pathology, but have less of an effect on AD risk in late-life. Identifying factors in mid-life that may modify the effects of APOE E4 in initiating AD pathology, and the subsequent emergence of symptoms, will uniquely contribute to the development of prevention strategies. The overall aim of the proposed study is to examine, in the 50-year Baltimore Longitudinal Study of Aging, whether plasma lipids measured in mid-life (sphingolipids, gangliosides, fatty acids, cholesterol and cholesterol esters) modify the association between APOE and cognitive decline, clinical onset of mild cognitive impairment or AD, and neuroimaging measures of brain atrophy and brain pathology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT Quantifying aging is a major goal in Geroscience research as the availability of a reliable marker of aging can facilitate understanding of the fundamental biology of aging, enable tracking of the aging process in different tissues and cell systems, and support identification and validation of interventions that extend lifespan and healthspan. Traditionally, aging has been monitored by following chronological age, mortality, age-related changes in gene expression, and/or other molecular features, however, there is currently no consensus on the best practices for quantitatively tracking progression through aging. The recent advent of biomarkers based on advanced omics approaches, such as DNA methylation, have provided some hope to support development of precise estimates of age, both in humans and mice. Nevertheless, the majority of such measures are trained as chronological age predictors, with little to no integration of biological, functional, or phenotypic data. Further, the modifiability of aging measures based on DNA methylation in response to lifespan and healthspan extending interventions is almost entirely unknown. We propose to address these challenges by developing a series of novel DNA methylation clocks by integrating information on phenotypic and functional aging, investigating links between DNA methylation and aging hallmarks, and evaluating DNA methylation responses to longevity interventions. We suggest that these clocks will offer a much-needed resource for the Geroscience community. We will develop these clocks using three general approaches. First, we will use cultured cells (MEFs) to induce or establish models of three well-known hallmarks of aging?cellular senescence, DNA damage, and mitochondrial dysregulation. We will then train epigenetic predictors of these hallmarks and validate them in vivo. We will also establish epigenetic alterations in response to novel and established longevity interventions. In doing so, we will develop biomarkers of intervention response that can be used to test mimetics, and/or optimize aging biomarkers. Finally, building on the highly characterized SLAM colony of C57Bl/6 and UM-HET3 animals, we will produce longitudinal methylation data across the lifespan that can be used to develop an epigenetic clock that can serve as a robust predictor of healthspan. We hypothesize that these new clocks will better capture biological age than chronological age trained clocks. Given that they were developed to capture different facets associated with the aging process, they can be combined to create a single aging measure that is more biologically informed and characterized compared to existing epigenetic clocks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "as fresh samples for isolation of normal organoids or breast stem cells. The pathology core will provide normal breast tissue for stem cell studies (Polyak) and for 3-D organoid cultures (Brugge). Freshtumor samples will be provided to establish primary tumor xenograftsin mice Weinberg). In addition, Dr. Richardson will participate in the pathologic comparisons, both molecular and histology, between the parent human tumors and the resultant xenografts. Gene expression array data obtained from human tumors will be used in system interaction analyses of BRCA1 relatedpathways (Livingston). Dr. Richardson will provide both tissue and scientific expertise to analyzefrozen tumor specimens and paraffin tissue microarrays for the expression of BRCA1gene products, including IRIS (Livingston). RELEVANCE (See instructions): Research using actual human breast tissue and breast tumor material will hopefully provide the most relevant answers to pathogenesis and tumor behavior of breast cancer in women. PROJECT/", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this study are 1) to test a group treatment program for women patients with borderline personality disorder (BPD) designed to decrease borderline symptoms and improve social functioning, and 2) to add to the understanding of the psychopathology of BPD by exploring a \"level of emotional awareness\" model to conceptualize its learning deficits. The specific aims of the proposed study are: 1) to test the efficacy of an 8-month, psychoeducation treatment program adjunctive to individual psychotherapy in reducing BPD symptoms, 2) to identify the characteristics of patients able to benefit from such treatments, 3) to explore treatment's mechanism of action, 4) to produce a treatment manual for the program, and 5) to provide training in the treatment of BPD. Forty-eight subjects who meet criteria for a BPD diagnosis using DSM-III-R, the Diagnostic Interview for Borderlines (DIB) and the Borderline Syndrome Index (BSI) will be randomly assigned to the treatment and waiting list control groups. The program's content is based on the deficits in emotional awareness, regulation, and expression and the consequent social, communication, and problem-solving skills like increased awareness of emotions, distress reduction, anger management, and need identification. It then moves on to interpersonal skills like emotional expression, communication, and conflict management. Treatment outcome measures include self-report (Symptom Checklist-90 SCL-90), Social Adjustment Scale-Revised), and structured interview measures DBI, Alexithymia Provoked Response Questionnaire). These measures will be given pre-treatment, post-treatment, and at a 6- month followup. A pilot study with nine subjects randomly assigned to treatment group and control provides strong support for the efficacy of this treatment program and the merit of a larger trial. Treatment group members improved significantly on all outcome measures (SCL-90, BSI, and Global Assessment of Functioning (GAF), while control subjects did not change significantly. This study will contribute to the much needed empirical evaluation of time-limited psychosocial group treatment for this difficult population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The specific aims of this Administrative Core include: 1) to provide the administrative structure to permit the effective day to day operation of the MCRC, 2) to provide the scientific expertise and leadership to identify and focus projects on clinically relevant issues and to insure that important unmet needs of underserved and minority patient groups are addressed, 3) to maintain and enhance internal communication and cooperation among MCRC members and to promote novel approaches to address clinically relevant issues, and 4) to expand the influence of the MCRC within the institution and to identify and secure institutional support to promote and enhance the activities of the Center. The Administrative Core will be housed in the Division of Rheumatology in the Department of Medicine. This Unit will be responsible for the overall direction, organization, operation and integration of the MCRC. The MCRC director, Dr. Richard Pope, will be assisted on a routine basis by the associate director, Dr. Rosalind Ramsey-Goldman, and the Methodology/Data Management Core director, Dr. Rowland Chang. The MCRC administrator, Mr. Christopher Loertscher, will assist in supervising and monitoring all budgetary issues and the scheduling of meetings by the executive committee, as well as MCRC sponsored research meetings and seminars, assisting in the preparation of reports and in dealing with regulatory matters. The scientific expertise and leadership within the Northwestern MCRC occurs at the level of the executive committee, that is chaired by Dr. Richard Pope and includes Dr. Roz Ramsey-Goldman, RowlandChang, Alan Dyer, Thomas Schnitzer, W. Zev Rymer, Lauren Pachman, and Rex Chisholm. A strength of this group is that it is composed of individuals with a variety of interests in clinical research on diseases relevant to the mission of the MCRC. The background that each member brings to the committee is quite different, which allows a truly multidisciplinary view of each issue. The committee meets bimonthly and considers all issues relevant to the scientific direction of the MCRC. The committee is responsible for identifying and pursuing new opportunities for interaction and development. These opportunities are particularly enhanced because of the roles of members of the executive committee both within the university community and nationally. It is through these interactions that the members of the committee are able to identify and promote important issues of national importance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose to study the development of drug resistance in the context of HIV protease inhibition to develop and test structural and synthetic strategies in response to this mechanism. The overall goal of this Program Project is to understand the mechanisms of viral resistance, enabling modeling and design of more sustainable anti-viral therapeutic strategies. The Program consist of four highly integrated Projects and three supporting Core facilities: Project 1, will enhance and extend a computational co-evolution approach to drug resistance by developing and applying detailed atomic models of drug/target interactions, modeling viral population dynamics and patient response under drug selection pressure, exploiting automated learning and hidden Markov modeling approaches to inform and refine these models; Project 2 will exploit the capabilities of high throughput crystallography to find and characterize novel binding sites on the protein target using fragment libraries to help construct new inhibitor leads to maintain efficacy against multi-site PR mutants, linking with optimization and synthetic efforts in Projects 1 and 3 respectively; Project 3 will utilize their \"Click Chemistry\" synthetic approaches for rapid development and evolution of novel fragment-based inhibitors in conjunction with Projects 1 and 2, and develop resistance probes with Project 4; Project 4 will experimentally characterize the evolution of HIV resistance in response to protease inhibition both within PR and in the rest of Gag-Pol, by exploiting tissue-culture time-course evaluation passaged virus in the presence of identified inhibitors, as well as from deep genetic analysis of selected patient samples; Core A will provide mutant and wildtype proteases, functional assays and chemical probes, and inhibitor analyses for the Program; Core B will provide the necessary x-ray structural data and computational analysis to integrate new information on protease mutants, and protease-inhibitor interactions; and Core C will assemble and make available to projects 1 and 4 time-course anti-retroviral treatment data on HIV infected patients as well as blood samples from highly resistant individuals for in-depth bioinformatic and viral genomic analyses. AIDS remains the major pandemic of our time. While patients infected with HIV can now be treated with drugs that enable them to live productive lives, the virus can subvert this treatment by developing resistance to these drugs. This study is aimed at a detailed understanding of HIV drug resistance, with the goal of developing new therapeutic strategies for more sustainable treatments to prevent AIDS. [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fear of intracranial hemorrhage (ICH) is the primary reason that the only proven therapy for acute ischemic stroke is not provided to otherwise eligible patients. ICH also deters the development of more effective stroke therapies. New reperfusion strategies, using an intra-arterial (IA)approach, which may restore blood flow more effectively by providing therapy directly at the site of an occluded artery, are accompanied by higher rates of the mildest forms of ICH than proven intravenous thrombolytic therapy. While ICHs of all types have been considered adverse events in every acute stroke trial to date, the mechanistic significance and clinical impact of mild ICH subtypes is not known. The primary hypothesis is that acute ischemic stroke patients treated with IA therapies who have the mildest radiological subtype of ICH will be more likely to have early reperfusion and good clinical outcome than those without any ICH. Using the largest available trials with revascularization status, the PI will pursue the following specific aims: (1) determine the relationship between radiological subtypes of ICH and timing of successful revascularization and (2) determine the relationship of ICH subtypes to clinical outcome. The proposed work would: (1) guide the clinician's perceptions of the interplay between risk and benefit in the setting of reperfusion, (2) impact the interpretation of future Phase I and II reperfusion trial safety data, and (3) inform the design of future reperfusion strategies using hemorrhage prediction and hemorrhage prevention strategies. This research project will position the PI to independently design and implement an acute stroke trial with the goal of minimizing clinically significant hemorrhage. With this career development award, the PI will gain practical experience with a large-scale Phase III acute stroke clinical trial, obtain exposure to innovative reperfusion strategies, and pursue formal training in biostatistics and clinical trial methodology through introductory and advanced coursework. The PI will become an expert on acute stroke reperfusion therapies, and particularly hemorrhagic transformation, with the skills to implement an acute stroke clinical trial independently. Relevance to Public Health: Despite the availability of an effective treatment for strokes due to blocked arteries (88% of all strokes) since 1996, stroke remains the third leading cause of death and the leading cause of disability in the United States. Therapies that restore blood flow to more patients, and do so more effectively and safely, are greatly needed to improve outcomes after stroke.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A group of seven qualifying Primary Users have detailed a broad range of experimental plans that specifically employ BIAcore analysis as summarized herein: (i) Characterization of association and dissociation rate constants for the interaction of immunoglobulin E (IgE) and the high affinity IgE receptor (FcepsilonRI) is a major goal of the PI. The effect of point mutations on alterations of k/on and/or k/off values will be determined by measurement of phage-displayed peptide binding to sensor chips C1 and/or F1 covalently derivatized with either recombinant IgE receptor ectodomain or IgE; (ii) Determination of the kinetics of the interaction between the T cell receptor (TCR) and its ligands, specifically MHC class I-peptide complexes. To extend our recently published study of peptide/Class I-TCR binding kinetics, which showed a correlation between the affinity of TCR for particular ligands and the biological outcome, we plan to determine TCR-ligand recognition in lipid monolayers using the HPA sensor chip to model cell-cell TCR-MHC/peptide mediated interactions; (iii) The superoxide generating system of neutrophils is a membrane-associated enzyme complex, NADPH oxidase, which consists of an integral membrane protein, flavocytochrome b245 and cytosolic proteins p47phox, p67phox and Rac. BIAcore analysis will be used to study the molecular interactions of the cytosolic and membrane factors, individually and in combination, and to determine how molecular interactions of the cytosolic and membrane factors, individually and in combination, and to determine how molecular interactions of the cytosolic and membrane factors, individually and in combination, and to determine how molecular interactions of the cytosolic and membrane factors, individually and in combination, and to determine how their affinity is affected by anionic lipids, ionic strength and GTP; (iv) The precise role of Rac in the foregoing system is still unknown, but one current hypothesis is that the association of GTP-bound Rac with p67phox increases the affinity of this protein for the flavocytochrome. This will be tested by determining the kinetics and affinity of binding of p67phox to immobilized, purified, relipidated flavocytochrome in the presence and absence of Rac-GTP in the analyze solution; (v) The energetic processes that drive the binding reactions for DNA and RNA will be determined using the zinc finger protein TFIIA, and truncated or mutated derivatives, in solution with either 5S DNA or 5S RNA immobilized on a BIAcore chip; (vi) Surface plasmon resonance technology will be utilized to study fibroblast growth factor (FGF) receptor- mediated signal transduction. Several proteins, identified using the two-hybrid system, show affinity for the cytoplasmic domain of FGFR-1. The binding components from the 2 hybrid analysis will now be analyzed using the BIAcore system to determine the kinetic binding parameters. (vii) Autoantibody binding to wildtype and mutant fibrillarin, in the presence and absence of mercury, using the BIAcore machine. Quantitation of Hg-dependent changes of antibody k/on and dissociation (k/off) from fibrillarin immobilized on the biosensor chip will then be assessed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Center engages in collaborative research that leverages the Center's molecular modeling software, expertise and facilities to bridge experiment and computation. While experimental techniques unveil critical aspects of molecular and cellular architecture, computational molecular modeling describes biomolecular systems in their native states at the atomic level. During the last funding period, the Center completed 35 Collaboration and Service projects with 41 joint publications on subjects such as the assembly mechanism and drug resistance of the ribosome, neurons' synaptic receptors and solid-state DNA sequencing nanopores. Currently, the Center is engaged in 42 ongoing collaboration projects with 56 joint publications so far. To make the Center's technological advances driven by the collaboration projects available to the biomedical community, the Center distributes its software for all major platforms, from laptops to supercomputers and cloud computing facilities. The Center ensures software usability by providing user support, maintaining up-to-date documentation, tutorials, and online discussion groups. Over the last funding period, the Center-developed molecular modeling and simulation packages VMD, NAMD, and Lattice Microbes served a community of over 100,000 users. Technical support for the community was provided mainly through the online discussion groups ?NAMD-L?, ?VMD-L? and ?LatticeMicrobes-Users?, where more than 17,000 software-related messages were submitted over the last funding periode. The Center also o?ers individual consultation for special cases requiring direct personal interaction, operating directly in its laboratory area a well-equipped Visitor Center to host visiting scholars and collaborators. 50 scholars/collaborators visited the Center for up to six months seeking technical and scienti?c expertise, and 107 leading scientists presented their work on the Center Seminar program at the host institution, the UIUC Beckman Institute. All the activities described above were possible only through the Center's Petascale Biology Gateway Facility, which provides the high-end computing and graphics resources required by the demanding preparation, visualization, and analysis tasks of advanced molecular modeling. This gateway also hosts the Center's website and all associated content, from scienti?c research-related material to software documentation and tutorials, as well as the online discussion platforms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT 1 (MONTINE): Abstract Genetic risk for PD-related cognitive impairment and its disease mechanisms The inaugural PANUC award was funded to pursue the association of three candidate genes with cognitive impairment and dementia in PD: APOE ?4 allele and variants in SNCA and MAPT; to these we added GBA and LRRK2 mutations. With our collaborators, we demonstrated that cognitive impairment and dementia are significantly more common among PD patients who carry APOE ?4 or GBA variants, less common in patients with LRRK2 mutations, and not associated with variants in MAPT or SNCA. In this project our approach will be to bring insight to disease mechanisms by determining the corresponding molecular pathology. During the current PANUC award, we and others used standard histopathologic criteria for Lewy bodies (LB) or Alzheimer's disease (AD). Unfortunately, this approach has led to unexpected and conflicting results for GBA variants and APOE ?4 in PDD in part because some studies were limited by low number of cases or inconsistent attention to genetic risk. Specific Aim 1 proposes to fill these gaps in knowledge by regional quantification of neurotoxic proteins and synaptic degeneration using a novel technique developed by our laboratory coupled with a large, multisite autopsy cohort assembled by PANUC. Specific Aim 2 reflects that although successful with our candidate gene approach during the inaugural PANUC award, we propose to expand gene discovery for cognitive impairment and dementia in PD by evaluating additional candidate genomic regions recently identified by the PDCGC, and by determining genetic influences on longitudinal change in cognition measured prospectively using consensus assessments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a response to Notice Number NOT-OD-09-058, NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. The goal of this revision supplement is to extend our studies of nicotine vaccine efficacy in rats by introducing methods for the delivery of nicotine through inhalation of cigarette smoke. Smoking cessation medications have added substantially to our ability to treat tobacco addiction, but their efficacy is limited and new types of medications are needed. Nicotine vaccines elicit nicotine-specific antibodies which bind nicotine and alter its access to brain. Three nicotine vaccines have shown preliminary efficacy in Phase I-II clinical trials, but efficacy is closely correlated with the serum antibody titer and current vaccines do not reliably produce sufficiently high titers in all individuals. The parent grant DA10714 is using rat models of nicotine addiction to study novel means of enhancing vaccine efficacy. Like essentially all current animal studies of nicotine addiction, DA10714 models tobacco addiction using parenteral (i.v. or s.c.) administration of pure nicotine. In contrast, smokers take in nicotine by inhalation and as one of over 4,000 chemicals present in cigarette smoke. The adequacy of using such artificial dosing paradigms to model cigarette smoking is unknown and largely untested. We have adapted and characterized methods for inhalation exposure of rats to smoke simulating the smoking of 1 cigarette or periods of heavier smoking. In this revision supplement we propose to use these methods to study the effects of nicotine vaccines on the absorption and distribution of nicotine inhaled in cigarette smoke. The purposes of doing so are to 1) expand the range of preclinical models available to study nicotine vaccines, 2) assess whether inhalation models provide novel information for vaccine evaluation, 3) examine the specific role of route-specific factors such as pulmonary antibody in mediating nicotine vaccine efficacy, and 3) develop quantitative models which can be more generally used to study the contributions of the inhaled route and other smoke constituents to tobacco addiction and treatment medications development. Nicotine vaccines are an attractive initial candidate for such study because vaccination is a pharmacokinetic intervention, and accurate pharmacokinetic modeling of nicotine intake may be important in understanding and exploiting its efficacy. Aim 1 will test the hypothesis that vaccination is effective in reducing nicotine distribution to brain over a range of clinically relevant dosing conditions. Aim 2 will test the hypotheses that vaccination is more effective in reducing the distribution to brain of inhaled compared to i.v. nicotine, and that such differences are in part mediated by the presence of pulmonary mucosal or tissue antibody. Because heroin and cocaine are also often smoked, the results of this study may inform ongoing efforts to develop vaccines for these addictions as well. PUBLIC HEALTH RELEVANCE: Cigarette smoking kills 5 million people worldwide yearly. Current medications are helpful for smoking cessation but are incompletely effective. We are studying the use of a nicotine vaccine to help smokers quit, which acts by binding nicotine in blood and reducing its access to brain. Rat models of tobacco addiction use nicotine delivered intravenously, which is informative but does not accurately model the human route of intake of nicotine by inhalation from cigarette smoke. This proposal will develop and study the utility of delivering nicotine to rats via exposure to cigarette smoke, and assess whether it allows better evaluation of nicotine vaccine efficacy and can expedite its further development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project is designed to investigate long-term regulation in brain of the enzyme choline acetyltransferase (CAT), catalyzing the biosynthesis of the neurotransmitter, acetylcholine. We shall study the biochemical basis for the observed multiplicity of molecular species of CAT and the mechanisms governing the regulation of its activity. To accomplish this, we shall (a) prepare and purify antibodies to the different forms of CAT found in rat brain; (b) analyze regionally in the brain the molecular nature of the different forms; (c) characterize the biochemical properties of each form; (d) determine their turnover rates; and (e) establish whether the mechanism of altered CAT activity affected by psychoactive drugs, hormones or brain lesions, as well as during growth and development, is one of induction (increase in enzyme protein) or activation (increased catalytic activity/molecule).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The 5-nitro drug, metronidazole (Mz), has been a mainstay of antimicrobial therapy for decades. Several of its simple derivatives such as tinidazole combine similar activity profiles with improved pharmacokinetic properties, but resistance to existing nitro drugs is increasing. Although commercial development of this drug class largely ceased decades ago, work by us and others over the last several years has shown that extensive modifications of the basic 5-nitroheterocyclic ring can lead to marked enhancement in activity against different microbes compared to existing drugs. These data suggest that Mz and other approved nitro drugs do not possess optimal activity in this drug class, yet important questions about the potential utility of novel nitro compounds must be addressed to advance their development as next-generation nitro drugs for clinical use: Is it possible to develop improved nitro drugs with broad-spectrum activity, or do enhanced activities exist only in microbe-specific fashion? Do new nitro drugs have different targets that can be exploited for overcoming resistance to existing drugs? What are the optimal pharmacokinetic properties of novel nitro drugs for maximal efficacy and potency against infections with different target microbes? Can new nitro drugs be developed with improved dosing regimens compared to existing drugs? Answers to these questions are not only critical for assessing the therapeutic potential of new nitro drugs, but are also key for identifying new leads for specific indications. The project will address these questions with a focus on two important protozoan pathogens, Trichomonas vaginalis and Giardia lamblia. We will evaluate a newly synthesized library of ~1,200 nitro drugs for activity against a broad range of drug-sensitive and drug-resistant strains of the target protozoa to identify library compounds more potent than Mz. Electrochemical approaches will be employed for determining the redox properties of the most potent nitro compounds to gain new fundamental clues about their mechanisms of action and potential toxicity. Subsequently, we will introduce new structural modifications into the top leads and evaluate them for bioactivity, cytotoxicity, electrochemical characteristics, and propensity to develop new drug resistance. Finally, we will evaluate the most promising nitro compounds for efficacy, potency, and pharmacokinetics in different murine models of protozoal infections. Upon completion of the proposed research, we expect to have elucidated broadly applicable principles that govern optimal efficacy of next-generation nitro-heterocyclic agents in the treatment of the clinically important parasitic diseases trichomoniasis and giardiasis. The comprehensive data sets to be generated will also be instrumental in selecting the most promising candidates as novel leads for the improved treatment of these infections, and potentially infections with other important pathogens, including Entamoeba histolytica, Trypanosoma cruzi, Helicobacter pylori, and Clostridium difficile, which can be treated with nitro antimicrobials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Antisense phosphodiester oligonucleotides, conjugated with acridine, and antisense methylphosphonate oligonucleotides, killed cultured Trypanosome brucei in the Phase I program. In the Phase II program, antisense olignucleotides will be edified at the base, sugar, and phosphodiester functional groups to increase their therapeutic potency. These modifications will be designed to increase oligonucleotide cell penetration, stability, and functional inhibition of target RNA or DNA. The nucleic acid target will be expanded beyond the 39 nucleotide spliced leader sequence of trypanosome mRNAs to include the small nuclear RNAs, the coding and non-coding of several mRNA sequences, and the actin gene promoter region. Modified oligonucleotides then will be tested against the trypanosome procyclic form in culture, and those with enhanced therapeutic activity against the bloodstream form. Active oligonucleotides will be tested for therapeutic efficacy using trypanosome infected mice. Positive results will be of interest to the pharmaceutical industry for the development of new antisense oligonucleotide chemotherapeutic agents for trypanosomes and other parasites.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The WHO Collaborating Center for Design, Methodology and Analysis of Epidemiological and Clinical Investigations in Diabetes was designated in 1986. The purposes of the Center are to collaborate with the World Health Organization in the implementation of the WHO/IDF action program to provide advice, consultation and collaboration with other investigators in the design, methodology and analysis of epidemiology and clinical investigations relating to the etiology and pathogenesis of non- insulin dependent diabetes (NIDDM) and its complications. The center will assist in the development and application of standardized methods for epidemiological and clinical investigations, and data analysis relating to diabetes and collaborate with those interested in applying such techniques elsewhere. The Center will advise and help in the design of new studies, including onsite assistance when necessary. The center serves as a central laboratory for the WHO Multicenter Study of Vascular Disease in Diabetes, as well as being a participating study center for this study which is examining the mortality and incidence of vascular complications of diabetes among different ethnic groups in different countries. In addition the center has initiated a collaborative study of impaired glucose tolerance in China, is collaborating in the preparation of a survey manual for diabetes mellitus on behalf of WHO Center personnel are participating in teaching a WHO sponsored course on Clinical Epidemiology and Public Health Aspects of Diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "As implemented, outside of a handful of reference institutions, electronic health record (EHR) systems have not been conclusively shown to improve health outcomes. The New York City Primary Care Information Project (PCIP) is the nation's largest community EHR extension project, with 148 primary care practices and over 1,000 providers in some of the city's poorest neighborhoods. The PCIP's goal is to maximize improvements in the quality of care through this investment of public funds. This public health perspective has led to development and implementation of clinical decision support and integrated registry tools, quality benchmarking, onsite technical assistance in quality improvement and practice redesign, and a novel reward and recognition program. In essence, the PCIP aims to provide even small independent practices with the health system advantages enjoyed by integrated health care organizations. The PCIP is evaluating trends in priority quality measures (e.g., blood pressure and lipid control) among 90 small practices randomized to receive clinical decision support tools and/or financial rewards. This proposed study of 60 comparison practices that have not adopted EHRs will provide a unique opportunity to demonstrate the potential of appropriately implemented health information technology for improving quality of care and reducing health disparities in a large community EHR program.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "(Revised Abstract) Description: The rapid spread of HIV in many Eastern European countries has been linked to an increase in injecting drug use among young people. Non-injecting drug use has also increased among young people in this region. Non-injecting drug users are both at risk of initiating injecting drug use and of acquiring through sexual transmission not only HIV but also other sexually transmitted infections. Research is needed into the factors that increase the risk of HIV transmission among young drug users in this region. Such research is needed not only in countries where there is considerable HIV infection among IDUs, but also in countries, such as Hungary, which may be in a pre-epidemic phase, i.e. where HIV prevalence is currently low, but where behavioral and network risk is common among young drug users. In the proposed research, we plan to conduct a pilot study of HIV risk among young drug users between the ages of 16 and 30 in Budapest, the largest city in and the capital of Hungary. The specific aims of the study are to: 1. use ethnographic methods to explore (i) the drug use and sexual risk behaviors and practices of young drug users, (ii) their risk and social networks, especially injecting and sexual mixing patterns with people from high HIV prevalence groups or countries; (iii) the knowledge, attitudes, beliefs and peer norms of young drug users about HIV risk and prevention and, (iv) among young drug injectors, the behaviors, network characteristics, personal biographies and circumstances that may be linked to their initiation of injecting drug use; 2. develop and administer a structured questionnaire to a small sample of young drug users that will utilize questions that are based on the findings from the ethnographic study and questionnaires used in our previous studies; 3. develop and test methods for sampling and recruiting \"hidden populations\" of young drug users. The ethnographic part of the study will utilize focus groups, in-depth ethnographic interviews, and observational techniques. In addition, existing secondary data will be collected and synthesized. A structured pilot questionnaire and methodology for the sampling and recruiting of study participants will be developed. Procedures for HIV, HBV and HCV testing and counseling will also be developed and assessed. All data collection instruments, protocols, reports, presentations and papers will be both in Hungarian and English. HIV prevention in Eastern Europe needs to target populations that are the most vulnerable for becoming infected, particularly young injecting and non-injecting drug users. This study will provide new knowledge on young drug users in Hungary and will also contribute to advancing research methods and capability in other Eastern European countries.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Examine differences between pre-term and full-term human milk. This study is also designed to identify material factorswhich may influence the concentrations and determine changes in concentrations which may result from storage of breast milk.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Children's Cancer Study Group is a multi-disciplinary organization formed to cooperate in research into malignant diseases in children: specifically, to study the factors which may cause or influence these diseases, and to develop, evaluate and improve methods of treatment. The primary objective of the cooperative group has been the evaluation of new biological and chemotherapeutic agents in the treatment of acute leukemia and solid tumors in childhood, and finding new and better ways of using the already known agents. This evaluation is done by pediatric oncologists, surgeons, radiotherapists, pathologists and immunologists within the cooperating institutions which make up the group. It is also our objective to provide supportive care for these children with leukemia and solid tumors which will enable all children who desire to do so to participate in our protocol studies. This supportive care includes financial supportive care (domiciliary care, travel assistance, outpatient costs, etc.), social service assistance, as well as medical supportive care.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies on long-term effects of two peroxisome proliferators inducing high and low carcinogenic activities respectively, indicated that factors other than oxidative injury may be important in the carcinogeneity of such chemicals. Speculation on the existence of specific receptors for mediating the action of peroxisome proliferators has recently been supported by the isolation of new members of the steroid hormone receptor superfamily which are activated by peroxisome proliferators: the mouse peroxisome proliferator-activated receptor (PPAR), the Xenopus laevis PPAR and the rat PPAR. Moreover, it has been shown that PPARs are capable of activating the promoter of the rat acyl coenzyme A oxidase gene, the key enzyme of peroxisomal fatty acid beta-oxidation, and mediates the induction of rabbit CYP4A6, a cytochrome P450 fatty acid omega- hydroxylase, by clofibric acid. This activation is carried out through specific response elements that have recently been identified. Our approach has been to examine whether differences between human and mouse PPAR could account for the marked differences in responsiveness to peroxisome proliferators on human hepatocarcinogenesis. To achieve this goal, we isolated the human PPAR from a human liver cDNA library and compared its structure and ability to activate the ACO response element with that of mouse PPAR. B-lymphoblastoid cells were used to construct a system which can be easily used to determine whether a chemical will be a peroxisome proliferator. The system uses an episomally-replicating plasmid containing the human PPAR cDNA under control of the thymidine kinase gene promoter, and a reporter gene having a PPAR response element driving expression of the CYP2A6 cDNA. A variety of constructs and promoters were tested to achieve maximal differences between untreated and peroxisome proliferator-treated cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Breathing is a remarkable behavior fundamental to life that mediates gas exchange to support metabolism and regulate pH. A reliable, non-stop, robust rhythmic pattern of respiratory muscle activity is essential for breathing in mammals. Failure to maintain a normal breathing pattern in humans suffering from sleep apnea, apnea of prematurity, congenital central hypoventilation syndrome, hyperventilation syndrome, Rett syndrome, and perhaps Sudden Infant Death Syndrome, leads to serious adverse health consequences, even death. Various neurodegenerative diseases, such as Parkinson's disease, multiple systems atrophy, and amyotrophic lateral sclerosis, are associated with sleep disordered breathing that we hypothesize results from the loss of neurons in brain areas controlling respiration. If breathing is to be understood in normal and in pathological conditions, the mechanisms for respiratory central pattern generation must be revealed. We focus on two brain sites essential for generation of the normal breathing pattern, the preBtzinger Complex and the retrotrapezoid nucleus/parafacial respiratory group. We propose a broad series of experiments both in vivo and in vitro in rodents using advanced techniques including: viral delivery to express genetically encoded opsins or DREADDs in key subpopulations of neurons in these regions; advanced optical techniques to determine the contributions of the preBtzinger Complex microcircuit to rhythm generation; state-of-the-art neuroanatomical techniques to establish, in appropriate and necessary detail, the interconnectivity of the brainstem respiratory pattern generator. The data from these experiments will provide an extraordinary window into the mechanisms underlying respiratory rhythm and pattern generation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We are entering an exciting era in pediatric rheumatology. New treatment approaches are improving the lives of children with juvenile idiopathic arthritis (JIA) to such a degree that it's now rare to see wheelchairs or crutches in our waiting rooms. Even splints, commonly used in the past to treat joint contractures, are seldom seen on our patients. Despite our progress, remission in JIA is rare. Recent work by our collaborator, Dr. Carol Wallace, has shown that only 5% of children with the polyarticular JIA (the most severe form of this disease) are in remission 5 years after diagnosis. Part of our problem in achieving remission is that, at the biological level, we don't really understand what \"remission\" is. It's a classic case of the difficulty of getting somewhere when you don't really know where you're trying to get. This grant application is about learning where we are trying to get. In this application, we aim to achieve a better understanding of what \"remission\" is using microarray-based biomarkers. Research from the Cobb (acute inflammation) and Jarvis (chronic inflammation) laboratories has demonstrated the feasibility of using genome-wide expression profiling can be used to define disease \"states\" (e.g., infected vs. not infected;in remission or not in remission). Furthermore, the Jarvis laboratory has demonstrated the promise of using these same technologies to predict clinical outcomes. For each group, these promising preliminary studies must be validated using larger patient populations and prospective study designs. In this application, we propose to validate peripheral blood biomarkers that already suggest that: (1) remission in juvenile arthritis can be identified at the molecular level through distinct gene expression signatures;(2) those signatures include the balance of both pro- and anti- inflammatory gene networks;and (3) the clinical course of children who reach an inactive disease state can be predicted based on molecular signatures that emerge in the peripheral blood mononuclear cells. Furthermore, we will take another step toward clinical application of this work by developing PCR-based whole blood assay to identify the most robust indicators of remission or clinical outcome. This project brings together two experienced investigators from two very different disciplines: pediatric rheumatology (Dr. Jarvis) and surgical intensive care (Dr. Cobb). Furthermore, the project brings together two computational biology groups spanning multiple disciplines, as well as other experienced pediatric rheumatology investigators. Thus, the project is highly responsive to the goals of the most recent NIH roadmap.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Adapted from the Applicant's Abstract.) This project is designed to describe some salient characteristics of mammalian cochlear hair cells. There are two types of such cells in our ears, inner hair cells and outer hair cells. The former are thought to be the true sensory receptors of the ear; they convey auditory information to the central nervous system. Outer hair cells, in contrast, may have primarily a mechanical effector (feedback) role in that they modify the mechanical input to the inner hair cells. To be studied in this project is the stimulus-response relations (electrical response versus hair deflection) for the two hair cell types in isolated organ of Corti segments. Also proposed is to study the mechanical motile response of outer hair cells to electrical stimulation and to hair deflection. These studies are conducted on single isolated outer hair cells for which a real life-like electrical-chemical-mechanical environment is provided. Hair cells are the key elements in the hearing process and their defects are the cause for the vast majority of hearing loss and deafness. Understanding their properties is a prerequisite for the eventual remediation of most hearing disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] The long-term goal of the proposed research is to develop an implantable polymeric device, which delivers gonadotropin releasing hormone(GnRH, aka leuteinizing hormone release hormone, or LHRH) in periodic pulses, mimicking the normal, ultradian endogenous rhythm. Rhythmic pulsed administration of GnRH has been used successfully in the induction of ovulation in amenorrhic women, and in the induction of puberty and maintenance of sexual maturity in both males and females with hypogonadotropic hypogonadism (HH), while continuous administration of GnRH fails. Present pulsatile treatment is administered using wearable pumps, with cutaneous breach by a catheter. For patients who must receive pulsatile GnRH for many years, this mode of delivery leads to significant inconvenience and danger of infection. The proposed device could be implanted subcutaneously or intraperitoneally, and may function for months or years. It is hoped that implantation will ameliorate the disadvantages of pump/catheter systems. The device concept features a hydrogel membrane, which undergoes periodic swelling-deswelling cycles, due to a feedback interaction between the membrane with the enzyme glucose oxidase. Release occurs during the swollen phase of the cycle. Oscillations are \"fueled\" by endogenous glucose at the site of implantation, whose level fluctuates only moderately in nondiabetics. (Rhythmic behavior of this device is distinguished from the glucose-responsive behavior of closed-loop insulin delivery devices.) While the feasibility of this concept has been confirmed in a model system, considerable work remains in converting the concept to a workable device that can be tested in patients. The specific goal of this proposal is to produce a prototype system whose properties can be tested in an in vitro environment that emulates in vivo conditions. In pursuing this goal, effects of the hydrogel membrane composition and thickness, the enzyme reaction, the device geometry, and the molecular properties of GnRH on the pulsatile release pattern will be studied. We will investigate device behavior in buffers and simulated plasma, when challenged with constant levels of glucose and programs of glucose fluctuations that are observed in nondiabetic individuals. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mouse leukemia cells bearing TL surface antigens escape from destruction in mice immunized against those antigens, but the same cells are effectively killed when other antigens (e.g., H-2) serve as targets. Correlating with escape is the tendency for TL plus tumor cells to acquire resistance to TL antibody-\\and guinea pig complement-mediated lysis following exposure to TL alloantiserum (antigen modulation or evasion). Evasion also affects some mouse lymphomas expressing murine leukemia virus major envelope glycoprotein, MuLV gp70. Evasion-positive tumors grow in all syngeneic mice immunized against gp70 by passive immunization with xenoantisera, whereas evasion-negative tumors are rejected by some immunized mice and growth is markedly suppressed in others. Evasion-positive and -negative variants of FLC745 and RBL-5 lymphomas are being studied to establish the relationship between antigen evasion and tumor escape. Xenoantisera, mouse antisera, and monoclonal antibodies to gp70 and other MuLV components expressed on the cell surface will be utilized, and evasion of cytolysis will be measured in a radiochromium release assay. Antigen lateral mobility, antibody-induced antigen aggregation, and steric hindrance of complement binding (guinea pig Clq) are apparently responsible for evasion. Factors not directly related to antigen evasion include: (1)\\quantitative representation of target antigens before and during antibody sensitization and quantitative aspects of antibody binding; (2)\\manner of antigen presentation at the cell surface, molecular associations, and molecular and ultrastructural organization; and (3)\\differences in detected antigen specificities. Defense mechanisms involved in rejection of evasion-negative tumors are being examined, and a fully homologous (mouse antisera and complement) antigen evasion assay is being developed. Short-term gp70 antigen evasion appears to result from lateral mobility/aggregation of gp70 molecules influenced by surface membrane dynamics, motile cell activity, and cytoskeletal elements as well as antibody binding. There is no apparent involvement of complement (C3), differences in gp70 antigenicity, surface representation or configuration, differential cell sensitivity to complement lysis, or different cell growth characteristics. (IS)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research seeks (1) to explore the development of auditory perceptual abilities, specifically pitch and timbre perception, during infancy, and (2) to delineate some of the acoustic cues infants can employ in those tasks. Both pitch and timbre perception appear to require the integration or synthesis of information gleaned from a peripheral analysis of the spectral characteristics of sound, whereas spectral analysis alone permits simple discrimination of sounds. On the basis of the animal literature, adult psychoacoustics work, and the available infancy research, it can be hypothesized that the synthesis of acoustic information will show dramatic developmental changes during early life. Furthermore, it appears that the ability to analyze sounds into their spectral components precedes the ability to integrate that information in the service of perception. The studies of pitch perception will attempt to establish the emergence of pitch for complex harmonic and inharmonic sounds, including the perception of the missing fundamental. Specifically, these studies will (1) evaluate pitch extraction from harmonic tonal complexes in 3 month olds, and (2) from inharmonic tonal complexes in 7 month olds; (3) determine whether 7 month olds can use spectral information to categorize tonal complexes, when they are unable to integrate that information; (4) determine whether 7 month old infants can use high frequency energy to extract pitch from tonal complexes; and (5) employ a psychophysical technique to assess the extent of the dominance region for pitch in 7 month olds, and (6) in adults. Since very little is known about timbre perception in infancy, an initial study will investigate infants' categorization of sounds on the basis of timbre, but in the presence of irrelevant variations in pitch. Infants from 3 to 8 months of age will be tested in an operant conditioning procedure. Since the proposed research will investigate both analytic and synthetic processing of sound, it is hoped that the general pattern of auditory development and the mechanisms underlying it can be uncovered. In doing so, the proposed research will provide a necessary preliminary step toward the formulation of a general model of auditory development. Furthermore, the brain-behavior relations uncovered will contribute to our understanding of the behavioral consequences of brain damage in human adults and children.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary objective of this study is to test whether the administration of supplemental oxygen to infants who develop moderate retinopathy of prematurity (ROP) will result in a reduction of at least one third in the number of infants with eyes progressing to threshold ROP. Infants receive continuous pulse oximetry saturation monitoring at one of two specified oxygen levels with weekly active at the Hospital of the University of Pennsylvania as part of a consortium of four intensive care units. Eighteen have been enrolled to date since recruitment began in 1994. Participation in this protocol will continue with enrollment of eligible patients. Ophthalomologists will continue ROP screening exams in the nursuries and will alert study personnel of babies who are at risk for developing pre-threshold ROP.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this study is to investigate the mechanisms by which oncostatin M (OSM) promotes vascular endothelial cell growth factor (VEGF) in human breast cancer. Oncostatin M is a member of the interleukin-6 (IL-6) family of cytokines and is produced by many immune cells and some tumor cells. Preliminary studies on human breast cancer cells indicate that OSM promotes expression of the potent proangiogenic factor VEGF. OSM also induces expression of the alpha subunit of hypoxia-inducible factor 1 (HIF1), a critical transcription factor for VEGF gene regulation. Our recent data and other published studies suggest that OSM may promote angiogenesis, tumor progression, and metastasis in vivo. In this grant, we propose to determine the multiple mechanisms by which OSM promotes VEGF expression in human breast cancer. To accomplish this, we will investigate OSM's induction of HIF11 in breast cancer cells and other cell types and confirm that OSM-induced VEGF expression is dependent on HIF11. Additionally, we will study the mechanism behind and the signaling pathway utilized by OSM to induce HIF11. We will also determine why, in contrast to OSM, another IL-6 family cytokine, leukemia inhibitory factor (LIF), induces HIF11 but does not promote VEGF expression. Results demonstrating that OSM activates multiple pathways to upregulate VEGF would provide insight into cytokine-induced angiogenesis in tumor cells and supply rationale for the design of breast cancer therapies that inhibit OSM expression, function, or signaling. The potential to develop experimental anti-OSM therapeutics is limitless: to date, there has been no attempt to inhibit OSM for the purpose of cancer therapy. PUBLIC HEALTH RELEVANCE: The proposed work could establish an important role for oncostatin M (OSM) in breast cancer progression and metastasis. This research could provide the rationale for the development breast cancer therapeutics that target OSM to prevent metastatic spread of the disease. In addition, this project represents the opportunity for undergraduate students at Boise State University to participate in biomedical research and gain a deeper understanding of the molecular and cellular events involved in breast cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hydrophobic membrane proteins perform a variety of functions in the cell, but their structures are notoriously difficult to solve. Thus, new strategies to obtain crystals of membrane proteins for structure determination are critical. The objectives of this proposal are to develop a toolbox of chaperones and use them to crystallize and solve the de novo, high resolution structure of two signal peptide peptidases (SPPs), which use catalytic aspartates to conduct hydrolysis within the lipid membrane. In contrast to work employing affinity reagents specific to the membrane protein of interest, our potentially transformative approach uses hypercrystallizable single chain antibody fragments (scFvs). Our chaperones are engineered for tight binding to a short epitope that can be inserted into any membrane protein. We expect that our tightly bound scFv chaperone will immobilize an SPP loop and provide a stable crystal lattice, leading to better diffracting crystals. SPPs trim signal peptides (SPs) to liberate them from the endoplasmic reticulum membrane. SPP substrates include SPs remnants derived from new histocompatibility complex 1b (MHC-1b) molecules. As a part of innate immunity, these processed peptides are presented on cell surfaces for recognition by Natural Killer cells to indicate that the cell is healthy. In addition, SPP substrates include SPs from proteins involved in immune response and muscle contraction. SPP is also hijacked by the Hepatitis C virus (HCV) for replication, and is related to presenilin, which uses similar chemistry to generate amyloidogenic peptides in Alzheimer Disease. SPP and presenilin comprise one of just three superfamilies of intramembrane proteases. The details of regulated intramembrane proteolysis, from cell biological signaling to active site chemistry, are of both fundamental biochemical importance and potential therapeutic application. How substrates are presented and hydrolyzed within the confines of the hydrophobic space of the lipid membrane, however, remain largely a mystery. At least 5 SPP variants have been sequenced, located in different regions of ER, and SPPs are conserved throughput biology, but there is no crystal structure yet. We will start by solving the structure an archeal homolog in complex with our chaperones as proof-of principle, and then expand to a eukaryotic SPP, whose biomedical relevant activity is known. To date, we have engineered our first chaperone and isolated an affinity complex with SPP by gel filtration. Independently, we have grown crystals of the chaperone and SPP. However, the crystals of SPP do not diffract well enough for structure determination, and thus the cocrystalllization technology is critical. The expected outcomes are a toolbox of crystallization chaperones as well as the first molecular picture of SPP, including the location of the active site and substrate-docking patches. Taken together, this project will contribute not only to the biology of immunoregulation and intramembrane proteolysis, but also broaden our knowledge of membrane proteins and enable other membrane protein structures to be solved.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Subjective well-being reflects an individual's overarching evaluation of the condition of her or his life. Researchers from economics to psychology to medicine have become increasingly interested in this construct. Health researchers have discovered connections between well-being and mortality, whereas policy-makers are debating how to best use information about well-being to inform policy decisions. The simplest approach to assessing well-being is to use global evaluative measures. Such measures are easily incorporated into large- scale surveys, which has allowed researchers to make substantial contributions to the science of well-being. However, there are pressing concerns about the validity of these evaluative measures. A major alternative is the use of experiential methods that track feelings of satisfaction and affect more or less in real-time. Proponents argue that these approaches are less cognitively demanding than evaluative measures, which makes it less likely that heuristics and biases will affect their validity. The gold standard experiential approach is the experience sampling method (ESM) in which a portable recording device is used to track feelings and behaviors in the moment. However, ESM research is expensive, time consuming, and difficult to implement on a large scale. A less intensive alternative is the Day Reconstruction Method (DRM) in which participants divide their previous day into psychologically-meaningful units and report what they were doing and how they were feeling during those different episodes. The DRM is more practical than ESM for large panel studies. The broad goals of this project are to critically evaluate the psychometric properties of these three approaches and to incorporate both experiential approaches into an on-going, nationally representative panel study that already includes evaluative measures. The first specific aim is to thoroughly evaluate the properties of the DRM by comparing ESM and DRM responses on the same day and by estimating the stability of DRM assessments using short- and long-term longitudinal studies and advanced psychometric models. The second specific aim is to precisely quantify the extent that mood, survey context, and priming issues compromise the validity of global measures of well-being. This information is essential for making informed judgments about evaluative measures which are the easiest to implement in large-scale survey research. The final specific aim is to incorporate both ESM and DRM assessments into an on-going, nationally representative panel study. This existing study already includes evaluative measures and has been used to make important discoveries regarding well-being over the lifespan. The goal of the final specific aim will be to evaluate how previous aging- related well-being findings obtained with evaluative measures generalize to experiential methods. More importantly, the data from this longitudinal study will be made publicly available so that other researchers can advance the science of well-being using all three approaches.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The fate and behavior of pollutants is often dictated by the coupling of chemical and biologically-mediated reactions. Both hydrophobic organic pollutants and toxic transition metals are strongly sorbed to soil surfaces at Superfund sites. Sorption reactions together with the hindered diffusion of pollutants. through small pores appear to control the concentrations to which biological populations (including indigenous soil microorganisms) are exposed. Reactions which act to enhance pollutant release kinetics can both exacerbate exposure and can be deliberately employed to enhance aquifer clean-up procedures. Bacterial polymers naturally occur in soil solution and have documented binding.properties for both transition metals and hydrophobic organic contaminants. Project research to date has demonstrated that bacterial polymers will act to enhance metal desorption and mobility in porous media, and related research has similarly shown that the bacterial polymers will act to enhanced sorption and mobility of hydrophobic organic pollutants in porous media. Therefore, bacterial polymers may enhance the bioavailability of both toxic transition metals and organic contaminants in soil. The proposed continuation of this research will quantify the desorption kinetics of a radio-labeled transition metal (109Cd) and a polynuclear aromatic hydrocarbon (PAH), 14C.phenanthrene, in the presence and absence of selected extracellular polymers, including several produced by isolates of indigenous soil bacteria. The research will use a novel kinetic model that employs a statistical distribution of release rates to reflect the multiplicity of binding site types and pore sizes that collectively act to control contaminant release. Techniques using fluorescent antibody (Fab) staining and microautoradiography (MARG), developed during prior phases of this NIEHS- sponsored research, will be employed to view the distribution of radiolabeled organic compounds, polymers, and bacterial cells at the microscale and will provide an improved understanding of polymer-mediated contaminant release mechanisms. MARG methods will also be adapted for use in assessing the bioavailability of sorbed metals and PAH to soil bacteria that adhere to soil surfaces through production of extracellular polymers. The project results will yield basic information on the micro- and macro-scale influence of extracellular polymers on the bioavailability of contaminants. in porous media. A range of experimental scales will be considered from the macroscopic behavior of pollutant sorbates in suspensions or columns of sorbent material to direct microscopic measurements at the scale of bacterial cells in their microhabitats. This range in experimental scale will facilitate construction and verification of environmentally relevant models that employ mechanistic descriptions of reaction and transport at the pore scale to make predictions of macroscopic behavior.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cryptococcal meningitis (CM) is a leading cause of death in AIDS patients in the developing world, responsible for up to 500,000 deaths each year in sub-Saharan Africa alone. In Uganda, cryptococcosis accounts for 30% of mortality in AIDS patients after starting ART. Many of these cases of CM may be preventable. Routine screening for sub-clinical infection, using a simple blood test (cryptococcal antigen or CRAG) in patients presenting to ART programmes, can identify which patients are at risk of developing CM. If identified, these persons could be given pre-emptive anti- fungal treatment to prevent development of overt meningitis and death. Our research team has demonstrated in a study of 311 persons with advanced HIV-infection in Kampala, Uganda, that such a screen and treat strategy would be highly cost-effective, at an estimated $21 per quality-adjusted life year (QALY) saved. We know that ART alone is insufficient to prevent the development of CM in patients with sub-clinical infection, and antifungal therapy is required. Yet, the optimal pre-emptive anti-fungal treatment dose and duration necessary to prevent CM in patients with sub-clinical infection have yet to be established. We propose to investigate how practical and effective CRAG screening and targeted pre-emptive fluconazole treatment would be in the operational setting of patients entering ART treatment programmes in Kampala City Council clinics in Uganda. In addition, a new point-of-care CRAG screening test will be evaluated in parallel with the currently used test. In the planned study, 1000 patients will be screened for cryptococcal antigenemia prior to starting ART. Those with a positive result will receive pre-emptive short course oral fluconazole therapy to treat sub-clinical cryptococcosis and determine if this is effective at preventing CM. All patients will then be started on standard ART and followed for 6 months to determine their overall outcome and incidence of clinical CM compared to historical cohort controls. The cost-effectiveness of the intervention with pre- emptive fluconazole therapy will be analyzed. The results of this operational research would be widely applicable to other regions in sub-Saharan Africa and could lead to improved public policies to prevent CM.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research seeks to understand determinants of maladaptive achievement patterns in children--patterns that may put children at risk for low self-esteem and negative affect. The first major issue examined is the potential adverse effects of certain kinds of teacher strategies and practices (i.e., controlling techniques) on the development of an extrinsic motivational orientation which is assumed to increase children's vulnerability to performance decrement, negative self-cognition and affect, and overall poor achievement. This issue will be examined in a longitudinal study to see the cumulative negative effects on children's propensity to maladaptive achievement patterns because of exposure to controlling teachers over consecutive years. By use of combined self-report, observational and experimental methods in a longitudinal design, we hope to examine both determinants of maladaptive achievement patterns and the severity of such patterns as a function of socialization practices in the classroom. The second aim focusses on an intervention strategy to \"immunize\" children from the proposed negative effects of controlling techniques on achievement patterns. Our goal, then, is to use a multiple method approach to provide converging evidence regarding socialization factors influencing the formation of maladaptive achievement patterns and intervention techniques that may produce more adaptive patterns.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have begun a clinical protocol to determine if resident lymphocytes in non-small cell lung cancer are reactive with tumor specific mutated neo-antigens and if so, whether they can be therapeutic if given patients using methods recently developed in the Surgery Branch (Tran et al, Science 2014). We are studying ways to sequence the genome of a patient's lung cancer, identify all mutated proteins, redisplay them effectively to the T-cells in their tumors and search for T-cells that can react to these mutated neo-antigens. We then wish to develop ways to isolate or enrich them, grow them and readminister them for therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Successful management of diabetes in youth is heavily dependent upon family adaptation to the affective, behavioral, and cognitive demands imposed by the disease. During pre and early adolescence, transition in responsibility for diabetes management, along with normal physiological and psychological developmental changes, create an especially challenging situation. Although many youths and parents negotiate this transition effectively, it is also a period when many other youths take costly, self-destructive paths resulting in preventable health care costs and psychological suffering in the short-term and accelerated onset and progression of long-term complications of the disease. Studies suggest that poor adaptation to diabetes during adolescence is likely to persist into early adulthood, accelerating the risks of long-term medical complications.[unreadable] [unreadable] Research to date suggest that adherence, quality of life, and glycemic control could be enhanced if behavioral interventions were routinely implemented as part of standard care. Yet there are many barriers to the translation of these interventions into routine clinical practice, including cost, access, third party coverage, availability of qualified clinicians, convenience, social stigma, and other such variables. A multi-component behavioral intervention that integrates psychological principles into medical management of diabetes is likely to enhance family management of diabetes during early adolescence in a practical, cost-effective and lasting manner.[unreadable] [unreadable] The goal of this multi-site study is to assess the efficacy of a clinic-integrated behavioral intervention for youth with type 1 diabetes and their parents. The study employs a randomized experimental design in which youth-parent dyads attending one of four clinical sites are stratified by degree of glycemic control and randomized to receive either standard care or a clinic-integrated behavioral intervention. The intervention is based on both individual and family system theoretical perspectives, including social cognitive theory, self-regulation, and authoritative parenting. It is designed to provide experiential training for families in the use of a problem solving approach to promote improved parent-child teamwork and more effective problem-solving skills for diabetes management. The intervention is designed to be applicable to the broad population of youth with diabetes and their families, flexibly implemented and tailored to the varying needs of families, and delivered at a low intensity over time to meet the changing needs and roles of families during the period in which responsibility for diabetes management typically undergoes transition. A combination of in-person assessments, telephone assessments, and in-clinic data collection will be utilized to assess glycemic control, adherence, quality of life, psychological status, and hypothesized mediators of these outcomes[unreadable] [unreadable] Unique aspects of the study include: (1) It is the largest to date, and the first multi-site study, to test the efficacy of behavioral interventions for improving adherence, glycemic control, and quality of life in youth with type 1 diabetes; (2) It is based on the concept that small changes across the distribution of risk factor may have a greater public health impact than individual or high-risk approaches, and as such is designed to address the population of youth with diabetes; (3) The sample is large enough to test intervention moderator effects including baseline level of glycemic control, socio-economic status, and family functioning; (4) 4 clinical sites are participating, 2 of which have substantial low socio-economic status and minority populations.[unreadable] [unreadable] Clinical Sites for this stucy include: Joslin Diabetes Center in Boston, MA; Nemours Childrens Clinic in Jacksonville, FL; Texas Childrens Hospital in Houston, TX; and Childrens Memorial Hospital in Chicago, IL. The coordinating center for this study is James Bell Associates.[unreadable] [unreadable] Several pilot studies informed this clinical trial. A longitudinal observational study, Developmental Influences on Management of Type 1 Diabetes, examined the influence of family, social, and behavioral variables on diabetes self-management behaviors with a particular focus on adolescent developmental transitions. A pilot intervention study, the Diabetes Personal Trainer Study, assessed the effectiveness of an individualized problem-solving approach, guided by principles of motivational interviewing and applied behavior analysis, and implemented by specially-trained undergraduate and graduate students, who served as diabetes personal trainers. A pilot study of an abbreviated form of the specific intervention approach used in the multi-site clinical trial assessed the feasibility of the study design and intervention approach.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Molecular biologists are increasingly realizing that computational modeling of sub-cellular mechanisms is crucial for making sense of human biology and advancing medical research in the context of the vast quantities of complex experimental data that are now being collected. The biology community needs agreed-upon information standards if models are to be shared, evaluated and developed co-operatively. The Systems Biology Markup Language (SBML) is becoming a de facto standard exchange format for representing formal qualitative and quantitative models of systems of biochemical reactions. The long-term objectives of this proposal are to support and further develop SBML and related software infrastructure for the benefit of the computational biology community. The specific aims are the following: 1. Work with the community of software developers and users to (a) continue evolving SBML and extending the language into SBML Level 3 with features developers have been requesting, and (b) formalize the SBML language development process and transfer its control to the community. 2. Maintain and extend existing software libraries for working with SBML with facilities for (a) reading and writing SBML Level 3, (b) reading model definitions in other formats including CelIML, and (c) transforming model definitions into other formats such as CelIML, MATLAB, Mathematica, HTML, and others. 3. Produce documentation, including a \"best practices\" guide to working with SBML, and performing training functions to help educate software developers and modelers in the use of SBML and the software tools available for working with the language.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Tuberculosis is an enormous global public health problem and the second leading cause of death due to an infectious disease. The global emergence of multidrug-resistant (MDR) TB is a significant barrier to effective TB control. Response rates to second-line anti-TB drugs (SLDs) used to treat MDR-TB are much lower than those for treatment of drug susceptible TB. Recent reports highlight the substantial risk of development of additional drug resistance during SLD treatment. Critical knowledge gaps exist regarding the mechanisms of amplified drug resistance during treatment of MDR-TB. Our preliminary data suggests that the TB cavity may be the site of further drug resistant development. With this background, we have hypothesized that pulmonary cavities in patients with MDR-TB facilitate the development and amplification of drug resistance as a result of sub-therapeutic intra-cavitary SLD concentrations. An enhanced understanding of SLD pharmacokinetics including tissue penetration properties and mechanisms of drug resistance will help guide improved MDR-TB treatment and prevention strategies. The Specific AIMs of this proposal include: 1) To determine the pharmacokinetics of key SLDs (levofloxacin and capreomycin) in plasma, pulmonary tissue, and cavitary lung among patients undergoing treatment for MDR-TB. Utilizing a unique patient cohort of MDR-TB patients undergoing adjunctive surgery in the Republic of Georgia and an innovative microdialysis method we will assess SLD concentrations among various compartments including pulmonary cavities, the site of the highest concentration of M. tuberculosis (MTB). 2) To investigate phenotypic and genotypic drug resistance profiles of MTB isolates recovered from sputum, lung, and cavitary lung tissue. We propose to perform whole genome sequencing and bioinformatics analysis comparing paired MTB isolates from different sites in the same patients (cavity, more normal lung tissue, sputum) to determine if cavities are sites of drug resistance development and amplification and to study population dynamics of MTB within the cavitary environment. 3) To determine the relationship between SLD tissue levels and the development of additional drug resistance mutations in MTB isolates from resected cavitary lung. We will use the pharmacokinetic and genomic sequencing data derived from AIMS 1 and 2 along with pharmacokinetic modeling to assess if low drug concentrations are associated with increasing drug resistance. The proposed investigations will provide novel data on mechanisms of amplified drug resistance and will challenge the way we currently dose SLDs and treat MDR-TB patients with cavitary disease. Candidate My long-term goal is to become an independent clinical and translational investigator and global leader in tuberculosis (TB) focusing on the mechanisms, transmission, and virulence of drug-resistant TB. A K23 award would be critical to my career development and provide protected time for research and research training. A Training Plan has been developed that links a training goal to each of the specific aims of the research proposal. During this K23 award period, I will build expertise in clinical research, pharmacokinetics study design and modeling, performing microdialysis, bacterial genomics including DNA sequencing, and performing bioinformatics analysis. My career development plan combines formal didactics, laboratory training, and hands on mentoring with relevant research experiences that address global TB priorities. A multi-disciplinary team of recognized leaders in clinical TB research, genomics, and pharmacokinetics will provide professional and scientific mentorship during the award period. The K23 award will allow for the investigation and accumulation of additional data that can subsequently serve as the basis for a subsequent R01 application focused on translational research involving investigation of the pulmonary cavity. Environment The substantial intellectual and physical resources of Emory University and established collaborations with partners in the country of Georgia at the National Center for Tuberculosis and Lung Disease (NCTBLD) provide a unique opportunity to carry out this translational research. The Georgian NCTBLD also has a long history of productive collaborations with Emory University including numerous ongoing funded research projects (including the site of a NIH Fogarty grant). Emory University is a research-intensive university with over half a billion dollars in research funding each year. The Emory Division of Infectious Diseases has 59 full time faculty members including leaders in clinical TB, pharmacokinetics and bacterial genomics research. The Emory Rollins School of Public Health, Emory Global Health Institute, and the Atlanta Clinical and Translational Science Institute (ACTSI, the NIH-funded CTSA) provide further excellent resources for career enhancement, development and research training. The proposal will also benefit from collaboration with the University of Florida Infectious Diseases Pharmacokinetics Laboratory (IDPL). The IDPL is a national reference laboratory for anti-mycobacterial drug concentrations and has extensive expertise in performing microdialysis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "BACKGROUND: Sixty percent of all cancer and seventy percent of cancer mortality occurs in people over age 65. As the population ages, there is an emerging need to develop a means for oncologists to characterize the \"functional age\" of older patients with cancer in order to tailor treatment decisions and stratify outcomes based on factors other than chronological age and to develop interventions to optimize cancer treatment. HYPOTHESIS: A geriatric instrument for older patients with cancer can be formulated to identify factors independent of age that predict cancer treatment morbidity and mortality and result in rationale interventions to optimize oncologic care. SPECIFIC AIMS: 1) To determine the generalized feasibility and psychometric properties of a newly developed geriatric instrument for older patients with cancer; 2) To assess the geriatric instrument's ability to predict which older patients are more vulnerable to toxicity from chemotherapy; and 3) To determine the impact of the geriatric instrument on clinical care and overall outcomes through specific interventions. CAREER DEVELOPMENT: Through this work, the candidate will meld her previous training in geriatrics and oncology and develop the skills needed to develop an independent research career in geriatric oncology. Five years of didactic coursework in clinical trial design, biostatistics, and measurement development are planned. The candidate will be mentored by experts in the fields of oncology (Dr. George Bosl, Mentor), geriatrics (Dr. Mark Lachs, Co-Mentor), psychosocial research (Dr. Alice Kornblith, Advisor), and biostatistics (Dr. Katherine Panageas and Dr. Bercedis Peterson, collaborators). Through this career development award, the applicant will acquire the skills necessary to pursue an independent research career in geriatric oncology. Through the support of this grant, the candidate aims to further knowledge about geriatric oncology, and thus enhance the health and independence as well as the quality of life of older Americans with cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose a summer clinical immersion program at the Weill Medical College of Cornell University that brings Biomedical Engineering (BME) PhD students from Ithaca Engineering College Campus to New York City Medical College Campus for dedicated intensive clinical exposure in the summer between their first and second year of graduate training. Each student will take bioethics lectures, shadow clinical practitioners and participate in clinical related research under the direction of an assigned clinician mentor. This clinical summer immersion is designed to connect BME to healthcare and society and to steer and enhance students'research interests towards medicine. It will immediately impact their PhD thesis research by adding a medical conscience and influence their long term career towards improving healthcare. PUBLIC HEALTH RELEVANCE: We propose to connect Biomedical Engineering Students'laboratory work with healthcare and help them give their discoveries the best chance to benefit clinical medicine by immersing them in a major medical center to shadow medical practitioners and participate in clinical research. When BME students see patients'suffering, they appreciate the urgency and importance of developing medical technology solutions to healthcare problems;when students watch in person their clinician mentor diagnosing and treating patients, students appreciate the utility of medical technology in medicine;when students see the limitations of the patients and the clinical environment, they develop a perspective on what kinds of technology solutions can be clinically workable;when students participate in clinical research, they appreciate the impact of medical technology", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite the use of highly sensitive serologic tests for screening blood donations for antibodies to HIV-1, HTLV-I/II, HCV and HBV in US blood centers, the delay between the onset of infection and the antibody response creates a \"window\" period during which the blood may be infectious but seronegative. As a result, a low risk of virus transmission due to contaminated blood products still exists. During PHASEI of this grant, we have developed a Multiplex RT-PCR based capture assay which permits the simultaneous direct detection of HTLV-I/II, HIV-1, HCV and HBV in a single assay. In the course of PHASE II, we propose to further refine this Multiplex RT-PCR assay, and to develop a kit format useful for routine screening of blood products in a clinical lab setting. The assay will be validated on a number of well characterized performance and seroconversion panels for all five viruses and on large numbers (>1,000) of blood samples from normal blood donors. This test will provide a highly sensitive, cost-effective direct measure of infectious virus in blood and help further reduce the antibody-negative \"window\" period between infectivity and detection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION The purpose of this CDA-1 proposal is to provide Dr. Naomi Bramhall with the support and mentorship necessary to make the transition from basic science research to clinical research of noise-induced hearing loss. Dr. Bramhall's previous research has focused on hereditary hearing loss and hair cell regeneration, but as a certified audiologist, she is eager to shift to clinical hearing research. This proposal will provide Dr. Bramhall with the tools and experience necessary to engage in clinical research and generate data which will be used in applying for future funding such as the CDA-2, will guide her path towards independence as a clinical researcher. Her primary mentor for this proposal is Dr. Dawn Konrad-Martin and Dr. M. Charles Liberman is a co-mentor. Loud noise exposure is routinely experienced during military service and hearing loss is the second most common service-related disability among Veterans. Individuals with hearing loss and poor speech discrimination ability often have little success with hearing aids because amplifying sound improves the audibility, but not the clarity, of the speech signal. Loud noise exposure may result not only in damage to outer hair cells, but also lead to auditory neuronal degeneration. Neural loss can result in decreased word recognition (Schuknecht 1994) and may explain why many Veterans have poorer speech discrimination than would be expected based on their ability to detect tones, particularly in noisy environments. Mice show rapid degeneration of auditory nerve peripheral terminals followed by a slow degeneration of spiral ganglion neurons over a period of several months following loud noise exposure, even when their pure tone auditory thresholds recover completely (Kujawa and Liberman 2009). The subpopulation of neurons most vulnerable to noise exposure consists of the low spontaneous rate fibers (Furman et al. 2013), which have a high sensitivity threshold and respond to higher intensity sounds. This neuronal loss is correlated with a decrease in the amplitude of the first wave of the auditory brainstem response (ABR). Age-related reduction in the amplitude of wave I has been demonstrated in humans (Konrad-Martin et al. 2012) and is consistent with temporal bone studies showing auditory neuronal loss with age (Makary et al. 2011), suggesting this metric of auditory neuronal survival is also applicable to humans. Previously, quantification of the degree to which auditory neuronal survival influences speech understanding has been difficult due to limitations in measuring auditory neuronal loss in humans, which could only be accomplished through post-mortem temporal bone studies. The main objectives of this proposal are to use electrophysiological methods to 1) determine if decreased ABR wave I amplitude is associated with noise exposure in humans and 2) examine the impact of noise exposure and decreased wave I amplitude on auditory perception. Our hypothesis is that noise exposure will be associated with decreased wave I amplitude, consistent with animal studies, and that this change will be correlated with decreased speech recognition at high sound levels, difficulty understanding speech in the presence of background noise, and decreased tolerance to loud sounds. Developing the ability to assess auditory neuronal survival with existing clinical tools and advancing our understanding of the specific auditory perceptual deficits resulting from neuronal loss will help guide future treatment options for Veterans suffering from hearing loss.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research is a study of the design and synthesis of agents useful primarily in the treatment of cancer. The approach used is to study the chemical and physical properties of the metabolic sequence that may be one of the controlling or limiting processes in the synthesis of deoxyribonucleic acid and ultimately cell division. The sequence is the reductive methylation of deoxyuridine-5'-phosphate catalyzed by the enzyme thymidylate synthetase. The research objective of this problem is a continuing study of the synthesis and in vitro testing of potential thymidylate synthetase inhibitors. Specifically the effective binding characteristics of the cofactor N5N10-Methylene-tetrahydrofolic acid, for thymidylate synthetase and dihydrofolate reductase enzyme will be examined. In this manner and by application of the chemical mechanism of the reaction and the physical properties directing enzyme-substrate or inhibitor affinity the nature of the groups important to binding can be defined and by design, attempts can thereby be made for specificity of action. Samples will also be submitted to CCNSC for testing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract The overarching goal of the Cancer Epidemiology Program is to investigate the causes of cancer through epidemiological studies incorporating genetic and molecular epidemiology approaches as well as risk factors/environmental and biomarker research that are strongly grounded in the relevant biology. A hallmark of this Program has been the integration of epidemiology and biology, along with the development and application of state-of-the-art biostatistical approaches and a defined focus on ethnic diversity that is consistent with the populations within the USC Norris catchment area. The Program is led by Graham Casey, PhD, a molecular geneticist who is interested in the integration of cancer epidemiology and molecular biology to develop novel insight into the complex etiology of cancers, and Duncan Thomas, PhD, a biostatistician with extensive contributions to design and analysis methods for genetic and environmental epidemiology studies and has had numerous cancer epidemiology collaborations. Drs. Casey (cancer genetics), Thomas (biostatistics), and Haiman (cancer epidemiology) comprise the Program's Executive Committee. The scientific aims of the Program are to: 1) elucidate the role of environmental and lifestyle factors (e.g., obesity, diabetes, radiation) in the etiology of cancer and study population cancer trends; 2) elucidate the role of genetic factors in the etiology of cancer with an emphasis on different racial/ethnic populations using existing and new cohorts; 3) determine the mechanistic and biological basis for genetic risk variants using large-scale fine-mapping and comprehensive cellular and biochemical approaches, and integrate biomarker and tumor biology studies into epidemiologic research; and 4) develop and apply novel study design and statistical analysis methodologies for environmental and genetic epidemiology research in cancer. Accomplishments during the project period include seminal contributions to the genetic etiology of the cancer field with over 40 GWAS-related manuscripts since 2010, leadership roles in several international genetics consortia, expansion of the genetics emphasis to study the biological implications of genetic inheritance through strategic recruitment, development of an integrative genomics theme to study the relationship between inherited and somatically acquired mutations in tumors, submission of several P01s, and increased collaboration with Cancer Control Research and other programs, that includes collaborative grant submissions. The Program is composed of 27 members from four departments within the Keck School of Medicine. Current grant funding totals are $12M in peer-reviewed funding (direct costs), of which 58% is from NCI, 25% from other NIH sources, and 8% in other peer-review funding sources. The Program is highly productive with 730 publications of which 26% are inter-programmatic, 47% intra-programmatic and 62% inter-institutional.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project #1. Impact of HIV-1 on the Rho GTPase-mediated neuronal cell development. It is now well accepted that the HIV-1 regulatory protein, Tat, has a neurotoxic effect and that treatment of both human and rodent neurons with Tat induces apoptosis by mechanisms that are not fully understood. The cellular protein, Pur-alpha, is a key target of Tat for directing the regulation of HIV-1 transcription, translation, and viral RNA transport. Indeed, both Pur-alpha and Tat have been detected in the same cells within HIVE infected lesions. Results in the Pur-alpha knockout mouse model have revealed a critical role for Pur-alpha during development, particularly in the coordinated development and differentiation of neuronal cells throughout the brain. Over the past several years, it has become clear that the RhoGTPases and related molecules play an important role in neuronal cell development, including neurite outgrowth, differentiation, axon pathfinding, dendritic spine formation, as well as neuronal cell maintenance. Results from our studies indicate that Tat has the ability to disturb this pathway by stimulating RhoA activation, an event that is accompanied by conversion of RhoAGDP to RhoAGTP by interacting with guanine nucleotide exchange factor (GEF) family members including PDZ-RhoGEF. Indeed, our preliminary results suggest that Tat may mediate RhoA activity via interaction with PDZ-RhoGEF. Upon its activation, RhoA is usually positioned in the plasma membrane, and via a series of signaling pathways that involve Rock, MAPK, and JNK1, NFkB induces neurite retraction. Moreover, activation of RhoA via heteromeric G-proteins including Ga12 and G<x13 can stimulate GSK3(3 which is also known to be involved in HIV-1 neurotoxicity. On the other hand, Tat can block activation of another key RhoGTPase, i.e. Rac1 and its partner p21-activated kinase 1 (PAK1), which appears to regulate actin cytoskeletal dynamics, maintain neuronal cell integrity, and promote neurite outgrowth. In addition, Tat has recently been shown to inhibit this pathway by acting through PAK1 and JNK1. Finally, Rac1 and RhoA were found to display aberrant expression and subcellular localization in the Pur-alpha knockout mouse model, suggesting that Pur-alpha is required for their proper function. Based on these preliminary observations, we hypothesize that Pur-alpha has a central role in the well-balanced activities of RhoA and Rac, and that its genetic ablation or its functional alteration by associating with Tat can derail several key parameters in RhoA and Rac signaling pathways including p115/PDZ and PAK, respectively, and results in pathological features that are commonly seen in neuronal cells of AIDS patients with CNS disorders. In this research project, we will perform a series of cell biological and molecular studies to unravel the molecular basis of Tat cross-talk with Rho GTPase and its impact on neuronal cell integrity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The University of New Mexico Health Sciences Center (UNM HSC) has developed an innovative and widely applicable model to provide treatment for patients with chronic, common and complex diseases who do not have direct access to specialty healthcare providers. This knowledge-on-demand model is called Project ECHO - Extension for Community Healthcare Outcomes. Conceived as a means to treat HCV-infected patients in New Mexico's rural communities and prison system, Project ECHO can assist in diagnosing and successfully treat many other chronic illnesses and conditions. Project ECHO connects urban medical center disease experts with rural general practitioners and community health representatives over a telehealth network. This enables them to effectively treat patients on site who would otherwise have to travel to urban healthcare facilities for specialty treatment. Project ECHO has enormous potential as an affordable healthcare intervention in rural communities where certain chronic diseases have reached epidemic levels and healthcare resources and personnel are scarce. The purpose of Project ECHO is to educate, train, and support rural general practitioners or other available healthcare representatives on the best practice treatment protocols for complex diseases they encounter in their communities. This model focuses on the principles of case-based learning and disease management using the telemedicine infrastructure and internet-based technologies to co-manage patients in community-based practices. The ultimate goal of Project ECHO is to provide the same level of healthcare to rural patients with chronic diseases as can be obtained in an urban setting. A secondary goal is to provide rural healthcare practitioners with a level of interaction and support commensurate with their urban counterparts to enhance their technical competence and decrease their feelings of professional isolation. Project ECHO is being field-tested in weekly, regularly scheduled telemedicine clinics hosted by UNM HSC specialists in the areas of hepatitis C. Through use of the UNM HSC Telehealth pathways and internet-based access, community healthcare providers around New Mexico are afforded the opportunity to present cases which are discussed among the network participants to jointly reach treatment decisions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Participants in the Baltimore Longitudinal Study of Aging aged 60 and older were examined to detect changes in psychological, neurological, and neuropsychological tests related to early signs of Alzheimer's disease. Six-year changes in immediate visual memory performance assessed by the Benton Visual Retention (BVR) test were used to predict Alzheimer's disease (AD). The subjects of this study were 279 community-dwelling adult participants in the Baltimore Longitudinal Study of Aging, eight of whom received AD diagnoses from a neurologist based on DSM III-R and NINCDS- ADRDA criteria. Subjects with diagnoses of AD had disproportionately larger changes in numbers of errors over the six-year retest interval prior to estimated onsets than subjects without AD diagnoses. In addition, significant risk for AD was associated with changes in BVR performance from a proportional hazards analysis. The relative risk for AD associated with six-year BVR changes of six errors was 5.9 (95% CI=5.0-7.1). These results suggest that symptoms of AD may be manifested by changes in immediate visual memory performance earlier than is detectable by clinical evaluation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lower levels of physical activity, an \"unhealthy\" diet and the presence of overweight/obesity are established risk factors for several types of cancer. There is a growing interest in exploring the potential links between environmental characteristics and chronic diseases, such as breast cancer in women. Previous research done by our group has demonstrated a link between neighborhood characteristics, such as walkability, and risk factors for chronic diseases. Therefore, using the San Diego cohort of the Women's Health Initiative (WHI), we propose to test the hypothesis that neighborhood characteristics would potentially moderate the association between common behavioral risk factors and incident breast, colorectal and total cancers. To do so, we propose to conduct a study employing the following specific aims: [unreadable] [unreadable] 1. To assess the baseline associations among the neighborhood environment factors (built, [unreadable] sociodemographic, and food) and common, behavioral cancer related risk factors (physical activity, diet, body mass index). [unreadable] [unreadable] 2. After adjustment for pertinent cancer risk factors, to determine the potential direct effects of the neighborhood environmental factors on incident cancer outcomes (breast, colorectal, total). [unreadable] [unreadable] 3. Using multivariable models, to ascertain the direct, mediated or moderated effects of both the neighborhood environment and cancer risk factors on incident cancer outcomes. Participants' neighborhood characteristics within a 1 kilometer buffer of their residence will be geocoded using geographic information systems (GIS) and linked to census, land use and business license data. This data will then be merged to the existing data from the San Diego WHI cohort, which includes extensive baseline characteristics (i.e. physical activity, dietary and body morphology measurements) as well as incident cancer events that were adjudicated using a standard protocol and central physician adjudicators. Once linked, appropriate analyses will be conducted to test the specific aims listed above. We believe the proposed study is innovative because of the prospective nature of the follow-up and it will test links between multiple neighborhood characteristics and not only cancer risk factors but multiple types of cancer. This research is important because it would extend the existing literature on neighborhood characteristics beyond common behavioral risk factors to incident cancers and it would inform and potentially influence theoretical models on the associations between neighborhood structure and cancer events. As such, this study could have implications for policy formulation on neighborhood design and development. PUBLIC HEALTH RELEVANCE: Lower levels of physical activity, an \"unhealthy\" diet and the presence of overweight/obesity are established risk factors for several types of cancer. There is a growing interest in determining if there are links between the environment in which one lives and chronic diseases, such as breast cancer in women. Therefore we propose to determine if the neighborhood environment influences the association between common behavioral risk factors and new breast, colorectal and total cancers. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Retinal pigmentation is one of several factors that profoundly influence visual system development. This is indicated by the fact that albino mammals (including humans), in addition to their external pigmentation deficits, have abnormalities of visual system development. Introduction of a functional tyrosinase gene into albino animals (mice and rabbits) corrects all of these abnormalities, proving that all aspects of this phenotype are due to the single gene defect in tyrosinase. Although it is known that the contribution of tyrosinase is crucial to visual system development, it is not known when or how tyrosinase influences development. I propose to determine the timepoints at which tyrosinase expression is necessary for the prevention of two albino abnormalities: decreased rod cell density, and a disruption of the retinogeniculate pathway through the regulated expression of tyrosinase during critical periods in the development of these two components of the visual system. Insight into the developmental function of tyrosinase will shed light on some fundamental issues of neuronal development, such as cell fate determination, axonal tracking, the timing of neurogenesis, and programmed cell death during development. I will control tyrosinase expression through the creation and use of a novel method of regulating transgenes in mice which involves modification of regulatory elements from the lac operon of E. coli for use in the mammalian genome. Tight regulation over the expression of transgenes will allow the investigation of a variety of previously unanswerable questions, including those addressed in the current proposal. This project will set the groundwork for a substantial broadening of the range of issues that can be addressed with transgenic mouse models.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: The research program completes a sample of infant twins for a multi-method longitudinal appraisal of temperament from the neonatal period through 30 months of age. Infant twins offer a powerful resource for detecting similarities and differences among genetically related pairs and examining the stability and organization of temperament characteristics over age. From the perspective of developmental behavior genetics, features of temperament may be examined for genetic influences on dimensions of temperament and their reorganization when transitions occur. Twins provide an opportunity to detect specific contributions of parents, family/home when two infants' temperament may differ markedly at a given time or over time. The principal aims of the program are to: (a) complete a sample of neonatal assessments of all twins in newborn nurseries for an initial measure of irritability, reactivity, and developmental status; (b) complete assessments of a longitudinal sample of twins so that 280 pairs have a multi-method appraisal of temperament at 3, 6, 9, 12, 18, 24, and 30 months, as based on direct observations of twins' behavior during standardized procedures in a structured laboratory setting, temperament questionnaires completed by parents, IBR ratings obtained during Bayley mental testing, and parental interviews; and (c) complete appraisals of the family/home environment and its contribution to temperament development. Laboratory sessions are videotaped. The results are ultimately addressed to the questions of origins, patterning among variables, sources of stability/instability over ages, and the joint collaboration of genetic and environmental factors for the determinants of temperament. This project, linked with other data sets, also demonstrates the potential relevance of newborn and infant temperament, examined longitudinally, for its contribution to preschoolers' liability for unintentional injuries.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite the prevalence of hyperactivity in school-age children, few attempts have been made to identify or treat this disorder prior to school entrance. Parent reports, however, suggest that in most cases hyperactivity and associated impulsivity and disciplinary problems are present in some form from infancy or toddlerhood onward. Follow-up studies of preschool and school-age hyperactive children further suggest that this disorder persists and is relatively resistant to treatment. Thus, the present research is aimed at the early identification and treatment of childhood hyperactivity. Hyperactive toddlers (2 1/2 - 3 1/2) are being compared with matched controls on a variety of parent report, family history, behavioral, and cognitive measures to determine the correlates of hyperactivity in toddlers and the usefulness of certain assessment devices with this age group. The efficiency of parent training groups as a treatment strategy is then being evaluated. Therapeutic effect is being assessed at the conclusion of eight weeks of structured group intervention and in a six-month post-treatment follow-up to evaluate short-term gains and maintenance of change in both child behavior and parenting skills.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this research are to investigate (1) effects of ionizing radiation in the immature spinal cord, (2) the reparative and regenerative capacities of the immature nervous system, and (3) the manner in which ionizing radiation alters these capacities. All experiments are carried out in rats because the developmental state of these animals at birth makes it possible to study the immature nervous system ex utero. Neurologic, histopathologic and autoradiographic methods are used in this research. The responses of the immature nervous system to injury are compared and contrasted with those reported by others in the adult nervous system; this, perhaps, will provide some insight into the reactions and limitations occurring in the mature state. In addition to the experimental data, the information derived from intact, control animals will contribute to the understanding of normal maturation of the nervous system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The design of a component to elicit broadly neutralizing antibodies is thought to be crucial for a successful HIV vaccine but is proving very difficult. In particular, immunogens designed from study of broadly neutralizing human monoclonal antibodies have generally failed to elicit antibodies with the appropriate specificities. A number of hypotheses have been advanced for these failures including restricted access to broadly neutralizing epitopes in recessed sites on the virus surface, proximity of the epitopes to viral membranes and epitope mimicry of self-antigens. In this new R01 proposal, we propose to generate transgenic mice carrying genes encoding two of the broadly neutralizing HIV antibodies 4E10 and b12 to investigate these hypotheses and to provide tools to study and rank potential HIV vaccine candidates. In the first instance, we propose to use mice carrying HIV antibody genes targeted to the physiological Ig loci to test if 4E10 and b12 have biologically relevant self-reactivity. If self-reactive, the B cells generated in 4E10 and b12 transgenic mice will be regulated by the immune tolerance mechanisms of receptor editing, anergy induction, or deletion and these mechanisms will be dissected. This may allow the design of immunogens in the future that do not evoke self-responses. If non-tolerant, transgenic mice carrying the 4E10 or b12 specificities should carry a greatly increased precursor frequency of B cells that are broadly neutralizing for HIV. We will then use an array of immunogens to explore the origins of any restrictions on eliciting 4E10 and b12 antibody specificities, to evaluate the most promising immunogens and ultimately to guide the design of modified and new immunogens to elicit broadly neutralizing antibodies. Relevance: To be successful, it is likely that an HIV vaccine must stimulate neutralizing antibodies. This proposal provides a mouse model to understand some of the problems in stimulating such antibodies by various vaccine candidates and to thereby suggest how better candidates can be designed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Behavioral & Social Sciences Core (BSSC) is structured to provide services that facilitate interdisciplinary AIDS research among established, new, and transitional investigators. Although the BSSC Core does not direct the science of the behavioral AIDS program at Penn, our Core services are guided by a set of scientific priorities and principals that program members have identified as crucial to scientific discovery, translation, and public health impact. These priorities include investigations that seek to define, measure^ahd^xparrdthe~understahdingof the role of contextual factors (e.g. social, sexual, and drug using networks; community/geography; Mental health and substance abuse) within which HIV transmission occurs and infection exists. These factors are viewed as critical to understanding risk of infection, access and adherence to treatment. The BSS Core at Penn is also well positioned to expand the role of behavioral sciences in the design and evaluation of clinical trials of biomedical interventions (microbicides, vaccines, and therapeutics) and to facilitate scientific collaborations with other investigators (behavioral and interdisciplinary), both domestically and internationally is critical to the development of sustainable programs of AIDS research. The Core plans to accomplish its objectives through the delivery of services which include: 1) Research application and implementation support.through the provision of consultation on AIDS research application as investigators conceptualize and prepare applications for funding; 2) Behavioral and biological assessment services to facilitate the expanded use of biological assays in behavioral research; 3) Target population services to identify and recruit populations and individuals of scientific relevance to AIDS research; 4) Support for the Penn CFAR Community Advisory Board (CAB) to develop a highly visible and active agenda of community education and research; and 5) Educational activities to stimulate integrated biological and behavioral research and advance our scientific priorities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The synthesis of ribosomes is a complex but tightly regulated process in which the cell synthesizes equimolar amounts of four ribosomal RNAs in the nucleus, and of seventy-five ribosomal proteins in the cytoplasm, and assembles them into a ribosome in the nucleolus. We propose to study in Saccharomyces cerevisiae the mechanisms controlling both the transcription of ribosomal RNA and the synthesis of ribosomal proteins. Ribosomal RNA: We have identified a sequence in the rDNA repeat with many of the properties of an enhancer element. We plan to identify the precise sequences involved in this effect. We will ask whether this enhancer element: a) binds a specific transcription factor? b) is involved in the regulation of transcription of 35S prerRNA? c) is responsible for the 1:1 ratio of the transcription of 35S prerRNA and 5S RNA, whose gene lies between the enhancer and the 35S initiation site? Ribosomal Protein: The rates of transcription of most, if not all, ribosomal protein genes are tightly coupled. To understand the mechanism behind this coupling we will identify the sequences essential for the transcription of one ribosomal protein gene and then determine if similar sequences are used for other ribosomal protein genes. We will ask if the same sequences are involved in regulation of their transcription and/or whether there is a transcription factor specific for these genes. Finally, we will analyze the mechanism for translational control of ribosomal protein synthesis, with emphasis on testing the hypothesis that a regulatory molecule interacts with the m7G cap and its adjacent sequences.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fetal alcohol syndrome (FAS) is an important cause of mental retardation. Exposure of the developing brain to alcohol can induce the death of neurons, which contributes strongly to learning deficits and neurological problems associated with FAS. Understanding the factors that influence neuronal vulnerability to alcohol induced loss in the developing brain is of considerable significance. We hypothesize that as neurons mature, they acquire a neuroprotective, signaling pathway that protects them against alcohol toxicity. We have discovered a neuroprotective pathway, which is mediated by nitric oxide (NO). This proposal will elucidate the function and molecular mechanisms of the NO-signaling pathway in protecting the developing brain against alcohol-induced neuronal loss. Our experiments will utilize a mouse strain carrying a null mutation for neuronal nitric oxide synthase (nNOS-/-), thus these animals lack NO-mediated neuroprotection. Specific Aim 1 includes morphological experiments examining the importance of NO-mediated neuroprotection on neuronal survival. Stereological methods will measure alcohol-induced neuronal losses in the hippocampus and cerebellum of mice that express or lack the protective pathway. Alcohol exposure will occur at a variety of ages in order to determine the impact of NO-mediated neuroprotection at different stages of brain development. Specific Aim 2 utilizes behavioral testing (eyeblink conditioning, Morris Water Maze) to determine whether the enhanced alcohol-induced neuronal losses in nNOS -/- mice are linked to greater functional deficits. Alcohol-induced deficits on these tests will be compared in nNOS / and nNOS+/+ mice and linked with neuronal losses. Specific Aim 3 includes molecular studies to determine the mechanism by which NO-signaling provides neuroprotection against alcohol toxicity. Experiments will determine whether nNOS is developmentally regulated in vivo and link alcohol vulnerability to expression levels and activity of nNOS. \"Rescue\" experiments in which the nNOS gene will be transfected into pathway-deficient neurons in culture derived from nNOS -/- mice will determine whether neuroprotection against alcohol toxicity can be restored. Immunohistochemistry and quantitative RT-PCR will explore the role of cAMP-responsive element binding protein (CREB) as a downstream effector molecule for the NO-signaling and elucidate CREB's role in regulating apoptotic proteins, which may underlie this neuroprotection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Picture Archiving and Communication Systems (PACS) are expected to replace the currently existing film systems within the decade. However, presently there are no formal measures to evaluate the cost-effectiveness of image management systems. The objective of this Program Project is to develop a methodology to compare the cost-effectiveness of a PACS and the existing film management system. We will measure cost-effectiveness in the Medical Intensive Care Unit, the Emergency Department, and in the outpatient Neurology suite. The effects of diagnostic outcomes on health outcomes will also be estimated. The first project develops and evaluates a model that describes the diagnostic value of an image management system. The model will be validated and tested to determine its predictive capability. The second project evaluates workstations for clinical effectiveness, analyzes the radiologist-referring physician interaction, and studies physician acceptance. Teleconsulting between the radiologist and the referring physician including the transmission of outside images from the referring physician to the radiologist will also be studied. The third project utilizes clinical-history derived rules to optimize the display with diagnostically-relevant information. The fourth project uses the results of the other projects to measure the cost-effectiveness of PACS. Both direct and indirect costs before and after implementing PACS will be combined with data from the diagnostic value model to derive the incremental cost-effectiveness and to compare the usefulness of PACS to other medical interventions. The Operations core is responsible for software development, data collection, and quality control. This Program Project will provide the methodology for other institutions to determine the value of implementing PACS in their environment. Additionally, it may be applicable for the assessment of other medical technologies and to aid in the comparison of investments in such technologies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Intercellular RNA transport is a potentially important means of cell-cell communication. In the nematode C. elegans the RNA channel SID-1 is essential for the cell-to-cell transport of silencing RNAs, SidTI and SidT2 (SidT1/2) are the highly conserved vertebrate homologs and published reports indicate that fish, mouse, and human versions of SidT1/2 have RNA transport activity. Analyses of RNA transport in cultured cells derived from SidT2 knockout mice confirm these reports and establish SidTI and SidT2 knockout mice and derived cultured cells as valuable tools for the identification and characterization of RNA signaling pathways in mammals. Extracellular RNAs are present in many body fluids and are being explored as disease indicators. How these RNAs are selected, transported, and whether they have a biological function is unknown. To address these issues the RNA transport activity of SidTI/2 will be exploited to selectively identify RNAs that are purposefully extracellular. These SidT1/2-dependent extracellular RNAs will be validated and then, to unambiguously establish the fact of intercellular RNA transport, their expression and transport will be rigorously examined. This analysis will be collaborative with consortium partners, whom have established powerful experimental systems to detect intercellular RNA transport. Importantly, the dependence of RNA mobility on SidTI/2 provides the means to disrupt their transport between cells, which will be important for deciphering the physiological and developmental of these mobile RNAs. Parallel to these discovery efforts, the RNA transport mechanism and the RNA features that permit or restrict RNA transport will be elucidated. These efforts will also be collaborative among the consortium partners. These findings will have bearings on how RNA transport is regulated and how the RNA transport pathway can be exploited to develop RNA-based therapies, including the safe and effective delivery of siRNAs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this project we will define a role for copper-zinc superoxide dismutase (SOD1) in the regulation of cell survival and death. While the wild-type (WT) SOD1 is a pro-survival protein, amyotrophic lateral sclerosis (ALS)-linked SOD1 mutants are toxic both in vitro and in vivo. We recently found that both WT and mutant SOD1 interact with the anti-apoptotic protein Bcl-2. However, the nature of the mutant SOD1 binding with Bcl-2 differs from WT SOD1. Contrary to WT SOD1, mutant SOD1 specifically localizes to spinal cord mitochondria where it forms SDS-resistant high molecular weight aggregates that bind and entrap Bcl-2. (Pasinelli et al, 2004, Neuron 43: 19-30). These studies suggest a potentially novel function for WT SOD1 in regulating cell survival and death, and a novel, toxic gain-of-function for mutant SOD1. Thus, while WT SOD1 may protect against cell death through its interaction with Bcl-2, mutant SOD1 may become toxic by aberrantly binding to Bcl-2 and converting Bcl-2 into a toxic or non-functional protein. In support of this hypothesis, we now have preliminary data indicating that both WT and mutant SOD1 might require Bcl-2 to exert their anti-and-pro apoptotic function respectively. With the present proposal we intend to characterize the anti and pro-death function of WT and mutant SOD1 and their respective interactions with Bcl-2. The ultimate goal is to understand the mechanism(s) of mutant SOD1-mediated toxicity and to define a potential role for the mitochondrial mutant SOD1/Bcl-2 complex in ALS pathogenesis. The specific aims are: 1) (A) To determine whether WT SOD1 pro-survival activity depends on its binding to Bcl-2, and (B) to determine whether mutant SOD 1-mediated toxicity depends on the aberrant interaction with Bcl-2. 2) (A) To identify the region(s) in SOD1 essential for the binding with Bcl-2, and (B) to study the difference in binding strength between WT SOD1 and Bcl-2 and mutant SOD1 and Bcl-2. 3) (A) To determine whether Bcl-2 undergoes conformational modifications upon binding with mutant SOD1 and.(B) to test the potential benefit of Bcl-2 and SOD1 like-peptides that abolish binding between Bcl-2 and mutant SOD1 on our cell culture model of mutant SOD1-linked ALS. 4) To determine whether Bcl-2 mediates mutant SOD1 mitochondrial translocation. 5) To study the correlation between mutant SOD1/Bcl-2-containing aggregates and ALS using transgenic ALS mice and patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this project, the investigators propose to study the pathophysiology of mitochondrial long-chain fatty acid ?-oxidation (FAO) disorders. Patients with a FAO disorder can present with fasting-induced hypoglycemia, cardiomyopathy, heart beat disorders, sudden infant death, myopathy and (exercise-induced) rhabdomyolysis. Current therapeutic interventions need to be improved, but development of novel therapies is hampered by limited insight into pathophysiological mechanisms as well as small patient groups and insufficiently developed animal models. Therefore the overall objective of this proposal is to increase our knowledge of the pathophysiology of FAO disorders by studying newly identified molecular mechanisms and metabolic pathways that contribute to the different disease presentations using well-established mouse and cell models. The investigators hypothesize that perturbations in metabolic signaling pathways and the resulting impaired protein homeostasis play crucial roles in the pathophysiology of FAO disorders and that peroxisomal metabolism is an important alternative pathway for fatty acid degradation when mitochondrial FAO is defective. They will test this hypothesis by addressing two specific aims. In AIM 1, the investigators will study the molecular mechanisms underlying perturbed protein homeostasis in a well-established mouse model for mitochondrial FAO disorders. For this they will determine the dynamics of metabolic signaling during fasting in liver, heart and skeletal muscle and will quantify metabolic fluxes in protein synthesis and degradation. They will use a specific therapeutic diet to assess whether the depletion of tricarboxylic acid cycle intermediates that results from impaired protein mobilization can be delayed. In AIM 2, the investigators will study peroxisomal metabolic pathways that are alternatives for fatty acid degradation when mitochondrial FAO is defective. Using genome editing technique in a cell line, they will elucidate the molecular players participating in one of these alternative pathways. They will also assess the physiological importance of these peroxisomal pathways by studying the biochemical and clinical features of a unique mouse model with a combined defect in mitochondrial FAO and peroxisomal ?-oxidation. Combined these two aims will yield not only the much needed refined mechanisms, but also novel pathophysiological insights in FAO disorders and thus novel potential targets for treatment of these diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research will investigate some of the biochemical and biophysical properties of structural proteins isolated from normal and cataractous human lenses. Gamma-crystallin, and other low molecular weight crystallins will be fractionated and characterized. The study will include: amino acid composition, molecular weight determination by ultracentrifugation and gel electrophoresis. Secondary and tertiary conformational parameters will be investigated by sensitive circular dichroism measurements. Effects of concentration, ionic strength, pH, and temperature on the conformation will be monitored by absorption, fluorescence and circular dichoism measurements. The properties of proteins isolated from normal and cataractous lenses will be compared. The properties of fluorescent pigments found in normal and cataractous lenses will be studied by using absorption and fluorescence measurements on whole lenses, lens thin-sections and lens homogenate. Low temperature (77 degrees K) absorption spectra will be utilized to increase the resolution. Plasma membrane proteins will be isolated from normal and cataractous human lenses. The study will include amino acid compositon, molecular weight determination, and carbohydrate analyses. Structural studies using circular dichroism and laser Raman spectroscopy will be performed. The effects of various detergents on the conformation of the membrane proteins will be investigated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Biliary atresia (BA) is a progressive, inflammatory, sclerosing cholangiopathy that presents in infancy and leads to bile duct obstruction, biliary cirrhosis and the need for liver transplantation in the majority of patients. The etiology of BA is not known; a proposed theory is that the bile duct injury is initiated by a viral infection, followd by a progressive, autoimmune-mediated response targeting bile duct epithelia. Our laboratory and others have established the contribution of T cell-mediated inflammation and autoimmunity to bile duct injury in the rotavirus-induced mouse model of BA and in limited human studies. Recent data from our laboratory reveals that B cell-deficient mice are protected from BA, suggesting that B cells are essential to the development of bile duct injury. Research in murine models and humans have demonstrated the significant contribution of B cells to the onset and progression of many different autoimmune diseases, despite the fact that the organ-specific injury in these diseases was traditionally thought to be solely due to T cell-mediated inflammation. The specific hypotheses to be tested in this proposal are two-fold: 1. B cells play a critical role in the development and progression of bile duct injury and obstruction in murine BA; and 2. BA patients have circulating serum autoantibodies that may provide clues to disease pathogenesis and serve as prognostic biomarkers of disease severity. Specific Aim 1: Establish the contribution of B cells to development of bile duct injury in murine BA through use of B cell knockout and transgenic mice. Investigations of knockout and transgenic mice will establish the B cell mechanism involved in bile duct injury, specifically B cell antigen presentation versus immunoglobulin production. Specific Aim 2: Determine the contribution of B cells to progression of bile duct injury in murine BA through administration of B cell-depleting agents. This aim has direct translational implications to potential new therapies for human BA. Specific Aim 3: Define serum autoantibodies in BA patients and determine correlation with disease severity. Serum autoantibodies will be identified from a protein autoantigen microarray. The utility of autoantibodies in BA as serum biomarkers of disease severity will also be assessed. Significance: These investigations will add a unique perspective and increase our understanding of how B cells function in the setting of virus-induced autoimmunity. Discovery of autoantibodies in BA would provide clues to autoimmune mechanisms of pathogenesis and function as useful biomarkers to gauge severity of disease or response to novel therapies. The potential benefit of B cell depleting agents in alleviating progression of disease could change the paradigm of how physicians care for BA patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The effects of lithium in emotionally disturbed children having at least one manic-depressive lithium responsive parent will be examined. Thirty children, ages 6-12, of both sexes, having psychiatric symptoms that are in some way incapacitating and are of major concern to the child, parent, or teacher will be selected. Children will be given lithium and placebo, in a double-blind, crossover design for 16-18 weeks. Behavior ratings will include: parent's and teacher's assessment, psychophysiological measures, child interview, and cognitive testing. The lithium administration will be monitored by an \"unblind\" coinvestigator. This project has been discontinued.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The grant proposes a program of basic and clinical research in stem cell transplantation. Preclinical studies in dogs will develop nonmyeloablative conditioning programs for patients with aplastic anemia, who only require immunosuppression for hematopoietic engraftment, and for patients with genetic diseases, e.g. sickle cell disease, where 20-30% donor-type hematopoiesis may suffice to relieve symptoms. The dog model, which allows for in vivo analysis of stem cell function, will be used to define the limits of cord blood transplantation, with emphasis on in vitro expansion that is needed to increase cell yield for transplantation into adults and for gene therapy. Additional studies will focus on improved retroviral delivery systems for gene therapy, to improve upon clinical gene therapy studies already begun in patients with Gaucher's disease. Clinical studies will address ways of increasing the application and efficacy of stem cell transplantation for treating aplastic anemia (AA), myelodysplastic syndromes (MDS), and autoimmune disease, as well as genetic diseases, in conjunction with these projects, in vitro studies of cellular interactions will define how diseased cells in MDS or AA may be capable of suppressing normal hematopoiesis, possibly leading to alternative forms of therapy for these diseases as well as other marrow failure syndromes. Marrow grafts from HLA-identical siblings will be continued with emphasis on developing new regimens for preventing graft rejection and acute GVHD. Efforts to increase the number of patients eligible for transplantation will focus on the use of less well matched family members and of unrelated individuals as donors. As more patients become long-term survivors, long- term follow-up studies have become imperative to understand, treat, and prevent secondary malignancies and chronic GVHD. All of the clinical and laboratory studies are supported by core components which provide sample acquisition and analysis, as well as study design, data processing, and statistical analyses. Other cores deal with the outpatient department and grant administration. The principles derived from the proposed studies will make it possible to provide more effective treatment to a greater number of patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Experiencing the empty nest during marriage is highly normative in middle adulthood in the United States, yet scant longitudinal marital research has been conducted during this period. Establishing how spouses and marriages fare during the empty nest is important because marital functioning predicts an extensive set of older adults' outcomes in the later years of life, including physical, psychological, and financial health, and mortality. As well, midlife health and well-being reliably predict the developmental transition to old age. In this proposal, I set forth two specific aims to address gaps in our understanding of the role of the empty nest in middle adulthood marital functioning and health. First, I propose to examine empty nest status as a predictor of both marital functioning (quality and course) and health (mental and physical) for men and women over time, and to then test moderators of the longitudinal associations. Second, I propose to use dyadic data obtained from participants and their spouses to test within-couple associations between concurrent empty nest status and husbands' and wives' marital quality and health, respectively, and to again test moderators of these linkages. This proposal seeks to conduct these aims with value-added secondary analysis of the Wisconsin Longitudinal Study (WLS). The WLS is uniquely suited to accomplish the proposed aims for several reasons. First, detailed interviews that spanned middle adulthood tracked participants' empty nest status, relationship functioning, and health over time. Here, sophisticated quantitative methods will appropriately handle longitudinal data. Additionally, the WLS incorporates marital functioning indicators of quality and course in a single study, offering the potential to reconcile disparate findings in the existing empty nest literature. Second, the WLS includes a spousal sub-sample, which will facilitate direct statistical tests comparing effects of the empty nest on marital quality and health for husbands versus wives. Again, appropriate dyadic quantitative modeling will be employed. Third, a broad array of theoretically-informed spouse and marriage characteristics will be tested as covariates and potential moderators, thereby elucidating protective and risk factors. Thus, the proposed research holds important implications for translational efforts designed to prevent and alleviate distress in partners and relationships. Identifying protective factors that encourage some spouses and marriages to thrive and endure in middle adulthood is critical to understanding the determinants of healthy aging and promoting public health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Psychophysical methods will be used to study perception of temporally ordered visual stimulation. Two novel techniques have been developed in the course of this project: one that employs temporal delays to mimic color differences resulting ordinarily from changes in the luminance ratio of complementary colors, and another which provides a measure of monocular sensitivity during binocular viewing. The program has two main objectives; one is an extensive investigation of spatiotemporal relationship existing in the visual field (specific questions include: the relationship between temporal asynchrony and spatial disparity of binocular stimulation, in the absence of stimulus velocity; the relationship between perceived vertical and the vertical horopter; an evaluation of sensory correlates of perceptual suppression during binocular rivalry); the other is development of the method of temporal visual resolution as an aid in neuro-ophthalmological diagnosis (the specific questions, here, are all addressed to anisotropies which may appear across the visual field; thus, hemispheric differences in visual-temporal resolution are expected to precede explicit field defects discovered, at a later stage, by traditional techniques such as campimetry). Data obtained so far support the conclusion that the central nervous system does not compensate for differences in conduction times.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although the human immune system has evolved to repel many forms of biological invasion, its ability to identify which cells are detrimental to the body is not perfect. The research proposed herein has two main objectives: to explore a new fundamental strategy for directing the human immune system to kill target cells and to apply that strategy to treating several forms of metastasis-prone cancer beginning with HT-29 human colon cancer cells. The strategy is partially inspired by the concept of prodrugs. A molecule will be made such that when it interacts with urokinase-type plasminogen activator (uPA, an enzyme overexpressed on the surface of many types of cancer cells, especially those that are prone to metastasize), it will be chemically transformed into an electrophile that will covalently attach to the cancer cell's surface. The molecule will be bifunctional, and the other end will bind naturally occurring human antibodies. Once antibodies have bound the cancer cell, the human immune system will be activated to kill the target. This general strategy should also be applicable to treating a variety of other medical conditions. The research is divided into the following specific aims. (1) To design and synthesize a substrate analogue that functions as a latent electrophile to be activated by uPA- mediated hydrolysis. (2) To use this enzyme-activated electrophile to covalently attach antigens to the surface of cancer cells. And, (3) to study the immunological response against human cancer cells labeled by this target-activated antigen. This research will employ various methods. Bifunctional molecules will be chemically synthesized using standard organic chemistry reactions. Analysis of uPA-mediated activation of these molecules will be assayed by high-performance liquid chromatography (HPLC). Analysis of covalent attachment of the activated electrophiles to the activating protein will be assayed by mass spectrometry. Noncovalent binding between the antigen-labeled protein and human antibodies will be assayed by enzyme- linked immunosorbent assay (ELISA). Binding of antibodies to cells will be assayed by immunofluorescence microscopy. Antibody-mediated cytotoxicity will be assayed in various cancer cell lines beginning with commercially available HT-29 colon cancer cells. PUBLIC HEALTH RELEVANCE: The research proposed herein offers a strategy to use small organic molecules to direct the natural human immune system to fight cancer. Specifically, this approach will use organic chemistry to make molecules that can selectively attach to cancer cells and direct the immune system to destroy those cells. Although the first application will be against the HT-29 human colon cancer cell line, the method proposed is broad enough in scope to target several types of cancer, especially those that are most likely to metastasize throughout the body.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite the fact that HIV infection has soared amongst heterosexual women of color, Latinas are consistently underrepresented in HIV research (Carmona, Romero & Loeb, 1999). Latinas are at increased risk for HIV due cultural inhibitions of condom use, idealized gender roles, and idealization of romantic relationships (East, 1998). Yet these relationships, when chronically stressful, can exacerbate mental health problems such as depression (Hammen, 2005), and long-term effects of poor immune response (Robles & Kiecolt-Glaser, 2003); it is thus imperative to examine the effects of couple conflict and depression on HIV+ Latinas' morbidity and mortality. This study aims to investigate the links between relationship functioning and immune functioning, as measured by CD4 cell count and prevalence of gynecological problems, over time and how depression may mediate these links, in both HIV+ and HIV- heterosexual Latinas. Its results will thus be useful in understanding and creating new public health prevention and intervention methods aimed at ethnic minority women. I am using a sub-sample of HIV-positive (N=97) and HIV-negative Latinas (N=52) from the University of California-Los Angeles Charles Drew Medical Center Women and Family Project, a longitudinal study examining the impact of HIV on women's lives (see Wyatt & Chin, 1992). The data has been collected and I will specifically analyze data spanning across two years, using hierarchical linear modeling (HLM) and path analysis and focusing on measures of relationship quality, depression, CD4 cell levels, and gynecological problems. In accordance with the stress/social support hypothesis, a theory that accounts for the varied effects of stress and social support in couple relationships on physiological health (Burman & Margolin, 1992), I predict that poor relationship functioning over time will be associated with decreased CD4 levels and a higher prevalence of gynecological problems, and that depression will mediate these associations. My proposed study is highly relevant to public health research in innovative and important ways. First, my project is unique in that I am looking at HIV-positive Latinas, a population that has been overlooked in HIV research despite the fact that HIV infection is the fourth leading cause of death amongst Hispanic women (CDC, 2004). Moreover, as there is a dearth of longitudinal studies in HIV research, my project will contribute a more comprehensive picture of the intricate interplay between relationships, immune regulation, and mental health. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A major theme of this SCCOR is discovery and elucidation of genetic defects that result in congenital cardiovascular malformations, particularly those affecting valvulogenesis. The objectives of Core B are twofold: 1. To ensure the availability of cohorts of patients with specific cardiovascular anomalies in whom phenotype has been well characterized by echocardiography. To catalogue and store echocardiographic and additional relevant clinical material along with DNA from both probands and parents so that the information is easily accessible, both by the SCCOR participants and other interested parties who satisfy the privacy governance issues that will be put in place. 2. To provide echocardiographic characterization of extended family members for probands with bicuspid or bicommisural aortic valve (BAV) (Project 1) or Noonan syndrome (NS) (Project 2). The resources obtained by these critical Core functions will facilitate identification of novel candidate genes and their genotype-phenotype correlations. As candidate genes become apparent, they can immediately be moved into mechanistically based studies by all of the SCCOR's participants.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Program in Cancer Biology is focused on investigations into mechanisms and signaling pathways involved in the malignant transformation of cells. The goals are to advance basic understanding of cancer pathogenesis and to translate basic science discoveries into the clinical arena to improve the outcome of patients with cancer. The Program encompasses three major thematic groups: 1) Mechanisms and Pathways of Oncogenesis;2) Cell Cycle, Cytoskeleton and Genomic Stability;and 3) Epithelial Neoplasia (including skin, prostate and lung cancer subgroups). Investigations are focused on mechanistic studies of oncogenes and tumor suppressor genes, the control of cell division, genomic stability and DNA damage checkpoints, as well as disease-specific interests in hematologic malignancies, skin, prostate and lung cancers. The 40 investigators represent two different schools (the School of Medicine and the School of Humanities &Sciences), 16 Departments, and five Divisions within the Department of Medicine. The research activities of the 40 investigators are supported by 73 peer-reviewed, investigator-initiated grants (R01s and others), and five T32 training awards. Total NCI support is $4.3 million, other NIH support is $7.2 million, and non-NIH support is $3.2 million. Since 2005, program investigators have published over 400 manuscripts relevant to cancer biology in peer-reviewed journals. The Cancer Center will continue to be invaluable in assisting with the translation of the basic science findings from this Program into new clinical approaches for the diagnosis and treatment of patients with cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The National Institutes of Health (NIH) has defined health literacy as the degree to which individuals have the capacity to obtain, process and understand basic health information and services needed to make appropriate health decisions. Unfortunately, fully 22% of US adults have only basic and an additional 14% have below basic health literacy. The relationship between limited literacy and adverse health outcomes has been well documented, and seminal reports about health literacy have been issued by the Institute of Medicine, Agency for Healthcare Research and Quality, and the American Medical Association among others. Each of these reports advocates further research to develop an effective response and research in health literacy has begun to flourish. An important next step to cultivate this emerging research field is to create a national professional interdisciplinary home for investigators dedicated to health literacy research. In this grant application we propose to deliver a sixth annual conference along a multi-year trajectory dedicated to the promotion of the field of health literacy research. This meeting will be designed to provide a venue to focus attention on health literacy research, emphasizing the role of health literacy research in the elimination of racial and ethnic health disparities and on patient empowerment. This venue serves crucial functions for the field of health literacy by providing an incubator for young investigators and encouraging a cross-disciplinary exchange of knowledge, collaboration, and creativity. We aim to create an environment in which controversial topics within the research community can be openly discussed. Dr. Paasche-Orlow has served as a guest editor for special journal issues emerging from each of the past meetings; we plan to continue our successful dissemination efforts with another special issue on Health Literacy to be published by the Journal of Health Communication. We also plan to improve the meeting's Web site so that it can serve as a public resource for sharing information about health literacy measurement instruments. This year the meeting will take place on November 3-4, 2014 in Bethesda, MD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A major goal of this proposal is to develop a novel experimental paradigm to engineer preventative HIV-1 vaccines. The V1/V2 domain of HIV-1 envelope is an important target for vaccine development for several reasons. V2 interaction with the 47 integrin of mucosal T cells may play a key role in the efficient capture of HIV-1 virus at the site of initial exposure during sexual transmission. The V1/V2 domain is also a key player in regulating neutralization sensitivity of HIV-1, and consists of epitopes recognized by some of the most potent broadly neutralizing antibodies reported to date. Here, we hypothesize that induction of antibodies by vaccination with V1/V2 immunogens that interferes, specifically with the V2 47 interactions, confers protection against HIV-1 acquisition. Since the conformation of V2 is highly variable, the V2 domain will be constrained by transplanting it into the compactly folded 9 kDa small outer capsid protein (Soc) from bacteriophage T4. To select for the functionally important V2 conformations, libraries of V2 domain variants consisting of founder/acute virus sequences, different glycosylation patterns, and random mutations will be constructed and expressed in CHO cells as secretory proteins. High throughput assays will be developed, one to select 47 binding conformations and another to select V2 conformational antibody binding variants. The biologically active Soc-V2 conformational variants will be arrayed on phage T4 nanoparticles by incubating the purified Soc-V2 proteins with Soc-minus Hoc-minus phage, up to 870 copies per particle. A Hoc- fused targeting ligand such as Dec205 mAb will also be assembled on the same capsid, up to 155 copies per capsid, to target the T4-V2 nanoparticles to the antigen-presenting dendritic cells. The immunogenicity of T4-V2 nanoparticles will be evaluated in mice by intramuscular route as well as transcutaneous (skin) route using heat labile enterotoxin as an adjuvant. The immune responses will be quantified for V2- 47 interfering antibodies, virus neutralizing antibodies and transmission-blocking antibodies. The best V2 variants down-selected from these assays will then be tested in the rabbit model. A novel HIV-1 transmission assay will be developed, which will precisely quantify the number of virus genomes that cross the host membrane after a few minutes of exposure to CD4+, 47+ and CCR5+ T cells. Based on real-time PCR, this assay would be extremely sensitive, rapid, and high throughput. Using this assay, V2 variants that can induce antibodies which block virus entry by interfering with the initial virus-host interactions will be selected. These might potentially serve as preventative HIV-1 vaccine candidates for further studies in nonhuman primates and clinical trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Surgical resection for cancer is becoming more minimally invasive with smaller incisions and less patient morbidity. However, this process switches the burden to the surgeon to remove all the disease with suboptimal visualization. This project proposes to develop a technique to employ pre- operative injection of ultra-sound contrast enhancement stationary microbubble and microshells to enable intraoperative detection of tumors even in cases with small skin incisions to allow surgeons to better identify and completely resect tumors and decease patient morbidty. Aims: To determine the optimal dose and lifetime of stationary microbubbles in a rabbit model as an ultrasound contrast agent in order to localize small breast tumors for surgical resection. To study the localization ability of a novel gas filled silica microshells compared to microbubbles as an ultrasound contrast agent and localization method for occult breast cancers using a rabbit model. PUBLIC HEALTH RELEVANCE: Because surgical resection of small early cancers remains so important in local control and prevention of metastases, techniques to improve its efficacy in visualizing and removing tumors completely can have a major impact on breast and other cancer treatment. We believe that improving targeting of nonpalpable cancers is crucial. This fact is particularly important in this era in which tiny cancers are detected by imaging and more advanced techniques are needed for localizing tumors. This project may be able to decrease positive margins, local recurrence and possible cure of early stage cancers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Taste controls an animals food intake. For the past fifteen years, our focus has been the isolation and characterization of genes encoding taste receptors and using these to mark the cells, define the corresponding signaling pathways, dissect receptor specificity, generate topographic maps, and trace the respective neuronal connectivity circuits. This research continues to involve a long standing and wide ranging collaboration with Charles Zuker and his groups at Columbia University. Together, we have now established that at the periphery there are 5 distinct classes of taste receptor cells that are selectively tuned to respond to sweet, bitter, sour, sodium salt and savory (umami) tastants and have demonstrated that activation of these cells elicit either appetitive or aversive responses accordingly. Taste information is transmitted to the nucleus of the solitary tract (NST) in the hind-brain through sensory neurons with cell bodies in the geniculate and petrosal ganglia. Projections from the NST diverge: conscious perception of taste quality is thought to involve a pathway that innervates the primary gustatory cortex in the insula; in addition, less well characterized circuits are believed to mediate immediate responses to tastants. Our current work focuses on understanding how taste information is transmitted to the brain and represented there to generate defined percepts and to guide behavior. We are using a battery of modern molecular genetic approaches to define and trace circuits involved and ultimately to relate these circuits to taste perception. These include: Ca-imaging techniques to assess the activity of neural ensembles; screening and sequencing approaches to identify markers for select subsets of taste responsive neuron; and optogenetic, pharmacogenetic, activation and silencing as well as other silencing and ablation approaches to modulate activity of specific parts of the taste circuitry. This reporting period has seen major progress in uncovering the logic for the transmission of taste information from the tongue to the brain (manuscript in press). Specifically, the tuning of taste primary afferents appears to closely match the tuning of the taste receptor cells pointing at a highly specific transfer of information. In mice, the odorant receptors are encoded by a family of more than 1000 genes. A fundamental feature of the mammalian olfactory system is that each olfactory sensory neuron expresses just a single member of this vast family of genes. However, the details of the control of odorant receptor gene expression remain unexplained. In a collaborative project with Leonardo Belluscio, we have demonstrated new aspects of regulation that contribute to the control of odorant gene expression and have devised a system that can reliably generate mice expressing a single odorant receptor in the vast majority of olfactory sensory neurons. In this reporting period we have continued to characterize the behavioral responses of a transgenic mouse that we generated that expresses an octanal receptor in almost all olfactory sensory neurons. Although this transgenic mouse line has the potential to express a second olfactory receptor from the same synthetic promoter as the octanal receptor, surprisingly it rarely does. Indeed, whereas the octanal receptor is found in 90% of all olfactory receptor neurons, the other receptor (that responds to acetophenone) is present in just 5% of cells and these do not overlap with the cells expressing the octanal receptor. We are therefore examining the mechanisms that account for this completely unexpected gene expression pattern and believe that they will provide insight into the unusual expression pattern of this remarkable gene family of sensory receptors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This protocol evalutes the growth, physical development and pituitary function in children surviving oncology diseases. Possible alterations of physiologic mechanisms resulting in hormonal failure, including treatment modalities will be studied.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overarching goal of the proposed research is to identify the underlying causes of speech understanding problems in older adults. Older listeners, with or without significant hearing loss, often have much greater difficulty understanding speech in the presence of competing sounds than do younger listeners (CHABA, 1988; Dubno et al., 1984; Duquesnoy, 1983). For hearing-impaired listeners, differences in degree of hearing loss (or audibility) account for a large portion of the variance in listeners' ability to understand speech in the presence of competing sounds, but other factors (e.g., cognitive or central auditory factors) play an important role for both hearing-impaired and normal-hearing listeners (see Houtgast & Festin, 2008; Akeroyd, 2008; Humes, 2007; Humes & Dubno, 2010; Humes, Kidd, & Lentz, 2013). One promising hypothesis, which has received relatively little attention to date, is that effective speech understanding in the presence of competing sounds depends strongly on the ability to follow the temporal structure of an utterance and to accurately anticipate the timing of speech events through a process of attentional entrainment. According to the entrainment hypothesis (see Large & Jones, 1999; McAuley et al., 2006) the ability to attentionally track the temporal patterns of speech helps listeners to perceptually isolate and resolve the details of a single talker's speech by enhancing attention to speech events that correspond to a talker's temporal pattern. Moreover, accurate extrapolation of temporal patterns through inaudible or partially audible portions of an utterance is proposed to facilitate the integration of intermittent glimpses of speech by incorporating them in a common temporal framework. This general hypothesis will be investigated through a series of experiments that test the predictions of entrainment theory in a speech context, in combination with a new test battery that assesses individual differences in entrainment ability. A sample of 1000 young and older listeners with and without hearing loss will be tested, using shaped amplification to ensure audibility for hearing- impaired listeners. The project will employ a combination of behavioral and neurophysiological measures to determine how entrainment ability is related to speech understanding, while controlling for individual differences in basic auditory capabilities and cognitive abilities previously shown to be important for speech perception.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Applicant's Description): The investigators hypothesize that the angiostatic factor IP-10 is an endogenous inhibitor of tumor-derived angiogenesis. In this proposal, they will test this hypothesis by characterizing the role of IP-10 in regulating overall tumor-derived angiogenic activity in an animal model of human non-small cell lung cancer (NSCLC). They will demonstrate that underexpression of IP-10 imparts increased angiogenic activity to NSCLC cell lines. Next, they will reconstitute IP-10 production in NSCLC cell lines, thus restoring the angiostatic balance, using two methods. (1) In vitro treatment of NSCLC cells with interferon gamma (IFNgamma) to induce IP-10 expression, and (2) Gene transfer technology to indirectly (IFNgamma) or directly (IP-10) augment IP-10 expression. They will then show that IFNgamma and IP-10 gene transfected cells have reduced in vitro angiogenic activity and reduced tumorigenicity in SCID mice. Finally, they will treat established NSCLC tumors in SCID mice with locally administered IFNgamma or IP-10 and show that IFNgamma inhibits tumor growth via an IP-10 dependent increase in angiostatic activity. These studies will lead to important new information about the pathogenesis of NSCLC, and enable the Principal Investigator to acquire the necessary skills to develop into an independent investigator.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The study of MHC restriction of T cell specificity and more particularly the relationship of this phenomenon to Ir gene function, has been a major concern of our laboratory. This project addresses three aspects of the specificity of T lymphocytes restricted to react with antigen and self Ia molecules. 1. The nature of Ia-antigen interactions, and the specificity of T cell receptors for self Ia. 2. The control of IL-1 production at the level of the antigen presenting cells and the role of Ia molecules in this process. 3. The selection of a differentiating T cell population in the thymus restricted to react specifically with self Ia molecules.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The paternal age effect (PAE) refers to an increased risk for a particular phenotype or mutation with increasing age of the father. This is of major relevance in psychiatry where three of the most severe disorders (autism, schizophrenia, and bipolar disorder) all show a PAE. These disorders are roughly twice as likely with fathers >40 years of age and three times more likely with fathers >50, compared with 20-25 year old fathers. The birth rate for males >40 has increased by 50% in the past 25 years (now 10% of all US births), so this is becoming a significant public health issue. Recent discoveries show an increased burden of de novo copy number variants (CNVs) in autism and schizophrenia, supporting the involvement of spontaneous mutations in severe mental illness and potentially helping to explain the PAE for these disorders. However, paternal age has not been explicitly shown to increase the mutational burden of offspring in any organism. Since a highly-powered search for a PAE on CNV and point mutation burden in humans is difficult to assess because of low numbers of offspring, we propose to use the laboratory mouse to answer this question. We already have DNA samples in hand to perform this experiment via next-generation sequencing and request funds to calculate the PAE on mutational burden. Furthermore, we have collected a range of fertility-related phenotypes in old and young males and psychiatrically-related behavioral phenotypes in the offspring of young and old fathers, allowing a direct comparison of genotype to phenotypes. We have collected all of the DNA samples and relevant phenotypes, and are ready to proceed with high-throughput sequencing in order to calculate mutation rates of great relevance to biomedicine. In Aim 1 we focus on CNVs, which are readily detected at low coverage sequencing due to our selection of mice for this purpose, and in Aim 2 we focus on point mutations, which require higher coverage. Clearly, the potential reward of this project is substantial. We know that PAE is a risk factor for multiple neuropsychiatric disorders but the mechanism underlying risk is unknown. In conclusion, if this project is successful, it would lay the foundation for extension to human psychiatric samples and detailed mechanistic studies within a next-generation mouse systems biology platform, the Collaborative Cross.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Three peptidic transition-state analogs have been proposed from highly-conserved polypeptide segments of the HIV-1 envelope glycoprotein and two of these analogs have been synthesized and used as immunogens for the production of murine monoclonal antibodies. Several hundred hybridomas have been shown to bind to the transition-state analog used for immunization and several of these show differential binding to the transition-state analog vs the original-sequence peptide.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Research within the Laboratory for Integrative Neuroscience, Section on Synaptic Pharmacology, continues to focus on mechanisms underlying neuromodulation and plasticity and the effects of alcohol and other drugs of abuse on these neuronal functions. Our main interest is the function of the dorsal striatum (DS), a brain region involved in action control and selection, as well as action learning. Striatal Synaptic Plasticity and Effects of Drugs of Abuse We have examined many forms of synaptic plasticity at synapses within the DS. One of the most prominent is long-term depression (LTD) at glutamatergic corticostriatal synapses produced via activation of the Gi/o class of G-protein-coupled receptors (GPCRs). In addition to LTD induced by activation of cannabinoid type 1 (CB1) and serotonin type 1B receptors, we have also observed that activation of the mu, delta and kappa receptors for opioid peptides can induce LTD. Likewise, inhibition of opioid peptide catabolism induces LTD, and this synaptic depression appears to involve actions of endogenous opioid peptides at the three receptor subtypes. The expression of the LTD induced by mu and delta opioid receptors involves decreased probability of glutamate release, resembling the mechanism of CB1-dependent LTD. Indeed, mu opioid receptor and CB1-dependent LTD can occlude on another, indicating that they share mechanisms at an overlapping population of synapses. In contrast, delta opioid receptor-dependent LTD cannot be occluded by mu-dependent LTD, indicating separate sites of action for these two opioid-mediated forms of synaptic plasticity. We have examined effects on striatal LTD of in vivo exposure to the widely-prescribed and heavily abused opioid receptor agonist oxycodone as well as ethanol. A single injection of oxycodone in vivo leads to loss of mu- and CB1-dependent LTD. The effects persist for up to 4 days following a single drug injection. The in vivo oxycodone injection has no effect on delta receptor-mediated LTD. Similar loss of mu- and CB1-dependent forms of LTD is observed following 2-4 weeks of in vivo exposure to intoxicating concentrations of ethanol. Thus, impairment of certain forms of presynaptic LTD appears to be a common impairment produced by different drugs of abuse. Loss of LTD likely alters cortical control of striatal medium spiny neurons, perhaps contributing to alterations in striatal-dependent learning produced by drugs such as ethanol, and perhaps contributing to habitual drug seeking. It will be important to figure out how mechanisms involved in LTD are impaired by in vivo drug exposure. The metabotropic glutamate receptor 2 (mGluR2) is another presynaptic receptor implicated in modulation and long-term depression of corticostriatal glutamatergic synpases. In collaboration with the laboratories of Drs. David Goldman and Markus Heilig here at NIAAA we have confirmed that a polymorphism that produces a premature stop codon in this gene (the grm2 gene) leads to loss of function of this receptor in alcohol-preferring (P) rats. Presynaptic receptor function is completely lost at corticostriatal synapses. Studies performed by the Goldman and Heilig laboratories indicate that loss of this receptor contributes to increased ethanol drinking and preference in both rat and mouse. It will be interesting to determine the contribution to these phenotypes of mGluR2 at corticostriatal synapses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Since graduating from medical school, my goal has been to become an academic surgeon with great emphasis on a career exploring basic science issues related to clinical problems. In vascular surgery, the need for improved small diameter vascular grafts has been recognized. Through the excellent research training I've received, I have developed an interest in the heterotypic endothelial (EC) and smooth muscle cell (SMC) interactions occurring within the vascular wall. I have used a SMC-EC co-culture system to model these interactions, and have developed a novel flow device to study the effect of flow on SMC-EC interactions. My objective is to further characterize the biologic mechanisms involved in this model, but I have found I lack the specific expertise in molecular biology to proceed. At Robert Wood Johnson Medical School, I have a unique opportunity to purse my career development by working with outstanding scientists in fields that will enhance my abilities and intellect. I plan to achieve my objective through course work at Rutgers University, attendance of basic science conferences, and regular meetings with my mentor and advisors designed to monitor and enhance my progress. This program will improve my fundamental knowledge base in basic science (molecular biology, biochemistry, and biomedical engineering), and allow me to expand the breadth of techniques used in my laboratory. This will allow me to continue to define the biologic mechanisms responsible for the complex cellular and cell-material interactions. Using this co-culture flow model to study heterotypic cellular and cell-matrix/biomaterial interactions in the presence of shear stress, we have observed that SMCs promote EC adhesion and adaptation to flow. We hypothesize the failure of biomaterials to support the essential processes of FC adhesion and migration is due to a lack of key matrix components, and the development of a matrix optimal for EC attachment and migration is dependent on SMC-EC communication. I now propose to study the mechanisms by which SMCs enhance EC adaptation to flow. Using BC detachment rates and migration in the co-culture flow model as bioassays, we will evaluate if SMCs enhance BC adhesion and migration by altering cell-matrix and cell-cell interactions. We will also determine if BC apoptosis is in involved in the detachment process, and whether BC apoptosis is modified by SMCs. We will partially characterize the factor(s) present in SMC conditioned media and determine by bioassays and immunodepletion if it is likely to correspond to a previously identified compound. If the factor(s) is not identified in this manner, we will continue the purification process to isolate the factor(s) and determine its identity. Conceivably, the responsible mediator(s) could be used as an adjunct in the development of new prosthetic or tissue- engineered biosynthetic grafts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Coxiella burnetii is the causative agent of both acute Q fever and chronic endocarditis in humans. Both plasmid and chromosomal differences correlate with the type of disease (acute or chronic) caused by an isolate suggesting that the different isolates have different virulence determinants. We have designed experiments to extend our knowledge of C. burnetii genomic and plasmid DNA and their relationship to disease. Unique plasmid regions will be analyzed by cloning, restriction enzyme, Southern hybridization and sequencing analyses. We will also identify the plasmid origin of replication for studies on transformation and mutagenesis of C. burnetii. Plasmid encoded surface proteins (possible virulence factors) will be identified by looking for protease-sensitive surface proteins produced by maxicells containing pUC/C. burnetii plasmid constructs and also by TnphoA mutagenesis. We will identify chromosomal encoded surface antigens employing both cosmid and 1 ZapII gene libraries in E. coli, by screening for colonies/plaques reacting with C. burnetii-specific antisera and monoclonal antibodies to isolates with differing disease potential. The distribution of antigens in C. burnetii isolates will be determined using surface iodination, PAGE, and western analyses. To understand the role of gene in virulence and intracellular growth of C. burnetii, we have designed experiments to evaluate gene function directly within the organism. We will use previously cloned C. burnetii chromosomal and plasmid genes as specific probes to assay their transcriptional activation at different times and under different conditions, in vitro and in vivo. Concurrently, we will examine qualitative differences in C. burnetii RNA production. Genes specifying unique or differentially expressed transcripts produced at different time points in vitro or post-infection will be cloned and sequenced. Proteins being synthesized under these conditions will also be evaluated by 1 and 2 -D PAGE. In this fashion we will develop an understanding of gene expression in C. burnetii during the host-parasite interaction. We also will examine plasmids and the chromosome for alternative virulence functions; factors that enhance intracellular survival and growth and thus pathogenesis. We will clone genes encoding key metabolic functions as well as those shown to be important for intracellular survival of other organisms. These genes will be cloned from C. burnetii employing the methods of PCR, mutant complementation and using homologous genes as probes. And because the development of genetic exchange systems is vital to an understanding of rickettsial genetic mechanisms, we will continue to develop transformation and mutagenesis systems for C. burnetii using electroporation and antibiotic selection in cell cultures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Stroke is the leading cause of long-term disability in the United States. Although acute revascularization therapies can be used to abort or reduce stroke burden, there are currently no drugs that improve recovery after a stroke has happened. The inflammatory response is a promising target for such therapies as it occurs in the days and weeks after a stroke and can be both detrimental and beneficial. A major unanswered question is how the injured brain modulates immune responses, and if there are molecular pathways that can be utilized to exert beneficial or limit detrimental effects on functional recovery via modulating the overall immune response. Astrocytes are a key component of the brain&#700;s injury response so-called reactive astrocytes are ubiquitous after brain injury. They are also increasingly recognized as key components of the brain&#700;s innate immune system. We propose to ask if Transforming Growth Factor Beta (TGFbeta) signaling in astrocytes modulates inflammation after stroke because it is a master regulator of immune responses. TGFbeta can resolve immune responses after injury and drive immune cell phenotypes towards less inflammatory states. Our preliminary experiments show that TGFbeta signaling is increased in the brain after stroke, persists for weeks, and occurs in reactive astrocytes. To test if TGFbeta&#700;s function in reactive astrocytes mirror its role in other types of immune cells we constructed mice in which TGFbeta signaling is decreased only in astrocytes. We have found that primary astrocytes from these mice exhibit a more pro-inflammatory phenotype after oxygenglucose deprivation, and the mice themselves demonstrate increased inflammatory responses after stroke. Based on this data we hypothesize that after stroke, TGFbeta signaling (1) occurs in reactive astrocytes, (2) limits the inflammatory response, and (3) improves functional recovery. We plan to test our hypothesis in three Specific Aims. In Aim 1 we will use reporter mice and immunohistochemistry to determine patterns of TGFbeta signaling after stroke. We hypothesize that there are increased responses to TGF-beta for weeks after stroke, and that reactive astrocytes are responding to TGFbeta after stroke. In Aim 2 we will test the function of astrocytic TGFbeta signaling in the neuroinflammatory response to ischemia, using genetic and pharmacological approaches and in vivo and in vitro experiments to target TGFbeta signaling in astrocytes. We hypothesize that astrocytic TGFbeta signaling drives resolution of the immune response to stroke. In Aim 3 we will use a genetic mouse model to ask if stroke-induced astrocytic TGFbeta signaling is beneficial or detrimental for functional recovery. We predict that astrocytic TGFbeta signaling improves recovery from stroke. With the completion of the proposed experiments we will have defined the length and cell specificity of TGFbeta responses after stroke. We will gain insight into how astrocytes influence the immune response to stroke, and into the functional diversity of reactive astrocytes. Our findings may lead to therapies that will target the brain&#700;s immune responses and benefit patients who present for medical care in the days after stroke.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract The evidence for the degree to which indirect effects (i.e. the protection conferred to unvaccinated individuals) of influenza vaccination reduce the risk of infection is lacking. Much of our current knowledge of indirect effects of influenza vaccination comes from ecologic studies comparing communities in which school children were vaccinated to those in which they were not. Subsequently, several randomized clinical trials vaccinated children and then compared the risk in their contacts to the risk in contacts of unvaccinated children. This previous work is generally limited to one year of follow-up and has not used laboratory-confirmed infection, limiting the quality of the evidence and the ability to draw conclusions about different influenza types and subtypes. The objective of this project is to determine the indirect effects of influenza vaccination in households over multiple years. We hypothesize that indirect vaccine effects can manifest in two ways at the household level. First, vaccinated individuals may be protected against community acquired infection thus preventing them from exposing unvaccinated household contacts. Second, unvaccinated household members can be protected through reduction in infectiousness of a vaccinated household index case, and a subsequent reduction in the secondary infection risk among household contacts. Using longitudinally collected data from the Household Influenza Vaccine Effectiveness (HIVE) cohort study we plan to conduct analyses to address the following specific aims: Aim 1) Quantify the indirect and total effects of influenza vaccination in reducing the risk of influenza infection; Aim 2) Estimate the indirect effect of influenza vaccination in reducing the risk of exposure to influenza at the household level; and Aim 3) Estimate the indirect effect of influenza vaccination in reducing the risk of influenza infection in unvaccinated members following introduction of influenza to the household. The expected outcome of the work proposed in these aims is to quantify the protection of unvaccinated individuals by influenza vaccination of household contacts by influenza type/subtype. We further expect to clarify how that protection is conferred, provide clearer evidence for targeting vaccination by age in the event of vaccine shortage, and to estimate the total effect of vaccination in preventing infection. Indirect effects of influenza vaccination are assumed, despite insufficient evidence. Thus, the impact of these findings will be to improve public health messaging by informing vaccine advisory bodies and other decision makers of the total benefits influenza vaccination.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This protocol will evaluate the antitumor activity of melphalan (L-PAM) and L-buthionine sulfoximine (BSO) in patients with metastatic melanoma. The effect of BSO pretreatment on glutathione metabolism in peripheral blood lymphocytes and tumor tissue, as well as BSO pharmacodynamics, will be studied.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of the proposed research project is to study the processes of ventilation of the middle ear and sinuses. The purpose is to obtain knowledge that will have direct application to clinical diagnosis. An approved dosage of Xenon-133, a radioactive gas, is administered to normal subjects by means of a bilateral negative-positive pressure air displacement technique that yields an initial concentration of the gas in the sinus and middle ear cavities. The rate of egress of the gas from the individual cavities is measured using a scintillation camera that is interfaced to a PDR-15 computer. Simultaneously, two small probes (6 mm diameter) are placed at counters. The computer data and the portable equipment data will be directly comparable, so the portable equipment can be assessed for its value in providing reliable rates of egress. Normal rates of ventilation of the mentioned cavities are being determined from the data. Subjects with confirmed pathology of the middle ear or sinuses will then be tested to determine if a desired correlation is obtained in relationship to normal standards. It is hoped that this research will help solve some of the most crucial problems of otology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Applicant's abstract): The elimination of neurotropic viruses such as herpes simplex virus (HSV) from the nervous system is likely to be complex, and to involve other mechanisms besides direct lysis of infected cells by cytotoxic T cells (CTL) because MHC antigens are not normally expressed in the nervous system. Preliminary studies have revealed an extensive and prolonged inflammatory response in the mouse trigeminal ganglion (TG) and brain stem during latency. T cells associated with secreted IFN-g were focused around neurons that appeared morphologically normal, some of which were latently infected with HSV. TNF-a and IL-2 transcripts were also detected during latency. Based on the investigators preliminary studies and published studies of others, it is hypothesized that IFN-g and TNF play an important role in suppressing HSV replication at the acute stage contributing to the survival of infected neurons, and it is speculated further that they might also function to modulate the frequency of detectable reactivation. In this application an immune-PCR assay will be used to determine whether HSV antigens are expressed in the ganglion and brain during latency and the cytokine profile in these tissues will be determined by RNAse protection assay. The investigators will determine the extent to which IFN-g and TNF-a synergize to control HSV replication in the nervous system by comparing the course of infection in control, IFN-g knockout (gko) and TNF receptor knockout (TNFR1,2) mice inoculated with wild type HSV, or a HSV recombinant (HSV-g) that expresses an IFN-g transgene. An HSV mutant (HSV-gR) engineered to express the IFN-g receptor (IFN-gR) will be used in studies with IFN-g receptor knockout (Rgko) mice to determine whether the effects of IFN-g are mediated through interaction with the receptor on infected neurons and other cells. The potential of IFN-g and TNF to modulate the frequency of detectable HSV reactivation once initiated will be examined in latently infected Rgko mice, gko mice, and control mice inoculated with type HSV-gR, and subjected to transient hypothermia as a reactivation stimulus. Studies proposed here aim to enhance our understanding of the role of IFN-g and TNFa/b in the control of acute HSV infections in the nervous system and clarify their role in latency. Since episodes of chronic IFN-g secretion have been demonstrated during HSV infection in the nervous system, HSV may be involved in a variety of neurological disorders associated with increased cytokine expression in the brain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The myeloproliferative neoplasms (MPN), originate at the level of the pluripotent hematopoietic stem cell. Each MPN can undergo clonal evolution and hematologic progression that terminates in bone marrow failure, marked by progressive man'ow fibrosis, extramedullary hematopoiesis, and/or transfonnation to acute leultemia (AL). The leading causes of morbidity and mortality in PV and ET are thrombotic complications, followed by myelofibrosis and AL. PMF has a much graver prognosis due to marrow failure, the development of massive hepato-splenomegaly and a higher rate of transformation to AL. The somatic mutation JAK2 V617F, present in between 30-95% of patients with MPN, confers constitutive tyrosine kinase activity and plays a role in the pathophysiology of MPN. MPN have not been the subject of an organized group of investigators focused on studying their pathophysiology in a manner which would lead to the development of translational clinical trials since the demise of the P Vera Study Group two decades ago. Although individual investigators have conducted clinical trials and two multi-institutional phase III studies have been completed establishing the efficacy of aspirin in reducing risk of thrombosis, and the utility of hydroxyurea in the long term in patients with ET none of these treatments has changed the natural history of the MPN. We have established the Myeloproliferative Disorders Research Consortium (MPD-RC) an intemationai group of 24 academic centers in the US and Europe to conduct translational clinical trials in MPN. The goals of this project are to conduct a series of hypothesis driven MPD-RC investigator initiated translational clinical trials, based on interactions with the laboratory based projects in the MPD-RC, in order to identify active new agents and approaches that alone or in combination, can change both the treatment paradigm and natural history of the MPN leading to improved outcomes for such patients. The specific aims include plans to conduct phase l/ll trials in patients with PMF testing such novel approaches as: CEP 701, a JAK2 inhibitor;azacitidine and vorinostat an epigenetic approach to disease modification based on observations made in Project 5: plitidepsin an inhibitor of protein tyrosine phosphatases known to change the natural history of MF in a Gata(low)'mouse model developed in project 4, and allogeneic stem cell transplant as a potential curative approach. In PV, the effects of pegylated interferon alpha-2a as well as anti-thrombotic agents will be studied in high risk PV and ET in phase 2/3 trials. These investigator initiated trials are completely dependent on the clinical research infrastructure of the MPD-RC", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary purpose of this project is to evaluate the effects of a culturally relevant genetic cancer risk assessment (GCRA) intervention on a population of underserved women of Hispanic descent, and examine a social-cognitive-cultural model of post-GCRA cancer screening and prevention behaviors that can guide future GCRA and psychosocial interventions for this population. The following aims will be addressed: 1) Assess cognitive, social and cultural factors influencing cancer screening and prevention behaviors following culturally relevant GCRA; 2) Conduct a focus group with Latina women who recently underwent GCRA to identify important cultural components in order to individualize GCRA to this population; and 3) Evaluate changes in breast and ovarian cancer screening and prevention behaviors after culturally relevant GCRA. Two focus groups will be conducted in separate single sessions consisting of Latina women that have recently undergone GCRA. The focus group will entail completion of researcher-developed instruments and participation in facilitated discussions of the instruments, key study variables, and other culturally relevant issues. Based on focus group qualitative data specific modifications will be made to assure that the GCRA protocol is culturally relevant. All participants will undergo culturally relevant GCRA following referral by their primary physicians. Genetic testing for a BRCA mutation will be offered when there is medical necessity and a reasonable calculated probability of a detectable mutation. Cancer screening and prevention recommendations will be given based on genetic test results or empiric risk estimates. Participants will complete a baseline medical history questionnaire and survey instruments measuring acculturation, fatalism, self-efficacy, expected outcomes, and social support pre-GCRA and 3-months post GCRA. A hereditary cancer knowledge survey will be completed by participants pre GCRA, immediately post GCRA and 6- months post GCRA. A follow-up questionnaire will be administered to participants 6-months post GCRA to assess adherence to risk management recommendations. The proposed pilot study is designed to develop and implement culturally relevant GCRA services for low income Latinas and to place the intervention within a theoretical context. The long-term objectives of this study are to utilize these pilot data to develop future interventional and longitudinal studies with experimental designs which can help reduce the disparity in the prevalence, recurrence, and mortality of cancer between the majority and minority population. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The potency and safety of TRAIL has prompted clinical trials with the recombinant protein as a novel treatment for human cancer. While TRAIL is very active in killing tumor cells, recombinant TRAIL possesses drug properties that limit its efficacy such as short serum half-life, instability, and the inability to cross the intact blood-brin barrier. To overcome these limitations we identified a small molecule inducer of the TRAIL gene, TIC10 that is superior to recombinant TRAIL in terms of stability, bioavailability, ability to cros the blood-brain barrier, cost of production, and spectrum of activity. Importantly, our data shows that TIC10 is highly active in several aggressive and therapy-resistant cancers. On a mechanistic level, TIC10 causes potent antitumor effects and TRAIL-induction that is mediated by the transcription factor Foxo3a, which directly regulates the TRAIL gene promoter. Furthermore, we found that TIC10 results in the dual inactivation of Akt and ERK, thereby inhibiting their constitutive phosphorylation of Fox3a and potentiating its translocation to the nucleus and binding to the TRAIL gene promoter. We hypothesize that TIC10 induces potent antitumor effects that require Akt- and ERK-mediated Foxo3a nuclear translocation and transcriptional activation of the TRAIL gene. To address the hypothesis we propose the following specific aims: Specific Aim #1: Identify TIC10-induced effects on Foxo3a expression, phosphorylation, and binding to 14-3-3; Specific Aim #2: Elucidate the role of Akt and ERK kinases in the mechanism of action of TIC10; Specific Aim #3: Determine the differential regulation of TRAIL gene transcription by FOXO family members. These studies will create a comprehensive molecular understanding of how TIC10 harnesses Foxo3a to achieve its potent antitumor activity and also has the potential to undercover novel regulatory mechanisms of Foxo3a activity that are biologically significant. Preclinical cancer models including orthotopic xenografts and transgenic mice along with primary human tumor specimens will substantiate the efficacy and validate the mechanistic findings regarding TIC10 and Foxo3a. Together, these studies will elucidate key and novel regulatory mechanisms involving Foxo3a with the first-in- class molecule TIC10 to yield insight regarding its mechanism of action as well as in clinical samples in the context of colon cancer disease progression. Our studies will facilitate the clinica translation of a novel anti- cancer therapeutic agent through further development in preclinical laboratory studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Age-related hearing loss (ARHL) is an epidemic influencing the lives of 48 million Americans6. Current clinical protocols to detect ARHL leave room for much improvement so that early diagnosis and intervention can be achieved. One way to improve current clinical protocols is through the implementation of fast, objective, and sensitive tests of cochlear function using tools such as stimulus frequency otoacoustic emissions (SFOAEs). SFOAEs are currently not utilized clinically, despite their potential for assessing the cochlea in a frequency- specific manner and sensitivity to minor insults to the cochlea. Before clinical implementation of SFOAEs can be pursued, three major gaps in knowledge must be addressed. First, the relationship between SFOAEs and psychophysical measures of hearing must be thoroughly examined across the entire range of human hearing. Second, the sensitivity of SFOAEs must be evaluated for detecting cochlear changes due to developmental and aging processes. Third, using SFOAEs, cochlear contributions in functionally relevant auditory deficits must be determined. Recent findings suggest that SFOAEs may be more sensitive than behavioral thresholds to age-related cochlear changes. However, a deeper examination is needed to determine how SFOAEs are influenced during a critical period of human life, which shows early aging effects in distortion product OAEs. In evaluating the potential of SFOAEs in the accurate and early detection of cochlear dysfunction due to ARHL, the specific aims of this proposal are to investigate the relationship between SFOAEs and psychophysical measures of hearing up to the highest measurable frequency (Aim 1), to examine the developmental and aging effects of OHC function using SFOAE amplitudes and phase slopes (Aim 2) and to apply this understanding to validate the clinical utility of SFOAEs in detecting OHC contributions to speech perception in noise (SPIN) deficits (Aim 3). In Aim 1, the relationship between thresholds derived from SFOAE input-output functions with those obtained behaviorally will be evaluated. Additionally, tuning estimates derived from SFOAE phase slopes will be compared to psychophysical tuning curves. In Aim 2, a cross-sectional investigation of cochlear function across a significant portion of human lifespan (7 - 45 years) will be performed using both behavioral thresholds and SFOAE amplitudes and phase spanning a significant range of human hearing (0.5 - 20 kHz) using state- of-the-art calibration techniques. In Aim 3, SFOAEs as well as neural measures will be examined in individuals with SPIN deficits. By addressing the aims of this proposal, the long-term objective to advance the clinical applicability of SFOAEs for detecting age-related cochlear dysfunction can be pursued.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our long term goal is to elucidate the principles by which cAMP controls cell development, proliferation and diffrentiation. cAMP regulates a striking number of physiologic processes, including intermediary metabolism, cellular proliferation, and neuronal signaling by altering basic pattrens of gene expression via activation of cAMP response element (CRE)-directed transcription. The mechanism of the CRE-directed transcription in cell proliferation, however, is largely unexplored. To elucidate the role of the cAMP enhancer (CRE) in the control of cell proliferation, we used transcription factor-decoy oligonucleotide approach. Our studies revealed that a synthetic single-stranded oligonucleotide composed of the CRE sequence, which self-hybridizes to form a duplex/hairpin, can penetrate into cells, compete with CRE enhancers for binding transcription factors and specifically interfere with CRE- and Ap-1-directed transcription in vivo . This oligonucleotide restrained tumor cell proliferation, without affecting the growth of non-cancerous cells. These results suggest that the role of PKA in cancergenesis may involve its transcription of array of genes. Recent development of high throughput DNA microarray enables parallel analysis of expression profiles of thousands of genes in a single hybridization for complex biological systems. We examined by the microarray analysis the expression of global cellular genes that are involved in PKA-cAMP-pathways by overexpression/suppression of the wild type and mutant PKA R and C subunit genes that cause changes in the growth patterns and phenotype of cancer cells. We found that approximately 240 cDNAs, representing 10% of the total DNA elements on the array, were found to have significantly altered levels of expression after treatment with RIa antisense oligonucleotide (ODN)(48hr, 0.2 mM) which produced 50% growth inhibition. The cDNA expression for these candidate genes all exhibited greater than 2.5 fold alterations (either up- or downregulated). Importantly our results showed that in comparison to cells treated with antisense ODN, the antisense gene overexpressing cells exhibited striking overlaps in the expression profiles. The same DNA elements up- or downregulated in the antisense ODN treated cells were similarly up- or downregulated in the antisense gene overexpressing cells. On further analysis by Scatter plots, we found that less than 2% of the DNA elements which exhibited altered expression were selectively up- or down-regulated in either antisense ODN treated cells or antisense gene overexpressing cells. The microarray analysis of gene profiling is also underway in cells treated with CRE-decoy oligonucleotides. This approach will lead us to potentially survey all the genetic pathways and also to discover hitherto unrecognized novel genes that may be involved in tumor growth and tumorigenesis. Furthermore, the discovery of novel genes by this approach coupled with the genetic and biochemical analyses may unravel the mechanism of cAMP-deregulation underlying cancergenesis and offer new targets for drug development and novel treatment strategies for cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is designed to elucidate some of the mechanisms by which differentiation occurs in animal cells. Animal cells with the same genetic information produce a variety of different proteins, and different amounts of the same proteins. I propose to investigate the mechanism by which the amount of ornithine transaminase (E.C.2.6.1. 13) is regulated in differentiating animal cells. By using cells in culture, it will be possible to use cells of a single, defined type, whose environment can be controlled. The Chinese hamster ovary cell line will be used.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Ohio Quality of Work Project is a social experiment which has been undertaken with the direction of representatives of labor, management and State government to improve the quality of work and productivity. The major objective of the Project is to set up throughout Ohio well-documented models of new work structures and cooperative labor/management/worker relationships in both goods manufacturing and service (including public sector). Each demonstration will be a three and a half year process including (1) an initial research portion measuring both human and economic factors of the work, (2) a continuing education process, (3) a work change program instituted by the demonstration participants and (4) remeasurements of human and economic indicators after 18 and 36 months. The instrument measuring the quality of work and its human outcomes will be used across all demonstrations. It will allow analyses to be made over time of the relationship between the quality of the working environment and a variety of outcomes. This proposal seeks funding for a portion of the research and evaluation component of this Project, to include outside evaluations of process, management systems and industrial relations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal is to develop a cell deposition system (CDS) prototype that can be used for slide preparation of diagnostic samples. The CDS has been constructed by interfacing the opto-electronics of a flow cytometer into a computer whose output can be data from analyzed cells or a word for each cell suitable for sorting that cell. The concept of CDS can be enlarged to include arbitrary sorting formats because the format is determined by a computer program. Hence, it is a matter of rewriting the computer program to alter the sorting format of the system and its associated data storage. The prototype will be used in several applications to determine its utility with fixed and living cells. Three useful classes of sorting have been identified and will be implemented. Applications in cancer detection include preparation of microscope slides by placement of cells in a highly ordered fashion. The cells are then identified morphologically and correlated with their two-dimensional histograms; other correlations include flow cytometry data with high-resolution image analysis data. In addition, there is the potential use of this device for obtaining high-frequency chromosome spreads on a microscope slide. Under computer control, the device performs the cell deposition function. With a recently purchased computer-controlled microscope, we are able to correlate each cell's visual characteristics with its flow cytometry parameters. Cells are deposited onto microscope slides covered with albumin; additionally, cells encased in a suitable growth medium can be grown on microscope slides. (3)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract This proposal details a 5-year training program for career development and advancement in academic cardiovascular medicine for Dr. Michael A. Burke, M.D., the principle investigator. Dr. Burke is a physician- scientist at Emory University (EU) School of Medicine. He completed clinical and research training in Internal Medicine and Cardiovascular Diseases at Northwestern University through the ABIM research pathway. He then completed subspecialty fellowship training in Advanced Heart Failure (HF) and Transplant Cardiology at Brigham and Women's Hospital (BWH). Finally, he completed a post-doctoral research fellowship in the lab of Drs. Christine E. (study co-mentor) and Jonathan G. Seidman in 2015. Dr. Burke has recently established his own laboratory at EU where he is embarking on a research and career development program under the combined mentorship of Drs. Ahsan Husain (EU) and Christine Seidman (BWH). Dr. Husain is a professor of medicine and expert in cardiomyocyte biology and Dr. Seidman is a physician-scientist and cardiovascular geneticist; both have an extensive track record of training future leaders in academic cardiology. Dr. Burke's research interest focuses on characterizing the epigenetic mechanisms that regulate gene expression with progression of dilated cardiomyopathy (DCM) to HF. His long-term career goals are to translate this research into clinical advances for patients with HF. He has published important research demonstrating temporal changes in cardiac transcription using a genetic model of DCM that suggests a key role for early activation of pro-fibrotic signaling. He has recently generated new evidence suggesting that epigenetic reader proteins are a key nodal point for pathologic gene transcription in the progression of DCM. The objectives of this research proposal are (1) to characterize the roles of specific TGF? isoforms and the bromodomain and extraterminal (BET) family of epigenetic reader proteins in DCM, (2) to establish a possible mechanistic link between TGF? signaling and BETs, and (3) to define the mechanism of BET recruitment to target genes. Understanding these mechanisms will provide important fundamental insight into the biology of HF and could unlock potential therapeutic targets for this common and morbid disease. This research will teach Dr. Burke the use of advanced molecular techniques including viral vector delivery in animals, chromatin immunoprecipitation with sequencing (ChIP-seq) and single-cell RNA-seq. Dr. Burke's career development plan also includes educational resources to further his scientific knowledge. Drs. Husain, Seidman and Burke have formulated a clear timeline for career development, including publication of research, presentations at national meetings and development of a plan for his subsequent transition to independent investigator. The support provided by EU and this comprehensive career development program will optimally position Dr. Burke to compete for independent grant funding by the end of the program period.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: The major goals of this project are: 1) To investigate the mechanisms underlying the anabolic effects of thalidomide, 2) To determine the dose of hGH which achieves the optimal anabolic effect, 3) To determine whether a combination and thalidomide act synergistically to improve the functional lean body mass (LBM). The applicant proposes that administration of a lower doses of hGH may avoid the metabolic and structural side effects of hGH and combination with thalidomide may be additive or synergistic. He proposes to study the effects of these two agents under metabolic ward conditions, using metabolic balance studies, whole body kinetic studies, direct determination of muscle protein synthesis, and body fluid compartments. The specific aims are: 1) to find an optimal dose of hGH which will achieve the protein-anabolic effects without causing (or minimizing), the hypermetabolism and accumulation of non-functional extracellular tissue. 2) to determine whether thalidomide decreases the net protein degradation without increasing lipid oxidation and REE. 3) to test whether administration of thalidomide in combination with lower doses of hGH achieve a selective increase in the functional, as opposed to non-functional, lean tissue and causes a smaller increase in the REE. Total number of 42 HIV+ patients and HIV- control subjects will be included over a period of 5 years. All subjects will participate in a 2 week inpatient study, which will involve treatments with hGH or thalidomide or both. Then the HIV+ patients will continue their treatments for another 10 weeks. At the end of these period HIV+ patients will be readmitted for a 5 day period for the repeat studies. The investigations will include 1) the measurements of body weight and composition (by DEXA, bioelectrical impedance, D20 and Na bromide dilution, and midarm circumference measurement techniques), 2) metabolic balance studies of nitrogen, K, Na, SO4, 3) determination of resting energy expenditure and substrate oxidation rates by indirect calorimetry, 4) protein and lipid metabolism studies using stable isotopes of leucine and glycerol, 5) determination of muscle protein synthesis by measuring the incorporation of leucine to muscle protein and abundance of myosin heavy chain mRNA, 6) measurements of hormones, metabolites, TNFalpha and other cytokines, immunologic and virologic parameters. This study will address the mechanisms of HIV-induced wasting and investigate the role of two potential medical treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Injection drug use is a serious public health problem, leading to blood-borne infections including HBC, HCV, and HIV. In spite of advances in the prevention of HIV/AIDS and hepatitis, injection drug use continues to contribute to new infections both directly through the sharing of injection equipment, and indirectly through sexual transmission from injection drug users (IDUs) to non-IDU sex partners. Behavioral interventions with IDUs are an important component of a comprehensive HIV prevention strategy. Enhanced interventions with IDUs have shown only modest effects over and above standard interventions. The key to improving interventions with IDUs may lie in understanding variations in responses to these interventions. Latent variable approaches (also known as mixture modeling) can be used to identify subgroups of individuals with similar response patterns. These approaches are often referred to as person-centered, in contrast to the traditional variable-centered approaches. To better understand how HIV prevention interventions influence injecting and sexual behavior of IDUs, we will conduct secondary analyses of data from the Third Collaborative Injection Drug Users Study / Drug Users Intervention Trial (CIDUS-3/DUIT). This study recruited young injection drug users in five U.S. cities to test a novel peer education intervention (PEI) against an attention control on changes in HIV/HCV risk behavior. We will use mixture modeling approaches, including latent class analysis, growth mixture modeling, and latent transition analysis, to analyze program outcomes. Our aim is to identify specific subgroups of IDUs for whom the intervention was most beneficial. Moreover, we seek to identify predictors of group membership which would enable us to assign individuals to the intervention that is most likely to have an impact. The proposed analyses will increase our understanding of the effects of the standard and enhanced interventions on injection and sexual risk behavior, and lead to recommendations for improving intervention efforts with injection drug users. The results may also have wider implications for strategizing prevention interventions with other populations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Vitamin A and its metabolites (retinoids) play a critical role in cardiac morphogenesis. Retinoids are potent human teratogens linked to several types of birth defects, including cardiovascular anomalies, while vitamin A deficiency can produce ventricular chamber hypoplasia, ventricular septal defects, and aortic abnormalities. A mutation in a single members of the retinoid receptor family (RCRalpha) reproduces many of the cardiovascular defects seen in vitamin A deficient embryos. The specific aims of the present proposal will capitalize on recent advances in CRE- lox technology ti specifically mutate the RXRalpha gene within specific cardiovascular cell type compartments during in vivo cardiac development: 1) to determine the role of RXRalpha pathways within atrial and ventricular muscle cells in control of cardiovascular morphogenesis by creating mice which harbor an early, complete atrial and ventricular muscle cell lineage restricted deficiency in RXRalpha via CRE-lox technology; 2) to determine the role of RXRalpha pathways within cardiac neural crest cell lineages in the control of cardiovascular morphogenesis by creating embryos which harbor a neural crest cell lineage restricted deficiency in RXRalpha; 3) To determine the role of RXRalpha pathways within endothelial/endocardial cushion cell lineages in cardiovascular morphogenesis by creating mice which harbor an endothelial cell lineage restricted deficiency in RXRalpha; 4) To determine the role of all retinoid receptor dependent pathways in specific cardiovascular cell types (atrial muscle, ventricular muscle, endothelial, and neural crest) in cardiovascular morphogenesis by creating mice which conditionally express a dominant negative retinoid receptor in specific cardiovascular cell types via a Cre-lox switch strategy. These studies should lead to the definitive identification of the specific cardiovascular cell types that generate the RXRalpha dependent signals required for specific steps in cardiac morphogenesis. Since mutations in a number of transcription factors have been implicated in human congenital heart defects, these studies of RXRalpha could lead to establishment of a new paradigm by which an individual transcription factor may be required in distinct cardiovascular cell types at precise stages of cardiogenesis to provide the molecular cues that lead to complex steps of cardiac morphogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Exposure to isocyanate, the essential cross-linker for making polyurethane, is a leading cause of occupational asthma world-wide. Three major types of isocyanate are used commercially, methylene-diphenyl diisocyanate (MDI) and toluene diisocyanate (TDI), or aliphatic, hexamethylene diisocyanate (HDI). Despite being the newest of the three, MDI has overtaken the isocyanate market in production and consumption, for many reasons, including its unique applications and properties. Importantly, at room temperature, MDI has a much lower vapor pressure than HDI or TDI, and exists as a solid, rather than a (volatile) liquid. Thus, MDI is presumed to be safer, because the potential for respiratory tract exposure is thought to be restricted to times when the chemical is heated or aerosolized (sprayed), i.e. during application. MDI is widely used in making flexible and rigid foams, coatings, elastomers and numerous other polyurethane-based products. Exposure to MDI is the best-recognized risk factor for the development of MDI asthma and exposure reduction is the primary strategy of disease prevention. Contemporary methods of monitoring MDI exposure are severely limited, and new approaches are needed to help protect millions of people exposed at work. In this new phase II application for the project Biomonitoring Methylene Diphenyl Diisocyanate (MDI) exposure and body burden, we aim to develop innovative biomonitoring approaches to exposure surveillance for MDI. The investigative team consists of scientists and physicians from Yale University School of Medicine and the small business, L2 Diagnostics, who are applying their expertise with immunoassays, to develop blood tests that measure (2) different MDI exposure biomarkers. One biomarker is MDI-specific antibodies (IgG), produced by the immune system in response to exposure. The second biomarker is the chemical (MDI) itself conjugated to albumin, the major protein adduct in vivo. To date, we have successfully completed the (4) Specific Aims of the projects 1st phase, and published supportive findings (Wisnewski et al Analytical Biochem 2010; PMID: 20123080), including the generation of key reagents, and their extensive characterization. Completion of these milestones demonstrates our readiness to embark on the specific aims of the project's 2nd phase (listed below), which should lead to new commercial assays for biomonitoring MDI exposure, which will help prevent ongoing exposures and new cases of MDI asthma. (Aim 1) Define the analytical performance characteristics of two new laboratory assays to measure (biomarkers of) MDI exposure. (Aim 2) Determine the clinical sensitivity of new MDI exposure (laboratory-based) assays, among MDI exposed workers and different populations of control unexposed individuals. (Aim 3) Using the new MDI exposure assays, establish the kinetics of MDI immune sensitization, and indications for use in evaluating exposed workers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Organic cation (OC) transporters play important roles in the disposition and clearance of many endogenous and foreign OCs in the body. In contrast to the kidney and liver, little is known about OC elimination mechanisms in the brain. This is rather unfortunate because many CNS active compounds, including monoamine neurotransmitters (e.g. dopamine, serotonin), CNS drugs (e.g. amantadine, nicotine) and neurotoxins (e.g. 1-methyl-4-phenylpyridinium (MPP+)) are small hydrophilic OCs that rely on transporters to regulate their brain levels. This proposal focuses on plasma membrane monoamine transporter (PMAT), a novel brain OC transporter first cloned in our laboratory. While structurally related to the equilibrative nucleoside transporter family (SLC29), PMAT possesses a unique and surprisingly diverse substrate specificity, transporting structurally heterogeneous OCs such as biogenic amines, clinically used drugs and neurotoxins. In humans and rodents, PMAT is most abundantly expressed in the brain and highly concentrated in the blood-cerebrospinal fluid (CSF) barrier (i.e. BCSFB or choroid plexus). We hypothesized that PMAT is the principal OC transporter at the BCSFB and is responsible for removing a variety of endogenous and xenobiotic OCs from the brain. Three Specific Aims (SAs) have been proposed. SA1 is focused on elucidating the molecular mechanisms governing transporter-substrate interactions to explain the unique and versatile substrate specificity of PMAT. SA2 is focused on elucidating the transport mechanism of PMAT and developing a cellular model for OC flux at the BCSFB. Lastly, in SA3, we will construct and validate a transgenic animal model, which will allow a variety of mechanistic studies to reveal the in vivo action of PMAT. We will apply several experimental methods, ranging from molecular biochemical techniques, computational biology, electrophysiology, immunohistochemistry to transgenic approach, to elucidate the structure, function and biological significance of PMAT in clearing neurotransmitters, drugs and toxins in the brain. The proposed studies have broad implications in our understanding of normal and pathophysiological functions of the brain. Detailed knowledge of OC transporters at the CNS barriers can also help to explain and predict the pharmacokinetics and pharmacodynamics of OC drugs and toxins in the CNS compartment and aid in the development of new strategies for drug targeting to the brain. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Stress echocardiography is a clinically established, cost-effective technique for detecting and characterizing coronary artery disease by imaging the left ventricle (LV) of the heart at rest and then after either exercise or pharmacologically-induce stress to reveal ischemia. However, acquisitions are heavily operator dependent, two-dimensional (2D), and interpretation is generally based on qualitative assessment. While a variety of quantitative 2D approaches have been proposed in the research literature, none have been shown to be superior to the still highly variable qualitative visual comparison of rest/stress echocardiographic image sequences for detecting ischemic disease. Here, we propose that the way forward must focus on a new computational image analysis paradigm for quantitative 4D (three spatial dimensions plus time) stress echocardiography. Our strategy integrates information derived from both radiofrequency (RF) and B-mode echocardiographic images acquired using a matrix array probe. The integrated analysis system will yield accurate and robust measures of strain and strain rate - at rest, stress and differentially between rest and stress - that will identify myocardial tissue at-risk after dobutamine-induced stress. This work wil involve the development of novel (1) phase-sensitive, correlation-based RF ultrasound speckle tracking to estimate mid-wall displacements, (2) ma- chine learning techniques to localize the LV bounding surfaces and their displacements from B-mode data, (3) a meshless integration approach based on radial basis functions (RBFs) and Bayesian reasoning/sparse coding to estimate dense spatiotemporal parameters of strain and strain rate and (4) non-rigid registration of rest and stress image sequences to develop unique, 3D differential deformation parameters. The quantitative approach will be validated with implanted sonomicrometers and microsphere-derived flows using an acute canine model of stenosis. The ability of deformation and differential deformation derived from 4D stress echocardiography to detect new myocardial tissue at-risk in the presence of existing infarction will then be determined in a hybrid acute/chronic canine model of infarction with superimposed ischemia. The technique will be translated to humans and evaluated by measuring the reproducibility of our deformation and differential deformation parameters in a small cohort of subjects. Three main collaborators will team on this work. A group led by Matthew O'Donnell from the University of Washington will develop the RF-based speckle tracking methods. An image analysis group led by the PI James Duncan at Yale University will develop methods for segmentation, shape tracking, dense displacement integration and strain computation. A cardiology/physiology group under Dr. Albert Sinusas at Yale will perform the acute and chronic canine studies and the human stress echo studies. A consultant from Philips Medical Systems will work with the entire team to bridge the ultrasound image acquisition technology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Farm workers living in the United States (US) carry a burden of HIV infection that is 10 times that of the general population. Migrant farm workers (MFW) who come from Mexico to the U live in rural areas, are marginalized from mainstream society, are commonly undocumented, and often lack access to necessary health and prevention services. Females MFW in particular comprise a unique group that experience high risk of HIV infection. This sexual risk is related to the threat of intimate partner violence, low social support, low HIV knowledge, high-risk partners, and in particular, cultural norms that may influence relationship power and the ability to negotiate for protective sex in the face of infidelty. HIV risk is closely tied to interpersonal power dynamics and patriarchal ideologies that can influence relationships. Moreover, migration to a new country is a life-changing event that can alter family structure and bring about changes in cultural values and beliefs, which could be harmful or protective in relation to sexual risk. Although a number of studies have investigated HIV risk among male MFW, very little research has approached the topic of sexual health for female MFW. Therefore, understanding the perceived sexual risks and strategies used to maintain sexual health by female MFW who were born in Mexico will assist in understanding what personal factors (intrinsic strengths) and which resources (physical resources, material resources, financial resources, supportive resources, relational resources, etc.) help recent migrants to maintain sexual health. Moreover, this research will form a foundation for planning population-specific and culture-appropriate interventions for female MFW. The specific aims of this study are: 1) To describe the perceptions and experiences of female MFW of Mexican descent in relation to their health risks: 2) To Identify the beliefs/values about culture, migration, gender roles, and power that are related to sexual health risks for female MFW of Mexican Descent: 3) To describe what resources and self-protective factors female MFW of Mexican descent use to mitigate these perceived risks: and 4) To generate a theoretical framework that is grounded in the voices and narratives of female MFW of Mexican descent regarding their experiences and perspectives on sexual health, power relations, and self-protective strategies. PUBLIC HEALTH RELEVANCE: Migrant farm-workers in the United States carry a burden of HIV infection that is 10 times that of the general population, and farm-worker women are particularly at risk for HIV in relation to interpersonal power dynamics, gendered social norms, and cultural beliefs about sexual health and nursing knowledge regarding appropriate interventions to increase the health and well being of this group. This qualitative study will focs on examining perceptions of risk and self-protective strategies and resources used by farm-workers women to maintain sexual health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (from Applicant's Abstract): The Ah receptor (AHR) is a ligand activated transcription factor that binds ligands such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and is thought to mediate many of the highly toxic and carcinogenic actions of TCDD. The AHR and its DNA binding partner ARNT (Ah receptor nuclear translocator) recognize specific DNA sequences, referred to as dioxin responsive element (DREs) resulting in the activation of gene transcription. Both the AHR and ARNT are basic/helix-loop-helix proteins. Basic/helix-loop-helix proteins are characterized by their ability to form homo- or hetero-dimers, contact DNA via their basic domains and form a variety of DNA binding species that recognize distinct DNA sequences and regulate specific sets of genes. Contrary to the generally accepted theory that TCDD exerts its adverse affects through the regulation of DRE responsive genes, we propose that TCDD actions include nuclear localization of the AHR, dimerization of ARNT and the subsequent removal of ARNT from the regulatory regions of other genes. This hypothesis suggests that ARNT, in the absence of the AHR regulates genes and that this regulation is adversely affected by the presence of TCDD and the AHR. Our preliminary data has shown that ARNT can form multiple partnerships. The ARNT homodimer recognizes the CACGTG sequence, the recognization site of the Myc/Max heterodimer involved in regulating cellular proliferation and differentiation. We will use Myc/Max gene regulation as a model of AHR-independent ARNT gene regulation. First, we will determine whether the ARNT homodimer will recognize the CACGTG site with high affinity required for biological significance and compare this binding affinity to that of Myc and Max. Then we will use transient transfection assays to determine whether theARNT homodimer formation may repress biological processes such as the induction of cytochrome P4501A1 and Myc induced cellular transformation. To determine why the AHR ARNT recognize different DNA half-sites, we will perform site-directed mutagenesis of their DNA binding regions followed by footprinting analysis. Finally, we will use cell-free transcription to identify the interacting transcription factors that facilitate AHR and ARNT gene regulation and determine whether different AHR and ARNT ratios found in vivo may affect the ability of the AHR and ARNT to heterodimerize and resulting differential gene regulation. The broad, long-term objectives of this proposal are to understand how the AHR and ARNT regulate and the mechanisms that dictate the tissue-specific effects of TCDD toxicity and carcinogenicity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Characterization of surface markers on B cells at successive stages along the differential pathway from early precursor to antibody forming cells (AFC); a) H chain class and allotypes of surface immunoglobulins (Ig): relationship to Ig produced by progeny AFC; ontogeny; functional role; b) VH markers: relationship to Ig produced by progeny AFC; genetic control; c) Non-Ig surface markers: correlations with H chain class; ontogeny; genetic control; functional role with respect to antibody production. Isolation and characterization of subsets of T or B cells and definition of their function(s) in humoral immunity. Development and study of Ig allotype (CH) congenic strains: a) New congenic strain development; maintenance and genetic testing of existing congenic strains; b) Study of Ig regulatory abnormalities in congenic strains. Production of (B cell myeloma) hybrid cell lines which survive indefinitely and produce a single species of Ig: a) Lines which produce antibody to Ig class, allotype or idiotype determinants; b) Lines producing antibody to non-Ig surface determinants on B or regulatory T cells; c) Lines which produce Ig of classes or allotypes where no myeloma tumors currently exist. BIBLIOGRAPHIC REFERENCES: Sato, Vicki L., S.D. Waksal, and Leonard A. Herzenberg. 1976. Identification and separation of pre T-cells from nu/nu mice: differentiation by preculture with thymic reticuloepithelial cells. Cell. Immunol. 24: 173-185. Stout, Robert D., Donal B. Murphy, Hugh O. McDevitt and Leonard A. Herzenberg. 1977. The Fc receptor on thymus-derived lymphocytes. IV. Inhibition of binding of antigen-antibody complexes to FcR-positive T cells by anti-Ia sera. J. Exp. Med. 145:187.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A recent appellate court ruling has vacated the FDA's Final Rule that implemented the 2009 Family Smoking Prevention and Tobacco Control Act (FTA) requirement of graphic warning labels (GWLs) on cigarette packs. According to the FTA, the GWLs are to include one of nine mandatory textual warnings and a picture illustrating the negative health effects of smoking. The court decided that the GWLs selected by the FDA infringed on the tobacco company plaintiffs' First Amendment rights. The court opined that FDA's evidence of effectiveness of GWLs in general and GWLs producing strong emotional arousal in particular was insufficient to justify the infringement. Specifically, the court noted te lack of data supporting the effectiveness of GWLs in reducing actual smoking. The ruling highlights the limitations of existing data on GWLs that have been obtained predominantly through large-scale surveys using self-report outcome measures. In addition, the ruling has touched on a more basic controversy on the mechanisms underlying the effectiveness of strongly salient format and content in persuasive health communications. This project will take a unique trans- disciplinary approach to address this issue from both a clinical and neurophysiological perspective. We shall use biochemical (urine cotinine levels) and neuroimaging (Functional Magnetic Resonance Imaging, fMRI) assays, as well as traditional measures of persuasion, to compare the brain and bio-behavioral (urine cotinine) impact of GWLs that were rated high or low on the emotional response (ER) scale, used by the FDA to select GWLs for its Final Rule. We shall study 150 non-treatment seeking smokers' brain response to GWLs and monitor their urine cotinine levels, before and after a 4-week long exposure to real-life cigarette packs carrying High or Low ER GWLs. Based on our preliminary data, we hypothesize that smokers exposed to high ER GWLs will exhibit larger reductions in UC than those exposed to low ER GWLs. Based on our preliminary data and prior literature pointing to the mPFC and the amygdala as key opponent brain regions mediating the brain response to audio-visual anti-smoking ads, we hypothesize that acute exposure to high ER GWLs will be associated with higher amygdala and mPFC response than low ER GWLs. In addition, we predict that the amygdala response to GWLs will decline after 4-week long naturalistic exposure to GWLs, while mPFC response will increase, with both effects more prominent in the High ER group. Secondary hypotheses will evaluate the predictive value of acute mPFC response on urine cotinine levels change, as well changes in cigarette craving, daily intake, and brain fMRI response to visual cigarette cues prior to and after the 4-week exposure period. By applying measures of brain (fMRI), behavior (cotinine levels), persuasion and motivation (attitudes and intentions towards smoking quitting) to a cohort of smokers exposed to different affective levels of GWLs in a real-life setting, the project will provide integrated results that would complement existing population-level data and inform science-driven regulation of cigarette packaging that balances constitutional law and public health interest.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is aimed at continuing our ongoing efforts to identify and characterize novel AD genes involved in presenilin-related pathways. AD candidate genes implicated in presenilin-related pathways will be derived from three different pools: Pool 1. Positional candidate genes mapping to established AD genetic linkage peaks, including UBQLN1, VPS26A, VPS35, VDAC1, VDAC2, NCSTN, PSEN1 and TFCP2; Pool 2. Candidate genes derived from systematic meta-analyses performed on ourAlzGene.org database, including CHRNB2, DAPK1, SORCS1, SORL1, TNK1, HMGCS2, CH25H, and SOAT1; and 3. Novel AD candidate genes from our ongoing whole-genome association (WGA) screens of the NIMH AD family sample in which >1400 subjects from 457 uniformly ascertained and evaluated AD families have been genotyped using three different Affymetrix genotyping arrays: 500K genomic single nucleotide polymorphisms (SNPs), 100K genomic SNPs, and 20K coding SNPS (cSNPs). Follow-up analyses of presenilin pathway-related AD candidate genes will include genetic confirmation/replication testing, linkage disequilibrium analyses, and mutation identification. In collaboration with the other P01 projects and cores, we will also carry out biological and functional validation studies of specific candidate genes based on our genetic results. In specific aim 1, genotyping of the NIMH sample will be completed for all genes in all three pools. In specific aim 2, SNPs exhibiting genome-wide significance for family-based association with AD in the NIMH sample will be subjected to replication testing in four independent AD family samples: CAG (224 families; 505 subjects; AD: 245), NIA (353 families; 1117 DNAs; AD: 815), NCRAD (369 families; 1266 DNAs: AD: 895), and NIMH African American (24 families, 58 subjects; AD: 49). For genes exhibiting the strongest association with AD, we will carry out extensive linkage disequilibrium mapping of additional SNPs and re-sequencing in probands and unaffected individuals of specific associated families for each locus. In specific aim 3., we will perform bioinformatic (in silico) analyses of AD candidate genes to identify which SNPs represent potentially pathogenic gene mutations/variants for AD. Finally, in specific aim 4., we will collaborate with the other P01 projects and cores to carry out biological validation and functional analyses of novel AD candidate genes, including effects of RNAi silencing and overexpression of wild-type and potentially pathogenic mutations/variants on presenilin function, e.g. APP trafficking/processing as well as A(3 and AICD generation, y-secretase activity, APP-PS1 interaction, and PS1 conformation. Lay Summary: The four known AD genes (APP, PSEN1, PSEN2, and APOE) are the subjects of the vast majority of current biological research on AD. Yet, these genes represent only ~30% of the genetic variance of AD. The goal of this project is identify the additional AD genes implicated in presenilin-related biological pathways to increase our knowledge of the causes of AD and the role of the presenilins in AD pathogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Malignant gliomas represent one of the most challenging tumors to treat. Gene-based therapeutics have been advocated as a novel treatment modality. Although initial results from clinical trials have revealed the safety of this approach, evidence for efficacy has been elusive. Analysis of tumor tissue has been shown that gene delivery mediated by replication-defective vectors occurred in the immediate vicinity of the needle infection delivery tract. A potential avenue that could increase the volumetric extension of gene delivery consists of using replication- conditional (tumor-selective, oncolytic viruses as gene delivery vehicles. Over the last four years, we have engineered herpes viruses (HSV) that are defective in the function of he viral ICP6 (encoding the viral ribonucleotide reductase function) and/or the ICP34.5 (encoding a virulence function) genes. Our studies have revealed that the viral oncolytic effect can produce tumor growth inhibition and regression and that and regression and that this effect can be augmented by delivery of pro-drug-activating genes. We thus propose to increase the oncolytic efficiency of these mutants by cloning into their genome the CYP2B1 and RED genes, responsible for the activation of pro-drugs of the oxazosphorine class, such as cyclophosphamide (CPA), and the CE gene, responsible for the activation of pro-drug, irinotecan (CPT11). This new viral mutant (MGH2) will be employed to test the following hypotheses: 1) Combined intratumoral transfect of two pro-drug-activating gene therapies enhances tumor cell oncolysis by MGH, 2) This strategy provides for effective treatment in animal models of herpes simplex encephalitis, and 3) Modifications in the function of viral genes responsible for host cell toxicity and production of viral progeny alter the extent of vector-mediated pro-drug activation. Taken in conjunction, these studies will provide not only justification for a treatment modality that we will be able to test in phase I clinical trials, but also a mechanistic insight into the variables affecting cytotoxicity by the viral and pro-drug activation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The field of evolutionary biology has long been interested in the molecular basis of morphological evolution; however a clear understandingremains elusive. Abdominal pigmentation in the subgroup Drosophilinae is a well studiedtrait suitable for studies to identify the genetic changes culminating in the emergence of similar and diversepigmentation patterns. In addition to diverse pigmentation patternsit is apparent, based on phylogeny, that similar patternsof pigmentation haveappeared independently. In the melanogaster speciesgroup sexually dimorphic, segmentspecific pigmentation has recently evolved. The terminal abdominal segmentsof males are dark, whereas all segmentsare lightly pigmentedin females. Closely related groups of species are monomorphic, i.e. all segments are lightly pigmented in both sexes. In all species, pigmentation is prevented where the gene products of bric-a-brac (bab), a represserof pigmentation, are expressed. In dimorphic species, a newly evolved circuitry involving homeotic and sex-determination inputs repressesbab expression in the terminal segments of the male, but not the female. This proposal will analyze how Drosophila melanogaster bab gene regulatory elements has evolvedto integrate responsiveness to homeotic and sex-determination inputs and the extent that both similar and diverse abdominal pigmentation patterns haveevolved through the modification of orthologous bab sequences. Insight from the mechanisms of enhancer evolution and function in Drosophilinae will very likely apply to the understandingof cis-regulatoryelements in vertebrates as similar sequence alterationshave significant medical implications inhumans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will investigate how Drosophila controls the expression of its Alpha-tubulin genes. The four genes of this small multigene family offer a good opportunity to investigate the control of tissue- and time-specific transcription and to investigate mRNA characteristics that determine mRNA stability and location. The issues to be dealt with in this investigation are: 1) the location and complexity of DNA sequences that cause one family member's transcription pattern to deiifer from that of another member; and 2) whether or not there are sequences in the mRNAs from different family members that cause them to differ in fate, that is to differ in intercellular transport, in cellular location and in stability. In pursing the first issue, regions and sub-regions will be exchanged between the Alpha-tubulin genes and the resulting genes used to transform Drosophila. The pattern of expression of these altered genes will indicate which sequences are necesssary for a particular tissue- or time- specific pattern of transcription. In pursuing the second issue, portions of the mRNA encoding regions of Alpha-tubulin genes will be excahanged with portions of other genes and then a transformation assay will be used to test the effect of 3 feet and 5 feet untranslated regions, introns and translated regions on the intercellular transport, the location and the stability of Alpha-tubulin mRNAs. The initial subjects for these exchanges will be an Alpha-tubulin gene which codes for a maternal mRNA, one which codes for an mRNA that first appears between egg laying and cellular blastoderm formation and one which appears to be transcribed at all times in all tissues.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term goal of the research in this proposal is to understand the neural mechanisms controlling the expression of arousal. The research will use combined behavioral, biochemical, and electrophysiological approaches to investigate the role of peptide transmitters and co-transmitters in the expression of arousal states. lt is hypothesized that intrinsic co-transmitter modulators in buccal motoneurons function to stabilize motor contractions under varying conditions. Extrinsic modulatory systems may override this autoregulation to produce plasticity in the system, which may be needed during different behavioral states or during different classes of behavior executed by the buccal mass. The proposed projects include: 1) Purification and sequencing of bioactive peptides In the ARC muscle. 2) Study of the characteristics of the release of peptides from motoneurons. 3) Localization and functional characterization of peptidergic neurons innervating the ARC muscle. 4) Analysis of a model of the functional organization of the motor control of the buccal mass.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Infections with Plasmodium parasites, the causative agents of malaria, constitute one of the world's largest disease burdens with up to 300 million infections and 1.2 million deaths each year. The parasite maintains an incredibly complex life cycle between mosquito vector and mammalian host. After transmission, it will pass through the skin and bloodstream as sporozoites, which infect liver hepatocytes, then develops as liver stages and emerges back into the blood as red blood cell-infective merozoites. The pre-erythrocytic (sporozoites and liver stages) phase is an ideal target for vaccine development as it is asymptomatic and has limited parasite numbers. However, as only one parasite can cause a fulminant blood stage infection, preventing blood stage disease with pre-erythrocytic immunity faces a challenge. Immunization with attenuated parasites which invade and infect the liver but fail to progress to blood stage have been highly effective in preclinical and clinical studies. The conventional understanding is that this protection relies predominantly by CD8+ T cells. However, I have shown that both monoclonal antibodies (mAb) against the sporozoite protein CSP and polyclonal antibodies (pAb) elicited by immunization with whole parasites are capable of providing robust protection against an infectious mosquito bite. The Abs elicited by whole parasite immunization (WPI) can also control a direct blood stage infection independent of T cell help. Little is known about the antibody effector mechanisms mediating this protection at either stage, but the cross-stage protection afforded by WPI provides an ideal platform on which to investigate these mechanisms. I propose to use this model to investigate the contributions of neutralization, complement-mediated lysis and opsonization underlying Ab-mediated protection in a rodent model of malaria. Aim 1 will focus on protection against sporozoite infection by passive transfer of mutant mAb which lack specific FC-mediated effector functions. These mutant mAb will be compared to WT mAb for their ability to reduce liver stage burden following infection by mosquito bite. Furthermore, passive transfer of WPI serum followed by mosquito bite challenge in mice deficient in complement, FC receptor binding or both will elucidate the role of each mechanism in the context of WPI. Aim 2 will expand this model to examine antibody-mediated protection in the blood stage of disease. Again, passive transfer of WPI serum to mice deficient in complement, FC receptor binding or both will be followed by direct blood stage challenge. By monitoring subsequent parasitemia, we will be able to determine the respective contributions (if any) of each effector mechanism to control of blood stage malaria. The studies proposed here represent the first comprehensive and exhaustive analysis of the mechanisms conferring antibody-mediated protection against both the sporozoite and blood stages of Plasmodium. Knowledge of the specific type of antibody response required for effective protection at these stages will guide the rational design of the next generation of malaria vaccines aimed at preventing infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term goal of this research is to understand the mechanisms by which cell adhesion to extracellular matrix regulate cell cycle progression. Dr. Guan and his colleagues have recently found that integrin signaling through FAK plays a major role in cell cycle regulation in cell adhesion. Their analyses using an inducible expression system suggested a critical role for focal adhesion localization of FAK and its associated signaling molecules, and identified Erks and cyclin D1 as important mediators of cell cycle regulation by the integrin-FAK signaling pathways. Studies from his laboratory and others have also suggested that FAK can interact with multiple intracellular signaling molecules to trigger several downstream signaling pathways. In addition, they have identified a novel cellular protein that can associate with both FAK and its related kinase Pyk2 and may function as an inhibitor of the FAK family kinases in intracellular signaling. In this proposal, the investigators plan to dissect the molecular mechanisms underlying cell cycle regulation by integrin-FAK signaling pathways. In Aim 1, they will first analyze the relative contributions of FAK/Src vs FAK/PI3K complexes in cell cycle regulation by FAK. They will then examine the roles of signaling molecules downstream from the FAK/Src and FAK/PI3K complexes including p130cas, Grb2 and Akt. Lastly, they will determine the role of focal adhesion localization of the FAK signaling complexes. In Aim 2, the group will examine FAK regulation of cyclin D1 at transcription level by analyzing transactivation of cyclin D1 promoter reporter by FAK signaling pathways as well as the effect of inducible expression of FAK and its mutants on endogenous cyclin D1. They will also investigate regulation of cyclin D1 protein synthesis and degradation and the potential role of eIF-4E, 4E-BPs and cyclin D1 phosphorylation as mediators of cyclin D1 regulation by FAK signaling pathways. Finally, they will determine whether overexpression of cyclin D1 can rescue cell cycle inhibition caused by disruption of FAK signaling. In the last Aim, they will identify the binding sites for both FAK and FAP200. They will then investigate the potential mechanism by which FAP200 may function as an inhibitor of FAK to regulate cell cycle progression, and the alternative possibility that FAP200 may serve as a novel FAK substrate which plays a role in cell cycle regulation by FAK. The investigators hope that these studies will provide new insights into the mechanisms by which cell adhesion to extracellular matrix regulate cell cycle progression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Development of an effective HIV vaccine is the long-term solution to control HIV/AIDS world-wide. The failure of adenovirus type 5 based HIV-1 vaccines in humans that was designed to elicit primarily antiviral T cells strongly suggests the need to develop novel vaccine approaches that generate high levels of antiviral T cells with improved function as well as protective antibody. Development of novel adjuvants is necessary to enhance the immunogenicity and efficacy of HIV vaccines. In this HIVRAD, we will use molecules that target mTOR (mammalian target of rapamycin) and CD40 pathways as novel adjuvants to elicit high levels of polyfunctional virus-specific CD8 T cells, CD4 T cells that are resistant to HIV infection and high avidity protective antibody that will lead to enhanced control of HIV infection. This PPG has two projects and two cores. Our project 1 will test the potential of CD40L expressed on the surface of virus-like particles as an adjuvant to enhance the immunogenicity and efficacy of DNA/MVA SIV vaccine. Our project 2 will test the safety, immunogenicity and efficacy of rapamycin (inhibitor of mTOR) as an adjuvant for DNA/MVA SIV vaccine. In addition, we will test for the effects of rapamycin on other replication defective and replication competent viral vectors. Finally, we will test the synergy between inhibiting mTOR and activating CD40 pathways for enhancing the immunogenicity and efficacy of DNA/MVA SIV vaccines. The macaque studies that will be conducted in projects 1 and 2 will be supported by a non-human primate core. An administrative core will provide coordination between projects 1 and 2, and core B, and help with data management and data analyses. RELEVANCE: WHO estimates that there are currently 32 million humans living with HIV/AIDS. Drugs and improved treatment regimens have successfully prolonged the lives of infected individuals in first world countries. However, these are not affordable for the vast majority of HIV-infected individuals. Even in developed nations these are limited by toxicity, affordability, the need for rigorous adherence to therapy and the emergence of drug resistant viruses. Thus, there is a great need to develop a safe and effective HIV vaccine that provides a low-cost, low toxicity solution to long-term control of viral replication - the main goal of this PPG. PROJECT 1: Title: CD40L-Adjuvanted Vaccines for HIV/AIDS Project Leader: AMARA, R PROJECT 1 DESCRIPTION (provided by applicant): The failure of Ad5 based HIV-1 vaccine in humans that is designed to elicit primarily antiviral T cells strongly suggests the need to develop novel vaccine approaches that generate high levels of anti-viral T cells with improved function as well as protective Ab. The goal of this project is to adjuvant the cellular and humoral immunity elicited by our DNA/MVA vaccine that has just entered phase II testing in humans in US. Specifically, we propose to target the CD40 pathway on dendritic cells (DC) and B cells using CD40L. Stimulation of CD40 on DC results in production of IFNa and IL-12 that are critical for generation of highly functional antiviral CD8 response. Activation of B cells through CD40 is necessary for germinal center formation where affinity maturation of B cells occurs leading to generation of high avidity Ab. Here, we will express CD40L on the surface of a SIV virus-like particle (CD40L-VLP). These CD40L-VLPs are potent activators of DC and B cells in vitro. In addition, they can be targeted to DC (through interaction between gpl20 on VLP and CD4 on DC) and allow presentation of Env in its native form that is critical for eliciting broadly cross-reactive neutralizing Ab. In specific aim 1, we will test the potential of CD40L-adjuvanted DNA/MVA SIV vaccine to enhance control of a pathogenic SIV challenge. In addition, we will test whether delivering DNA by electroporation enhances the immunogenicity of the adjuvanted and non-adjuvanted DN/VMVA vaccines. Recent studies from Dr. Rafi Ahmed's lab (PI of project 2) demonstrated that mTOR regulates memory T cell development and inhibition of this pathway following infection or vaccination using rapamycin enhances the magnitude and functional quality of antigen-specific CD8 T cells. Rapamycin has also been shown to down regulate expression of CCR5 on CD4 T cells that results in marked reduction of HIV replication. This could be an added advantage for HIV vaccines, because the vaccine-elicited CCR5'virus-specific CD4 T cells may not be infected by the virus. Essentially we may be reducing the frequency of potential virus target cells while preserving the much-needed CD4 T cell help following infection. In specific aim 2, we will test the synergy between inhibiting mTOR and activating CD40 pathways for adjuvanting the immunogenicity and efficacy of DNA/MVA vaccines. RELEVANCE: WHO estimates that there are currently 32 million humans living with HIV/AIDS. Drugs and improved treatment regimens have successfully prolonged the lives of infected individuals in first world countries. However, these are not affordable for the vast majority of HIV-infected individuals. Even in developed nations these are limited by toxicity, affordability, the need for rigorous adherence to therapy and the emergence of drug resistant viruses. Thus, there is a great need to develop a safe and effective HIV vaccine that provides a low-cost, low-toxicity solution to long-term control of viral replication - the main goal of this PPG.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In collaboration with Dr. Michael Tarlov, we have further investigated the optimal sensing layer design for the characterization of the carbohydrate moiety of glycoproteins. This resulted in the recognition that multi-valency of lectins is an important factor in the choice of immobilization strategy. Previously we had recognized surface site heterogeneity is a key problem of surface binding analysis, and developed a method for extracting affinity and kinetic rate distributions from experimental data. For this purpose we had programmed a software tool that is now widely used. We have now further improved this program by facilitating the application of corrections for mass transport limited binding, broadening the applicability of this approach.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This 3-site collaborative U01 aims to improve identification of individuals who will develop schizophrenic psychosis (including brief psychotic disorder, schizophreniform disorder, schizophrenia, or schizoaffective disorder) at the initial prodromal stage of illness, prior to the onset of the full schizophrenic syndrome. Accurate identification of schizophrenic psychosis risk offers what may be the field's best hope for developing more effective treatment strategies, including secondary prevention of this typically devastating disorder. Without sensitive and specific prodromal diagnosis strategies, intervention studies are controversial, and the results of any studies will have limited impact on clinical practice. Identification efforts to date have focused on attenuated positive symptoms, but these criteria do not consider negative symptoms that occur in the prodromal stages of psychosis and are fundamental to schizophrenia. To enhance the potential sensitivity of prodrome evaluation we have developed a modified version of the \"Criteria of Prodromal Syndrome\" (COPS) that retains attenuated positive symptoms, but also considers selected negative symptoms in the diagnosis of prodromal state. We propose to develop a schizophrenic psychosis risk prediction model, and our proposed risk factors are selected based on the hypothesis that schizophrenia results from a pathological neurodevelopmental process that occurs during a critical stage of forebrain development in gestation and affects the development of neurons primarily in the thalamic, prefrontal and frontal cortical, and limbic regions of the brain (thalamolimbic- cortical circuitry [TLCC]). These neurodevelopmental abnormalities are likely to be expressed premorbidly by subtle behavioral, cognitive, and structural \"vulnerability markers\". In most cases, these abnormalities require specific maturational processes (i.e., synaptic elimination, myelination), which occur around puberty, to unmask the vulnerability and trigger dysfunction, resulting in the development or worsening of attenuated positive and negative symptoms (clinically defining the \"at risk\" state), as well as diverse but specific impairments in social function, social cognition, neurocognitive function, olfaction, and motor function. We hypothesize that as connectivity of the TLCC becomes more dysfunctional, a consequence will be increased severity of measurable impairments with more domains being affected to a greater extent. Thus, the number and severity of symptomatic manifestations of TECC circuit impairment are indicators of a biologically high-risk state for schizophrenic psychosis. Furthermore, we hypothesize that these vulnerable neural circuits may be further perturbed by environmental events that typically occur during adolescence, such as stressful life events or drug abuse. Such stressors may exceed the adaptive capacity of relevant circuits producing the characteristic symptoms that signal the onset of the illness. To develop the schizophrenic psychosis risk assessment model we propose a 3-site prospective study of 180 individuals meeting modified \"Criteria for Prodromal Syndrome\", and 80 help-seeking control subjects who will be prospectively evaluated over 2-5 years for risk of developing schizophrenic psychosis. The collaborative team has developed leading instruments in this field and has substantial expertise in social cognition, neurocognition, developmental psychopathology, statistics and data management. Each site has provefi its ability to recruit prodromal patients in a previous collaboration.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Graded changes in light intensity at dawn and dusk are a hallmark of the natural illumination cycle, yet their contribution to circadian entrainment has received little experimental attention. The program described below will make use of recently developed twilight simulation technology, for systematic investigation of the role of dawn/dusk transitions in the entrainment of circadian rhythms of nocturnal fossorial rodents (rats and hamsters) and of diurnal primates (squirrel monkeys)--animals adapted to vastly different lighting environments. In keeping with our long-term goal of elucidating the process of photic entrainment under naturalistic, ecologically valid conditions, the rodents will be provided with a dark nest compartment and will thus have the opportunity to self-select their daily light exposure. Experiments are designed to test a number of specific hypotheses, formulated on the basis of preliminary evidence from our laboratories and those of others. We aim (1) to examine entrainment and light self-exposure patterns under twilight cycles with seasonally-varied photoperiods and with different cycle lengths, and under corresponding rectangular light-dark (LD) cycles; (2) to compare the zeitgeber strength of these two cycles, as well as any after-effects they might exert on circadian period measured in constant darkness (DD): (3) to identify distinctive features of the self- selected daily illumination pattern and determine their contributions to period and phase control; (4) to evluate potential light adaptation effects occurring during the early portion of the dawn twilight signal; and (5) to analyze the temporal relations between the circadian rhythm of visual sensitivity, as measured by psychophysical signal-detection methods, and dawn/dusk illumination cycles. Recent observations in winter-depressed patients suggest that low-intensity twilight signals can have marked therapeutic consequences, and elicit shifts in sleep phase and in melatonin secretion patterns previously thought to require bright-light interventions. This, in seeking a systematic account of twilight-induced circadian effects in both nocturnal and diurnal mammals, within the present proposal, there is already a strong basis for expecting that refined clinical treatments of sleep phase and seasonal affective disorders will ensure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The polyamines, putrescine, spermidine, and spermine, are major polybasic compounds in all living cells. These amines are important for many systems related to growth and differentiation. For many years we have been studying how these polyamines are synthesized, how their biosynthesis and degradation are regulated, their physiologic functions, how they act in vivo, and the structure of the various biosynthetic enzymes. For this purpose we have constructed null mutants in each of the biosynthetic steps in both Escherichia coli and Saccharomyces cerevisiae, and have prepared overexpression systems for the biosynthetic enzymes. Our overall studies have aimed at the use of these mutants plus those of S. pombe to elucidate the physiological functions of the polyamines, and, in particular, to ascertain the physiological effects of polyamine deprivation. In our last report we reported our studies on the effects of polyamine-deprivation on cell-cycle progression in S. pombe; we showed that in the absence of polyamines the cell cycle stops in the G1-S phase, and that these cells showed profound morphological changes. During the current year we have completed our studies showing that polyamines are important in protecting E. coli cells from the toxic effects of oxygen and hydrogen peroxide. In these studies we showed that, whereas wild type cells can be grown in an 100% oxygen atmosphere, polyamine-deficient cells are killed under these conditions. Addition of polyamines to the medium prevents this toxicity. The involvement of superoxide in this toxicity was shown by the ability of the Mn-SOD plasmid to protect the polyamine-deficient cells from the oxygen toxicity. Partial protection was also achieved by the addition of certain amino acid mixtures or by addition of sucrose or sorbitol. In our most recent work we have studied the relative importance of spermidine and spermine for the growth of S. cerevisiae. In these studies we have shown that spermidine is specifically required for the growth of S. cerevisiae, and that spermine will not substitute for this requirement unless it is first converted to spermidine by the FMS1 encoded amine oxidase. In these in vivo studies we have shown that in yeast, as in other cells, spermidine, and not spermine, is required for the hypusine modification of the protein initiation factor eIF5a, and that presumably this is one reason for the specific requirement of spermidine for growth. In addition to the in vivo studies we have purified the FMS1 encoded amine oxidase from an overexpressed yeast strain, and studied the amine oxidase activity of the purified protein in vitro. Current work is directed at extending the use of the spe3 ( delta) fms1 (delta) mutants to elucidate the relative importance of spermidine and spermine in a variety of physiological functions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of the proposed research is to study the steps in the enzymatic formation of conjugated bile acids in order to enhance understanding of the control of bile acid conjugation. Using methods developed in this laboratory, it is now possible to independently assay each of the enzymatic reactions necessary for conjugation. These assays will be used to study the subcellular distribution of the enzymes involved. The enzyme proteins involved in each reaction will be purified and the substrate affinities and substrate range will be studied for each step. Physiologic and pharmacologic controls of conjugation will be studied by assaying the individual reactions. Initial work will be performed in animal models as a prerequisite for future study in human tissue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is well known that Ca2 ion is required for normal cell growth and is involved in the regulation of a number of cell functions (i.e., cell division, motility, secretion, and maintenance of cell shape). In order to achieve a better understanding of the precise role of Ca2 ion, it is first necessary to determine the specific interaction of Ca2 ion with intracellular components. The specific tasks of this proposal are to study the biochemical mechanisms of Ca2 ion in cells by studying the intracellular Ca2 ion-binding proteins. This will be accomplished by using, in a complementary manner, biochemical and cell morphological techniques. The expected common denominator for mediating Ca2 ion stimulated processes in a ubiquitous low molecular weight protein which binds Ca2 ion with high affinity. Current data suggest that this protein may prove to be the intracellular Ca2 ion-receptor protein. The general approach of this proposal is to study this intracellular Ca2 ion-receptor with regard to its biological synthesis, intracellular localization, and involvement in cell division and motility. In particular, the Ca2 ion regulation of microtubules and microfilaments will be investigated. It this manner it is hoped that further insight will be gained regarding the specific mediation of calcium regulated events in the cell.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract This is a revised renewal application for years 11-16 of the University of Florida ?Training Program in Regenerative Medicine? T32 Institutional Research Training Grant DK074367. The current renewal application seeks an additional five years of funding for a multi-disciplinary training program in Stem Cell Biology & Regenerative Medicine to support fuor pre-doctoral trainees and two post-doctoral fellows. This represents the addition of one additional pre-doctoral training slot over our current grant award, as recommended by the previous reviewers. The goal of this training program is to produce independent investigators capable of sustaining productive research programs studying the molecular and cellular aspects of stem cell biology and the use of stem cell products in patient care. Stem cell biology, outside of hematopoietic stem cells, is a relatively new area with great potential for development of novel treatments for both human and animal diseases ? Regenerative Medicine. With the emerging interest in stem cell biology and Regenerative Medicine, the need for trained researchers cannot be overstated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Epidemiological studies of US nuclear weapons workers allow evaluation of the effects of low dose, low dose rate radiation exposures accrued in an environment of mixed radiological and non-radiological exposures associated with the USDOE weapons complex. In the proposed study, we will examine a cohort of nearly 22,000 badge-monitored workers at the Savannah River Site (SRS) who will be followed over a fifty-year period. Past research on this epidemiologically important population has focused on analyses of standardized mortality ratios. The proposed study, in contrast, will focus on radiation-mortality associations in this cohort while investigating potential sources of bias and effect modification. We will examine radiation-mortality associations under varying lag assumptions, and investigate potential changes with age-at-exposure in susceptibility to the carcinogenic effects of radiation. Next, we will investigate differences between workers in the carcinogenic effects of radiation exposure due to non-radiological exposures accrued at SRS. Mechanistic models of carcinogenesis suggest that initiating exposure to some non-radiological carcinogens may modify the effects of subsequent ionizing radiation exposures. We will use a job-exposure matrix to identify workers with routine potential for exposure to several non-radiological agents, and apply innovative methods to examine the joint effects of radiological and non-radiological exposures. Finally, we will examine the role of tritium and neutron exposures in these analyses of radiation-mortality associations. We will investigate whether variation in radiation risk estimates between subgroups of workers reflects heterogeneity in radiological exposures. Study results for this large USDOE cohort will be evaluated in relation to observations from studies of other DOE facilities; and, study data will be compiled in a manner that will facilitate future pooled analyses. In this way, the proposed work will substantially strengthen the available epidemiological information about low level radiation effects in USDOE cohorts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The novel neoclerodane diterpene Salvinorin A is the most potent naturally occurring hallucinogen. It has been reported to be the first subtype selective opioid ligand. In a screening against an array of receptors, Salvinorin A inhibited only kappa-opioid receptors (KORs). The activation of KORs has been documented to induce a wide range of behavioral effects, including sedation and perceptual distortions. Given its high selectivity, Salvinorin A could be used to investigate the precise role of KORs in the modulation of human perception. This has promoted interest in the development of a concise chemical synthesis of Salvinorin A. The synthetic scheme proposed for Salvinorin A should construct the molecule in an efficient manner. The key step is an intramolecular double Michael reaction, which assembles the tricyclic core and five of the seven stereocenters. A chiral bis(oxazolinyl) copper(II)-catalyzed vinylogous aldol condensation and a chirl auxiliary-mediated aldol reaction will be used to install the remaining two stereogenic centers. A late stage intermediate is readily amenable toward the preparation of analogues for use in a structure-activity relationship study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "HIV is unable to establish infections in macaque monkeys. We previously reported that an HIV-1 derivative, designated HIV 1NL-DT5R, containing a 7 amino acid segment from SIV CA and the entire SIV vif gene was able to initiate spreading infections in cultured macaque PBMC whereas wild type HIV-1 could not. Those experiments indicated that the presence of 666 SIV nucleotide sequences (6.7%) at these two specific locations within the full-length 9894 nucleotide HIV-1 genome was sufficient to counteract innate restriction factors residing in simian cells such as APOBEC3 and TRIM5alpha family members, which otherwise block HIV-1 replication. These results have now been extended to the organismal level by showing that HIV 1NL-DT5R, which is more than 93% HIV, was able to establish infections in all 5 pig-tailed macaques inoculated and elicited humoral responses against all of the HIV-1 structural proteins using commercially available Western blotting strips.[unreadable] [unreadable] In other studies, we have continued investigations of CCR5-utilizing SHIVs that are able to consistently replicate to high titers in inoculated macaques and cause immunodeficiency in a timely manner. Our approach has been to serially passage such R5 SHIV candidates animal-to-animal and introduce genetic changes associated with augmented replicative properties into the starting molecularly cloned virus. Presently we have generated R5 SHIVs exhibiting peak plasma viremia levels reaching 7 to 8 logs of viral RNA copies/ml and set points of 4 logs of viral RNA copies/ml. In most cases, memory CD4+ T lymphocytes are exclusively targeted for infection and depletion. These R5 SHIVs can be used in vaccine experiments in which efficacy is measured from virus load determinations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The mechanisms by which the complement system regulates the metabolism of connective tissue continue to be investigated. In organ cultures of fetal rat bones, complement activation initiated by antibodies reactive with cell surface antigens results in the enhanced synthesis of prostaglandins which, in turn, causes resorption of the bone. A cell similar to a monocyte may be responsible for the elevated levels of prostaglandins found in these cultures since complement activation by immunoglobulins stimulates prostaglandin production by macrophages in culture. Immunoglobulin fragments which are incapable of directly activating these cells but which activate the alternative complement pathway are effective in this regard. Following interaction of the components of the complement system with macrophage membranes, enhanced incorporation of prostaglandin precursors into prostaglandins occurs. These studies demonstrate that both steroid as well as non-steroid inhibitors of prostaglandin biosynthesis abrogate this response. Complement activation also inhibits the growth of bone in organ cultures of calvaria. Although prostaglandin production is stimulated in these cultures, this event is unrelated to bone growth since indomethacin inhibits prostaglandin production but does not restore bone growth.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The project on Behavioral Pharmacology is primarily aimed at elucidating interactions between functions of (a) the glutamate-mediated corticostriatal system and related anatomical systems and (b) dopaminergic systems, including the nigrostriatal system, and functional effects of stimulant and neuroleptic drugs. Related to this primary aim are several related goals: 1. Elucidation of the functional effects of the \"AA2\" or quisqualic acid excitatory amino acid receptor, in relationship to stimulants, compounds with possible anticonvulsant effects, and alcohol withdrawal. 2. A search for more effective antagonists of the AA2 receptor. 3. Studies on the hypothetical non-specificity of postsynaptic denervation supersensitivity in the central nervous system. 4. Studies of CNS effects of sulfur-containing amino acids. The ultimate purpose of these studies is to enhance our understanding of the mode of action of neuroleptics and possibly brain dysfunction in schizophrenia as well. Secondary purposes are to enhance our understanding of mechanisms of stimulant drug effects, alcohol addiction, to develop compounds with potential utility for the acute treatment of stimulant overdose, schizophrenia, and seizure disorders, and to understand the possible forms of plasticity in the corticostriatal system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Animal Models Core provides critical support for the animal research proposed in all 4 projects. Support is provided for the maintenance of the transgenic mouse colonies used in the Program Project Grant. These colonies provide necessary models for the study of erythropoiesis, hemoglobin switching, and for vector-induced insertional mutagenesis. This Core provides funds for cage costs, and a research technician for genotype and breeding services. This Core also provides the expertise and resources necessary for the rhesus macaque autologous transplantation model used in projects 3 and 4. This model is essential for the further development of safe and effective gene therapy approaches to sickle cell disease. For example, the rhesus macaque model provides an accurate assessment of the efficiency by which lentiviral vectors can transduce human hematopoietic stem cells (HSCs), and whether expression of the transferred globin gene will occur at potentially therapeutic levels. Primates are also necessary to develop in vivo HSC selection strategies, given our prior experience that mouse models alone are insufficient for these studies. The Core provides expert technical services for the entire transplantation procedure, the supplies necessary to accomplish the transplant and the support the animals will require through the period of immunosuppression. We have developed a state of the art primate facility, with particular emphasis on the humane care and psychological well being of the animals, and have the necessary expertise to perform these experiments at a level commensurate with human stem cell transplantation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Integrative molecular biology requires understanding interactions of large numbers of pathways. Similarly molecular medicine increasingly relies on complex macromolecular diagnostics to guide therapeutic choices. A fundamental argument for laser capture microdissection (LCM) of tissues is that without separation of specific cell populations from complex tissues, we will miss critical control functions of thousands of regulated transcription factors, cell regulators, and receptors that are expressed at low copy number. Without detecting changes in many of these critical effectors, the integrative understanding of tissue function and pathology will not proceed effectively. In complex tissues - particularly among pathological variations - it is exceptionally difficult to measure the majority of molecules that are at low copy number per cell without first isolating specific cell populations. For example, among Cancer Genome Anatomy Project partially sequenced cDNA libraries, only LCM-dissected ovarian cancer cDNA libraries are exceptionally informative about ovarian cancer biology. After LCM isolation of pure target cells, the library construction protocol used had selectively amplified a small number of rarer transcripts to the level that allowed statistical comparison of their expression between highly purified low and high malignant potential cancer cells. Many of these ?overamplified? genes overexpressed in the high malignant potential compared to low malignant potential ovarian cancer libraries are known to be oncogenes, and genes associated with invasion and metastatic processes in other tissues. The LCM techniques that we started developing ten years ago are now widely used in molecular analysis of genetics and gene expression changes within target cells within complex tissues. However, in global proteomic and lipid studies without molecular amplification methods, the quantity of isolated cells sufficient to perform accurate characterization of less abundant species is problematic as the microscopic visualization, targeting, and isolation in laser microdissection has a maximal rate of about 1 to 20 cells per second depending on the cells? microscopic distribution within the tissues. Recently, in collaboration with NCI and CIT, we invented and are now refining an automatic ?target-directed microtransfer? technique based on macromolecule-specific labeling of cells not requiring user visualization or microscopic targeting and capable of much higher throughput rates. This technique (patent pending) is built on our solid physical understanding of thermoplastic microtransfer and uses a much simpler device and transfer films than commercial laser microdissection instruments. Our current prototype is capable of isolating all specifically immunolabeled cells or organelles within 1 cm2 region of a standard immunostained tissue section in about five seconds, which corresponds to specific separation from approximately 50,000 cells per second. With this technique we can exceed the cell separation rates of standard technologies such as fluorescence-activated cell sorting while preserving our ability to harvest directly from standard sections of complex tissues. This rapid, automated microtransfer method has improved spatial resolution (~1 micron) and is consequently particularly well-suited to isolate highly dispersed, specific cell populations (e.g., stem cells or only those neurons in the supra-optic nucleus that express vasopressin) or specific organelles (e.g., neuronal nuclei in the brain). The spatial relationships (morphology) among the specific cells in the tissue are preserved on the transfer film. As this technology becomes more robust, we will seek to integrate the microtransfer with molecular profiling of specific cells within tissues, including routine proteomic and lipidomic analyses, particularly for the large number of less abundant molecular species. If microdissection and molecular analysis can be made clinically practical, the expression levels of sets of approximately 20 to 100 critical, stage-specific disease markers within a selected cell population might provide reliable diagnosis and intermediate endpoints of response to molecular therapies in individual patients. Our analysis of large gene expression and protein databases suggests that a significant fraction of all genes is expressed in any specific cell type and that the levels of gene products universally exhibit a highly skewed power-law distribution similar to those characterizing many other complex systems. We have developed mathematical models for the evolution of such distributions that predict the observed distributions of genes, protein domains, and gene expression observed in species of increasing biological complexity. We foresee an evolution of molecular diagnosis from one based on the qualitative or quantitative analysis of a few key biomarker macromolecules to one in which special clustering algorithms analyze complex multivariate databases. Such analyses should permit a more complete identification of highly correlated clinical cases and allow us to characterize their response to molecular therapies specifically designed to prevent progression. We are attempting to develop new approaches for better integration of our thermoplastic microtransfer methods of microdissection with downstream macromolecular analysis to permit more routine and simpler multiplex molecular diagnostics. A key feature is using the polymer matrix in which target cells are embedded for affinity purification and then for direct optical detection within the transparent polymer. We are using a variety of microscopy techniques in our lab to quantitatively characterize protocols for incorporating affinity nanoparticles in the tissue and polymer matrix. In the longer term, we foresee using in situ optical labels to quantify the spatial distributions of specific molecules captured within the microtransfer and retained following simple purification steps. Coupling the robust and simple automatic microdissection with rapid purification and detection of species might provide unique abilities to follow macromolecular changes in normal tissue development and in pathologies such as cancer progression within prostate, colon, breast, lung, and ovary tissues. In continuing collaborations with NCI, we have developed standard procedures for the isolation of normal and pathological cells from clinical specimens. We have used our models of the statistics of expression levels in cell populations to identify genes differentially expressed in cancer progression. With future integration of microdissection and macromolecular analysis, we believe the critical role for many less abundantly expressed genes in determining normal function and pathological changes will be more easily studied and integrated into molecular diagnostics and selection of clinical therapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Human apolipoprotein A-IV (apo-IV) is a plasma protein whose most distinctive property is its labile affinity for the surface of plasma lipoprotein. Although the specific function of apo A-IV in lipid metabolism has not yet been determined, studies, conducted by the Principal Investigator have contributed to a growing body of evidence which implicates apo A-IV as an important factor in the intravascular metabolism of high density lipoprotein (HDL). Recent studies now suggest that the labile binding of apo A-IV to lipid surfaces may be central to its physiological function. Specifically, we have proposed that the labile binding of apo A-IV to HDL constitutes a barostatic mechanism which maintains the surface pressure of HDL in a critical range required for optimal activity of lecithin-cholesterol acyltransferase (LCAT) and cholesterol ester transfer protein (CETP). The purpose of this proposal is to investigate the impact of molecular structure on the interaction of apo A-IV and phospholipid, with particular regard tot he molecular mechanism of the CETP reaction. We therefore propose four specific aims: 1) Recombinant apo A-IV deletion mutants will be studied by spectroscopic and surface balance techniques to examine the effect of specific amino acid domains on its ordered structure, thermodynamic stability, and lipid affinity; 2) The interaction of recombinant apo A-IV deletion mutants and phospholipid will be studied by physico-chemical techniques to examine the impact of protein structure on the properties and LCAT reactivity of apo A-IV/lipid complexes; 3) The interaction of CETP and HDL will be investigated using fluorescent labelled recombinant and native HDL to determine the mechanisms by which apo A-IV facilitates CETP-catalyzed lipid exchanges and the interconversion of HDL subspecies; 4) CETP will be studied using surface balance techniques to elucidate the mechanisms of the reaction and the role of apo A-IV facilitates CETP-catalyzed lipid exchanges and the interconversion of HDL subspecies; 4) CETP will be studied using surface balance techniques to elucidate the mechanisms of the reaction and the role of apo A-IV in lipid exchange phenomena. The investigation of the structure and function of the plasma apolipoproteins has led to major advances in the diagnosis, treatment, and prevention to atherosclerotic cardiovascular disease, the leading cause of death in the USA. The long term goal of this proposal is to delineate the physiological functions of human apo A-IV.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is an urgent need to develop novel approaches to accelerate osteogenesis for the treatment of skeletal injury and disease. Combat troops and aging Veterans alike suffer from devastating orthopaedic injuries. While the young soldier suffering from a blast injury requires difficult limb reconstruction due to massive bone and soft tissue loss, the senior Veteran suffering from age-related bone loss is incapacitated by osteoporotic fractures, which are often associated with diminished osteogenic capacity, delayed healing, and non-union. Challenges in both acute trauma- and age-related fracture repairs converge on one critical deficiency: diminished ability to make bone due to a depleted stem cell pool. Autologous bone grafting, the gold standard for replenishing the stem cell pool in young healthy patients, can be unpredictable in cases of extreme tissue loss and in older patients. This is where our motivation lies: to mobilize and recruit a patient's own stem cells to the site of injury, and then activate these cells to produce bone in a timely manner. Our studies focus on a soluble stem cell recruitment factor, SDF-1, and its role as a paracrine signal regulating bone homeostasis and regeneration in response to mechanical loading and injury. The overall objective of this work is to determine the role of SDF-1 signaling during osteogenesis and regeneration using both loss-of-function models and exogenous SDF-1 treatment. Our central hypothesis is that SDF-1 acts as a paracrine mediator of progenitor recruitment and osteogenic differentiation in load- and injury-induced osteogenesis. We further hypothesize that SDF-1 treatment can accelerate bone repair in aged mice, a model of diminished regenerative capacity, possibly by enhancing cellular recruitment. To our hypotheses, we propose the following Specific Aims: (1) Determine the influence of osteoblast- and osteocyte-expressed SDF-1 on skeletal homeostasis and load-induced osteogenesis; (2) Determine the influence of osteoblast- and osteocyte-expressed SDF-1 on bone regeneration; and (3) Demonstrate that locally delivered SDF-1 can augment bone regeneration in aged mice, a model of suboptimal healing. If our hypotheses are correct, our studies will provide convincing evidence that SDF-1 works in a paracrine fashion in bone, a finding that would significantly advance our understanding of skeletal homeostasis and serve as a basis for developing therapeutic protocols to treat devastating orthopaedic injuries and disease in our Veterans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The program aims to examine the significance of changes that phagocytic leukocytes undergo when they ingest bacteria, with respect to the role those changes play in protecting the host. The leukocytes may be stimulated by solid particles, or by some more soluble substances, to produce and release compounds of both leukocytic and bacterial origin. These phenomena are relevant to maintenance of health, because they provide a bulwark against infection. On the other hand, some biochemical sequelae of this stimulation may injure host cells. Generation and release of activated oxygen species is a key phenomenon in these contexts. Studies of the nature of the enzymes involved, of the \"trigger\" mechanism for stimulation of phagocytic cells, and of morphological changes the leukocytes undergo, have been made, and will be extended. The focus will be on the effects of some to the very molecules that phagocytes release from bacteria upon the phagocytic leukocytes themselves. Some other cells may also be affected. For example, monomeric breakdown-products of bacterial cells walls are well known as immuno-adjuvants and pyrogens, and have now been found to be somnogens. Binding sites (receptors?) for these substances have been found on macrophages, and, in preliminary work, on glial but not neuronal cells. These binding sites will be characterized, their variation followed as a function of the condition of the animal, and physiological effects ascribed to the ligand related to occupancy of the cellular binding sites. Radioactive (125I) muramyl ligands have been made and the conditions for binding studies are established. Attempts to isolate the receptor (binding) proteins will employ methods that include affinity columns with immobilized peptidoglycans. Muramyl peptides (exogenous substances) and an endogenous substance (serotonin) mimic each other \"pharmacologically.\" Thus, muramyl peptides induce slow- wave sleep in which serotonin has been implicated. Indications are that the latter substance \"activates\" macrophages, e.g., by conditioning them to release activated oxygen species upon later stimulation - a function of muramyl peptides. Work will examine aspects of the synergism between stimuli and ionophores (for Ca++) and possible physiologically significant synergistic situations. Further, environmental (bodily) components may affect the lipid matrix of the phagocytic cell - membrane, triggering release of active oxygen species as shown in vitro. Both these matters could be of importance in vivo in bacterial infections, as may be the implications of the somnogenic activity of muramyl peptides.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This consortium proposal represents a joint effort to study the Omega- and (Omega-1)-hydroxylation of prostaglandins E, A, and F2Alpha by the microsomal fractions of liver, lung, kidney and male sex accessory organs of humans, rabbits, and pigs. The multi-faceted approach outlined in this proposal is intended to determine the sites of formation of hydroxylated prostaglandins, the physiological states regulating their levels, and their mode of metabolism by cytochrome(s) P-450. Human male accessory sex organs and seminal fluid will be examined for metabolic capacity or hydroxy prostaglandin content. These studies will utilize autopsy material, organs obtained surgically, and seminal fluid derived from patients seen for infertility for a variety of reasons. Rabbit lung and kidney will be studied from pregnant or progesterone-treated animals which are induced in their hydroxylation capacities toward prostaglandins E and F2Alpha. The transcriptional aspects of these induction processes will be examined by isolating mRNA from lung tissue induced 100-fold by pregnancy or progesterone treatment and translating in a cell-free system to determine the amount of mRNA synthesized with time after treatment or pregnancy. Pig kidney and lung tissue will be utilized as sources for purification of the cytochromes P-450 involved in prostaglandin Omega- or (Omega-1)-hydroxylation in untreated and progesterone-treated animals in order to study the physicochemical properties of these cytochromes P-450. By correlating the results of these studies in the various species, it is hoped that some insight will be gained with regard to the metabolic sources, fate, and significance of these hydroxylated prostaglandins.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sepsis is the leading cause of deaths in US hospitals contributing to or causing 52% of all US hospital deaths and is the most expensive condition treated in US hospitals, costing more than $17 billion annually. More than 300 million blood culture bottles are produced annually, with a market size greater than $1.2B in the US and EU alone. The two-year phase II period spanned February 2013 to February 2015 (5 R44 AI094781-03). The developments, funded in part by this grant, have created a new paradigm to diagnose blood stream infection causing sepsis. The work of this grant has assisted in creating the SpecID blood culture system, which the company is now commercializing. The SpecID solution replaces the present 3-step process for characterization of blood samples, currently divided into 1) detection of infection; 2) an obligatory Gram stain for all positive cultures performed on the bench by trained technician; and then 3) species ID by MALDI-TOF, (again requiring skilled sample preparation), The SpecID solution combines faster detection, more accurate Gram status determination than the benchtop stain, and ID more accurate than MALDI, in a single fully automated step which reduces the time to ID, streamlines lab workflow, and saves significant cost. The result is improved patient outcomes and improved lab operating performance. Phase IIB aims address application to clinical human specimens along with development of instruments and media appropriate for commercial use. The objectives of this proposal reflect the major goals required to achieve commercial entry of the SpecID smart culture system into commercial, high volume laboratories including development of the SpecID blood culture instrument, blood culture bottle sensor attachment, and the clinical and analytical studies to prove performance, gain regulatory approval and make first sales.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application is submitted is in response to PAR-09-173: Pilot Intervention and Services Research Grants (R34). We propose to use this grant mechanism to address three priority areas of the NIMH including decreasing the morbidity associated with depression, improving the mental health of females, and reducing the racial disparity in mental health. We address these issues by developing a novel approach to preventing depression in preadolescent African American girls growing up in urban, low-income environments. Aims: The specific aims for the proposed application are to: 1) Develop methods and infrastructure for carrying out tests of proof-of-principle, and 2) Establish proof-of principle through evidence of feasibility and acceptability and preliminary studies of efficacy and determination of settings for deployment. Approach: Our approach will be to target individual and interpersonal vulnerabilities that have been shown to confer risk for depression in our existing research on African American girls living in low- income, urban settings. Specifically, we aim to increase assertiveness and active coping in order to reduce early emerging depressive symptoms during the preadolescent period. In addition, we propose that increasing interpersonal connectedness with the mother via maternal engagement in the prevention program is necessary to yield effects that generalize and are maintained. Investigators: The Principal Investigators, Drs. Kathryn Keenan and Kathryn Grant, are among the leading researchers in the field of developmental psychopathology who focus on children living in urban poverty. They are currently testing developmental models of depression in girls growing up in poverty. The present application is a joint venture in which their collective expertise in depression research and recent interest in translating their empirical findings to the prevention of depression will be applied. Innovation: The three novel aspects of our approach to preventing depression in females are targeting the preadolescent period, deriving individual vulnerabilities from existing datasets to target in the prevention program, using the mother as the method of delivery of the preventive intervention, and addressing barriers that have limited participation of low-income urban African American families in treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It has repeatedly been reported that dietary Na + induces an increase in blood pressure (BP) i.e. elicits \"salt-sensitivity\" (SS) only when loaded as NaCI. However, in both normotensive and mildly hypertensive, middle-aged African-Americans (blacks), we find that: 1) In the great majority of those with SS, dietary NaHCQloading induces a robust increase in blood pressure that is fully two-thirds that induced by dietary NaCI loading. 2) In those with such \"selective Na + sensitivity\" (sNaS), NaHCO-Ioading and NaCl-loading induce similarly robust increases in renal vascular resistance (RVR). We will test this hypothesis: In most blacks, the Na + component of NaCI selectively elicits SS mainly by inducing an increase in peripheral vasoconstriction that increases RVR and is mediated by, and varies in extent directly with, the plasma concentration of Na +, but does not require, as the hypothesis of Guyton does, a NaCI-mediated increase in plasma volume that entrains an increase in cardiac output. We will study healthy, salt-sensitive and salt-resistant blacks with normal to mildly increased blood pressures. Under controlled metabolic conditions, dietary NaCI and K + will be set at 30 and 50 mmol/70kg/d, respectively. Immediately after a 7-day control period, a dietary Na-salt will be loaded, 250 mmol/d, over a 7-day period. In each subject, we will measure the effects both of NaCI and of NaHCO3-1oading on: 1) BP and Na + balance (NAB), throughout; 2) extracellular volume (ECV), plasma volume (PV) and RVR, on the 7th day of the control period and on the 2nd and 7th days of Na+-Ioading; 3) cardiac output (CO) and total peripheral resistance (TPR), daily. The hypothesis predicts that in those with sNaS: 1) Oral Na+-Ioading with either salt will induce robust increases in BP, RVR and TPR that precede any increases in CO. 2) The preceding increases will persist, and they, and the absolute values each attains, will vary directly with the concomitant plasma concentrations of Na +induced, but not with the concomitant values of CO. 3) NaHCO3-1oading will not induce greater, and may induce lesser, increases in PV, CO, ECV and NaB than those induced by NaCl-loading in salt-resistant subjects. 4) Either hypertonic NaHCQ or NaCI intravenously administered over a 2 hr-period, will induce acute increases in BP, RVR and TPR greater than those induced by an equimolar but larger volumes of hypotonic NaCI that are more PV-expanding. 5) The acute increases in plasma Na + concentration will predict the extent of acute increases in BP, RVR and TPR.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Non-uniformity of myosin ATPase activity has been observed within single cells of rat ventricles during reperfusion following a brief period of ischemia and in old rats. In the reperfused hearts the non-uniformity is transitory. If the period of ischemia is sufficiently brief the contractility of the heart returns to pre-ischemic levels and the non-uniformity in myosin ATPase activity disappears as the contractility recovers. The specific aim of this project is to determine the cause of this non-uniformity and the conditions that either predispose to it or make it less likely. The abnormality of the myofibrils may have important clinical implications because non-uniformity of myofibrillar function within a single cell may impair the ability of these cells to shorten uniformly and consequently may hamper mechanical performance of the heart. There are two possible explanations for the non-uniformity in myofibrils of old hearts: 1) non-uniform distribution of myosin isozymes; and 2) non-uniform specific activity of a single myosin isozyme. In the reperfused heart, the more likely cause is non-uniform specific activity because the rapidity of appearance is probably too great for change in myosin isozyme content of myofibrils. Ca-activated and actin-activated myosin ATPase will be determined by a microphotometrical technique that can localize and measure ATPase activity in a single A-band. The concentration of a specific isozyme of myosin will be measured in the adjacent serial sections by quantitative immunocytology. Since sections can be as thin as 3u, isozyme distribution in a cell and specific activity of myosin ATPase in a cell can be resolved within 2 sarcomere lengths. The relation of non-uniformity to activity of a cAMP dependent system that regulates myosin will be examined. The spatial relationship of the myofibrils with different myosin ATPase activity to other organelles and of cells with non-uniformity to tissue vasculature will be examined to determine if some causal relationship might exist. The effect of the duration of ischemia, the conditions of reperfusion and the degree of catecholamine stimulation on the appearance and disappearance of non-uniformity will be studied.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Marval Biosciences Inc. has developed a novel liposomal X-ray contrast agent (NCTX)1. With prior SBIR funding (1R43EB04700), we demonstrated facile blood pool imaging, and enabled the simultaneous imaging of cardiac and pulmonary features, something that is not possible to do with conventional contrast agents. Marval is currently developing large scale production facilities for NCTX, and has demonstrated 4 liter production of the agent. We anticipate producing NCTX at the 50L scale under GLP conditions within the next 12 months, and pursuing an IND application shortly afterwards. In a recent, highly significant discovery it was shown that NCTX can predict the outcome of chemotherapy using Stealth Liposomal Doxorubicin (SLD). NCTX consists of a clinically used Iodinated contrast agent encapsulated in a Pegylated liposome. One expects therefore, that NCTX will extravasate in tumors in the same way that SLD (e.g. Doxil(r)) does, by the so-called Enhanced Permeation and Retention (EPR) effect. In addition to the molecular effect of doxorubicin in inhibiting topoisomerase-II, the efficacy of SLD in treating a tumor is fundamentally dependent on (1) the extent of its localization in the tumor and (2) the release of the active drug from the liposomes after localization. It is well-known that the degree of tumor vasculature leakiness differs not only among same type tumors but even spatially within the same tumor. It was therefore hypothesized that the extent of extravasation of NCTX in individual tumors would be a predictor of the extravasation of SLD, and therefore of the efficacy of SLD in treating a specific tumor. To test these hypotheses, NCTX was used to image rat mammary adenocarcinomas using a clinical mammography instrument. It was demonstrated that in individual rats bearing tumors of identical size and morphology, those specific tumors that exhibited high NCTX uptake as visualized by X-ray imaging were the most susceptible to treatment by SLD. There are currently 128 ongoing clinical trials in the US (www.ClinicalTrials.gov), of Stealth Liposomal Doxorubicin (Doxil(r) and various generic equivalence candidates). Even if a small fraction of these trials are successful, one anticipates that the number of patients treated with SLD will dramatically increase in coming years. While liposomal doxorubicin does reduce the side effects of chemotherapy, it still has significant side effects of its own, including cardiotoxicity and hand-foot syndrome12. The ability to predict the efficacy of liposomal doxorubicin in treating a specific tumor, in a patient-specific manner, would therefore be of enormous clinical utility. In Phase 1 of this SBIR project therefore, we seek to develop NCTX as a prognosticator of SLD efficacy. If successful, this project will lead to a new indication for NCTX. To date, we have demonstrated the prognostication ability in a syngeneic rat mammary tumor, the MAT B-III adenocarcinoma. Yet, liposomal doxorubicin (specifically Doxil) is primarily used for Ovarian cancer. In this project, we therefore propose to test the prognostic ability of NCTX in relevant ovarian cancer mouse models. PUBLIC HEALTH RELEVANCE: Marval Biosciences Inc. has developed a novel liposomal X-ray contrast agent (NCTX)1. With prior SBIR funding (1R43EB04700), we demonstrated facile blood pool imaging, and enabled the simultaneous imaging of cardiac and pulmonary features, something that is not possible to do with conventional contrast agents. Marval is currently developing large scale production facilities for NCTX, and has demonstrated 4 liter production of the agent. We anticipate producing NCTX at the 50L scale under GLP conditions within the next 12 months, and pursuing an IND application shortly afterwards. In a recent, highly significant discovery it was shown that NCTX can predict the outcome of chemotherapy using Stealth Liposomal Doxorubicin (SLD). NCTX consists of a clinically used Iodinated contrast agent encapsulated in a Pegylated liposome. One expects therefore, that NCTX will extravasate in tumors in the same way that SLD (e.g. Doxil(r)) does, by the so-called Enhanced Permeation and Retention (EPR) effect. In addition to the molecular effect of doxorubicin in inhibiting topoisomerase-II, the efficacy of SLD in treating a tumor is fundamentally dependent on (1) the extent of its localization in the tumor and (2) the release of the active drug from the liposomes after localization. It is well-known that the degree of tumor vasculature leakiness differs not only among same type tumors but even spatially within the same tumor. It was therefore hypothesized that the extent of extravasation of NCTX in individual tumors would be a predictor of the extravasation of SLD, and therefore of the efficacy of SLD in treating a specific tumor. To test these hypotheses, NCTX was used to image rat mammary adenocarcinomas using a clinical mammography instrument. It was demonstrated that in individual rats bearing tumors of identical size and morphology, those specific tumors that exhibited high NCTX uptake as visualized by X-ray imaging were the most susceptible to treatment by SLD. There are currently 128 ongoing clinical trials in the US (www.ClinicalTrials.gov), of Stealth Liposomal Doxorubicin (Doxil(r) and various generic equivalence candidates). Even if a small fraction of these trials are successful, one anticipates that the number of patients treated with SLD will dramatically increase in coming years. While liposomal doxorubicin does reduce the side effects of chemotherapy, it still has significant side effects of its own, including cardiotoxicity and hand-foot syndrome12. The ability to predict the efficacy of liposomal doxorubicin in treating a specific tumor, in a patient-specific manner, would therefore be of enormous clinical utility. In Phase 1 of this SBIR project therefore, we seek to develop NCTX as a prognosticator of SLD efficacy. If successful, this project will lead to a new indication for NCTX. To date, we have demonstrated the prognostication ability in a syngeneic rat mammary tumor, the MAT B-III adenocarcinoma. Yet, liposomal doxorubicin (specifically Doxil) is primarily used for Ovarian cancer. In this project, we therefore propose to test the prognostic ability of NCTX in relevant ovarian cancer mouse models.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/ Abstract Bioassay-guided fractionation of cells often uncovers small molecules that bind macromolecular targets in new and unexpected ways. Exploration of the chemical reactivity and target selectivity of these metabolites has laid the chemical foundation for the development of new biological tools and therapeutics. The molecular architecture of secondary metabolites is challenging and different than the chemical space explored by most medicinal chemistry campaigns: there is recognized `natural product-like' space and we lack the tools to explore it with the same depth as `drug-like' space. Our lab has developed new tools to more easily access natural products, and has focused our efforts on secondary metabolite families that appear to covalently modify their targets as the basis for their phenotypic effects. Our chemical syntheses are deliberately concise and easily-scaled to enable subsequent investigation into reactivity and biological activity. The current application significantly advances these efforts, provides compelling preliminary data as a foundation for the proposed work, and delves into new areas of chemistry. In this proposal, two areas of research are described: 1. the identification and investigation of covalently-reactive pharmacophores associated with the asmarine alkaloids, Nuphar dimers and isocyanoterpenes; and 2. the development of cross-coupling technology to access `natural product-space' more generally. In the first area, we disclose a preliminary cellular target of the unusual N-hydroxydiazepine purine (HAP) pharmacophore of the asmarine alkaloids and address unsolved problems posed by this motif. As part of a theme that runs throughout our work, we show how the aims of chemistry and biology intersect in the study of covalently reactive secondary metabolites. We also investigate the sulfur-electrophilicity of the Nuphar dimers, a property recently demonstrated by our lab to operate in organic solvent and cellular environments. We propose solutions to the challenging stereochemical problems posed by the monohydroxy dimers and show how chemistry developed in our lab can generate a combinatorial library of sulfur electrophiles. In a third area, we investigate the challenging architecture and reactivity of the isocyanoterpenes, which we recently demonstrated can kill Plasmodia by an alternative mechanism to the heme detoxification pathway often suggested. We also propose a novel caging strategy to render the isonitriles systemically-viable. In part two, we investigate a bimetallic catalytic cycle capable of solving long-standing problems in chemical synthesis. The proposed methodology is supported by proof-of-principle examples and provides chemists the reaction vehicles necessary to `escape from flatland.'", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Methods and Analysis Core will have three functions: 1. to supply, operate and maintain the specialized equipment used in individual Research Projects for monitoring a variety of overt circadian rhythms in rodents and humans and to implement standard procedures for their computer storage. 2. to provide investigators in the individual Research Projects with standard methods for reporting and analyzing temporal profiles of activity, temperature, sleep stages, hormonal levels, and behavioral variables. In particular, the Core will provide novel methods for statistical analysis that are specially designed for studies with multiple outcome measures, as are all studies proposed in the four individual Projects. 3. to organize and operate a centralized data bank in order to allow for comparisons between standardized recordings of sleep, body temperature, wrist activity and performance tests among subject groups in individual Projects #1 and #2 as well as for comparisons between standardized recordings of activity and other behavioral measures in rodents in individual Projects #3 and #4. The services provided by this Core Facility will be used by all four individual Research Projects. The availability of a centralized pool of monitoring equipment will prevent unnecessary equipment duplication and provide the individual investigators with expert assistance for optimal operation and trouble-shooting of the equipment. The availability of standardized procedures for data reporting and analysis will enable each investigator to extract as much information as possible from the collected data and facilitate the integration of individual projects as well as communication between investigators in the various Research Projects. The operations of a centralized data bank will permit to perform additional comparisons of various study groups across Projects and thus derive more information from the Program Project as a whole than from the sum of each of its components.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fertility in females requires controlled maturation of the oocyte, supporting granulosa cells (GCs), and thecal cells that comprise the ovarian follicle. Follicle growth is a dynamic process that demands exquisite regulation. Follicles are restrained at the preantral stage until they are stimulated by the pituitary hormone follicle stimulating hormone (FSH). In response to FSH GCs produce steroid and protein hormones and growth factors that regulate the hypothalamic/pituitary axis and uterine receptivity, and promote oocyte maturation and development of the follicle to a preovulatory phenotype. All of the documented responses to FSH are mediated via cAMP and it's predominate intracellular target, cAMP-dependent protein kinase (PKA). Indeed, GCs offer one of the best examples of a cellular model whose responses are orchestrated by PKA. Signaling by PKA is confined to specific locations in cells by virtue of a family of A-kinase anchoring proteins (AKAPs) that localize pools of PKA, their substrates, and interconnected signaling enzymes. This application focuses on the mechanisms by which PKA integrates transcriptional networks to imitate maturation of GCs. PKA accomplishes this integrating function by phosphorylating substrates that directly regulate transcription or by regulating pathways whose targets regulate transcription. The co-activator beta-catenin is emerging as one potential PKA substrate necessary for activation of a subset of FSH target genes. PKA also phosphorylates an unidentified substrate that directs activation of the phosphatidylinositol-3 kinase (PI-3K) pathway fundamental to GC survival, proliferation, and differentiation. Among the many PI-3K pathway targets, the transcriptional factor FOXO1 (forkhead box O factor 1) requires phosphorylation/inactivation to permit induction of at least a subset of FSH target genes. We postulate that PKA phosphorylates both beta-catenin to activate its co- activator activity and a substrate to direct activation of the PI-3K pathway to activate and inactivate a network of FOXO1-regulated target genes. Aims test the following hypotheses: that a specific AKAP targets a pool of PKA to a multi-enzyme complex that directs activation of PI-3K;that induction of FSH target genes like Lhcgr requires activation of beta-catenin;and that the PI-3K pathway target FOXO1 regulates a network of direct target genes that maintain GCs in an immature stage. Understanding how FSH signals to direct follicular maturation can translate into safer and more effective treatments of infertility and early pregnancy loss as well as new approaches for contraceptive drugs. PUBLIC HEALTH RELEVANCE: FSH signaling to mature follicles to the preovulatory phenotype is required for fertility. Understanding how FSH signals to direct follicular maturation can translate into safer and more effective treatments of infertility and early pregnancy loss as well as new approaches for contraceptive drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fetal alcohol syndrome is a significant preventable health problem world wide. Studies in humans and animals suggest that prenatal ethanol exposure (E) also disrupts the normal development of the immune system, resulting in immune disfunction that may persist well into adult life. Data also indicate that stress can have a profound negative impact on immunocompetence. The proposed research is the first to investigate the interactive effects of prenatal ethanol and postweaning stress on immunocompetence. We have shown that E results in: 1) long-term effects on offspring immunocompetence; 2) alterations in the hypothalamic-pituitary-adrenal (HPA) response to stress, Our preliminary studies demonstrate interactive effects of prenatal ethanol and stress in adulthood on lymphocyte populations. The proposed research will focus on one critical aspen of the immune response, T cell--B cell interactions. The ability of T helper cells to facilitate immunoglobulin production by B cells, and conversely, the ability of B cells to respond to T lymphocyte-derived activation and proliferation signals, will be assessed. In adulthood, offspring will be subjected to a 3 week chronic intermittent stress regimen or will remain undisturbed. Experiments in which mitogen-activated T cell supernatants from both prenatally treated and control animals are cultured with activated B cells from both prenatally treated and control animals will allow us to determine if the potential defect lies on the T cell or the B cell side of this system, or if both cell types are functioning abnormally. Our working hypothesis is that prenatal ethanol will cause abnormal T cell--B cell interactions, via either defective T cell activation or aberrant lymphokine production by activated T cells. Our experimental paradigm will also enable us to determine whether B cells are affected. We also hypothesize that stress will exacerbate the immunosuppressive effects of prenatal ethanol, and based on our previous data, that males will be more affected than females by exposure to prenatal ethanol and stress. This approach will increase our understanding of two fundamentally important issues: the nature of the immunoteratogenic effects of ethanol, and the possible role of stress in exacerbating these effects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lyme disease was once restricted to coastal North Atlantic and selected Upper Midwest communities, but the distribution and prevalence of this zoonosis has greatly expanded in the last two decades. The aggressively human biting deer tick vector of the agent of Lyme disease was first recognized to maintain a distinct lineage of Powassan virus (POW) in 1997, but human encephalitis cases attributable to ?deer tick virus? was recognized only in 2006. Severe neurologic disease, a hallmark of classical Powassan fever, remained rare in residents of Lyme-endemic sites until recently. Encephalitis cases attributed to Powassan virus are now being increasingly reported from New England and the upper Midwest. The biological basis for the recent zoonotic emergence of POW requires analysis. The Eurasian tick borne encephalitis virus complex (TBEV) comprises diverse species (including POW), subtypes and geographic isolates that vary in their capacity to cause human disease, and there is a rich literature concerning variation in the genetic, phenotypic and clinical characteristics of Eurasian TBEV that should guide our analysis of the potential public health burden of POW in North America. Our overaraching hypothesis is that Powassan virus, like TBEV, also comprises lineages, genotypes, or populations that differ in capacity to cause human disease. We propose to test this hypothesis using our existing geographic isolates of POW as well as additional isolates that we shall collect for their capacity to cause neurologic disease using a published mouse model for TBEV neurotropism. In addition, it may be that certain lineages, genotypes, or populations of POW are more efficiently transmitted by the human biting deer tick vector of Lyme disease, and this helps to explain the emerging epidemiological situation. Accordingly, we shall also determine whether geographic isolates may differ in their capacity to infect and be transmitted by ticks. Finally, we shall apply the powerful tools of whole genome sequencing to analyze the population structure of Powassan virus as well as identify any genetic correlates of neurotropic capacity. Taken together, these observations may help explain the changing epidemiology of an arbovirus that has been silently enzootic for over 2 decades in Lyme disease endemic sites, but is now apparently an emerging zoonosis. Ultimately, by describing the pathobiological correlates of Powassan genetic diversity, we can better define the potential for tick borne encephalitis to continue to emerge as a public health burden in North America.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proximal action of parathyroid hormone (PTH appears to be activation of cyclic AMP formation in its target tissues, bone and kidney. Prostaglandin E1, a potent stimulator of bone resorption, also activates cAMP production. In addition, there are a number of other hormonal and pharmacologic agents which affect calcium metabolism, but do not directly activate adenylate cyclase or stimulate cAMP accumulation in bone or kidney. These include thyroid hormone, gonadal steroids, adrenocortical steroids, and benzothiadiazine diuretics. The proposed studies investigate the possibility that these agents modulate the endorgan responses to PTH and PGE1 through an interaction on cAMP formation. BIBLIOGRAPHIC REFERENCES: Marcus, R. Cyclic Nucleotide Phosphodiesterase from Bone: Characterization of the Enzyme and Studies of Inhibition by Thyroid Hormones Endocrinology 96:400, 1975. Marcus, R., C. Lundquist, and I.J. Chopra. In vitro inhibition of cyclic nucleotide phosphodiesterase by thyroid hormones. Abstract #470 Program of the 57th Annual Meeting, The Endocrine Society, New York, June 18-20, 1975.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Reactive hyperemia is a normal physiologic event in response to an abrupt total obstruction of blood flow. In the myocardium this event has been noted for years but only recently has the technique been developed that allows the time related transmural blood flow pattern to this insult to be documented. Two groups of dogs were studied using radioactive microspheres to determine the tramsmural myocardial blood flow during reactive hyperemia. The unrestrictive group, group I, underwent acute occlusion of the circumflex, coronary artery for 15 seconds and transmural flow was determined at 5 seconds, 45 seconds, and 90 seconds after release of the occlusion. The restrictive group, group II, was the same as group I, but in addition had a partial occlusion of the circumflex coronary artery that maintained the distal coronary pressure at approximately 80 mm. Hg mean pressure. Transmural blood flow pattern of group I revealed that both epicardium and endocardium underwent reactive hyperemia of identical magnitude and that the time of peak flow in each was identical. Furthermore, the endocardial/epicardial flow ratio remained normal throughout the reactive hyperemic response. Preliminary data from group II would indicate that the epicardial flow is greater in magnitude and earlier in time than that of the endocardium which is of lesser magnitude and later in time in the reactive hyperemia response.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A hallmark of immunity is its longevity, manifested by the durability of memory B cells (Bmem ) and plasma cells (PC).Through the analysis and isolation of extremely rare antigen-specific Bmem populations, we have scrutinized their functional responsiveness and their transcriptome. Such genetic analyses and functional studies in vivo have opened up new biological leads into the cells and factors that govern the fate of Bmem. The future five years will be devoted to new exciting initiatives in Bmem biology. Over the last funding period, we have: 1) Defined the role of BCR affinity in B cell fate, 2) Developed new strategies to isolate and study Bmem, 3) Resolved the role of BAFF family members in Bmem and PC survival;4) Shown that Bmem can be selectively primed relative to PCs, 5) Identified a lineage commitment factor, Id2, for Bmem, 6) Discounted the contribution of TLR stimulation in Bmem persistence and 7) Identified CCR6 as an important chemokine receptor in Bmem migration and function. Given these findings, the Specific Aims for the next funding period are: 1) To define the role of TLR signaling and bystander T cell help in Bmem survival. The prevailing paradigm is that TLR signaling or bystander T cell help sustains Bmem longevity. Our preliminary data shows that TLR agonists in vivo do not induce Bmem cycling, but instead induce terminal Bmem differentiation. Studies are proposed to comprehensively evaluate TLR and T cell signaling on Bmem fate. 2) Continue studies to understand the role of Id2 as a lineage commitment factor in the differentiation of Bmem. Transcriptional profiling has shown that Bmem over-express Id2. Id2 is a nuclear factor that antagonizes transcription. Studies in ld-2-deficient mice (B-restricted) confirm a selective defect in the Bmem compartment. Studies are proposed to fully evaluate the role of Id2 in post-germinal center B cell differentiation and Bmem in mice where Id2 can be conditionally regulated. 3) Resolve the functional significance that Bmem are selectively responsive to CCR6 stimulation. An hypothesis is presented that selective responsiveness to CCR6 may be central to the longevity of Bmem. Our understanding of Bmem and PC biology is central to the development and improvement of vaccines used to enhance the public health. While vaccines are undoubtedly the most successful form of immunotherapy to date, a broader and more molecular understanding of the factors and cells that govern the durability of immunity is well warranted.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cancer cells exhibit alterations in parameters of nuclear architecture that control cell fate and compromise control of cell groyvth. Our Program has established new biological paradigms by showing that gene regulatory factors integrate cell signaling at chromatin microenvironments ('subnuclear foci') and support epigenetic mechanisms through association with mitotic chromosomes. In collaboration with other Program Project investigators. Project 1 will now establish new dimensions in gene regulation by defining perturbations in architecturally linked regulatory mechanisms during interphase and mitosis in AML and breast cancer cells. Our central hypothesis is that (i) subnuclear targeting of transcription factors to gene regulatory foci during interphase and (ii) the association of transcription factors wifh their target genes in mitotic chromosomes are fundamental to the retention of biological states of normal and cancer cells. Therefore, we will use IF microscopy, biochemical, genomic and proteomic approaches (i) to characterize modificafions in architectural epigenetics and molecular pathological consequences of expressing the translocation-related t(8;21) AML-ETO fusion protein (Aim 1), (ii) to analyze genes that are transcriptionally and spatially controlled by Runx2 in chromatin micro-environments ('subnuclear foci') during interphase in breast cancer ceils (Aim 2), and (iii) to examine Runx2 mediated architectural epigenetics in breast cancer cells by characterization of Runx2 and cognate gene regulatory factors that associate with mitotic chromosomes (Aim 3). By investigating the functional role of Runx2 in establishing chromatin micro- environments ('subnuclear foci') during interphase and architectural epigenetics in cancer cells during mitosis, we will challenge traditional biochemical views of gene regulation by defining the pathological linkages between modifications in nuclear architecture and gene expression that are fundamental to the molecular etiology of tumorigenesis. RELEVANCE (See instructions): Changes in the overall shape and structure ofthe nucleus are pathological hallmarks of cancer cells that are linked to cellular transformation. This study will use state-of-the-art methods to characterize how targeting of oncogenic transcription factors to specific subnuclear structures and mitotic chromosomes supports gene regulation as components of a novel epigenetic mechanism ('architectural epigenetics').", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Certain neuron classes in the cerebral cortex exhibit heightened vulnerability in Alzheimer's Disease, whereas others are relatively resistant to pathology. For example quantitative neuroanatomic analyses suggest that large pyramidal cells that furnish long corticocortical projections and reside in layers III and V may be particularly vulnerable to degeneration and/or neurofibrillary tangle and neuritic plaque formation. The experiments outlined in this project are directed at the development of a detailed molecular/anatomic profile of these pyramidal neurons. In addition, investigation of the cellular pathology of cerebral cortex in other dementing illnesses will be initiated, in order to determine the degree to which other dementias can be traced to the loss of the same subset of cortical cells and connections, regardless of the etiology. A common neuroanatomic denominator for dementia would greatly facilitate attempts to understand the cellular basis of dementing illnesses, and in turn, aid in directing therapeutic and preventative efforts to the appropriate cell class. These studies will involve detailed quantitative neuroanatomic analyses of non-human primate as well as neurologically normal and diseased human cortices. The anatomic methods employed will be chemically-specific histochemical procedures that allow for the correlation between biochemical or molecular data and cellular localization. The biochemical systems to be investigated include, cytoskeletal proteins, calcium-binding proteins, amyloid-beta-protein, and the excitatory neurotransmitter glutamate. The quantitative neuroanatomic analyses rely on computer-assisted microscopy and involve studies of cell distribution as well as morphologic and intracellular characteristics of individual neurons. An important underlying assumption in this proposal is that the selective vulnerability in Alzheimer's Disease is related to the molecular and anatomic heterogeneity of the neuronal constituents of cortex, and that a specific anatomic/molecular profile renders a cell vulnerable. If the elements of the biochemical and anatomic phenotype that are most clearly linked to differential cellular vulnerability in AD can be determined then it will be possible to develop means to protect those neurons that degenerate in AD. The protection of these neurons must be the paramount goal in developing a strategy for the management of AD, since prevention of a neurodegenerative disease is much more likely to be achievable than the development of a cure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Photodynamic therapy (PDT) with the phthalocyanine photosensitize PC 4 causes rapid generation of reactive oxygen species (ROS), mitochondrial permeability transition (MPT), depolarization and swelling, release of cytochrome c, and activation of both necrosis and caspase-dependent apoptosis in tumor cells. However, PC 4 and a recently synthesized PC 4 derivative PC 181 also localize to endoplasmic reticulum (ER) and lysosomes. We hypothesize that damage to these organelles by PC 4 and PC 181 leads to additional perturbations, such as Ca2+, iron and protease release that ultimately promote MPT-dependent cell killing after PDT. Our goal is to further characterize the role of the MPT in PDT-induced killing of cancer cells and to determine the interactions of damage to ER and lysosomes in promotion of death pathways. In Aim 1, we will characterize changes of cytosolic, mitochondrial and ER Ca2+ after PDT using compartmentally localized fluorescence indicators and Confocal microscopy. Using specific interventions, we will determine whether depletion of ER Ca2+ stores, inhibition of mitochondrial Ca2+ uptake and intramitochondrial chelation act to suppress MPT after PDT and prevent subsequent cell killing. We also determine whether increased cytosolic Ca2+ may activate Ca2+-dependent calpains, to cause Bid processing and translocation to mitochondria with con- sequent mitochondrial dysfunction, MPT, and cell death. In Aim 2, we will investigate the contribution of lysosomal iron and protease release in relation to the MPT and cell death. We will measure cytosolic and mitochondrial chelatable iron using specific fluorescent indicator and determine the effects of compartmentally loaded iron chelators. Similarly, we will assess protease release into the cytosol and investigate the protection conferred by specific cathepsin indicators and knock-out cells. We expect iron and protease release from lysosomes to synergistically promote mitochondrial permeabilization by activating Bid cleavage and translocation to mitochondria. In Aim 3, we will investigate strategies to enhance PDT toxicity in cultured cells and in isolated mitochondria. We will evaluate the ability of salicylate to decrease the threshold of MPT onset and to enhance tumor killing after PDT. The proposed studies will enhance our understanding how to enhance PDT-induced killing of cancer cells and further increase the efficacy of PDT translation to the benefit for cancer patients. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goals of the proposed work are (1) to establish a role of cortico-cortical feedback in visual decision- making, (2) to examine a potential link between top-down cortico-cortical feedback such as feature-based attention and decision-related activity in visual neurons and (3) to expand the candidate's experimental expertise by optogenetic techniques in behaving primates. Achieving the third objective will complement the candidate's previous training in awake behaving primate visual neurophysiology. Achieving the first and second objective will provide a launching point for the candidate to develop an independent research career, toward a long-term goal of an improved understanding of the role of feedback in perceptual decision-making. These objectives will be accomplished in two phases. In the 2-year mentored phase, the candidate will conduct supervised research in Dr. Callaway's and Dr. Reynolds'laboratory at the Salk Institute to develop and use optogenetic techniques to characterize cortico-cortical feedback connections to V1 (Aim 1) in fixating monkeys. This training environment is uniquely suitable because of Dr. Callaway's expertise in developing genetic techniques for neurobiology combined with Dr. Reynolds'expertise in the study of attention in primates, the immediate availability of monkeys and the intellectual resources available at the Salk Institute, in molecular neurobiology, genetic techniques, viral vectors, neuro-anatomy, visual neuroscience and theoretical neuroscience. In the next 3-year phase, the candidate's independent research will test the hypothesis that decision-related activity in V1, and modulation by feature-based attention in these areas, share a common mechanism, using single unit neurophysiological recordings (Aim 2), and optogenetic techniques (Aim 3). Aim 3 will at the same time allow the candidate to examine the role of cortico-cortical feedback from V2 to V1 in visual decision-making. It represents an innovative approach to study the role of cortical circuits, a merging of established lines of research (attention and perceptual decision-making) and will advance our understanding of the neural circuitry involved in the dynamic processes of cortico-cortical feedback underlying decision making. Impaired control of cortico-cortical feedback appears to be involved in brain disorders such as schizophrenia. Understanding the role of cortico-cortical feedback may therefore inform clinical treatments for patients with schizophrenia. PUBLIC HEALTH RELEVANCE: Relevance to public health The \"read-out\" of visual cortex serves as a model system for interpretation, decision-making and reasoning. This area therefore sheds light on brain functions central to brain disorders affecting higher mental function, such as Alzheimer's disease, Schizophrenia and neglect. Ultimately, a better understanding of the role of cortico-cortical feedback and of how activity in cortical areas is related to perception, decisions and behavior may also allow the development of more sophisticated neural prostheses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Drug and alcohol abusing and dependent delinquents represent a large and greatly underserved population that is at high risk of presenting significant deleterious outcomes and long-term costs for themselves, their families and communities, and society (Belenko & Dembo, 2003). As described by recent reviewers (Belenko & Logan, 2003; Cooper, 2002), juvenile drug courts began emerging in the 1980s to address these problems, and 268 juvenile drug courts were in operation by December, 2003 (Huddleston, Freeman-Wilson, & Boone, 2004). Although juvenile drug courts have continued to proliferate in recent years, evaluation of their capacity to reduce offender substance use and criminal activity has lagged. The proposed study builds on our recent research findings supporting the effectiveness of juvenile drug court and the capacity of evidence-based treatments to enhance juvenile drug court outcomes (Henggeler et al., in press) as well as taking advantage of our extensive experience in the transport of evidence-based practices to community treatment settings. The overriding purpose of this Stage 1-B Treatment Development Study is to develop and test a relatively flexible and low cost strategy for enhancing the outcomes of juvenile drug courts by integrating components of evidence-based treatments into existing substance abuse and drug court services. Specific aims include: Aim 1: Adapt existing intervention and training protocols from evidence-based practices (i.e., contingency management for treating adolescent substance abuse; family engagement strategies from evidence-based treatments of juvenile offenders) for integration into juvenile drug court sites. Aim 2: Conduct a study to examine youth (e.g., substance use and criminal behavior) and system level (e.g., feasibility, implementation fidelity, retention and completion rates, consumer satisfaction, cost estimates) effects of implementing the intervention protocols in six juvenile drug courts. Aim 3: Revise intervention and training protocols in preparation for Stage II study if findings are supportive. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Matrix Biochemistry Section focuses its research on the functions of five major noncollagenous proteins first found associated with the mineralized matrix of bones and teeth but that we later showed are also made by many metabolically active ductal epithelial cells. The five proteins are bone sialoprotein (BSP), osteopontin (OPN), dentin matrix protein-1 (DMP1), dentin sialophosphoprotein (DSPP), and matrix extracellular phosphoglycoprotein (MEPE). We have made a strong case for the genetic relatedness of these seemingly different proteins and there is increasing acceptance of the SIBLING (Small Integrin-Binding LIgand, N-linked Glycoprotein) family concept. The genes encoding these proteins are all clustered in a tandem fashion within a short (400,000 base pairs) region of human chromosome 4 and similarly on all other mammals studied to date. After comparing the intron-exon structures and conserved motifs of their respective protein-encoding exons, we proposed that the five genes might be the result of ancient gene duplication and subsequent divergence. The most recent event appears to be a duplication of the DMP1 gene in the common ancestor of mammals and reptiles. Both lines then independently modified a different DMP1 gene to become DSPP-like during the evolution of modern dentin. We and others have shown that all known cases of non syndromic Dentin Dysplasia (DD) and Dentinogenesis Imperfecta (DGI) are the result of either a variety of point mutations at the very beginning of the DSPP gene or deletions later in the gene that result in frameshift mutations within the long repeat domain. In the past we proposed that all known mutations have dominant negative effects (mutations in a single copy of the gene cause the diseases but complete loss of one copy does not) but the mechanisms for this remained unexplored. Recently we have shown that all known mutations (except Y6D) cause the retention of the mutant DSPP proteins in the endoplasmic reticulum of our model system. Furthermore, we have shown that the retained mutant proteins cause the loss in the DSPP protein made from the normal allele. Mutations causing only small amounts of the normal DSPP protein to be secreted out of the cells cause the more severe disease, DGI. Our current research involves a focus on the trafficking of both normal and mutated acidic proteins within the endoplasmic reticulum (ER) as well as how the cells naturally destroy disordered proteins that fail to traffic out of the ER. Searching databases we have found that many secreted, acidic proteins encode similar peptide motifs that we propose are used to interact with a conserved ER cargo receptor. This proposed cargo receptor then traffics negatively charged proteins (as well as a variety of other proteins that have tendencies to either self-aggregate or form complexes with co-translated proteins) out of the ER before they can reach high enough concentrations to form aggregates that cause cells to malfunction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The proposed research is designed to determine if increased dietary iron and iron stores contribute to the development of alcholic liver disease in African Americans. Our preliminary studies indicate that high dietary iron leads to increased hepatic iron stores in both Africans and African Americans, and that incrreased hepatic iron stores are associated with hepatic dysfunction in Africans who consume alcohol. We postulate that the oxidative environment induced by alcohol in heptocytes and Kupffer cell &#40;hepatic macrophages&#41;leads to diassembly of the ironfur cluster of cytocolic aconitase/IRP 1 and conversion of this enzyme to an apoprotein that binds to RNA stem loops &#40;iron repsonisve elements-IREs&#41;in iron metabolism transcripts. This non-physiologic increase in IRE-binding activity of IRP1 in turn leads to abnormal repression of ferritin synthesis and abnormal increases in transferrin receptor synthesis and potentially toxic cytosolic labile iron concentrations. Based on our cell culture and animal model studies, we further postulate that increased non-heme iron content in Kupffer cells primes these cell for NF-kB activation and proinflammatory gene expression and thereby contributes to the pathogenesis of alcoholic liver disease in African Americans. We aim to test two central hypotheses: i&#41;high dietary iron contributes to an alcohol induced tendency for abnormal iron-loading of cells, and ii&#41;increased hepatic iron contrributes to liver damage in the setting of alcoholic liver disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This institutional T32 training application titled: \"Clinical Research Training in Oral Diseases for Future Academicians\" represents a response to RFA DE-05-008 from the University of North Carolina at Chapel Hill. Our Clinical Research Scholars Program (CRSP) is our School's response to the perceived shortage of dental school faculty who are qualified to conduct research that will advance knowledge in the field. The program primarily focuses on training dentists in clinical research who are also enrolled in advanced clinical education programs to prepare to become academicians who can function as interactive scientists who have the ability to address the expanding opportunities in dental, oral and craniofacial clinical research. As designed, our program embraces the full spectrum of translational clinical research including fundamental mechanisms of human disease, therapeutic intervention, clinical trials, and oral epidemiology. It includes 3 major options: 1) a 2-year postdoctoral fellowship in clinical research, 2) a 3-year PhD program in Oral Epidemiology, and 3) a 1-year Visiting Scientist (mid-career) fellowship. The program integrates Mentorship and curricula primarily from the UNC Schools of Dentistry, Medicine and Public Health. Each student is assigned multiple Mentors from all schools and is expected to write a grant application by the end of their program. The 2-year program is based on our Medical School's K30 curriculum and joint dental and medical school GCRC experiences. The PhD program earns a PhD from the Department of Epidemiology. The Visiting Scholars Program is based on the Robert Wood Johnson Clinical Scholars Program with a second year of Mentorship at their home institution. The CRSP will draw upon the existing interdisciplinary research activities within UNC that currently unites basic scientists with clinicians, clinical investigators and epidemiologists to provide a comprehensive training environment for clinical researchers. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is the goal of this program to develop efficient, practical (within the context of the complexity of the molecules involved) syntheses of the polyether ionophores. These antibiotics function as agents for ion/amine transport and through their ability to facilitate the transport of biologically relevant ions (Na+, K+, CA2+, etc.) or amines have the capacity to alter significantly cell metabolism. Commercially valuable as coccidiostats and anabolic agents, the polyether antibiotics are also proven cardiotonic agents. Therapeutic utilization of the ionophores as human cardiotonic agents is, however, severely limited by their toxicity and residual character that prevents effective dose regimes. Access to related chemical structures that still retain the ability to chelate metal ions/amines reversibly, but can overcome some of the toxicity and metabolic stability problems associated with the natural products may lead to useful therapeutic agents. It is the purpose of this effort to develop technology that can be generally applied to the synthesis of the characteristic polyether structure and design analogs of these ionophores that through biological testing will help define the requisite structural parameters for activity. Initial targets for the synthetic methodology part of the work are monensin and nigericin and then analogs related to these antibiotics will be prepared by that technology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Plasmacytoid dendritic cells (pDCs) represent a unique immune cell lineage that plays an essential role in innate immune responses against viruses. pDCs recognize virus-associated nucleic acids and respond by rapidly secreting massive amounts of type I interferons (IFN), hence their other name: type I interferon- producing cells (IPCs). pDCs can also differentiate, upon activation, into conventional dendritic cells that are capable of presenting antigens to the adaptive arm of the immune system. Thus, pDC are critical for the body's control of viral infections. On the other hand, abnormal pDC hyperactivity has been associated with increased type I IFN levels in autoimmune diseases such as psoriasis and systemic lupus erythematosus. Therefore, pDCs do play important roles in both normal and aberrant immune responses and represent potential useful targets in the treatment of some diseases. pDCs possess unique cellular and molecular characteristics and gene expression profiles that enable them to directly recognize viruses, secrete massive amounts of type I IFN, and differentiate into conventional dendritic cells. Over the last few years, these molecular and cellular features of pDCs have been well characterized; however, the genetic basis of pDC lineage commitment, maturation, and function still remains poorly understood. The overall objective of our research project is to study the role of basic helix-loop-helix transcription factors in the development and function pDCs, and elucidate the pathways that these transcription factors control in pDCs. PUBLIC HEALTH RELEVANCE: These studies will give us insights into the genetic network which controls the development and function of pDC, and may pave the way for the development of new approaches in the treatment of viral infections and some autoimmune disease. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This contract is designed to attain extramural support for developing, designing, interpreting, and evaluating clinical trials, epidemiologic and natural history studies. In addition, it will provide for outcomes research involving eye diseases and visual disorders and some preclinical studies. The focus shall be on the design of studies and the collection, analysis, and interpretation of data emanating from these studies, as well as support, and monitoring patient safety and follow-up. Contractor shall also provide analytical and data management support, as described in the work statement, for specified clinical research data bases, cost-effectiveness and economic analyses, quality of life assessment and outcomes research. This will include, but not be limited to, the following areas: analysis of Medicare and other health care databases; evaluation of existing NEI databases such as, centralized NEI Intramural Research database, the Eye Disease Case Control Study, Early Treatment Diabetic Retinopathy Study, Framingham Eye Study, and intramural AIDS and uveitis databases. The 46 active/follow-up trials in 2015 are listed below: For more info: http://clinicalstudies.info.nih.gov/cgi/protinstitute.cgi?NEI.0.html 15-EI-0202: Evaluation of Oral Minocycline in the Treatment of Geographic Atrophy Associated with Age-Related Macular Degeneration * 15-EI-0128: The Genetics of Inherited Eye Disease * 15-EI-0052: Pilot Phase I/II Study of the Evaluation of Interferon Gamma-1b Administered Topically for Macular Edema/Intraretinal Schisis Cysts in Rod-Cone Dystrophy (RCD) and Enhanced S-Cone Syndrome (ESCS) * 15-EI-0041: Patient Expectations When Enrolling in an Early-Phase Clinical Trial * 15-EI-0038: A Phase I/IIa Study of RS1 Ocular Gene Transfer for X-linked Retinoschisis * 15-EI-0020: Adaptive Optics Retinal Imaging * 14-EI-0173: An Investigation of Retinal Findings in Patients with Signs and Symptoms of Alzheimer's Disease Enrolled in 09-M-0198 * 14-EI-0171: The Focal Electro-Oculogram in Macular Disease * 14-EI-0108: Pilot Study to Evaluate Oral Minocycline in the Treatment of Cystoid Macular Edema Associated with Retinitis Pigmentosa * 14-EI-0078: A Phase 2 Multicenter Randomized Clinical Trial of Ciliary Neurotrophic Factor (CNTF) for Macular Telangiectasia Type 2 (MacTel) * 14-EI-0064: Whole Exome and Whole Genome Sequencing for Genotyping of Inherited and Congenital Eye Conditions * 13-EI-0154: Color Vision as an Outcome Measure for Clinical Trials of Inherited Retinal Degenerations * 13-EI-0124: A Pilot Study of Nitisinone in the Treatment of Oculocutaneous Albinism, Type 1B * 13-EI-0072: Microbiome and Ocular Inflammatory Disease * 13-EI-0049: Genetics of Uveal Coloboma * 12-EI-0203: Natural History of ABCA4-Related Retinopathies * 12-EI-0042: NEI Intramural Biorepository for Retinal Diseases * 11-EI-0264: A Pilot Study for the Evaluation of Minocycline as a Microglia Inhibitor in the Treatment of Central Retinal Vein Occlusions * 11-EI-0263: A Pilot Study for the Evaluation of Minocycline as a Microglia Inhibitor in the Treatment of Branch Retinal Vein Occlusions * 11-EI-0245: Generation of Induced Pluripotent Stem (iPS) Cell Lines from Somatic Cells of Participants with Eye Diseases and from Somatic Cells of Matched Controls * 11-EI-0173: The Natural History of Ocular Graft-Versus Host Disease * 11-EI-0147: Longitudinal Investigation of Dark Adaptation in Participants with Age-Related Macular Degeneration * 10-EI-0140: Genotype-Phenotype Study of Patients with Plaquenil -Induced Retinal Toxicity, with Evaluation of the ABCA4 Gene * 10-EI-0093: Immunogenetic Mechanisms in Behcet's Disease * 10-EI-0016: Internal Monitoring of Eye Movement in Schizophrenia * 08-EI-0169: Evaluation and Treatment Protocol for Potential Research Participants with Ocular Diseases * 08-EI-0102: Screening Study for the Evaluation and Diagnosis of Potential Research Participants * 08-EI-0099: Epigenetics, Molecular Genetics, and Biomarkers of Degenerative and Inflammatory Ocular Diseases * 08-EI-0043: Age-Related Eye Disease Study (AREDS) and AREDS2 Follow-Up * 08-EI-0031: Visual Motor Coordination * 06-EI-0236: National Ophthalmic Genotyping and Phenotyping Network, Stage 1 - Creation of DNA Repository for Inherited Ophthalmic Diseases * 06-EI-0068: The Vitreous Proteome and Inflammatory Mediators in Ocular Inflammatory Disease * 06-EI-0059: Clinical and Molecular Studies in Families with Glaucoma and Related Diseases * 06-EI-0058: Clinical and Molecular Studies in Families with Myopia and Related Diseases * 05-EI-0143: Molecular Genetics of Retinal Degenerations * 04-EI-0008: Clinical and Molecular Studies in Families with Corneal Dystrophy or Other Inherited Corneal Diseases * 03-EI-0123: Clinical and Molecular Studies in Families with Congenital or Hereditary Cataracts * 03-EI-0122: Evaluation of Immune Responses to Different Antigens in Non Infectious Ocular Inflammatory Diseases * 03-EI-0033: X-Linked Juvenile Retinoschisis - Clinical and Molecular Studies 13-EI-0206: A Randomized, Controlled, Double-Masked, Clinical Trial of Autologous Serum Eye Drops for Severe Ocular Chronic Graft-versus-Host Disease (GVHD) in Hematopoietic Stem Cell Transplant (HSCT) Patients * 13-EI-0102: Gevokizumab Treatment for Active Scleritis By IL-1 Inhibition (GATSBY) * 12-EI-0167: A Phase I/II Study of the NT-501 Intraocular Implant Releasing Ciliary Neurotrophic Factor (CNTF) in Participants with CNGB3 Achromatopsia * 11-EI-0244: A Pilot Phase I/II Study for the Evaluation of Dextromethorphan as a Microglia Inhibitor in the Treatment of Diabetic Macular Edema (MiDME2) * 07-EI-0213: Laboratory of Immunology/National Eye Institute Repository * 06-EI-0050: Multicenter Uveitis Steroid Treatment (MUST) Trial Protocol * 05-EI-0096: Natural History and Genetic Studies of Usher Syndrome", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Incomplete respiratory neuron maturation causes significant morbidity during the perinatal period, yet the mechanisms by which respiratory neuron maturation occurs during this vulnerable time window is not understood. Thus, there is a critical need to identify these basic neural mechanisms of perinatal respiratory control. The objectives of the proposed research are to elucidate developmental processes of respiratory neuron network maturation and to identify brainstem respiratory centers/circuits necessary for perinatal breathing. The central hypothesis is that hindbrain respiratory neuron networks undergo critical developmental maturation during the late embryonic, perinatal, and post-natal periods in mammals, and that developmental abnormalities in neuronal and glial maturation contribute to the pathophysiology of autonomic respiratory neuron dysfunction. The proposed research is inspired by our group's findings of Central Congenital Hypoventilation Syndrome (CCHS), a rare human disorder characterized by an inability to sense CO2 and which is linked to PHOX2B poly-alanine repeat and non-polyalanine repeat (NPARM) mutations. The rationale for the proposed research is that the lack of a basic fundamental understanding of which autonomic neural circuits are required for perinatal breathing represents a barrier to the ultimate implementation of interventions aimed at improving morbidity for premature infants. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: 1) Determine the extent to which selective expression of a dominant negative NPARM-PHOX2B mutation regulates perinatal chemosensation-induced respiratory drive, 2) Determine which brainstem circuits are lost in NPARM-CCHS, and 3) Determine the extent to which selected ablation of brainstem astrocyte population promote congenital hypoventilation. Under the first aim, we will test the effects on ventilation control and brainstem anatomy after targeted brainstem expression of a dominant negative NPARM PHOX2B mutation using an already proven conditional transgenic mouse approach. In the second aim, we will combine an innovative transgenic approach to identify which brainstem circuits are lost in congenital hypoventilation. In the third aim, we will determine the extent to which neuronal-glial interaction are necessary for appropriate autonomic respiratory control in the perinatal and post-natal period. The approach is innovative because it uses novel and validated tools, techniques, and reagents from distinct disciplines that allow us to address previously unanswerable questions. The proposed research is significant, because it is expected to vertically advance and expand understanding of which neuronal-glial circuits are required for proper control of autonomic regulation of breathing at birth. The tools and basic knowledge gained from these studies will form the foundation of future studies where interventions to improve autonomic respiratory neuron function in premature babies are designed and validated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Adapted from the application) Critical to a better understanding of the pathogenesis of HIV-associated lung disease is an animal lentivirus model that parallels HIV-associated immunodeficiency. Evidence suggests that the feline AIDS-causing lentivirus, Feline Immunodeficiency Virus (FIV) causes serious pulmonary immunodeficiency characterized by increased susceptibility to Toxoplasma gondii pneumonia. The proposed research will employ the use of FIV for examining the defects in alveolar macrophage (AM) function. The application describes experiments designed to test two hypotheses: first, that the constitutive level of activation of AM from HIV/FIV patients is a direct result of viral infection of the AM independent of lymphokine conditioning; secondly, that HIV/FIV infection inhibits an autocrine interferon (IFN)-gamma loop necessary to prime AM for IL-12 production. a. Career Development Plan: The plan consists of two phases. During the first Phase, Dr. Ritchey will complete his Ph.D. training. He entered the Ph.D. program in July of 1993, and has completed the didactic training as well as the preliminary examination. In phase two (years three and four), Dr. Ritchey will remain at North Carolina State University, in a postdoctoral position. b. Research Plan: Feline Immunodeficiency Virus (FIV) will be used to examine the defects in alveolar macrophage (AM) function. Experiments are planned to test two hypotheses: first, that the constitutive level of activation of AM from HIV/FIV patients is a direct result of viral infection of the AM independent of lymphokine conditioning; secondly, that HIV/FIV infection inhibits an autocrine IFN-gamma loop necessary to prime AM for IL-12 production. Studies during the first three years of this program will test these hypotheses by evaluating cytokine expression patterns of AM collected in a temporal fashion following in vivo FIV infection. The last year of this research will correlate the results of the in vitro studies with in vivo challenge of FIV-infected cats with T. gondii.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Scott &White Memorial Hospital provides comprehensive, high quality health care for a 38 county area in Central Texas. Since the early 1900s, Scott &White has demonstrated leadership in providing innovative cancer care emphasized by medical education and research. Continuing with this tradition of quality care for the community, Scott &White seeks to continue its funding for the Community Clinical Oncology Program (CCOP) to support these objectives. The mission of the Scott &White Cancer Institute is \"to provide the most comprehensive, highest quality cancer care, focusing on the prevention and treatment of cancer, enhanced by the application of scientific discoveries to ultimately improve the care of those living with cancer at Scott &White and throughout the world.\" Funding from the NCI since 1997 for our CCOP has enabled the Cancer Institute to evolve into a recognized leader in clinical cancer research and treatment for our service area. The proposed CCOP consists of the Scott &White component and Southwest Regional Cancer Center affiliate. The component and affiliate will work together in promoting innovative cooperative group studies in the community and contributing to quality research through refinement of processes to identify patients for protocol enrollment. The objectives for the CCOP will be to further support the commitment to delivering research options to the community that include cancer control, prevention, and treatment studies. The CCOP will continue to build on the existing strategies to recruit minority participants onto NCI-sponsored protocols by establishing relationships and by providing access to services in underserved areas. The disease-specific teams were developed to expedite patient discussion and treatment plan decisions, will continue to improve processes with a patient-centered approach to treatment. Community-based involvement will be strengthened by increasing participation in cancer-related organizations that support the goal of eradicating life-threatening cancers through education, research, and providing access for underserved areas. A consistent growth in the Central Texas area, increased health plan participation and the expansion of outreach clinics has placed an increasing demand for cancer services at Scott and White.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Preliminary data showed that the relationship between the PAT index and IR signal, calculated as a ratio between temperature before and during the first 100 sec of occlusion, holds promise to simplify and substantially shorten the procedure of endothelial assessment. Thus, peripheral vascular endothelial function testing with both PAT and IR methods correlates with the presence or absence of Sickle Cell Disease. These data suggest that peripheral vascular endothelial function testing with IR imaging is feasible, and this is an important next step in development of this non-invasive imaging technology capable of identifying, differentiating, and locating peripheral vascular beds with endothelial dysfunction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The project is related to the isolation of proforms of fibrous elastin from lung tissue. A fibrous elastin precursor which is approximately 70,000 daltons in size has been isolated from the lung tissue of the copper-deficient chick. This protein is similar in property to tropoelastin which has been isolated from the aortas of copper-deficient swine and chicks. In addition, a protein, which has an amino acid composition similar to tropoelastin (i.e., eighty-five percent of the total residues as gly, ala, val, pro and over 40 residues/1000 residues lys), has also been isolated with an apparent molecular weight of 100,000 plus daltons. The protein appears to be a proform of tropoelastin. Selective isolation of the \"proelastin\" is achieved by isoelectro focusing tissue extracts. Studies are currently being conducted to establish more clearly a precursor-product relationship with respect to the putative proform and tropoelastin and their roles in the final formation of fibrous elastin. In addition, dietary copper-deficiency and cadmium intoxication appear to result in severe alterations in lung elastin metabolism.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A method has been developed to measure rapidly and accurately three important material constants of a hydrogel sample. The shear modulus, m, the bulk modulus, k, and the hydraulic permeability, 1/f, can be determined from a single stress relaxation experiment. The material constants are treated as free parameters whose values are optimally estimated X)y minimizing the variance between predicted and empirical force relaxation waveforms. Values of the three constants obtained by this method agree with values obtained by independent free-swelling and permeability measurements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this UH2 and UH3 is to study how exosomes can deliver siRNAs across the blood brain barrier to enter neurons and other brain cells. The immediate target is the mutant huntingtin mRNA. Huntington's disease (HD) is caused by an increase in the CAG trinucleotide repeats to ? 36 in series; it necessitates years in a high level nursing facility because of neurodegeneration first in striatum and cortex and then to other brain structures. HD patients have cognitive impairment, depression and aberrant movements. Most HD patient present by 30 to 40 years of age; a few have a juvenile onset. A rational treatment is to decrease expression of mutant huntingtin mRNA; this therapeutic can be accomplished in HD mouse models by siRNA, antisense oligonucleotides (ASO) and adeno-associated virus (AAV) with shRNAmir directed against huntingtin mRNA. However, delivery remains a pitfall to practical implementation of the therapeutics. siRNA and ASO require long-term infusion. In non-human primates, ASO administered to spinal fluid does not reach the striatum and spread of siRNA is limited in brain. Although promising, AAV-shRNA requires several injections into brain areas and the shRNAmir is unregulated. A gap in HD therapeutics can be filled by microvesicles normally extruded by cells, exosomes. Exosomes with rabies virus glycoprotein (RVG) on their surface can be injected into the blood, cross the blood brain barrier, and enter neurons and glia. RVG-exosomes can carry siRNA cargo. Delivered into the blood circulation, the exosomes deposit siRNA in neurons to engage in RNA interference. Our purpose is to develop exosomes as a therapeutic in HD. The UH2 examines the ability of RVG-exosomes carrying siRNA against huntingtin mRNA to cross the blood brain barrier to enter neurons. Localization in brain and RNAi dependent knock down will be studied. Hyper-functional siRNAs will be sought. Because exosomes are made from cytoplasm of cells, exosome mRNA, miRNA, and implaced siRNA will be identified by deep sequencing. Immune reactivity and immune-neutralization will be studied, since exosomes have potential antigens, like RVG, and will need to be administrated often. The UH3 further establishes exosome-based therapeutics, by study of reversal or prevention of neuropathology and aberrant movement in HD mouse models. Dosing of exosomes will be secured. A team of experts in HD pathogenesis, siRNA development, RNA identification and measurement, RNAi mechanisms and exosome production and brain delivery will carry out the studies. Harnessing exosomes for brain delivery is expected to form a viable therapeutic to reduce expression of mutant huntingtin in patients with HD. Patients with other genetically- based neurodegeneration will benefit.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Human plasma high density lipoproteins (HDL) from normolipemic sera were analyzed as a distribution of at least three major components, each with characteristic particle size, hydrated density, and consituent apolipoprotein composition. The plasma levels of two components, HDL2b and HDL2a, were variable among 160 normal individuals while HDL3 levels remained essentially constant. Polydispersity of apolipoprotein composition was demonstratable in all three HDL components from both normal and abetalipoproteinemic individuals. In addition to these three components, the HDL from cord blood plasma of neonates was found to contain additional components which accompany a threefold decreased activity of the plasma enzyme lecithin: cholesterol acyl transferase. This work suggests that HDL2 (HDL2b plus HDL2a) is the major contributor to the inverse correlation of HDL cholesterol with the prevalence of coronary heart disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Computational methods for mechanistic understanding of inter-sample variability The overall goal of this R21 application is to develop a computational framework that will allow for the prediction of physiological differences between experimental samples. Differences between individual samples can be catalogued and described at the physiological level, in terms of properties such as action potentials, and also at the molecular level, in terms of measurements such as gene expression. Linking variability at one level to variability at another level in a quantitative manner, however, is not straightforward. Here, through an innovative combination of experimental studies, mathematical modeling, and statistical analyses, we will develop methods that allow for molecular-level differences between samples to be translated into quantitative predictions of physiological differences. This Multiple Principal Investigator proposal utilizes the complementary expertise of the two PIs. Dr. Eric Sobie is expert in cardiac physiology, mathematical modeling, and computational approaches for understanding variability~ Dr. Christoph Schaniel is expert in stem cell biology, differentiation of pluirpotent cells into specific cell types, and high-throughput methods. The combined efforts of the two PIs will generate new quantitative data and will yield new computational methods that can be applied broadly to understand variability in different contexts. To achieve the overall project goals, we propose to: 1. Collect measurements of cardiac physiology and expression of relevant ion channels, pumps, and transporters. These measurements will be matched on a sample-by-sample basis. 2. Perform population-based simulations with dynamical mathematical models to develop quantitative and mechanistic predictions regarding how differences between samples in expression of important genes are translated into physiological differences. 3. Use regression-based statistical methods to analyze the experimental and simulation results, and to relate the two sets of predictions to each other. Not only is this exploratory research likely to provide insight into the physiology of cardiac myocytes derived from stem cells, it is also likely to demonstrate a novel computational framework that can be used for quantitative treatments of variability between samples in many biological contexts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The quest to characterize the human microbiome is a daunting goal, but one that promises to enhance significantly our understanding of health and our management of a wide variety of disease states. In this quest, two features of the human microbiota in particular, pose major challenges: the large proportion and number of as-yet uncultivated species, and the extreme unevenness of the microbial communities, with a resulting large number of potentially important community members that fail to be \"seen\" in routine surveys. The ability to identify, isolate, and sequence the genome of single bacterial cells would allow us to characterize and understand both rare and uncultivated microbial species, and materially advance our understanding of the human microbiome. In recent work, a microfluidic device has been designed and fabricated, with features that mimic an integrated electrical circuit; this device isolates individual bacterial cells, and allows their genome to be amplified in nanoliter volumes. In this Application, a plan is proposed for optimization and augmentation of this microfluidics device, so that environmental contamination is reduced, rare cell types are more easily captured, larger numbers of cells are screened more quickly, and gene expression is more easily measured from single cells. The long-term objectives of this work are to enhance our understanding of the human microbial communities, and in particular, of novel or poorly-characterized, uncultivated microbial community members. This proposal responds to critical unmet needs posed by the NIH Human Microbiome Project. The following are the Specific Aims of this proposal: Aim 1. To reduce the contribution of environmental DNA to single cell genomic sequence data, and increase the \"signal-to-noise\" ratio of the sequence data obtained with our cell- sorting, genome amplification microfluidics device. The experimental approach involves the integration of optical (laser) tweezers into the device. Aim 2. To improve the ability to detect and capture rare microbial community members with the microfluidics device. The experimental approach involves the integration of fluorescence in situ hybridization techniques, specific probes, and fluorescence imaging with the microfluidics device. Aim 3. To increase the speed of single cell selection and isolation with the microfluidics device. The experimental approach involves more highly parallel microdevice designs, optimization of laser power and laser optical path, and further automation of cell manipulations. Aim 4. To enhance the capability for gene expression analysis in single bacterial cells. The experimental approach involves the development of on-chip protocols for RNA isolation, reverse transcription, and use of digital PCR to quantify transcript abundance from single cells. PUBLIC HEALTH RELEVANCE: This research is relevant to public health because it will lead to a better understanding of the microorganisms that live on and in the human body. Because the human indigenous microbial communities help to maintain health and when disturbed, contribute to disease, this research may lead to new tests for diagnosing or predicting disease associated with microbial community disturbance, and new strategies for maintaining or restoring health. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neuropathologically, Alzheimer's disease (AD) is defined by the presence of plaques composed of the amyloid-beta (Abeta) protein. This alone makes definition of the natural history of amyloid deposition in living subjects an important goal, but this longitudinal information is not obtainable through post-mortem studies. This information will be essential in the assessment of anti-amyloid drugs intended to alter the natural history of amyloid deposition. Our group has developed a novel amyloid-imaging positron emission tomography (PET) radiotracer termed \"Pittsburgh compound-B\" (PIB) that, in initial proof-of-concept studies, performed well to distinguish AD from control subjects and localized in brain with a regional distribution consistent with that of the post-mortem distribution of plaques. While these results are exciting, certain basic and fundamental characteristics of this PIB PET technology must be further defined before this amyloid imaging technology can reach its potential as diagnostic tool or a surrogate marker of efficacy for anti-amyloid therapies. One of these characteristics is the variation of the quantity and regional distribution of PIB retention over time in individuals. This will be the focus of Project 2 of this Program Project (P01). To address this question, we will perform 12- and 24-month follow-up e studies on 15 control subjects, 15 MCI patients, 15 mild and 15 moderate AD patients who had identical bas line studies in Project 1 of this P01 (or in pilot studies during the year prior to initiation of this P01). All will have been clinically evaluated and diagnosed at the University of Pittsburgh Alzheimer Disease Research Center and further evaluated at each study point with a neuropsychological battery through the P01 Clinical Core. All subjects will be studied longitudinally with fully dynamic PIB PET scans at 12- and 24-months after their baseline evaluation in Project 1. In addition, volumetric MRI and FDG PET will be performed so we can directly compare changes in PIB retention to changes in hippocampal volume and cerebral metabolic rate. Image analysis will be performed in the P01 Imaging, Methodology and Statistics Core. The quantitative PIB PET data will be compared to neuropsychological measures to explore correlations between changes in regional amyloid load and changes in cognition. The association of apolipoprotein-E genotype and education with PIB retention also will be explored. We hypothesize that PIB retention in individual subjects will increase through moderate stages of AD and then plateau and that the rate of increase in PIB retention in individual subjects will be of larger magnitude and be detectable earlier than the rate of decrease in hippocampal volume or cerebral metabolic rate.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The volume and electrolyte composition of saliva are dependent on the coordinated activity of ion transport proteins including multiple Na+-dependent mechanisms. Acinar cells employ a combination of Na+/H+ exchangers and Na+/K+/2Cl- cotransporters to drive fluid secretion and related processes such as cell volume maintenance and intracellular pH regulation. Ductal cells then act on acinar cell secretions to conserve NaCl utilizing Na+/H+ exchangers, Na+ channels and other ion transport proteins. The general functional properties of most Na+ transport proteins are understood. However, it is not clear what contribution individual Na+ transport proteins make to the overall secretion process, or which compensatory mechanisms may arise if the activity of a given transporter is perturbed. Thus, Aim 1 will determine which Na+ transport mechanisms are expressed in mouse salivary glands. The level of expression and the localization of different isoforms will be assessed by northern hybridization blots and immunohistochemistry, respectively. Aim 2 will examine the regulation of the different Na+ transport proteins by secretagogues. It is predicted that functional activity will be modulated for those transporters involved in the formation of saliva during stimulation. The results from the studies proposed in Aims 1 and 2 will provide essential information for predicting the role each transporter plays in the overall fluid secretion process. However, a definitive test of the importance of a given Na+ transport protein can most easily be obtained by studying an animal defective in the expression of that transporter. In Aim 3 the effects of gene disruption will be examined to determine the functional significance of individual Na+ transport proteins. Mice lacking expression of four different Na+ transport proteins are currently available for study (Na+/H+ exchanger isoforms NHE1, NHE2 and NHE3; Na+/K+/2Cl- cotransporter NKCC1). Mice expressing mutations for three additional Na+ transport proteins present in salivary glands are under development (epithelial Na+ channel ENaC; Na+/H+ exchangers NHE4 and NHE5). This multidisciplinary approach to the functional characterization of Na+ transport proteins will define the mechanisms involved in the production of saliva. The results of these studies will provide a foundation for future studies to analyze the in vivo structure/function relationship of a given Na+ transporter, which may ultimately aid in the development of treatments for various forms of salivary gland dysfunction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research addresses a fundamental aspect of children's cognitive and social development -- their understanding of the mind. Understanding of the mind plays a critical role in the ability to function as effective social beings. Without an understanding that others have mental states, like beliefs and ideas, people would be unable to communicate and interact sensibly. The proposed work will examine an important yet unexplored aspect of this understanding -- children's knowledge about imagination. Children's understanding of imagination is important to study for several reasons. First, as a component of pretense, it serves as a vehicle for exploration of adult experiences and identities. Second, recent theorizing grants imagination a central role in the emergence of an understanding of human behavior. Finally, for adults and children, imagination serves as a source of much creative thinking and new discoveries. Despite the importance of an understanding of imagination, this question has been neglected in research on children's understanding of mind. Most studies have focused on aspects of children's understanding of epistemic mental states -- those mental states, like beliefs, that originate in direct interaction with physical reality and purport to represent reality truthfully. The conclusions drawn from the research are broader, however, encompassing children's understanding of representational mental states generally. This raises several important questions about children's understanding of fictional mental representations--those mental states, like imagination, that are representational, but do not purport to represent reality accurately. The proposed studies will examine the development of young children's understanding of the fictional mental state of imagination. The proposal addresses questions in three areas: (l) what is children's understanding of the correspondence between imagination and physical reality? (2) are there differences in children's ability to conceive of multiple representations with regard to fictional versus epistemic mental states, and (3) what do children understand about the origin of imagination and other mental states? Answers to these questions will fill a gap in our knowledge of children's understanding of imagination specifically, and will aide in developing more comprehensive theories about children's understanding of the mind more generally.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to examine the developmental course of alcohol-related problems by focusing on the dynamic associations among family relationships, financial well-being, and drinking behavior from adolescence through young adulthood. To achieve this goal, we propose using four waves of data from the National Longitudinal Study of Adolescent Health (the Add Health Study) in response to PA-08-167. An important predictor of alcohol problems in both adolescence and young adulthood is compromised parent-adolescent relationships. Another predictor is financial stress which may be particularly relevant during times of economic downturn. However, little is known about the dynamics of financial stress and its association with alcohol problems in adolescence and young adulthood. Testing an extension of The Family Stress Model (FSM), this research examines the potential long-term effects of family financial stress and parent-child relationship quality in adolescence on young adult family relationships, financial stress, and alcohol problems. The research will contribute to several areas of basic and practical knowledge relevant to alcohol- related problems, prevention and treatment, and to programs and policies related to reducing financial stress and debt levels during young adulthood. The specific research aims are: Aim 1 (Adolescence and the Transition to Adulthood): To test an extension of the Family Stress Model in which adolescent family financial stress predicts compromised parent- adolescent relationships, which in turn influence alcohol problems in adolescence and into young adulthood; Aim 2 (Young Adulthood): To examine the dynamic associations in young adulthood between relationships with parents, financial stress, and alcohol problems;and Aim 3 (The Full Model): To test an elaborated model of the associations between adolescent family financial stress, relationships with parents, and young adult financial stress and alcohol problems. This combined model posits that family financial stress in adolescence is linked to young adult alcohol problems through its potential lasting influence on relationships with parents and financial stress. In addition to the full model, alternate models will be explored to test alternative pathways of influence: in particular, adolescent and young adult alcohol problems may predict compromised relationships with parents in young adulthood as well as financial stress. Once the suitability of these models has been established relative to alcohol problems, we will extend the analyses to the use of illegal substances. Because alcohol problems and problem trajectories vary by gender and race/ethnicity, analyses of potential gender and ethnic differences, or combinations of gender and ethnicity, will be integrated within each aim. PUBLIC HEALTH RELEVANCE: This research examines the long-term effects of family financial stress and parent-child relationships during adolescence on young adult family relationships, financial stress, and alcohol problems. By using a national study that follows participants from adolescence through young adulthood, we can track developing patterns of alcohol problems and identify factors that contribute to these patterns. This study provides a unique opportunity to extend research in financial stress and alcohol problems in young adulthood, to test multiple models for understanding the mechanisms and directions of influence among these key constructs, and to identify possible points of prevention and intervention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "NOT-OD-09-058: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Application The immune system has the potential to eliminate altered neoplastic cells with incredible specificity. A consistent in-frame deletion in the extra-cellular domain of the epidermal growth factor receptor (EGFRvIII) represents a truly tumor-specific target amenable to immunotherapeutic attack. Our multi-institutional Phase II study demonstrated that vaccination with an EGFRvIII-specific peptide in patients with newly-diagnosed glioblastoma multiforme (GBM) induces potent T- and B-cell immunity, produces nearly complete radiographic responses in all patients with residual tumor, and universally eliminates EGFRvIII-expressing cells. Recurrent tumors, however, continue to express wild-type EGFR suggesting that the immune response is specific, but productive intra-molecular cross-priming against other potential tumor-associated antigens is incomplete. We believe that productive extension of such secondary immune responses is hindered by the presence of regulatory T-cells (T ) which are disproportionately represented within the peripheral blood and tumors of Regs patients with GBM. T are characterized by constitutive expression of the high affinity interleukin (IL)-2 Regs receptor (IL-2R1)(CD25) and are uniquely dependent on IL-2R1 signaling for their function and survival. In the context of the existing grant, we conducted a randomized trial demonstrating that an IL-2R1-blocking antibody, daclizumab, significantly reduces TReg levels in patients with GBM with a nadir at 5 weeks without reducing overall CD8+ or CD4+ T-cell counts. Preliminary analysis also suggests that daclizumab enhances EGFRvIII-specific cellular (P=0.01) and humoral (P=0.003) immune responses compared to the saline treated group. The effect of a single dose of daclizumab wanes after 12 weeks consistent with its known half-life, and TRegs recover, however. With this supplement, we seek to extend our results by examining the effects of serial administration of daclizumab in this same patient cohort. We HYPOTHESIZE that serial doses of daclizumab therapy in patients with newly-diagnosed GBM will extend the duration of functional TRegs inhibition and further enhance vaccine-induced immune responses. Consistent with the goals of the American Recovery and Reinvestment Act, this Supplement would accelerate the tempo of our research in this area and allow for job creation and retention. PUBLIC HEALTH RELEVANCE: Treatment for malignant primary brain tumors, which are the most common cause of death among children and account for more deaths in adults than melanoma, currently represents the most expensive medical therapy per quality-adjusted life-year saved currently provided in the United States. We have developed a vaccine that eliminates tumor cells containing a tumor-specific protein (EGFRvIII) in patients, but tumor cells that express related normal proteins survive. In this proposal, we will see if prolonged elimination of immunosuppressive \"regulatory\" T-cells that inhibit immune responses to these related normal proteins will enhance the effectiveness of the vaccine without inducing deleterious autoimmunity. PHS 398/2590 (Rev. 11/07) Page 1 Continuation Format Page", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To investigate the physiological function of nonmuscle myosin heavy chain II-B (II-B), we generated a point mutant II-B mouse where amino acid arginine-709 was replaced by cysteine (R709C). Due to the presence of the Neo cassette in the targeted allele intron, both heterozygous (B-R/CN) and homozygous mutant (B-CN/CN) mice showed decreased expression of mutant II-B. The B-R/CN mice were generally healthy and fertile. However, most of the B-CN/CN mice died within the first two days after birth, and the remainder survived for up to 15 days suffering from cardiac and brain abnormalities similar to the B-/B- mice which die prior to birth. The B-CN/CN mice developed an early postnatal ataxia coupled with abnormal development of the cerebellum, which was remarkably reduced in size and had abnormal foliation. Detailed histological examination and neuronal migration assays revealed significant neuronal migration defects in the B-CN/CN mouse. These included an abnormal protrusion of the facial nuclei into the 4th ventricle, premature arrest of the pontine migrational stream, and slowed migration of cerebellar granule cells. This study provides evidence that NMHC II-B is required for migration of certain groups of neuronal cells. Removal of the Neo cassette from B-R/CN mice using Cre-recombinase rescued the decrease in II-B expression (i.e., increased the mutant protein) and generated B-R/C mice, which revealed a novel phenotype. Most of the B-R/C mice (90%) died shortly after birth showing absence of the small intestines from the duodenum through the sigmoid colon and collapsed lung alveoli. The unexpected phenotype was due to the herniation of the liver and intestines during embryonic development and the ingestion of these organs by the mother. Present studies are directed at understanding the role of NMHC II-B in generating this abnormal herniation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study will investigate the effect of dietary carbohydrate and aerobic exercise in a 2X2 design on body composition, energy expenditure, insulin secretion and catecholamine excretion in obese subjects receiving a hypo-caloric diet.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Proposed projects include: An extension of studies on the effect of ions, in particular potassium and calcium, on sensitivity of the neuromuscular junction to neuromuscular blocking agents. An examination of the effect of anesthetics on drug-induced membrane electrical noise. Determination of the effect of barbiturates on sensitivity of the neuromuscular junction to competitive neuromuscular blocking agents. Examination of the relationship of physical chemical properties, in particular, pKA and solubility in non-polar solvents, to potency of barbiturates. Determination of the extent of post synaptic effects to \"run-down\" of end-plate potential trains seen with some \"competitive\" neuromuscular blocking agents. Refinement of a heuristic model to serve as a basis for determining clinical dosage regimens of competitive neuromuscular blocking agents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 20.6 % of the general population in the United States smokes cigarettes and this group may be comprised of smokers with severe nicotine dependence who are resistant to smoking cessation pharmacotherapies and treatments (Hughes, 2011). Varenicline is a first-line medication for smoking cessation that has been shown in meta-analytic reviews to be superior to other smoking cessation treatments (Cahill et al., 2010), however 56% of patients who take varenicline do not achieve smoking abstinence. One strategy to increase quit rates may be to administer a complimentary medication to augment the efficacy of varenicline. The anti-epileptic medication zonisamide is a good candidate for adjunct treatment because it increases dopaminergic tone, normalizes glutamate homeostasis, and potentiates GABA release. Zonisamide also improves sleep and promotes weight loss, two prominent issues not addressed by varenicline. Finally, the PI of this proposal has documented unpleasant changes in the taste of cigarettes and reductions in nicotine withdrawal among patients receiving zonisamide for the treatment of cocaine dependence. The proposed study will explore the efficacy of varenicline + zonisamide for smoking cessation using a controlled, clinical trial. Eligible participants (n=60) will be smokers (>10 cigarettes per day or >1 year) seeking treatment. They will be randomly assigned to receive zonisamide + varenicline or placebo + varenicline under double-blind conditions for a 10-week period. Participants will visit the clinic weekly to receive medication and smoking cessation counseling and also to complete self-report ratings of smoking, nicotine withdrawal and other smoking-related indices. Smoking status will be assessed via urinalysis testing for the nicotine metabolite cotinine (<200ng/ml will be considered abstinent) from specimens obtained at each study visit. Cotinine is a sensitive indicator of smoking status with a longer half-life then carbon monoxide (CO) and is therefore more likely to detect low or intermittent smoking. The study hypothesis is that participants who receive the combination zonisamide + varenicline will achieve greater smoking abstinence when compared to varenicline alone. The primary outcome measure for this study will be the 4-week rate of biochemically-confirmed continuous smoking abstinence during weeks 7-10 of the intervention. Secondary outcomes will include self-reported rates of smoking, subjective effects of cigarettes, mean body weight change from baseline to week 10, self-reported sleep quality, and nicotine withdrawal symptom severity. Results from this study will contribute important information on successful and cost-effective smoking cessation interventions for individuals who do not respond optimally to current smoking cessation medications. This study will advance the science and clinical treatment of smoking cessation, and will provide the prerequisite data to develop a larger scale clinical trial evaluation of the combination zonisamide + varenicline for smoking cessation. PUBLIC HEALTH RELEVANCE: Varenicline (Chantix) is a successful smoking cessation medication, yet 56% of smokers are unable to quit smoking despite using varenicline. Combining varenicline with zonisamide, an antiepileptic medication that has been associated with smoking reduction, may produce higher smoking quit rates compared to varenicline alone. The information collected from this study will be important for understanding whether combination medication treatments (like varenicline & zonisamide) produce more robust smoking cessation outcomes when compared to medications administered independently, and may be used to guide the development of smoking cessation studies for smokers who have not responded optimally to other currently available smoking cessation pharmacotherapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Because Crohn's Disease (CD) has an unpredictable course, past research attempted to understand the psychosocial factors that impact disease relapse and exacerbation of symptoms (Searle & Bennett, 2001). However, this literature presents equivocal results, which may result from differing experiment designs. The present study attempts to examine the types of stressors (life events, daily, perceived) that predict relapse and define the within-person temporal relationship between stress and symptom exacerbation. In a sample of 25 CD patients undergoing a biologic treatment, an electronic diary will be used to examine the within-person variations of symptoms and stress within a day and over 8 weeks (time from initial treatment to subsequent treatment). The hypotheses are: 1. Patients who report more stress will relapse sooner compared to those who report less stress. 2. Previous reports of stress will be associated with subsequent reports of symptom exacerbation. Thus, this study will be able to not only identify if stress and disease activity are related, but also the type of stress that most impacts the clinical course of CD. In addition, within-person analyses of the temporal relationship between stress and symptoms may aid in the development of future treatment studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY There is a fundamental gap in how the presence of family companions in medical visits influences indicators of quality of care, such as patient-centered communication (e.g., communication that reflects respect for patients'values and incorporates patient's concerns and preferences into decision-making), specifically for patients with heart failure - a particularly vulnerable population by virtue of its high morbidity and mortality. The long-term goals of this project are to improve the quality and delivery of care and subsequent health outcomes for Heart Failure (HF patients). The overall objective here, which is our next step in pursuit of our long-term goal, is to determine whether family accompaniment influences processes of care within the medical visit and use this information to inform intervention development. The rationale for the proposed research is that once we know how family members influence quality and delivery of care we can fulfill the call to action of policy makers and health care models to \"create an integrated, coherent plan for ongoing medical care in partnership with patients and their families.\" The research strategy is built around the central hypothesis that involvement of family members in clinical care will facilitate patient-provider communication and potentially improve subsequent HF self-management behaviors. The specific aims to achieve our overall objective are: 1) To investigate the impact of family companion's presence on patient centered communication as assessed by the following audio-tape measures: the patient-centeredness ratio (ratio of communication that furthers the patient's agenda to communication that furthers the provider's agenda), patient activation and engagement behaviors, and physician facilitation and patient activation behaviors;2) To assess the impact of family companion's presence on provider communication related to patient education and counseling about HF self-management behaviors (e.g., weighing, diet, symptom management, physical activity, and medication adherence);and 3) To use intervention mapping (IM) to design a family-involved HF intervention based on qualitative data and results of Aims 1 and 2, and pre-test the intervention. The mentored research, formal didactics, and planned presentations/publications will set the stage for independent funding as a clinician-scientist. PUBLIC HEALTH RELEVANCE: PROJECT NARRATIVE: The proposed research is relevant to public health by seeking to improve the delivery of patient/family centered care for patients with chronic illness by understanding how the patient-physician interaction is influenced by family member accompaniment to medical visits. Thus, the proposed research is relevant to NHLBI's mission to enhance the health of patients living with heart failure so that they can live longer and more fulfilling lives.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a competing renewal application that requests continued support for providing pre- and post-doctoral trainees with strong methodological and practical training in quantitative cancer research. The application leverages a unique combination of strengths from the Harvard T.H. Chan School of Public Health, Dana-Farber Cancer Institute (DFCI) and Dana-Farber / Harvard Cancer Center. This training program, now in its 34th year, draws upon a distinguished faculty, consisting of biostatisticians and computational biologists, as well as world renowned experts in cancer treatment and research. Its overarching goal is to provide the trainees with all essential elements of training needed to successfully undertake modern cancer research. The specific goals of this training program are to train students and postdoctoral fellows to be (1) quantitative scientists in cancer research, who are capable of using probability, statistics, computer science and mathematics to increase our knowledge and understanding of cancer; (2) strong team leaders/players as well as excellent communicators in a cancer research environment, who can effectively disseminate their research results and assume active roles in the design, analysis and interpretation of cancer clinical trials, cancer population studies and cancer genomic studies. All predoctoral students supported by this training grant are required to take a concentration in cancer-related courses. During their first and second summer periods in the program, students are required to participate in research activities of the DFCI or DF/HCC, performed under the supervision of faculty mentor/trainers affiliated in the program. Afterwards, many of these students will take up residence at the DFCI or elsewhere at DF/HCC and continue their research in cancer, which eventually evolves into their dissertation projects. All the postdoctoral fellows are closely involved with the practice of quantitative sciences in cancer and are typically in residence at the DFCI. All trainees are required to actively participate in the a working group seminar series on quantitative issues in cancer research, which serves as a primary forum at Harvard to discuss current issues and challenges on this topic, as well as in several of the DF/HCC sponsored symposia and event. This proposal requests 5 years of funding to support 10 pre-doctoral students and 1 post-doctoral fellow annually.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development of drugs that inhibit the K-Ras oncogene represents one of the greatest unmet needs in the treatment of human cancer. The Ras gene is the most frequently mutated oncogene in cancer, with a greater than 30% cumulative mutation frequency across all cancer types. Cancers with Ras mutations are aggressive and respond poorly to standard therapies. Previous attempts to target K-Ras have failed due to the difficulty of competing with the picomolar nucleotide affinity for the active sit and due to the high similarity of most GTPases. Our scientific co-founder, Professor Kevan Shokat, has discovered a novel, small molecule approach to target the most chemically tractable K-Ras mutant that contains a glycine-12 to cysteine mutation. The G12C mutation is the most common K-Ras mutation in lung cancer. Indeed, 43% of all lung cancers with K-Ras mutations contain the G12C mutation. This mutation positions a chemically reactive sulfhydryl group on the surface of K-Ras. We have carried out a preliminary 500 compound library screen based on mass spectrometry for molecules which bind covalently to K-rasG12C, H-rasG12C and counterscreened against WT K-Ras. 17 hits were identified from the screening library, and the first round optimization of the initial hits led to the discovery of a potent inhibitor JO-01-18. We have now solved the crystal structure of JO-01-148 bound to K-Ras G12C and identified a previously undescribed allosteric pocket on the surface of the protein adjacent to the cysteine moiety. This pocket makes it possible to identify irreversible inhibitors that bind in the pocket and selectively target the cysteine at position 12. Importantly, these small molecules inhibit only mutant K-Ras and not the normal protein. We have now solved more than 10 X-ray crystal structures of irreversible inhibitors bound to this allosteric pocket and synthesized more than 120 compounds. A clear SAR has been established. We are now proposing to further validate our lead G12C compounds in biochemical and cellular assays. The Phase I specific aims are: (1) Develop assays to evaluate K-Ras effector binding; (2) Demonstrate that the G12C irreversible binders can disrupt K-Ras effector binding; and (3) Demonstrate that the G12C irreversible binders can differentially affect tumor cells with G12C mutation compared to cells with wild type K-Ras and other K-Ras mutations. The Phase I milestone is the identification of K-ras G12C inhibitors that suppress proliferation of tumor cells with G12C K-ras mutation five-fold more potently (as measured by IC50 values) relative to tumor cells with other K-Ras mutations or wild type Ras. Collectively, we expect the Phase I results to demonstrate that we can generate a small molecule inhibitor that will specifically inhibit the growth of tumor cells wih K-Ras G12C mutation. If our approach is successful, our Phase II studies will more fully examine the safety, efficacy, and PK/biodistribution of a lead formulation for advancement to an IND application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed study will investigate modifications of the sympathetic responses of the uterine circulation due to changes in blood gases and temperature in the pregnant sheep. The distal end of the severed right sympathetic chain will be stimulated at varying frequencies as the pregnant uterus is perfused at constant flow with blood whose gas tensions and temperature is altered in the extracorporeal circuit. Alterations in uterine conductance will be expressed as function of frequency of stimulation and a family of curves generated. The decreased responsiveness of the pregnant animal to carotid sinus occlusion will be studied in the pregnant uterus perfused at constant flow to ascertain the role of the uteroplacental circulaton in damping the response. The pressure regulated circuit will be used to perfuse the uterus with mildly hypoxic blood, and after severing of the right sympathetic chain, flow-pressure curves will be recorded in effort to elicit autoregulation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will support the training of six predoctoral students each year in the area of biostatistics, with an emphasis on applications to modern problems in genomic science. The training combines rigorous coursework in statistical methods and theory, additional courses in bioinformatics and genomic science, and an extensive laboratory training experience. For the latter, trainees will begin as supervised statistical consultants for a matched genomics lab, then over the course of a year progress into active collaborators in one or more lab projects. Most students will be supported for the first three years of their graduate programs. The scientific training will be supplemented with training in the responsible conduct of research developed specifically to meet the needs of researchers in this area. The training involves collaboration among biostatistics, genomics, and philosophy faculty members. An active recruiting plan is described for enhancing the diversity of our training and graduate programs, including a summer program bringing faculty and undergraduate students from minority serving institutions to NC State during the summer to initiate collaborative work with training faculty.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have developed databases and software useful for comparative analysis of protein three-dimensional structure. These tools are distributed freely to biologists and developers of biotechnology software. MMDB (Molecular Modeling DataBase) is the 3D-structure component of the Entrez molecular biology retrieval system. MMDB is an ASN.1 database where all data items describing macromolecular structure are validated and explicitly listed, so that application software need not contain the complex logic required to retrieve this information from text formats such as PDB files. Work has concentrated on addition of accurate taxonomy assignments for macromolecular structures within MMDB, creation of new message and data types for transmission of structure-structure alignment data to local viewers, and on construction of an automated monthly update and indexing system, Pubstruct. CN3D (\"see in three dimensions\") is a multi-structure visualization program distributed as part to the Entrez client software and in a stand-alone version lauchable via the MIME protocol in World-Wide-Web Entrez. The software differs from other public domain viewers in supporting display of multiple aligned structures from Entrez's \"structure neighbor\" database, and in supporting simultaneous highlighting/picking of multiple sequence and multiple structure alignments. Other features added this year are on-the-fly alignment of the sequences of homologs, so that an Entrez user may easily map conserved sequence features onto the know 3D structure. These software features are intended to facilitate molecular biologist's identification of important structure-function relationships within protein families. Work this year has concentrated on improvements to CN3D. The software has been modified to use an industry-standard 3D graphics library, OpenGL, which provides much better quality molecular graphics rendering. We have also added core-structure alignment editing and threading tools to the sequence display windows, to support curation of CDD (a Conserved Domain Database). Work is in progress to revise and simplify the data structures underlying CN3D, so that further improvments in graphcis presentation, specific to describing conserved features in protein families, may be added to future versions of CN3D. A new version of Cn3D incorporating these changes was released in June, 2002 and downloaded by over 50,000 users as of October, 2002. This version provides sophisticated alignment editing tools, in addition to greatly improved molecular graphics performance on popular computing platforms. As of October, 2003, over 150,000 copies of CN3D have been downloaded. A new \"related structures\" link has been added to NCBI BLAST servers, to provide easy-to use mapping to 3D structure whenever possible.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pulmonary hypertension (PHN) in respiratory distress syndrome (RDS) evolving to bronchopulmonary dysplasia (BPD) results acutely from poor lung inflation and increased blood flow through the patent ductus arteriosus, and chronically from altered reactivity, vascular remodeling, and hypoplasia of the pulmonary vascular bed. The lung develops in relative hypoxia compared to 21% 02 seen at normal term delivery and, especially, to hyperoxia that preterm neonates commonly face. Hypoxia-inducible factors (HIFs) could lower pulmonary artery pressures by modulating surfactant, ductus arteriosus, and pulmonary vascular development. Preliminary data show that HIFs can impact lung expansion at birth, expression of surfactant proteins and lipids, patency of the ductus arteriosus, and angiogenic factors and processes in developing lungs - all of which canmodify pulmonary hypertension. Further data show that HIFs are highly expressed and stable in third trimester fetal primate lungs, while one of them, HIF-1a, declines dramatically after preterm birth. Using cofactors 02, Fe2+, oxoglutarate, and ascorbate, HIF prolyl- hydroxylases (PHDs) specifically regulate HIFstability. We reported that PHDinhibitors (PHDi) profoundly alter stability of HIFs and downstream gene expression (VEGF and its receptors) in lung endothelial and epithelial cells, and fetal lung explants, even in extreme hyperoxia. We hypothesize that PHDi can decrease PHNby restoring the fetal VEGF/eNOS axis and improving perinatal surfactant homeostasis. We propose to examine effects of PHDi delivered antenatally or postnatally, and by differing routes, in ovine models of RDS and persistent PHN in preterm and term ovine models, respectively. Chronically instrumented fetal/neonatal lambs will have pre- and/or postnatal hemodynamic, blood gas, and respiratory physiology measurements. In lung tissue from these animals, we will measure HIFs, HIF- dependent gene products, surfactant proteins and lipids, inflammatory cytokines and cells, and lung morphometry to assess vascular development and remodeling. Results will provide valuable information about pathophysiologic roles of HIFs in perinatal PHN and potential therapeutic uses of PHDi in RDS and PHN.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The subunit composition of electrophoretically isolated human haptoglobin 2-1 polymers was elucidated by 1) determination of the ratio of alpha 2 to alpha 1 polypeptide chains using gel densitometry, 2) analysis of the beta chain composition by enumeration of fractionally saturated hemoglobin-haptoglobin intermediates, and 3) determination of molecular weight of each polymer by sodium dodecyl sulfate (SDS) gel electrophoresis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "font color=\"#990000\";b>Benign and Malignant Thyroid Disease Among Those Exposed as Children to Fallout from the Chornobyl Accident</b;/font;br>The Chernobyl nuclear reactor accident in Ukraine in 1986 contaminated large parts of Belarus, northwestern Ukraine, and bordering Russian provinces. A case-control study in Belarus, with individual dose estimates, demonstrated a statistically significant link between thyroid cancer and environmental radiation dose from fallout related to the Chornobyl accident. With the assistance of the Department of Energy (DOE) and the Nuclear Regulatory Commission (NRC), NCI organized follow-up screening studies of benign and malignant thyroid disease among those exposed as children to fallout from the Chornobyl accident in Belarus and in Ukraine. The thyroid glands of approximately 12,000 subjects in each country have been screened biennially for four cycles in Ukraine and three cycles in Belarus. A prevalence analysis of thyroid cancer based on data from the first screening in Ukraine has been published in JNCI, and a descriptive paper on the pathology of the tumors has appeared in Cancer. A paper on autoimmune thyroiditis has been published in JCEM, a paper on antibody levels among the low-exposed group has appeared in Clinical Endocrinology, and dose-response analysis of follicular adenoma is in the American Journal of Epidemiology. A number of other manuscripts, including one on incident thyroid cancers on the in utero exposed and on hypothyroidism are close to submission. The projects are currently preparing to transition from active screening to other forms of follow-up widening the existing national cancer registries.<br;br><font color=\"#990000\";b>Cancer Mortality Among the Population Exposed to Radioactive Waste Released in to the Techa River Adjacent to the Mayak Nuclear Facility in Russia</b;/font;br>Villagers living along the banks of the Techa River in Russia were exposed to chronic external and internal environmental radiation from radioactive waste released into the river by the Mayak nuclear facility. Cancer mortality is being evaluated in a cohort of approximately 30,000 people who received low to moderate radiation doses from the contaminated river. Current efforts focus on improving data quality, tracing the population for vital status information, and on statistical analyses and manuscript preparation. Associations between solid cancer and leukemia mortality and radiation dose have been observed.<br;br><font color=\"#990000\";b>Thyroid Nodules as a Measure of Radiation Risk Among a Population of 3000 Residents in Kazakhstan Exposed as Children to Fallout from Nuclear Testing at the Semipalatinsk Test Site</b;/font;/br>Thyroid nodule prevalence, determined by ultrasound screening, was used as the main outcome variable for evaluating the effects of environmental radiation fallout exposure in a population of 3000 village residents in Kazakhstan affected by radioactive fallout from nuclear bomb tests at the nearby Semipalatinsk Test Site during 1949-1962. A new dose-reconstruction algorithm, developed jointly by NCI and the Institute of Biophysics in Moscow, was used to estimate individual thyroid doses from external and internal sources of fallout-derived radiation, based mainly upon residential history and estimated consumption of milk from dairy animals grazing on contaminated pasture. Both external and internal dose were found to be significantly and independently associated with nodule prevalence after adjustment for sex and for age at screening. Current efforts, which include obtaining new information through the use of focus group interviews and application of statistical methods that take account of random errors in estimated dose, are focused on sources of uncertainty, especially those involving internal dose from radioactive isotopes of iodine ingested through the pasture-to-milk food chain. Polymorphic variants in 13 genes were studied for effect modification of the radiation dose-response relation and main effects on thyroid nodule risk. A variant in XRCCI was found to interact with radiation dose and thyroid nodule risk.<br;br><font color=\"#990000\";b>Radiation Biodosimetry</b;/font;br>Periodic reviews of the scientific literature are conducted to identify the most current techniques used to estimate doses from past exposure to occupational and environmental radiation. These data are used to inform current epidemiologic studies of irradiated populations (eg. Chernobyl, Techa River, X-ray technologists) where doses may be missing, to estimate dose uncertainty or validate estimated doses.<br;br><font color=\"#990000\";b>Ultraviolet Sunlight Exposure, Ionizing Radiation, and Risk of Skin and Other Cancers</b;/font;br>Exposure to ultraviolet (UV) radiation from sunlight plays a predominant role in the etiology of melanoma and non-melanoma skin cancer. There is accumulating evidence linking melanoma and two major non-melanoma skin cancers (i.e., basal cell carcinoma and squamous cell carcinoma) to different patterns of UV exposure (i.e., total, chronic and acute intermittent), but the nature of the relationship is complex and not fully understood. We have recently completed a study to evaluate the validity of questionnaire-based sun exposure data using histologically assessed sun-related skin damage. Furthermore, it has also been suggested that sunlight exposure may have a protective effect for non-Hodgkin lymphoma (NHL) and some other forms of cancer. We recently completed questionnaire-based collection of lifetime UV sun-related radiation exposure from more than 70,000 U.S. radiologic technologists and have undertaken analyses to assess risks of specific forms of cancer in relation to this exposure. We also plan to evaluate whether skin cancer risks associated with occupational exposures to ionizing radiation might be modified by questionnaire-derived estimates of UV sun-related radiation exposures. <br /;br /><font color=\"#990000\";b>NHANES III: Prospective Vitamin Study</b;/font;br>The NHANES III data set with measured vitamin D levels on about 20,000 people is a unique resource that allows investigation of many issues on the relationship between vitamin D and specific causes of mortality. This cohort includes actual vitamin D measurements and about 2253 deaths, of which 1072 are circulatory disease deaths and 536 are all cancer deaths. We have completed analysis of the relationship between baseline vitamin D and all cancer mortality, with a manuscript published. The Follow-up has been extended- with 60% more deaths. Analyses of cancer outcomes are being under taken in the extended dataset.<br /;br /", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this study is to assess the role that contextual factors (e.g., community-level health and social resources, community socioeconomic status) play in shaping mental health and recovery in the aftermath of a disaster. The notion that community characteristics are important determinants of post disaster functioning was dramatically brought home in much of the public discourse about Hurricane Katrina that focused extensively on the role that underlying socio-economic position, resource availability, and preparedness may have played in the aftermath of this disaster. The proposed research would be the first study, to our knowledge, to provide a full, rigorous, and theoretically-based test of the various pathways through which community-level factors shape individual risk and community rates of psychopathology and adverse behavioral outcomes after disasters. Our broad aim is to demonstrate the utility of an ecological conceptualization of disaster recovery, focusing on individuals' health outcomes such as the occurrence/severity of posttraumatic stress disorder (PTSD), depressive symptoms, and chronic stress. To achieve these aims we will collect both secondary and primary data to describe (A) community characteristics of Mississippi counties before and after the disasters, and (B) sociodemographics, event exposures, and mental health among 1,000 residents of Mississippi. To characterize pre-disaster community resources in ways that are not subject to recall bias, we will assemble a contextual database from publicly available sources, such as the US Census and the Mississippi Development Authority. We will also assemble objective information about community-level exposure and postdisaster community resources, including estimates of need and amounts of FEMA monies awarded. In order to obtain individual-level information, we will select a random sample of 1,000 adults from three geographic areas formed by levels of damage caused by Hurricane Katrina. Using multiple sampling modalities we will sample persons proportional to their representation in the general population before the hurricane and include both persons who were and those who were not evacuated as a result of the hurricane. Key hypotheses will be tested using multi-level hierarchical models and ecologic analyses. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our long term objective is to develop a detailed understanding of globin gene activation during erythroid cell differentiation. To model these events we are characterizing globin gene expression during the differentiation of murine erythroleukemia (MEL) cells. We have identified three proteins that bind specifically to the promoter of the alpha-globin gene DNase I footprinting and electrophoretic shift assays were used to monitor the enrichment of each protein through several chromatographic steps. DNA sequence affinity chromatography was then used to purify each protein. We propose to employ these purification protocols to accumulate a sufficient amount of each factor to obtain amino acid sequence data. These data will be used to prepare oligonucleotide probes that will be used to screen MEL cell cDNA libraries. In addition, direct screens of MEL cell cDNA expression libraries will be initiated. For these experiments, double-stranded oligonucleotide comprised of the binding site of each protein will be synthesized, ligated, nick-translated, and used to probe cDNA expression libraries. The identification of clone that encode alpha-globin specific DNA binding proteins will allow us to compare these proteins to other DNA binding factors, to characterize the functional domains of each protein, and to investigate their tissue distribution, synthesis, and turnover. Antisera could also be prepared against synthetic peptides derived from the deduced amino acid sequence of each protein. This analysis will help us to obtain a deeper understanding of how nuclear proteins regulate cell growth and differentiation. In this regard it is of interest to note that MEL cells are transformed erythroid precursors arrested in differentiation at the CFU-e state. We have shown, however, that MEL cell differentiation leads to characteristic changes in the activities of the identified alpha-specific binding proteins. It is not unreasonable to speculate that an understanding of how these factors function and how their activities are modified during differentiation might lead to the development of novel strategies for selectively altering factor activities. Because these factors appear to play important roles in mediating growth and differentiation, these insights could have important implications for approaches aimed at altering the uncontrolled growth and lack of differentiation characteristic of many tumor cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gonorrhea is the second most common bacterial sexually transmitted infection ? the most common is chlamydia, which often coinfects with gonorrhea. About 80 million new cases of gonorrhea occur worldwide annually. Over 555,000 cases were reported in the U.S. in 2017. Serious sequelae of gonorrhea in women include infertility, ectopic pregnancy and chronic pelvic pain. Neisseria gonorrhoeae (Ng), the causative agent of gonorrhea, has become resistant to almost every antibiotic in clinical use. Resistance to ceftriaxone and azithromycin ? the recommended first-line of treatment ? portends an era of untreatable gonorrhea. The CDC has listed Ng as a microorganism with a threat level of ?Urgent.? Development of a safe and effective vaccine against gonorrhea is a public health priority. Our group has shown that a monoclonal antibody (mAb) called 2C7 that targets a widely-expressed Ng lipooligosaccharide (LOS) epitope (the 2C7 epitope) is bactericidal. A peptide mimic (mimitope) of the 2C7 epitope, when configured as a polymer (a ?multiantigen peptide?) elicits bactericidal Abs and attenuates Ng vaginal colonization in mice. Encouraged by the recent retrospective observation that a group B meningococcal vaccine composed of detergent-extracted outer membrane vesicles (MeNZB) designed to curtail a meningococcal epidemic in New Zealand also reduced incident gonococcal disease by 31%, we will incorporate the gonococcal 2C7 epitope into meningococcal outer membrane vesicles to target both meningococci and gonococci. This innovative approach is advantageous because both diseases are highly prevalent in adolescents and young adults. In addition to the economic advantages, a ?pan- Neisseria? vaccine would help address an already crowded vaccination schedule and may mitigate the stigma associated with vaccines against STIs. In Aim 1, we will develop meningococcal native outer membrane vesicles (NOMVs) containing LOS that displays the 2C7 epitope. Meningococcal FH-binding protein (FHbp) will be mutated to abrogate binding of human FH and thereby elicit higher bactericidal Ab against Nm than wild- type FHbp. We will also delete Rmp and H.8, which may elicit blocking Ab against Ng and Nm, respectively. Elicited Abs will be tested for complement-dependent bactericidal activity against homologous and heterologous Ng strains and tested for efficacy versus Ng in transgenic mice we have developed that express the human complement inhibitors, FH and C4BP, to better simulate the complement environment in humans. We will use mice deficient in C9 (the last step in formation of the bacteriolytic pore) or mice depleted of PMNs to elucidate the mechanism of action of the candidate vaccine, which will also define correlates of protection. In Aim 2, we will develop detergent-extracted outer membrane vesicle vaccines (dOMVs) that display the 2C7 mimitope peptide in the framework of the major outer membrane protein, PorA. Immunogenicity and efficacy of this vaccine will be evaluated as described in Aim 1.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT TITLE: Southwest Tribal NARCH X Administrative Core PROGRAM DIRECTOR/PRINCIPAL INVESTIGATOR: Dr. Kevin English Project Summary/Abstract The Southwest Tribal NARCH X Administrative Core will provide overall leadership to the Center and will coordinate, integrate, and support the various Center components and activities in order to fulfill the overall NARCH mission, which is to implement health research projects prioritized by area tribes, reduce distrust of research among American Indian and Alaska Native (AI/AN) communities, and enhance health research partnerships to reduce AI/AN health disparities, while promoting a cadre of AI/AN scientific and health research professionals. The Southwest Tribal NARCH X Administrative Core will be based at the Albuquerque Area Indian Health Board, Inc. (AAIHB), an intertribal organization, which has extensive experience in the oversight of prior NARCH grants (NARCH I, NARCH III, NARCH V), and currently administers a NARCH VII grant. The Southwest Tribal NARCH X Administrative Core will help fulfill the major aims and goals of all Center components by pursuing the following specific aims: 1) establish a sustainable structure which will facilitate the conduct of Southwest Tribal NARCH X work by providing administrative and technical support as well as by stimulating exchange among all proposed Center components in collaboration with academic and tribal partners; 2) maintain the Southwest Tribal NARCH Community and Scientific Advisory Council (CSAC) to ensure rigorous tribal oversight of all Southwest Tribal NARCH components and activities and foster sustained alignment with tribal priorities and key cultural considerations; and 3) support the continued operation of the Southwest Tribal Institutional Review Board (IRB) to advance the benefits of ethical health research to area tribes and reduce AI/AN community distrust of research and researchers by giving tribes greater control over the research process. Expected outcomes of the Southwest Tribal NARCH X Administrative Core include rigorous tribal oversight of all Southwest Tribal NARCH X components, continued alignment of Southwest Tribal NARCH X components and activities with tribal priorities and important cultural considerations, enhanced protection for individual and community research participants, streamlined IRB review processes, enhanced research alliances among all partners (AAIHB, tribes, and academic partners), and manuscripts and professional presentations highlighting key findings of all Southwest Tribal NARCH X research and innovations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Carcinoma of the esophagus and carcinoma of the pancreas are among the most lethal of human neoplasms. Efforts to devise successful therapeutic regimens for the primary neoplasm are hampered by lack of an animal model suitable for the study of surgical procedures or radiation therapies appropriate for use in man. Using carcinogens with proven effect in small animals, we have successfully induced invasive bronchogenic carcinoma in dogs. In this application, we propose to extend this approach to the development of carcinoma of the esophagus and carcinoma of the pancreas in dogs. Achievement of a predictable model of esophageal and pancreatic cancer, in which the lesion is reproducible at pre-selected localized sites, will provide 1) opportunities for studying the transition of normal tissues into pre-neoplastic and neoplastic lesions, 2) a preparation for serial diagnostic studies utilizing methods feasible in man but not possible in the small animals usually employed in carcinogenesis research, and 3) a potential for meaningful therapeutic investigations using this large animal model for esophageal and pancreatic carcinoma. As the work progresses and neoplastic lesions are produced, collaborative projects with other members of the City of Hope Cancer Research Center will be developed. These will include basic immunologic and cytogenetic studies of the evolving tumors, and multimodality therapeutic approaches involving radiation and medical oncology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The mucopolysaccharidoses (MPS) are a group of hereditary diseases characterized by defective metabolism of glycosaminoglycans (GAGs). The disorders are usually associated with severe dysfunction of the nervous system as well as of liver, spleen, heart, bone, and other tissues. Objective of this project is the study of mechanism of pathogenesis of these diseases with emphasis on brain involvement and mental retardation. We are using a comparative approach. For this purpose we study the changes, in GAGs, sphingolipids, and pertinent lysosomal enzymes in tissues of patients with various types of MPS and we make correlation in terms of clinical and ultrastructural findings. Our laboratory contributed significantly in understanding the chemical pathology and in particular the neurochemistry of MPS IH, MPS IS, MPS II, MPS III A and MPS III B. To complement the studies with human subjects, a drug (suramin) induced animal model of MPS has been developed and a canine model, (natural), of MPS I (alpha-L-iduronidase deficiency), has been fully characterized. In an attempt to reverse the progressive deterioration caused by this incurable metabolic disorder, five doges with MPS I were transplanted with marrow from normal or heterozygous littermates. Transplanted bone marrow provides the affected dogs with self-renewing source of cells that produce the enzyme needed to complete the metabolic process. Two of these recipients were killed on post-bone marrow transplantation (BMT) day 628 and 594 at which time both were clinically healthy. Large reduction of stored GAGs, from all tissues including the brain, appears to be consistent consequence of BMT therapy in canine MPS I.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Multiple Sclerosis (MS) is an autoimmune disorder of the central nervous system typified by axonal demyelination and neuronal death. To date, there is no effective treatment to cure the disease, and the available therapies do not alter the outcome of the disease. The development of better therapeutic options requires in-depth understanding of the molecular mechanisms leading to disease development and progression. Our group previously discovered a single nucleotide polymorphism (SNP, rs6897932) within exon 6 of the IL-7 receptor alpha chain (IL7R) that is strongly associated with the risk of developing MS. Our group further showed that the MS-associated allele of the SNP increases skipping of exon 6 both in vitro and in vivo, leading to increased production of a secreted receptor (sIL7R) that is unable to activate the IL-7 signaling pathway. Importantly, sIL7R has been linked to the disease in both human patients and animal models. These results directly implicate splicing of IL7R to the pathogenesis of MS, and posit the trans-acting factors controlling its splicing as candidate MS susceptibility genes. I have uncovered two of the trans-factors controlling IL7R exon 6 splicing: the RNA helicase DDX39B, which activates exon inclusion, and the polypyrimidine tract binding protein (PTBP1), which represses it. Furthermore, we uncovered several SNPs in the DDX39B gene region that are associated with MS, thereby establishing DDX39B is itself a risk factor for MS. None of the associated SNPs are located within the coding sequence of the gene, suggesting that one or more of these SNPs may contribute to the disease association by modifying DDX39B expression. Through bioinformatics and gene expression analyses, I have uncovered mRNA isoforms encoding either the full-length protein or a novel short protein, and several mRNA isoforms that are candidate targets for nonsense-mediated decay. Importantly, I have established two of the SNPs could alter expression levels of these isoforms. The goal of this proposal is to provide functional links connecting DDX39B to the pathogenesis of MS. Specifically, I aim to: 1) elucidate its role in the regulation of IL7R exon 6 splicing; 2) understand the functional roles of the different protein isoforms; and 3) uncover the SNPs responsible for its association with MS. I will combine biochemical and genetic approaches to elucidate the mechanism by which DDX39B activates exon 6 splicing, and to characterize the functional roles of the protein isoforms. I will test the impact of selected SNPs on DDX39B expression by combining in vivo gene expression analysis and functional studies using reporter minigenes. Successful completion of this research will advance our current understanding of the molecular underpinnings of MS, in particular by functionally linking DDX39B to the pathogenesis of MS, and providing a functional characterization of the DDX39B gene. Given that DDX39B has been associated with numerous autoimmune disorders, our results could be relevant to the mechanistic etiology of other autoimmune diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The secretory pathway in the guinea pig exocrine pancreas has been defined in the past in terms of 3H-leucine incorporated into secretory protein (autoradiography and cell fractionation studies). Using a two-dimesional gel technique (isoelectric focusing followed by SDS gradient gel electrophoresis), the secretory pathway will be redefined on a protein specific basis. This will involve studies on (1) cell fractions taken at varying time points during a pulse-chase protocol using a mixture of fifteen 14C amino acids, and (2) radiolabled secretion discharged into incubation medium from pancreatic lobules under varying conditions designed to favor evaluation of (a) discharge, (b) intracellular transport, and (c) the secretory pathway taken as a whole. Using mRNA derived from puromycin-high salt disassembly of bound polysomes from dog pancreas, the primary translation products obtained in vitro will be compared to radiolabeled secretion derived from pancreatic slices by SDS gradient gel electrophoresis and radioautography. Since preliminary studies suggest the presence of pancreatic precursors, attempts will be made using selective immunoprecipitation methods, to isolate several specific precursor-product pairs. Tryptic mapping and sequencing studies will be done on these pairs to determine if the additional peptide sequence is attached to the NH2 terminal end of the precursor molecule, as postulated by the \"signal\" hypothesis. BIBLIOGRAPIC REFERENCES: Cellular cyclic nucleotides and enzyme secretion in the pancreatic acinar cell. Haymovits, A., and G. Scheele. Proc. Nat. Acad. Sci. USA 73, 156-160 1976. Secretory Proteins of the Exocrine Pancreas. Scheele, G.A., in Cell Biology (P.L. Altman and D.D. Katz, eds.), Federation of American Societies for Experimental Biology, Bethesda, Maryland, 334-336, 1976.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We are conducting studies to determine the function of the Mi-Tfe family of basic helix-loop-helix leucine zipper (bHLH-Zip) transcription factors in mammalian development. The Mi-Tfe family consists of four related genes- Mitf, Tfe3, Tfeb, and Tfec. The Mitf gene encodes the microphthalmia transcription factor (Mift). Mutations in the human MITF gene are responsible for a common human pigmentation and hearing disorder, Waardenburg syndrome type 2 (WS2). More than 20 independent Mitf mutations have been isolated in the mouse and shown to affect a number of cell types, including melanocytes, osteoclasts, and mast cells. In contrast, Tfeb, Tfe3, and Tfec were isolated by biochemical means and little is known about their function in vivo. In collaborative studies, we showed that the Mitf-Tfe proteins can bind the E-box sequence as homodimers or as heterodimers with other family members. To investigate the function of the Tfe genes in mammalian development, we used embryonic stem (ES) cell knockout technology to make germline null mutations in the three mouse Tfe genes. The effect of each mutation on development was then measured alone or in combination with mutations in other Mitf-Tfe family members. Surprisingly, these studies did not identify any essential functions for Mitf-Tfe heterodimers in development. This is in marked contrast to what has been observed for the Myc/Max/Mad family of bHLH-Zip proteins, where the heterodimers are thought to have essential functions. Future studies will attempt to use chemical mutagenesis and genetic modifier screens to identify other proteins in the Mitf signal transduction pathway.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this project is to determine in molecular detain the energetic-structure-function relationship in exchangeable apolipoproteins, thereby providing an insight into molecular mechanisms of their action in the pathogenesis of atherosclerosis and other apolipoprotein-related disorders. Exchangeable apolipoproteins are water- soluble protein components of lipoproteins that mediate lipid and cholesterol transport and metabolism and play crucial roles in the pathogenesis of atherosclerosis, coronary heart disease, stroke, and other major human disorders including several forms of systemic and cerebral amyloidosis. Structural adaptability of apolipoproteins to heterogenous lipoprotein complexes and to plasma is absolutely essential for their functions, and has to be understood in detain in order to elucidate molecular mechanisms of apolipoprotein action in normal and in diseased states. The proposed work addresses this long-term goal through studies of the energetics, structure and folding pathway of the smallest human plasma apolipoprotein C-1 (apoC-1, 6 kDa). The ability of apoC-1 to activate lecitin: cholesterol acyltransferase (LCAT) may account for normal plasma levels of cholesterol esters in subjects with deficiency of the major LCAT activator, apoA-1. ApoC-1 delays the clearance of potentially atherogenic triglyceride-rich particles by inhibiting their uptake via the apoE-mediated low-density low-density lipoprotein receptor- related pathway. Thermodynamic and structural analyses of synthetic human apoC-1 and a series of its site-specific mutants targeted towards the predicted amphipathic alpha-helical regions will be carried out by using a combination of far-UV circular dichroism spectroscopy, differential scanning calorimetry, and x-ray diffraction methods. Such analysis will 1) dissect the folding pathway of lipid-free apoC-1 in solution, from partly folded monomeric to fully folded oligomeric or lipid-bound state; 2) determine, at the level of individual amino acids, the critical factors for the stability and cooperatively of the amphipathic alpha-helical structure in apoC-1; 3) determine the conformation apoC-1 in various self- associated status, such as 2D filaments and 3D crystals, to provide models fir a variety of functional apolipoprotein conformations. The results of this analysis will provide the energetic and structural basis for understanding mechanisms of functional apolipoprotein reactions and will help to understand their amyloidogenic properties, thereby leading to identification of rational therapeutic targets in a variety of apolipoprotein-related disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this application we argue that mechanisms of transition from acute to chronic pain critically depend on brain emotional and motivational learning and memory circuit, namely the properties of the corticostriatal system. This is a radical departure from the classic pain research tradition, which emphasizes mechanisms of nociceptive encoding and representation. We provide human and animal model data consistent with the idea, and propose unraveling underlying mechanisms and translating this information to clinical application by testing a drug treatment strategy for preventing transition to chronic pain We recruit Dr. Surmeier (a world renowned expert on the physiology and reorganization of the striatum) and Dr. Martina (a young scientist with expertise in molecular and electrophysiological studies of the brain), to collaborate with Apkarian on this project, and Drs. Fields and Zubieta act as consultants for the project. In Aim 1 we combine the expertise of the three collaborating labs (Apkarian, Martina, Surmeier) to link large-scale brain imaging outcomes to cellular, molecular, and electrophysiological changes that we predict the corticostriatal undergoes in the transition from peripheral nerve injury to neuropathic pain-like behavior in rodents. In Aim 2 we compare among potential drugs for preventing transition to chronic pain-like behavior in a rat model. In Aim 3 we use the best candidate drug in a human early phase clinical trial, combined with brain imaging, for prevention of transition to chronic pain. The successful completion of the study should dramatically change current notions regarding brain mechanisms of pain chronification, and provide a new concept of treatment strategy for preventing the transition to chronic pain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract In recent years, the demand for home hospice care has grown rapidly. Family members and friends who act as informal caregivers are essential to the provision of palliative care services;however, this role is not without adverse effects on the caregivers themselves. It is well documented that emotional needs of individuals caring for dying persons in their home are not well attended, and interventions aiming to provide support to informal hospice caregivers are notably lacking. In this context, problem solving therapy (PST) provides an overall coping process that fosters adaptive situational coping and behavioral competence. This reduces and prevents the negative effects of stressful life events with regard to both psychological and physical well-being. We propose a randomized controlled trial to fully evaluate the PST intervention for informal hospice caregivers. Additionally, we aim to evaluate how the modality of the intervention (face to face vs video) impacts its effectiveness. We propose a 4-year randomized trial study in which hospice caregivers will be randomly assigned to a group receiving standard hospice care with the addition of social support visits (attention control group) or a group receiving standard hospice care with the addition of the problem solving intervention delivered face to face (intervention group 1) or a group receiving standard hospice care with the addition of the problem solving intervention delivered via video (intervention group 2). The specific aims include an assessment of the impact of PST on caregiver quality of life, problem solving ability, caregiver anxiety, and caregiver perceptions. Furthermore, we aim to evaluate the cost effectiveness of the PST intervention. PUBLIC HEALTH RELEVANCE: Narrative The proposed research study explores a problem solving intervention that will improve caregivers'quality of life, and cost-effective innovative tools to support its delivery. Thus, the study is highly relevant to public health as it affects the health of a continuously growing segment of our population, namely these who are called to assume the essential role of informal caregiving, and aims to demonstrate the value of a feasible and sustainable problem solving intervention for hospice caregivers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Gastric atrophy and metaplasia are critical epithelial cell changes that promote gastric carcinogenesis. Gastric cancer is a major public health issue world-wide, being the third leading cause of cancer-related death in the world with nearly one million victims every year. Chronic inflammation is a major predisposing risk factor for the development of gastric cancer, demonstrated by the fact that patients with chronic atrophic gastritis have significantly higher risk of cancer than their healthy peers. While the association between chronic inflammation and cancer risk is clear, the exact mechanisms by which the inflammatory process in the stomach leads to carcinogenesis in some patients but not others are not understood. There is a complex cytokine milieu associated with any chronic inflammatory process, and polymorphisms in cytokine genes in human patients have been shown to have a significant effect on the risk of gastric cancer development. Due to the strong association between the prolonged cytokine production during inflammation and increased gastric cancer risk, we are interested in determining the mechanistic role that cytokines serve in regulating the development of gastric atrophy and metaplasia during chronic gastritis. Epstein-Barr Virus-Induced Gene 3 (Ebi3) encodes a protein subunit (EBI3) of the cytokine IL-27, which is well- characterized and has documented effects on CD4+ T cells during inflammation. EBI3 is also a component of the poorly understood immunosuppressive cytokine IL-35 thought to be made by regulatory T cells (Tregs). We use a novel murine model of inflammation-induced gastric atrophy and metaplasia, TxA23. In this model we have discovered that mice deficient in the production of Ebi3 have accelerated development of atrophy and spasmolytic polypeptide expressing metaplasia (SPEM), important preneoplastic epithelial cell lesions. We have also observed that Tregs, which are critical suppressors of gastritis and resulting lesions, exhibit a defective phenotype in Ebi3-/- mice. However, it is unclear whether this defect is due to the extrinsic lack of IL- 27 signaling or the intrinsic inability to express Ebi3, and thereby IL-35. Our hypothesis is that EBI3 expression is critical for suppressing the development of gastritis, gastric atrophy, and gastric metaplasia due in part to the effects of IL-27 acting on Tregs and increasing their suppressive functions. In Aim 1 we will determine whether IL-27 is the critical EBI3 containing cytokine responsible for suppressing disease progression. In Aim 2, we will determine the effect of IL-27 signaling into Tregs during the development of gastric atrophy and metaplasia.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objectives of this research are to identify and characterize the cellular mechanisms underlying ethanol induced changes in neuronal development, and to assess the role of these changes in the etiology of CNS abnormalities associated with Fetal Alcohol Syndrome (FAS). Key neuropathologic features of FAS include altered neuronal morphogenesis and synapse formation in the hippocampus. Recent studies have elucidated some of the molecules and processes responsible for the distinct growth characteristics of axons and dendrites, thus providing the basis for novel experiments to determine the mechanisms underlying ethanol's disruption of neuronal development. The use of primary cultures of embryonic rat hippocampal pyramidal neurons is integral to the objectives of the project. Neurons in these cultures develop axons, dendrites, and synapses in a sequence of events that mimics their development in vivo. Experiments in this proposal are designed to use immunofluorescent cytochemistry coupled with quantitative morphometric analysis, time lapse videomicroscopy, and Fura-2 intracellular free calcium measurements in the following specific aims: (1) Compare the sensitivity of cultured hippocampal neurons exposed to ethanol at different times relative to development of axons, dendrites and synapses, (2) Distinguish whether ethanol induced changes in neuronal development result from direct effects of ethanol on neurons, or indirect effects on neurons mediated by astrocytes, (3) Determine whether ethanol's effects on process outgrowth involve altered regulation of intracellular calcium levels. The results of these studies will establish whether a disruption of process outgrowth and molecular compartmentalization is a key aspect of ethanol's neurodevelopmental toxicity and will provide important insight into the mechanism(s) underlying these actions. This fundamental knowledge can be expected to provide a basis for improving the identification and treatment of affected individuals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (From the Applicant?s Abstract): The studies proposed herein will: (i) determine the roles of membrane FasL (mFasL) and soluble FasL(sFasL) in neutrophil-mediated inflammation within the eye and most importantly, (ii) determine whether sFasL can be used to downregulate neutrophil-mediated inflammation and prevent neutrophil-mediated ocular damage. Preliminary studies demonstrate that within the immune privileged eye, mFasL activates neutrophils and initiates innate immunity, while sFasL inhibits neutrophil activation ad prevents innate immunity. Most compelling though, was the ability of sFasL to block the pro-inflammatory effects of mFasL when both sFasL when both sFasL and mFasL were equally expressed within the eye. The proposed studies will utilize a novel method of expressing DNA directly within the cornea to characterize, in vivo, the inflammatory response induced by the different forms of FasL expressed within the corneal stroma. The ability of both mFasL and sFasL to directly activate neutrophils will be assessed in vitro using stromal cells transfected with the different FasL cDNAs. The final set of experiments will determine, in vivo, whether sFasL blocks neutrophil mediated inflammation in the cornea, and to elucidate, in vitro, the mechanism by which sFasL blocks mFasL induced neutrophil activation. We anticipate that these studies will yield new information as to how FasL regulates inflammation within the eye, and possibly lead to the development of new treatments using the soluble form of FasL to prevent ocular damage secondary to neutrophil mediated inflammation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The 42 kDa processed fragment of the Plasmodium falciparum Merozoite Surface Protein 1, MSP1-42, is a leading protective human malaria vaccine currently undergoing clinical studies. Protective immunity is antibody-mediated, and directed to the C-terminal, immunodominant 19 kDa region of MSP1-42, or MSP1-19. Available evidence indicates that the vaccine-induced immunity to MSP1-42 or MSP-19 relies on the use of very powerful adjuvants (i.e. CFA) and on repeated hyper-immunizations to elicit high levels of protective antibodies. These are formidable obstacles in the way of field deployment of the malaria vaccine. Recently, we have shown that by refocusing the antibody responses to the N-terminal immuno-silent region of MSP1-42, MSP1-33, biologically active antibodies can be produced when combined with low concentrations of anti-MSP1-42 antibodies. This creates an opportunity to devise new vaccine constructs and/or immunization regimens to efficiently induce protective antibodies under the more favorable logistics of a reduced immunization schedule and the use of readily acceptable adjuvants. Accordingly, Specific Aim 1 will explore strategies to develop bivalent MSP1 vaccines as well as prime/boost immunization regimens, based on MSP1-33 and MSP1-42, in order to achieve a balanced induction of protective antibody responses. The focus will be on the use of less powerful adjuvants and a reduced immunization schedule. Specific Aim 2 will define experimental conditions in which combinations of anti-MSP1-33 and anti-MSP1-42 antibodies will inhibit parasites carrying different allelic and variant forms of MSP-42. This information, together with the down-selected vaccination strategies from Specific Aim 1, will be used to formulate MSP1-33/MSP1-42 vaccines to elicit strain-transcending, parasite inhibitory antibody responses. Significance: This exploratory project provides the critical proof-of- principle on which further development and validation of the dual MSP1-33/MSP1-42 vaccine strategy can be based. The strategy will rapidly lead to a new generation of MSP1-based malaria vaccines deployable with significantly improved efficacy and logistical feasibility, while utilizing the already existing technologies/platforms of demonstrated safety. PUBLIC HEALTH RELEVANCE - PROJECT NARRATIVE: Malaria is a major killer in developing countries and the development of effective vaccines is critical in controlling this deadly disease. This exploratory application seeks to improve the efficacy and the logistical feasibility of deployment of a leading candidate human malaria vaccine, Merozoite Surface Protein 1, MSP1. This is based on new and significant discovery of the immune responses to the MSP1 vaccine, which allows for novel re-design of the vaccine. Success in this project will move this vaccine forward in terms of significantly improving its potential to be effective in large populations in the field.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During mammalian development, the heart is the first organ to develop; the product of exquisite spatial and temporal control over cardiogenic mesodermal cell fate specification. From heart looping to cardiomyocyte cell fate determination, multiple transcription factors play critical roles during the development of the heart. Genetic silencing of Nkx2.5 in mice, the mammalian homologue of D. melanogaster tinman, results in embryonic lethality between 9.0 and 10.0 days post coitus due to abnormal heart morphogenesis. Clinically, missense and nonsense mutation in Nkx2.5 are highly conserved across multiple generations of families with high incidences of congenital heart defects. In Nkx2.5-/- mice, the cardiac specific isoform of the DEAD-box RNA helicase Mov10l1 (Csm) was found to be absent. DEAD-box RNA helicases are involved in virtually all biological processes involving RNA; specifically the nuclear DEAD-box RNA helicases p68 and p72 have been shown to be required for the processing of microRNA primary transcripts. MicroRNAs are critical for normal developmental processes are disregulated under pathological conditions. We demonstrate that Csm physically interacts with argonaute-2, a member of the RNA induced silencing complex, at P-bodies which are known to be sites of mRNA degradation. In addition, morpholino knockdown of Csm in the Danio rerio model system completely abrogates the heart looping process inhibiting cellular proliferation, whereas alpha-MHC promoter FLAG- Csm transgenic mice exhibit a hyperplastic heart. Transgenic animals have a significantly reduced survival rate. FLAG-Csm was found to co-immunoprecipitate a subset of cardiac microRNAs suggesting that it is actively involved in microRNA regulatory pathway. The overall hypothesis is that Csm is critical to both the normal development and function of the heart by facilitating the maturation of mircroRNAs throughout development and into adulthood. To test our hypothesis, we will determine the biochemical function of Csm and its interacting partners. Finally we will determine the biological and physiological effects Csm overexpression, and in conjunction with our biochemical and molecular analyses, determine the functional significance of Csm in the regulation of cardiac gene expression and the maintenance and function of the normal myocardium.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Active transport of vesicular cargoes is vital to the targeted delivery of organelles, proteins, and signaling mol- ecules in the complex and crowded cellular environment. Accordingly, defects are linked to developmental, neurodegenerative, pigmentation, immunological, and other diseases. Knowing the detailed mechano- chemistry and structural dynamics of isolated motor proteins is essential for integrating the divergent mechani- cal and kinetic properties of cytoskeletal motor families. We have developed a number of powerful new bio- physical tools that reveal the modulation of mechanical and ATPase reaction kinetics under applied mechani- cal force, elucidate the essential rotational conformational transitions of specific domains within the motor pro- teins and produce reliable force, stepping dynamics, and local viscoelastic parameters of the cellular environ- ment. We will apply these unique tools to investigate the divergent biochemical and mechanical properties of myosin-I (Myo1) and dynein isoforms that have that have not yet been approached at the mechanistic detail now possible. Aim 1: Using simultaneous optical trapping and TIRF microscopy, determine stress- and strain-dependence of the binding and dissociation of ATP and ADP that fuel motion and control motor stepping; Aim 1A: in myosins 1b and 1c; Aim 1B: in budding yeast and mammalian cytoplasmic dynein. Aim 2: To determine the rotational motions of motor heads and lever arms that generate force and cargo translocation using single molecule polarized TIRF (polTIRF) microscopy; 2A: in myosins 1b and 1c; 2B: in yeast cytoplasmic dynein. We discovered that Myo1b and Myo1c have markedly different strain-dependent attachment lifetime, even though their unloaded kinetics and protein sequences are quite similar. We will apply our formidable single-molecule mechanical and fluorescence technologies to understand how these closely related motors have optimized their kinetic and structural variations for their different cellular roles. Mammalian and Saccharomyces cerevisiae cytoplasmic dynein have overlapping roles in their respec- tive cells, but their properties are very different. We are in the unique position to compare them using advanced biophysical methods. We will determine their strain-dependent ATPase dynamics and rotational motions. We anticipate that these studies will provide insight into the motors' mechanisms, and will also reveal biochemical and mechanical adaptations to their distinct functions. Aim 3: We will use our unique new technology for measuring and calibrating optical trap signals within live cells to 3A: determine the functional speciali- zations that vary the complement of motors among early and late endosomes, and small and large la- tex bead compartments (LBCs) endocytosed or phagocytosed into live cells. 3B: measure forces and motor dynamics during remodeling of endoplasmic reticulum (ER) by interaction with endosomal vesi- cles. All three aims represent close, essential collaborations with Drs. Ostap, Holzbaur and Shuman. Many cargos have opposing motors bound simultaneously; these motors may operate in teams functioning either cooperatively or competitively. We will focus on the role of oppositely directed motors at critical junctures in cellular organelle trafficking: early and late endosomes and the remodeling of endoplasmic reticulum we have observed when transport vesicles interact with the ER. Together, these studies will provide insights into the roles of the molecular motor families in regulating organelle motility, morphology and remodeling. These stud- ies will lead to a much improved understanding of myosin I and dynein isoforms that operate very differently from the better understood kinesin and myosin motors, leading to a more rigorous understanding of their func- tions in cell biology and disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Marfan syndrome is a systemic disorder of connective tissue with autosomal dominant inheritance and a prevalence of at least 1 in 10,000 individuals. Cardinal manifestations involve the ocular, skeletal and cardiovascular systems. While all clinical features impair quality of life, cardiovascular features are uniquely associated with early mortality due to myocardial failure or aortic rupture. Variability in the age of onset, tissue distribution, and severity of manifestations is common both in and among families. Despite major advances in the medical management of Marfan syndrome, the effectiveness of such therapies is compromised by the need for accurate and early diagnosis. It is now known the mutations in the gene on chromosome 15 encoding fibrillin (FBN1), a major glycoprotein component of the extracellular microfibril, can cause Marfan syndrome. Despite this understanding, gaps in knowledge include the molecular basis for clinical variability, the functional significance of various domains and residues of fibrillin, whether MFS truly lacks genetic heterogeneity, and the clinical spectrum that can be associated with fibrillin gene defects. The overall goal of this proposal is to gain insight into the normal and aberrant biology of the fibrillin protein (and of microfibrils) by analyzing the effects of naturally occurring mutations on clinical, cellular, and biochemical phenotypes. This will require comprehensive study of the FBN1 gene in patients with Marfan syndrome and clinically related disorders. Using refined mutation defection techniques and a novel panel of highly informative intragenic microsattelite polymorphisms for haplotype segregation analysis, it will be possible to determine whether FBN1 gene defects are associated with all cases of Marfan syndrome, to what degree allelic and/or genetic heterogeneity account for interfamilial clinical variability, and whether FBN1 gene defects underly divergent phenotypes that appear to manifest primary disruption of the elastic fiber-microfibrillar array system. By similar methods, the genes encoding other constitutive elements of extracellular microfibrils, or proteins known to associate with microfibrils, can be scrutinized as potential disease loci or as the site of sequence variants able to modify the clinical expression of mutant fibrillin alleles. Immunofluorescence and quantitative pulse-chase methods will allow determination of the cellular and protein phenotypes. In addition, much can be learned about the fundamental structural and functional properties of FBN1 domains by the study of wild-type and mutant forms using NMR spectroscopy. Finally, expression studies in cell culture and transgenic mice will allow the examination of the pathobiology of fibrillin in models that mimic the physiologic complexity of the human system. Correlation of mutant genotype to clinical, cellular, and biochemical phenotype will yield insight into the normal functions of specific fibrillin domains and residues, will enhance our understanding of pathogenesis, and should have both diagnostic and prognostic significance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Latex rubber articles in the form of condoms and gloves are used as barriers against the transmission of infectious diseases. The medical community have identified the use of condoms as an effective means for preventing the transmission of the human immunodeficiency virus (HIV) and other sexually transmitted diseases. Universal precaution procedures for health care workers requires wearing, and changing, latex examination gloves. Formed rubber latex items frequently reduce but do not eliminate completely the possibility of transmission of viruses such as HIV. This proposal suggests a method for coating latex articles with a thin, biologically bland, non-porous polymer to eliminate the possibility of virus transport via micropores in the latex membrane.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed Michigan Regional Comprehensive Metabolomics Resource Core (MRCMRC or MRC2) is a fully integrated program that will provide researchers nation-wide with the expertise and infrastructure to determine the levels of known and unknown metabolites in cells, tissues and biological fluids. In addition, the MRC will provide opportunities for training in the technology of metabolomic analysis, statistical analysis and bioinformatic evaluation of metabolite data as well as approaches to Incorporation of metabolomics into basic, preclinical, translational and clinical research. Incorporated into this service component will be a robust research component directed towards improving the breadth of metabolite detection, the quality and efficiency of metabolomic analysis and importantly, tools to turn spectral data into knowledge to for the researcher The MRC2 will contain an Administrative Core to oversee and integrate operations; an Analytical Core to help design experiments for directed quantitative measure of metabolites or high-throughput evaluation of large numbers of known and unknown metabolites; a Data and Information Technology Core which will perform primary data processing or re-mining of archival data and will serve as a data repository; a Statistics and Bioinformatics Core which will assist researchers in the statistical evaluation of metabolomic data and integration with other 'omics or phenotypic data; and a Promotion and Outreach Core which will organize and provide seminars, symposia, workshops and web-based videos to increase the lay and research communities knowledge and use of metabolomic data. PUBLIC HEALTH RELEVANCE: The RCMRC grant is directed towards providing increasing the technology to measure small molecules (metabolites) found in the body. These metabolites are the building blocks of all cells and participate in all body processes. By providing researchers with ways to measure the metabolites accurately from cells and tissues and in the blood, new insights into the ways In which changes in metabolism can contribute to diseases, such as diabetes and caner will be found. With this knowledge, new ways to prevent and treat a variety of diseases may be possible.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recently investigators from several countries have reported an increased rate of birth of malformed or maldeveloped infants to alcoholic mothers. These authors suggest that the observed defects are the direct result of maternal alcoholism. The observation of withdrawal symptoms in neonates whose alcoholic mothers had been intoxicated preceding (just before) delivery, raises the additional question whether such children may have an increased sensitivity to alcohol in later life. The present series of studies using a rat model are aimed at investigating the effects of ethanol during the period of gestation on the prenatal development and later behaviors of the offspring. The proposed research has 3 objectives: 1) At maturity, to compare the two groups of offspring for increased sensitivity to alcohol using intragastric self-administration techniques, 2) To compare the offspring of alcohol treated and control mothers with a battery of development and behavioral tests including measures of learning, 3) To isolate the effects of the administration of ethanol from nutritional stress by the isocaloric replacement of ethanol by carbohydrate and by pair feeding of control and experimental animals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Whether by administration of a vaccine or anti-pathogenesis substances, intervention of schistosomiasis by immunologic means is a major goal of schistosomiasis research efforts today. This application is to renew our program examining the role Schistosoma mansoni irradiated cercariae play as a model for inducing anti-schistosome immunity and anti-pathogenesis, and as candidates for a live vaccine. To determine vaccine effectiveness in situations approximating that to be encountered in the field, immunity will be assessed aginst multiple 'trickle' cercarial challenges. Using mice immunized with irradiated Schistosma japonicum cercariae, the specificity of vaccine induced resistance will be more thoroughly examined by challenge with the homologous or a heterologous schistsome species (S. mansoni). Anti-pathogenic effects of live vaccine administration will be examined in the mouse, along with the possibility that immunologic suppression of protection develops in cases where suboptimal levels of immunity are stimulated. Basic issues related to a recently developed cryopreserved live vaccine will also be addressed. These are: (1) by autoradiographic techniques determine the vaccine's migratory properties in the naive host, and (2) Attempt to enhance the vaccine's efficiency by concurrent administration of immunopotentiators and the cryopreserved vaccine. These experiments should answer pressing questions related to anti-schistosome immune mechanisms, while evaluating more critically the feasibility of irradiated larve as components of a live vaccine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed work addresses several of the Institute of Medicine's (IOM's) national priorities in clinical effectiveness research: disparities in oral health and health care. Combining extant dental data with other components of the rich VA patient databases, we will be able to examine the effectiveness of fluoride in preventing caries in patients with five of the 14 priority conditions specified by AHRQ: medically compromised veterans with dementia, depression and bipolar disorder, diabetes, HIV-Aids and persons with alcohol dependence. Recent data suggest that dental caries is as much of a problem in adults as children (Dye, et al., 2007). However, significant knowledge gaps remain in our knowledge base regarding prevention of caries in adults (Griffin et al., 2007). Using an extant, rich electronic data base from a health care system providing dental treatment to over 342,000 veterans annually, this proposed retrospective, longitudinal study takes maximum advantage of a unique opportunity to examine the following specific aims in a population of medically complex veterans. Aim 1. Examine the effectiveness of prescription-strength, self-applied fluoride and professionally applied fluoride in the prevention of caries in medically compromised veterans who are at high risk for caries. Veterans will be defined as at risk for caries if they have had two or more restorations in the previous year. Aim 2. While recent evidence suggests that fluoride is effective in preventing caries in adults, and multiple exposures to fluoride reduces caries risk in children (Weintraub, 2006), limited data exist showing that multiple exposures or modalities are more effective than a single modality in adults. Thus, we have a unique opportunity to examine whether multiple exposures to fluoride will be more effective than a single exposure. Aim 3. Over the past year, the Department of Veterans Affairs introduced a quality measure (the fluoride monitor) that examines, on a quarterly basis, the percent of veterans at high risk for caries who receive fluoride treatments. This project will examine the effectiveness of the introduction of this fluoride monitor in reducing the rate of restorations in medically compromised veterans at high risk for caries. PUBLIC HEALTH RELEVANCE: Despite dramatic gains in oral health and access to dental care during the past 50 years, dental decay remains a problem in medically complex older adults. The aging of the 76 million baby boomers, who are retaining more teeth than ever before, makes this a compelling problem. The purpose of this study is to examine the effects of prescription strength, self-applied, brush on fluorides and in-office fluoride treatments in preventing decay in a real-world setting of medically compromised older adults at high risk for dental decay.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this project is to describe the ensemble of afferent neural responses to intracochlear electrical stimulation as a function of stimulation variables. Several types of ensemble models will be investigated, including combinations of the various field and neural models from Projects II and III and the addition of stochastic properties to such combined models. Each ensemble model will predict both the spatial and temporal patterns of neural excitation. Conditions will include manipulations in (a) electrode coupling configuration [e.g., monopolar, radial bipolar, longitudinal bipolar], (b) stimulus waveform [e.g., sinusoidal, pulsatile], and (c) the number and geometry of electrode pairs in multichannel electrode arrays. Predictions from models spanning these conditions will test our understanding of the mechanisms underlying psychophysical and behavioral findings with respect to threshold, dynamic range, intensity discrimination, frequency discrimination, channel interactions, and channel discrimination. These predictions will be evaluated directly within this project using single-unit experiments. Such evaluations will, in turn, allow further refinement of the ensemble models and their constituent parts. Iterations of this model/test cycle will periodically refocus or efforts toward key gaps in existing knowledge.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to develop an Early Vocalization Analyzer (EVA), a computer program to automatically analyze recorded samples of infant vocalizations. This program will be the basis of a screening test, for use in hospitals and clinics, to evaluate which infants are at-risk for later communication or other developmental problems. The major purpose of this Phase-I project is to determine whether the landmark detection theory of Stevens, as implemented for the recognition of adult speech, can be adapted to the analysis of syllabic pre-speech vocalizations produced by typically developing infants. We will also merge this landmark- detection algorithm with a kernel version of EVA, developed by a team at Northeastern University, to produce a proof-of-concept prototype that will count the number of utterances, detect and count syllable-like vocalizations, and classify them according to fundamental frequency and duration. The goal of Phase II is to develop a commercial software program to automatically analyze infant utterances from audio recordings, as a non-invasive research and clinical-screening instrument. PROPOSED COMMERCIAL APPLICATIONS: The product would be useful for hospitals and infant-screening clinics assessing developmental risks, as well as hearing deficits. It could also be used by treatment centers to the efficacy of early intervention. Finally, it would provide a research tool for investigations of the earliest stages of infant speech and of age-specific control of the speech articulators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Articular cartilage is the remarkably durable tissue that covers the articulating bone surfaces in our joints and permits pain-free movement by greatly reducing friction between bones and distributing stress. Unfortunately, when cartilage is damaged due to injury or disease, it has a limited capacity to heal and can lead to premature arthritis. Arthritis is a major health issue that is predicted to increase as our population ages. As such, there is considerable interest in the development of techniques to repair or reconstruct damaged articular cartilage. The implications of a successful method for cartilage repair would be great in terms of the number of patients affected and their quality of life. Importantly, in this era of escalating health care costs, successful cartilage repair would decrease the long-term costs of health care related to joint replacement and multiple revisions thereof. Our long-term objective is to develop a tissue engineering approach for cartilage repair using periosteum. Periosteal grafts have the potential to contribute to each of the three requirements for tissue engineering (scaffold, cells, growth factors), and have been used successfully in biological resurfacing for the repair of damaged articular cartilage. The principle obstacle to overcome for the use of periosteal tissue grafts for tissue engineering is the declining chondrogenic potential of periosteum with increasing age of the patient. We hypothesize that subperiosteal injection of specific growth factors can be used as a pretreatlnent to \"activate\" periosteum for articular cartilage tissue engineering. In essence, the engineered tissue will be prepared in vivo for transplantation. In this proposal, we plan to examine the potential for subperiosteal injection of known chondrogenic growth factors to enhance (1) the number of chondrogenic precursor cells in periosteum in vivo (Aim 1), (2) the in vitro chondrogenic potential of periosteum (Aim 2) and (3) the potential of periosteum to repair articular cartilage defects (Aim 3). Beyond articular cartilage repair, these studies have the potential to impact the use of periosteum for other applications such as tissue engineering of bone. In addition, by eliminating the need for cell culture expansion facilities and expertise, the cost of this approach should be considerably less thereby making cartilage repair more globally accessible.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Liver fibrosis is the excessive accumulation of extracellular matrix as a wound healing response to chronic injuries from a broad range of liver diseases, irrespective of the underlying etiology. Advanced liver fibrosis may lead to cirrhosis, liver failure, and neoplasia. Extracellular matrix deposition is crucial to wound healing by promoting tissue formation and integrity, but commonly becomes uncontrolled in chronic injuries and leads to fibrosis. Ductular reaction is part of the injury response in the liver occurring at te interphase of the parenchymal and portal compartments that promotes bile duct regeneration. Although prominent in response to biliary and cholestatic damages, ductular reaction is observed in virtually all chronic human liver diseases. Recent studies have revealed that the matricellular protein CCN1 induces cholangiocyte proliferation and ductular reaction in response to cholestatic injuries through integrin ?v?5-mediated activation of NF?B, leading to Jag1 expression and Jag1/Notch signaling. Furthermore, CCN1 is able to restrict and resolve liver fibrosis induced by either hepatotoxin (carbon tetrachloride) or cholestasis (bile duct ligation) b inducing cellular senescence in myofibroblasts derived from hepatic stellate cells and portal fibroblasts. CCN1 also appears to regulate myofibroblast activation and resolution of inflammation. Based on these observations, we hypothesize that CCN1 serves multiple functions in different stages of hepatobiliary injury repair through binding distinct integrins in different cell types. We will scrutinize this hypothesis in three specific aims: (1) to conduct genetic and functional analyses of the heretofore unexplored CCN1-?v?5-NF?B axis in ductular reaction; (2) to examine how CCN1 induces the reactive cholangiocyte phenotype and differentiation of hepatic progenitor cells in the ductular reaction niche; and (3) to investigate how CCN1 regulates the initiation and termination of fibrogenesis in injury repair. These studies will elucidate the diverse roles of CCN1 as a critical regulator of injury repair in chronic liver diseases, and may lead to new therapeutic targets and treatment strategies for the prevention and treatment of liver fibrosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose to investigate the impact of the built environment characteristics on body size, physical activity and diet in residents of New York City (NYC). We will use data from three large human health studies. The first data set will be from 1000 Black and Hispanic children enrolled in prospective cohort study of Head Start families. From all of the children we will have complete demographic data, and mother's reports on the height and weight and physical activity of the child. From 500 of the children data will be available on skin fold thickness, body mass index (BMI) and 6 days of physical activity monitoring. Data will be available at baseline and from two annual follow-ups of the cohort. The second data set will be from 18,000 adult New Yorkers from whom we will have demographic data and objectively measured BMI. The third will be from 2,400 healthy Black and Caucasian. From these women we will have demographic data, questionniare data on diet and physical activity and objective measures of BMI, waist and hip circumference and body fat by bio-impedence. From all of these subjects we will have a home address which we will geocode into a CIS model of New York City. Multi-level modeling approaches will be used to determine whether neighborhoods characterisitcs (for example; land use, street design, availability and quality of parks and retail food stores, zoning, public transport, farmers markets) predict body size, physical activity and diet. Because of the very large immigrant population in NYC we will conduct qualitative research on how acculturation to the built environment of NYC impacts body size. Prior work suggests that acculturation to main stream American norms leads to weight increases in immigrants and their children. We do not see this effect in our data and propose that NYC is not \"main stream America\". We propose to conduct qualitative research on Hispanic mothers and children in the Head Start study and investigate how they relate to the built environment of NYC. Our goal is to develop a new measure of acculturation that includes adaption to the built environment of NYC and will be useful in studies of obesity. The quantitative research will also be used to develop new GIS based measures of aspects of the City that are important to the mothers. This research on the phsyical environment and adaption to it, will identify strategies for building and organizing communites that increase physical activity, improve diets and reduce obesity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Funds are requested to support six (6) full-time predoctoral trainees per year. The program seeks to develop researchers able to contribute to analysis of social phenomena and problems and an understanding of underlying processes. A foundation of \"core\" courses in the major subdisciplines of our field, regular research meetings and special topic seminars, and an intensive research apprenticeship are employed to enhance trainees' skills in conceptual analysis, theory development, and research. The overlapping interests and shared commitment of our faculty naturally lead our students to emphasize work that speaks to important social problems and their amelioration. Ongoing work involves the role of basic cognitive, motivational, and affective processes in intergroup conflict in the under-mining of minority students' self-confidence and aspirations, and various social and health problems such as interpersonal violence, debilitating shyness, and depression. Three themes-the role of self and self-reflective processes, the significance of emotion and emotional regulation, and the need for a broader cultural perspective-have played a particularly important role in guiding our efforts. Approximately 85 students apply for admission to the Stanford social program each year, of whom 3 or 4 especially promising candidates enter the program and plan a course of study suited to meet the Department's formal requirements and their own interests and professional aspirations. As they progress, students take increasing responsibility for originating and pursuing their research. Students are encouraged to work with multiple advisors, and to cross traditional disciplinary borders. Numerous interdisciplinary programs at Stanford offer additional research and educational opportunities. Most of our graduates take academic positions, but significant number elect positions in applied settings concerned with health care, education, or new communication technologies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A role for vitamin D in the pathogenesis and treatment of Mycobacterium leprae (mLEP) has been presumed for many years. Pafients suffering from the progressive, bacilli-abundant lepromatous form (L-lep) ofthe disease are more likely to be vitamin D-deficient and benefit clinically from ultraviolet B (sunlight) irradiafion or dietary supplementafion vitamin D. Such pafients are also at risk for developing dysregulated over- producfion of the acfive vitamin D metabolite 1,25-dihydroxyvitamin D (1,25D) from circulafing 25- hydroxyvitamin D (25D) by disease-activated macrophages. On the other hand, intracellular 1,25D synthesis and acfion at the level of the vitamin D receptor (VDR) is crucial for mounfing an anfimicrobial response to mLEP. Therefore, the mechanism(s) that govern vitamin D metabolism and acfion in leprosy is a key component to the disease. Our recent studies In vitro clearly demonstrate the differenfial expression of the funcfional elements ofthe vitamin D system, including CYP27B1-hydroxylase and the VDR in Type I and II interferon-driven, T-lep and L-lep granulomas, respecfively. Taken together, these clinical and laboratory findings lead us to theorize that mLEP Infection and the respective T-lep or L-lep downstream interferon- directed pathways, will differentially Impact the synthesis, metabolism and function of active vitamin D metabolites in the macrophage and other elicited, VDR-expressing inflammatory cells in the infectious microenvironment ofthe host with leprosy. To test this hypothesis, we will undertake three conceptually novel, mechanisfic aims. First, the vitamin D system components (CYP2R1, vitamin D hydroxylase; CYP27B1; CYP24A1, 24-hydroxylase; and VDR) will be quantitafively mapped in T-lep and L-lep granulomas at the single cell level. Second, the orchestrated effects of T-lep or L-lep immune response secretomes, and associated downstream interferon responses, on the metabolism and immunoacfion of vitamin D in human inflammatory cells will be characterized using innovafive molecular tools developed in Projects 1 and 3. Third, using recently-conceived RNA sequencing technologies, the funcfional consequences ofthe human host vitamin D deficient state, and its rescue In vitro and in vivo, on immune responses to and killing of mLEP will be probed. When analyzed in concert with the experimental results of the other CORT projects, it is anficipated that this work will set the stage for the pracfice of manipulating human vitamin D balance in promofion ofthe innate and adapfive immune response in leprosy specifically and in granuloma-forming diseases in general.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Inductive mechanisms play a major role in the development of the central nervous system as well as of other tissues in the organism. In particular, given the importance of the visual system in vertebrates, induction of eye development is one of the most extensively studied and serves at a paradigm for the inductive process. However, little is known about the induction of the neuroretina. The proposed research will utilize a combination of molecular, biochemical and histological approaches to identify factors that induce formation of the retina. We will undertake two parallel strategies. Firstly, we will investigate the effect of candidate retinal inducers identified in other systems. In addition, we will also perform an expression screen of Xenopus cDNA library for transcripts that induce formation of the retina. We believe that the combination of these two approaches will successfully identify genes that play a role in retinal development. These experiments will also further the understanding of general inductive mechanisms and their regulation. In addition, further comprehension of the inductive process in the eye will advance the prospects for regeneration and repair of ophtamological damage.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Tests whether bisphosphonate (daily oral alendronate) therapy maintains femoral and vertebral bone mass in women over 65 years old.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Cullin-RING ubiquitin E3 ligase (CRLs) super-family is responsible for much of the signal-dependent protein turnover in eukaryotes. CRL specificity derives from the identity of the substrate adaptor, which often interacts with the substrate in a modification-dependent manner. With more than 200 distinct CRL substrate adaptor proteins for the 7 human cullins, the CRL system controls many facets of biology that impinge on disease and aging. However, our understanding of CRL-substrate relationships is largely limited to a few well- studied adaptors, with many CRL adaptors remaining unstudied. We have taken a multi-pronged approach that uses Global Protein Stability (GPS) profiling, quantitative diGLY capture proteomics, and substrate capture by interaction proteomics using specific CRL substrate adaptors to identify more than 600 high priority candidate CRL substrates. Nevertheless, thus far only a portion of the human proteome has been sampled for substrates and that vast majority of candidate CRL substrates have yet to be paired with the appropriate CRL substrate adaptor protein (such as an F-box protein). To address these major limitations, we propose the following aims: 1) AIM 1 will provide a more comprehensive database of candidate CRL substrates in human cells through further development and screening of a v3.0 GPS system, thereby accessing components of the proteome that are not accessible through the current v2.0 system. Improvements will include new C-terminally tagged GPS libraries, bar-coding, as well as Next-gen sequencing of sorted libraries, thereby avoiding limitations of microarray hybridization methods currently employed. 2) AIM 2 will utilize focused high throughput screening and quantitative diGLY proteomics to specifically link high priority candidate substrates to specific cullins and will use both GPS and RNAi screening, as well as interaction proteomics, to link CRL1 substrates with specific F-box protein adaptors, followed by extensive validation. This analysis will provide the first large-scale analysis of the CRL1-F-box system in mammals. 3) AIM 3 will produce a web-based database for dissemination and analysis of CRL-substrate relationships by the community. In addition, selected high priority candidate substrates and the corresponding F-box adaptor proteins will be analyzed through biochemical and cell biological methods to place these proteins into regulatory and physiological pathways with an initial focus on a BACH1, a signal-dependent transcriptional repressor of NRF2-dependent genes and a novel substrate of SCFFBXL17 discovered via our proteomics platform. Together, this work will substantially improve our understanding of the CRL-substrate landscape and will set the stage for in-depth studies that further define signals that dynamically control the proteome via CRLs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract Marijuana is the most commonly used illicit drug in the United States, and prevalence rates have increased over the past several years after a period of relative stability. Currently there is no approved medication to treat cannabis use disorder, and relapse rates following behavioral interventions remain unacceptably high. There is an urgent need for pharmacological and psychosocial interventions, but optimal treatment approaches cannot be developed or implemented without understanding mechanisms that underlie and maintain marijuana use. Exposures to drug-related stimuli and stress have been found to increase craving (humans) and drug-maintained responding (mostly in laboratory animals). Given that continued use and relapse likely result from multiple factors, exposure to drug-related cues during periods of stress may increase the likelihood and intensity of drug seeking behavior. Few clinical studies have focused on the interaction of stress and cues on craving and drug-seeking; thus our understanding of the separate and combined effects of stress and cue exposure on drug seeking is currently limited. In a double-blind, placebo controlled cross-over study, 14 males and females with mild-to-moderate cannabis use disorder will receive the pharmacologic stressor, yohimbine (0, 20, and 40 mg), prior to marijuana and neutral cue exposure. Our primary outcome measure is marijuana-seeking behavior assessed using a progressive ratio marijuana choice task. Participants will be given 11 choice opportunities on the choice task to work for either puffs from a marijuana cigarette (6.0% THC; max 11 puffs) or money alternative (max $22). Specific aims are to determine whether: (1) presentation of polysensory marijuana cues increases marijuana craving and choice to self-administer marijuana; (2) placebo-controlled administration of a pharmacological stressor (yohimbine: 0-mg, 20-mg, and 40-mg, PO) alters marijuana craving and choice to self-administer marijuana; (3) marijuana-related cues and yohimbine have additive effects on marijuana craving and choice to self-administer marijuana; and, (4) craving elicited by marijuana-related cues, yohimbine, or both in combination mediates (within subject) marijuana puff-maintained responding. Our overarching hypothesis is that a sustained pharmacological stressor will potentiate cue-induced marijuana craving and seeking. The objective is to understand cue- and stress-induced cannabis abuse to effectively target these risk factors for intervention. To the best of our knowledge, this is the first study to investigate subjective, physiologic, and endocrine responses to a pharmacologic stressor combined with drug-related cues in chronic cannabis users, and to investigate their additive effects on marijuana craving and seeking. Results from this project will advance our understanding of mechanisms that underlie drug seeking behavior and relapse, as well as provide a test for compounds that reduce stress or craving and thus may prove beneficial for treatment of cannabis and other drug use disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this merit extension proposal, we are requesting funds to extend the critical findings of S1P release and signaling in the vasculature. The first major aim of the research is to understand how S1P metabolic enzymes are regulated in the vascular endothelial cells to impact local (autocrine) as well as systemic S1P signaling. Secondly, we will fully explore the basic mechanisms and physiological impact of the S1P signaling system as a sensor of biomechanical shear stress on the endothelial cells. First, we hypothesize that Sphk1 in the endothelial cell is primarily involved in local/autocrine signaling mode important for endothelial cell homeostatic and angiogenic reactions. We will derive an endothelial cell-specific inducible knockout of Sphk1 and examine angiogenesis, endothelial cell patterning, pathologic angiogenesis, vascular permeability and inflammation. To examine the effect of systemic S1P, which is derived primarily by red blood cells, we will examine endothelial cell functions in the red blood cell-specific Sphk1 knockout. Localized S1P signaling will be defined at the morphologic and ultrastructural level by immunolocalization studies in vivo as well as in primary cells. Second, we will explore the novel hypothesis that S1P metabolism and signaling constitutes a novel shear sensor/ signaling module in the endothelial cells. Mechanisms of how SIP is formed and released will be explored in detail in cultured vascular endothelial cells. In particular, we will focus on mechanisms that down regulate SIP lyase enzyme.\\Interestingly, laminar shear-induced downstream signaling mechanisms are critically dependent on S1P1 receptor signaling, suggesting that this receptor is a sensor of physiological shear stress. We will explore this mechanism in detail using cultured endothelial cells. We anticipate that these experiments will allow us to better understand the role of sphingolipid metabolism and signaling in vascular endothelial cells. Since SIP receptor modulators have entered the therapeutic era, this knowledge is likely to have important clinical utility.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Over the past eleven years, we have worked as a Program Project team consisting of four investigators with diverse expertise in the areas of epithelial cell biology, structural biology and membrane trafficking to study, in several closely interrelated projects, the cell and molecular biology and diseases of mammalian urothelium. Our research focuses on, as a central and unifying theme, a group of integral membrane proteins called uroplakins that represent major differentiation markers of mammalian urothelium. During the last granting period (2004-2009), our team has demonstrated that abrogation of uroplakins in transgenic mice resulted in compromised urothelial barrier function and overactive bladder; that defects in Rab27 and Vps33a lead to a depletion of fusiform vesicles and an accumulation of multivesicular bodies, respectively, thus establishing their involvement in uroplakin trafficking; that FimH can induce transmembrane conformational changes in the uroplakin receptor complex thus providing a novel mechanism for the bacterium-induced host cell changes; and that distinctive molecular alterations in genetically engineered mice underlie the two pathways of urothelial tumorigenesis. Our team has therefore functioned well in pursuing biologically important problems related to urothelial growth, differentiation and diseases; in having synergetic interactions and extensive collaborations; in effectively sharing resources; and in having made significant progress advancing the urothelial biology field. During the next five-year grant period, we will continue to work as a team to ask the following questions: What are the roles of uroplakins in the stabilization, enlargement and repair of the urothelial apical surface (Project 1)? What are the roles of molecular machineries including MAL and Rab27b in regulating uroplakin trafficking (Project 2)? What is the structure of uroplakins and how does the uroplakin complex anchor into an underlying cytoskeleton (Project 3)? And what are the roles of individual uroplakins and their subdomains in the uroplakin receptor complex in mediating the bacterial binding-induced signals in host umbrella cells (Project 4)? Results from this highly collaborative and synergetic team effort will lead to a better understanding of urothelial function, and have implications on a number of important urological problems including bladder outlet obstruction and urinary tract infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The epidermal growth factor receptor tyrosine kinase (EGFR) is one of the most commonly activated oncoproteins in lung adenocarcinoma, glioblastoma and other cancers. The identification of cancer-associated EGFR mutants, and their predictive power to select patients for treatment with EGFR inhibitors including erlotinib and gefitinib, has led to a major advance in cancer treatment. The collaborative efforts of the Eck and Meyerson laboratories for the past seven years have focused on understanding the structure-function relationship of cancer-derived EGFR mutants with the overarching goal of improved targeted therapies for cancers bearing these mutations. Refinement of our understanding of the mechanisms of action of mutant EGFR, and the spectrum of response of specific mutants to EGFR inhibitors, will permit more efficient development and application of EGFR-directed therapies. In this renewal, we will undertake the following Specific Aims: Aim 1) Characterize the response of novel EGFR mutants, identified by genomic studies, to low- molecular weight ATP-competitive enzymatic inhibitors and to antibody therapies, Aim 2) Perform structural and functional studies of the exon 20 insertion mutants of EGFR, which are resistant to gefitinib, erlotinib and cetuximab, to ascertain potential therapeutic approaches, and Aim 3) Analyze the substrate specificity of wild- type and cancer-derived EGFR mutants through peptide library array experiments, mass spectrometry-based phosphoproteomics, and crystal structure determination. Execution of these aims will help define the pathogenesis of EGFR-driven tumors and will thus provide a critically important mechanistic foundation for clinical trias targeting EGFR in a variety of cancer types using known agents. Additionally, our efforts will structural and mechanistic foundation for development of next-generation EGFR inhibitors that can be effective in patients whose tumors carry EGFR mutants that are resistant to currently available inhibitors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this stud yis to estimate the effectiveness of AR-623 inducing a complete remission in first relapse acute promyocytic leukemia (APL) patients who have not received oral all-trans-retinoic acid (ATRA) within one year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Myocardial infarction (MI) has emerged as a major health problem during the past two decades. In the infarcted myocardium, loss of contractile myocardium and blood vessels is induced by MI. Cardiac repair at the site of myocyte loss preserves structural integrity and is integral to the heart's recovery. Angiogenesis, the growth of new blood vessels, is critical for cardiac repair following infarction. Impaired angiogenesis in the infarcted heart can lead to rupture and immature/weakened scar tissue. Stimulation of angiogenesis is beneficial in the treatment of coronary artery disease and cardiac repair. Angiogenesis is a tightly regulated process and numerous inducers of angiogenesis have been identified. There is growing recognition and experimental evidence that reactive oxygen species (ROS) plays an important role in stimulating cardiac angiogenesis. However, its underlying molecular mechanisms remain to be elucidated. Oxidative stress is developed in the infarcted heart, particularly in the infarcted myocardium during the early stage of MI, and is temporally and anatomically coincident with the formation of new vessels. The overall objective of this proposal is to explore the underlying mechanisms by which ROS promotes angiogenesis in the infarcted heart. Angiogenesis occurs through degradation of basement membranes/extracellular matrix (ECM) followed by endothelial cell proliferation and migration, tube formation and vessel maturation. Using an experimental MI model created by coronary artery ligation in Sprague-Dawley (SD) rats with pharmacological intervention; inducible nitric oxide synthase (iNOS) gene knockout mice; and cultured endothelial cells, we intend to fulfill the following specific aims: Specific Aim 1: To explore whether ROS promote angiogenesis by regulating endothelial cell migration and adhesion in the infarcted myocardium. Our hypotheses are: 1) ROS regulate the balance of matrix metalloproteinase (MMPs) and tissue inhibitors of MMPs (TIMPs), enhancing degradation of basement membrane and other ECM components, thus allowing endothelial cells to detach and migrate into the infarcted myocardium; and 2) ROS stimulate the expression of angiogenic integrins, thereby activating the integrin signaling pathway and promoting endothelial cell adhesion and migration. Specific Aim 2: To determine whether ROS elevate angiogenesis through triggering of endothelial cell proliferation and tube formation in the infarcted heart. Our hypothesis is that ROS activate the expressions of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF), the major angiogenic mediators, and their signaling pathways in the infarcted myocardium, which, in turn, initiate endothelial cell proliferation and tube formation. Specific Aim 3: To examine whether ROS regulate angiogenesis via promoting vessel maturation and survival in the infarcted myocardium. Our hypothesis is that ROS activate the angiopoietin (Ang) signaling pathway in the infarcted myocardium, thus stimulating vessel maturation and stabilization. PUBLIC HEALTH RELEVANCE: The objective of this proposal is to investigate the cellular and molecular mechanisms regulating myocardial repair/remodeling following infarction (heart attack), particularly the role of reactive oxygen species on angiogenesis (new vessel formation) in the infarcted myocardium.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Complex persistent pain conditions (CPPCs) such as headache conditions, fibromyalgia, temporomandibular disorders, irritable bowel syndrome, and vulvar vestibulitis are high prevalent and shared or comorbid chronic pain conditions. There are two features of CPPCs that are fundamental to the aims and goals of this proposal: 1) the etiology of CPPCs is multifactorial and 2) the clinical manifestations of CPPCs are diverse. In this Program Project, we expect to identify a mosaic of risk factors for each of five CPPCs: fibromyalgia (FM), episodic migraine (EM), vulvar vestibulitis (VVS), irritable bowel syndrome (IBS), and temporomandibular joint disorders (TMD). Furthermore, we expect to characterize clusters of patients within each CPPC that vary significantly according to manifestations of their condition in addition to its painful characteristics (e.g., fatigue, dysfunction, sleep loss). Importantly, we expect some clusters of patients to be more alike across CPPCs than within any single CPPC, consistent with our view that there is some overlap in the manifestations of CPPCs. A unifying hypothesis integrating this Program is that multiple genetic factors, when coupled with environmental exposures (e.g. injury, infections, physical and psychological stress), increase the susceptibility to highly prevalent CPPCs by enhancing pain sensitivity and/or increasing psychological distress. To address the aims and goals of the subprojects and cores described in this application, a group of accomplished pain clinicians, pain researchers, psychophysiologists, molecular and cellular geneticists, biostatisticians and epidemiologists have been brought together to form this Program. Studies proposed in this Program Project application seek to identify the psychological and physiological risk factors, clusters, and associated genetic polymorphisms, that influence pain amplification and psychological profiles in enrollees who have established CPPDs. Additionally, the proposed studies seek to characterize the biological pathways through which these genetic variations causally influence CPPCs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Hypertension is a chronic condition associated with increased risk of mortality and morbidity from stroke, coronary heart disease, congestive heart failure, and chronic renal disease. Difficulty in controlling hypertension is due, in part, to the complexity of the pathophysiology of hypertension. In addition to the involvement of multiple pathways and feedback mechanisms, hypertension is often associated with other pathological conditions such as renal dysfunction, which is tightly linked to the renin-angiotensin system (RAS). Angiotensin (AngII), an important regulator of RAS, plays a critical role in regulation of blood pressure and volume homeostasis. These effects are mediated mainly via Ca2+ mobilization. Investigators reported the involvement of sphingosine kinase1 (SK1) in the mobilization of intracellular Ca2+. However, the exact mechanism is poorly understood. Here, we tested whether SK1 plays a role in AngII-stimulated Ca2+ release and regulating vascular tone. Our early observation using FLIPR to measure the intracellular Ca2+ levels indicated a characteristic pattern of AngII induced increase in intracellular Ca2+ level, an immediate peak followed by a sustained Ca2+ release. Additionally, selective inhibition of EGFR by AG1478 or the SK1 inhibitor, (DMS) inhibited the second phase. Also, AngII and EGF increased endogenous SK1 activity and enhanced production of S1P. Deletion of SK1 or DMS treatment abrogated EGF-induced intracellular Ca2+ elevation. Moreover, perfusion of the arteriole of the isolated rabbit glomerular structure with either AG1478 or DMS blocked the sustained elevation of intracellular Ca2+ and pressor response of Ang II. In vivo experiments performed on wild type and SK1-/- mice revealed that deletion of SK1 reverses the effect of AngII on GFR. Based on our data we propose to elucidate the role of EGFR transactivation and SK1/S1P in AngII-dependent intracellular Ca2+ mobilization, vascular tone, and renal hemodynamics. This study will enable us to gain important insight into the pathophysiology of hypertension and may provide novel therapeutic interventions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract The mucopolysaccharidoses (MPS) are a family of genetic, lysosomal storage disorders characterized by deficiencies in enzymes that degrade glycosaminoglycans (GAGs). Patients with MPS suffer from crippling skeletal abnormalities that are unresponsive to current treatments. MPS VII presents with a particularly severe skeletal phenotype, where patients exhibit progressive kyphoscoliotic deformity and spinal cord compression resulting in chronic pain and paralysis. MPS VII is caused by deficient beta-glucuronidase activity, leading to accumulation of multiple GAG types. The molecular mechanisms linking this GAG accumulation to cellular dysfunction and skeletal disease are poorly understood, impeding development of effective therapies. Our laboratory uses a clinically-relevant, naturally-occurring canine model of MPS VII that closely mimics the progression of skeletal disease that occurs in human patients. In previous work we demonstrated that MPS VII dogs have cartilaginous lesions in the vertebrae that compromise the stability of the intervertebral joint. These lesions are caused by failed conversion of cartilage to bone during postnatal growth. In preliminary studies, we have identified the precise developmental window when abnormal ossification first manifests in MPS VII dogs and that this can be traced to a failure of resident chondrocytes to progress through hypertrophic maturation. We have also shown that there is abnormal GAG accumulation in MPS VII epiphyseal cartilage from an early age, and, using whole transcriptome sequencing, that there is dysregulation of the Wnt/?-catenin signaling pathway at this crucial juncture in the disease progression. Wnt growth factors are critical regulators of chondrocyte differentiation, and GAGs are known to be important regulators of Wnt distribution and activity, suggesting a link between GAG accumulation and dysregulation of this pathway. The objectives of this proposal are to investigate mechanisms of failed bone formation in MPS VII and establish improved treatment paradigms using a clinically-relevant canine model. Our central hypothesis is that abnormal accumulation of GAGs in MPS VII epiphyseal cartilage disrupts the signaling pathways necessary to initiate and sustain chondrocyte hypertrophic differentiation. Further, we hypothesize that to effectively treat bone disease in MPS VII, it will be necessary to both normalize GAG turnover and activate requisite osteogenic signaling pathways in epiphyseal cartilage. In Aim 1 we will define temporal and spatial relationships between GAG accumulation and epiphyseal chondrocyte differentiation potential at key stages of bone disease progression in MPS VII dogs, from birth to skeletal maturity. In Aim 2 we will establish the critical role of Wnt/?-catenin signaling dysregulation in delayed epiphyseal bone formation in MPS VII dogs. In Aim 3 we will establish if therapeutic targeting of Wnt/?-catenin signaling, alone and in combination with enzyme replacement therapy, can enhance bone formation in MPS VII dogs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Based on successful pursuit of overall goals we will extend in vivo DTH and in vitro cell-mediated immunity tests of patients with breast disease, persons with other diseases and healthy individuals with T antigen. Further development of our quantitative assay to measure alterations in anti-T IgM, IgA and IgG serum levels to establish a sensitive, specific in vitro immunoassay to screen, diagnose and monitor breast cancer accurately and economically. Monitoring is an important aspect as we have now greater than 100 patients under continuous surveillance for up to 6 years. Clear histopathologic definition of the antibody, which clearly delineates benign from malignant breast lesions histochemically.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our previous work demonstrated that the KSHV-encoded K8.1A virion glycoprotein is essential for infection of B cells (MC116 human B cell line, human tonsillar B cells). This requirement stands in marked contrast to other non-B cell types, for which our results confirm published findings that K8.1A is dispensable for infection of other cell types (the 293 human embryonic kidney epithelium cell line, the African green monkey Vero cell line, human microvascular endothelial cells. In FY2016 we extended this work by demonstrating that another anti-K8.1A monoclonal antibody inhibits KSHV infection selectively for B cells. Thus we have identified a specific virus-encoded tropism determinant for the major cell type harboring KSHV in infected persons, and associated with two KSHV-associated lymphoproliferative diseases. We have extended studies on the effects of producer cell type on KSHV tropism, by expanding the number and types of cell lines constitutively infected with the KSHV.219 reporter virus (GFP linked to a constitutive cellular promoter; RFP linked to the KSHV lytic phase PAN promoter). In these inducible cells, the KSHV lytic phase can be induced by the lytic switch protein RTA (replication and transcription activator), linked to the tet-operator and thereby inducible with doxycycline. We now have inducible variants of the unusually KSHV-susceptible human B cell line MC116, the telomerase-immortalized human microvascular endothelium cell line TIME, the 293 cell line and the Vero cell line. We have characterized in detail the effects of specific activation with doxycycline versus nonspecific activation with BUdR, by measuring both RFP (fluorescence microscopy, flow cytometry) and the production of infectious KSHV (titration on fresh Vero cells). These inducible cell lines will be most valuable in our study of the impact of producer cell type and mode of activation on the target cell tropism of KSHV virions. We have also found a second mode for KSHV infection B cells, namely antibody-dependent entry. This mechanism is K8.1A independent, and in fact was first observed when the anti-K8.1A mAbs were found to greatly stimulate infection of Raji cells, which are otherwise refractory to KSHV. Preliminary data suggest that the antibody-dependent infection pathway is mediated by Fc receptors on the B cell surface. In collaboration with Dr. Robert Yarchoan, NCI, we are analyzing the sera from infected people with different KSHV-mediated syndromes. Preliminary data suggest that sera from MCD and PEL subjects have antibodies that greatly stimulate KSHV infection of Raji cells, whereas sera from KS subjects have lower levels of KSHV-stimulating antibodies. As yet, we do not know the relationships between the stimulating activities and the total levels of anti-KSHV antibodies or antibodies against any specific KSHV-encoded proteins.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development of movement depends on the maturation of the spinal cord motor circuits. Embryonic motor output is characterized first by spontaneous activity resulting in fetal jerk movements that later become more rhythmic and show limb alternations. In newborns motor development then undergoes a period of reflex and postural maturation leading into coordinated weigh-bearing locomotion. All these changes occur as a consequence of different maturation phases of the spinal synaptic networks and more specifically of local interneurons that ultimately control motor neuron firing patterns and motor output. Our long term objective is to understand the development of these local spinal circuits. Advances in this field were hampered in the past by the diversity and complexity of spinal interneurons. Fortunately, the recent development of mouse models expressing genetically-encoded reporters to identify interneuronal lineages now permits their study through development and allowed new understanding of the principles that govern spinal interneuron development. Our work has been focusing on one lineage of embryonic interneurons, named V1, that provide inhibitory control to ipsilateral motoneurons. Previously, we showed that this group diversifies into different types of adult interneurons, including Renshaw cells and Ia inhibitory interneurons (IaINs) that provide respectively recurrent inhibition to the same motoneurons and reciprocal inhibition between motoneurons with antagonistic actions around single joints. These two interneurons thus perform critical, but different roles in motor control. A basic question is then what mechanisms diversify single embryological groups of interneurons into distinct functional classes in adult. Previous analyses on Renshaw cells provided some important insights, including that this specification might occur early in embryological development and that individual cell types acquire different synaptology and functions through the different phases of spinal cord development. In this proposal we aim to investigate the development of the large group of V1 interneurons, including IaINs, that are interposed in proprioceptive reflexes. The underlying hypotheses are that IaINs are a group of V1-interneurons specified by their early birth (simultaneously with Renshaw cells) to acquire in the embryo transient motor axon inputs and functions that are quite distinct from those in adult. We also hypothesize that later they shed these embryonic inputs and develop connectivity that allows them to mediate reciprocal inhibition. Therefore we propose three aims to find out their birth dates (aim1), their major synaptic inputs and outputs in embryo (aim2) and the maturation of the reciprocal inhibitory circuit postnatally (aim 3). A corollary of this work is that congenital deficits that result in arrested spinal network development do not necessary imply a network with immature adult connectivity, but more likely networks of different connectivity appropriate to an earlier developmental point. Therefore it is important to understand the nature of this earlier connectivity to better appreciate the diversity of neurological motor deficits expressed in newborns. PUBLIC HEALTH RELEVENCE The work proposed will study spinal cord development focusing on the neuronal networks that mediate the maturation of coordinate movements and locomotion. The study will use powerful new mouse genetic methodologies to analyze the cellular elements that organize this network during development. The research is thus guided towards understanding normal and abnormal motor development in newborn and infants [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Personal health records (PHRs) integrated with electronic medical records (EMRs) are an essential tool to promote patient-centered care and ultimately improve health outcomes. Although adoption and use of integrated PHRs is increasing, it typically occurs within large integrated healthcare systems that have business models supporting their use. Whether such systems can have broad uptake in typical primary care practices remains unknown. In a previous AHRQ-funded R18, we created a PHR for prevention that is integrated with a clinician's EMR, which we term an \"interactive preventive healthcare record\" (IPHR). The IPHR also incorporates clinical decision support software, a reminder system, tailored educational materials, and decision aids into one actionable package for both patients and clinicians. In a study that tested IPHR feasibility and effect, the IPHR enhanced clinician-patient communication and promoted the delivery of recommended preventive services. Although these findings are encouraging, the study design permitted only small subsets of the study practices'patients to be included and patients were introduced to the IPHR with only a low- intensity invitational letter. The next logical step is to conduct a feasibility trial to assess whether practices can, under usual practice conditions, sustainably and more intensively integrate the IPHR into care. We propose to test the question in eight practices in the Virginia Ambulatory Care Outcomes Research Network (ACORN) that participated in the first study. All adult patients presenting for care will be considered eligible for IPHR use. Guided by organizational change theory, we will engage practices to create a shared vision on how to integrate the IPHR into care delivery using practice champions, learning collaboratives, and a patient-centered communications strategy. From the IPHR and EMR databases, we will apply the RE-AIM model to measure Reach, Effectiveness, Implementation, and Maintenance. We define Reach as the proportion and characteristics of patients seen for an office visit who establish an IPHR account and receive prevention recommendations, Effectiveness as change in documented preventive service delivery after patients use the IPHR, Implementation as clinician response to IPHR-identified patient needs, practice-level Maintenance as the practice's ability to sustain Reach 6 to 12 months after initial PHR fielding, and patient-level Maintenance as whether patients continue to use the IPHR after 6 months. We will compare the Reach in practices promoting the IPHR to patients using the integrated approach to the historical control rate of the less intensive invitational letter;the eight study sites will provide 80% power, using a one-sided 0.05-level test to detect a 10% incremental increase. We will conduct key informant interviews and record and analyze learning collaboratives to understand how practices integrated the IPHR and mediators and moderators to use. Findings from this study will assist in the design of a future practice-level randomized controlled trial, and inform practices, policymakers, and payers about how to integrate a PHR in typical primary care practices. Patient-centered health information technology systems, such as integrated personal health records, have great potential to improve the quality of care by providing centralized medical information, patient education and activation, enhanced patient and clinician communication, decision support, and reminder systems. However, these systems cannot improve health if they are not used by patients and clinicians. We seek to learn whether primary care practices can encourage patients to use such a system by integrating it into routine care and to determine how the intervention influences healthcare delivery", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": ". National Ovarian Cancer Prevention and Detection Study: The National Prospective Cohort Study of Risk-Reducing Surgery and Ovarian Screening among Women at Increased Genetic Risk of Ovarian Cancer (also known as \"GOG 0199\") is the cornerstone of CGB\"s intervention studies research portfolio. Developed and chaired by Dr. Greene, it is a non-randomized natural history study of risk-reducing salpingo-oophorectomy versus a novel ovarian cancer screening strategy (the ROCA algorithm) [NCI Protocol #02-C-0268]. This study opened in July 2003, and closed to new patient enrollment in November 2006, after having accrued 2605 high-risk women (1029 surgery arm; 1576 screening arm). Follow-up will be completed in November 2011. This complex, multi-institution international study exemplifies the type of project that would be difficult to mount through extramural funding mechanisms. Accomplishments to date include successfully completing accrual to this study, completing the research-based BRCA1/2 mutation testing for the 1,500 mutation-unknown study participants (including both full sequencing and testing for large deletions, rearrangements), nearly completing the central pathology review of 1037 RRSO samples, publishing a report detailing the rationale and design of the study, along with baseline characteristics of the women in each cohort, developing a model of the factors which influence the choice of RRSO versus screening, contributing 1576 screening subjects to a pooled analysis (with the Cancer Genetics Network) of determinants of baseline CA-125 levels (Skates et al., unpublished data), and negotiating a collaboration between GOG and CIMBA under which DNA and clinical data from 199\"s 1400 mutation carriers will be contributed to pooled multiple candidate gene association studies of breast cancer risk, and two genome-wide association studies (GWAS), one for each BRCA gene. Breast Imaging Pilot Study in Women from BRCA Mutation-Positive Families (NCI Protocol 01-C-0009): is the second major intervention studies programmatic component. It has reached its accrual goal of 200 women from BRCA1/2 mutation-positive families, 170 of whom are known mutation carriers. Prospective follow-up is ongoing, and scheduled to end in January 2010; 25% of subjects have completed the study. To date, breast cancer and ovarian cancer have developed in 14 and 1 participant, respectively. Analyses of baseline mammographic density and MRI volume in carriers versus non-carriers (the latter including 100 low-risk women from NCI/CCR\"s Breast Cancer Susceptibility Study) are well underway. These include a collaboration with Dr. Funmi Olopade and colleagues at the University of Chicago, in which our digitized mammographic images are being used to evaluate, and possibly validate, various novel imaging characteristics that may prove to be better predictors of breast cancer risk than standard mammographic density. We have published the first formal method for enumerating cells in breast duct lavage (BDL) fluid, described the results of BDL in the largest cohort of BRCA mutation carriers yet reported, and quantified the tolerability of the BDL procedure. DNA samples from mutation-positive DNA from BI participants has also been added to CGB\"s collection of samples being utilized in our genetic modifiers collaboration with CIMBA. Preliminary data document that fentogram quantities of estrogen metabolites are detectable in both NAF and BDL fluid, with levels in the former much higher than the latter. A Phase II Randomized Trial of Zoledronic Acid versus Standard Care to Prevent Post-RRSO Osteopenia/Osteoporosis (GOG-215): was designed as a companion study to GOG-199, targeting one of the major complications of RRSO-related premature menopause, i.e., significant bone loss, increased risk of osteoporosis and fracture. Participants are enrolled in GOG-215 just prior to or immediately following RRSO; all participants receive oral calcium/vitamin D supplementation, proscription of tobacco exposure, and exercise recommendations. Half are also randomly allocated to receive 3 zoledronic acid infusions, one every 6 months. Bone mineral density and serologic markers of bone metabolism are the primary study endpoints. GOG-199 participants who cross-over from screening to surgery remain eligible for this study. The entry criteria have been relaxed to include all premenopausal women undergoing RRSO. This study is currently open at 50 sites nationwide, and has enrolled 18 participants. Dr. Larissa Korde is Co-PI of the national protocol, and also the local PI of this study which has been opened at the NIH Clinical Center [NCI Protocol #06-C-0204]. A Pilot Study of a 3-month Intervention for Increasing Physical Activity in Sedentary Women at Risk of Breast Cancer: is also being run by Dr. Korde. She designed and initiated this project while still an oncology fellow in the Medical Oncology Branch/CCR, and has continued to run the study since joining CGB. It asks whether a physician recommendation for increasing physical activity along with the use of a pedometer will be effective in increasing physical activity in a sedentary population of breast cancer survivors and women at increased genetic risk of breast cancer [NCI Protocol #04-C-0276]. Changes in mammographic density, total body fat and selected hormones will be explored as surrogate biomarkers. This study has closed to accrual, and a preliminary report has been submitted for publication. Inherited Bone Marrow Failure Syndromes: Our study of persons from families with one of a variety of inherited bone marrow failure syndromes (e.g., Fanconi's anemia) continues steady accrual [NCI Protocol #02-C-0052]. Squamous cell carcinomas of the oral cavity, esophagus, labia and cervix are among the non-hematologic malignancies which occur excessively in patients with these syndromes. Participants in this study are undergoing intensive evaluation in search of both clinical and molecular abnormalities which might represent cancer precursors, particularly with regard to cancers of the head/neck and female genitals. The role of the human papilloma virus (HPV) in the etiology of these cancers is being explored. If suitable clinical or molecular endpoints can be identified, consideration will be given to the development of an intervention program which would target persons from these high-risk families. An HPV vaccine trial among persons with FA is presently under consideration.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Noninsulin dependent diabetes mellitus is associated with increased risk for coronary artery peripheral vascular and cerebrovascular disease (CVD). This could be attributed to metabolic derangements typical of diabetes, disorders of platelet function and coagulation mechanisms, and hypertension. However, the major part of excess risk of CVD in NIDDM cannot be attributed to diabetes per se, because the risk precedes NIDDM, and is uncorrelated with the degree of hyperglycemia. Because insulin resistance is a major cause of NIDDM and appears to precede overt diabetes, it is hypothesized that insulin resistance, per se, and/or the hyperinsulinemia which characterizes resistance without full blown diabetes may be the factor(s) which are responsible for CVD in individuals at risk for NIDDM. This application is for a Central Laboratory Facility, and is responsive to an NIH Request for Cooperative Agreement (RFA NIH-91-HL-03-P). It is submitted in cooperation with a Field Center Application also submitted from the University of Southern California (M. Saad, P.I.). In this application, we propose to make a series of measurements which will establish the degree of insulin resistance as well as cardiovascular risk factors in a group of 1600 subjects to be tested in specific field centers. In this application we propose the formation of two cores: an Assay Core, to carry out a series of biochemical measurements, and a Kinetic Analysis Core, to calculate insulin sensitivity (SI) and glucose effectiveness (SG) from the assay results. This laboratory conceived, validated and developed the minimal model approach which is proposed to be used to assess insulin sensitivity (i.e., resistance). Assays will be performed in 4 laboratories at USC: Bergman (insulin, glucose, C-peptide), Meiselman (blood and plasma viscosity, red blood cell aggregation, zeta sedimentation ratio, and hematological indices), Francis (Fibrinogen, factor VIIc, plasminogen activator inhibitor a, prothrombin factor F1.2, and fibrin D-dimer fragment). In addition, lipoprotein risk factors for CVD will be measured off-site by Dr. Barbara Howard and Dr. Linda Curtiss. Additionally, we propose a Kinetic Analysis Core, which will calculate SI and SG from our assay results. Minimal model calculations will additionally be compared to the same parameters calculated by different software. Results of these determinations will be transferred electronically to a Coordinating Center for analysis. It is believed that this combined Assay/Kinetic Analysis facility will provide an efficient, accurate and precise assessment of insulin resistance in different subject groups, and as such will yield an accurate assessment of the importance of insulin resistance and hyperinsulinemia in the development of cardiovascular disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is based on a fundamentally new approach to the isolation of bacterial virulence genes involved in pathogenesis, termed IVET (in vivo expression technology), that allows for the positive selection of bacterial genes that are specifically induced in host tissues. This methodology will be used to characterize the genetic and environmental factors that regulate the expression of in vivo induced genes (ivi) in the human pathogen, Salmonella typhimurium. Such studies will provide insights into the molecular mechanisms governing the regulation of pathogenicity, which is fundamental to the understanding of infectious disease. Genetic analysis of genes that answer the IVET selection is facilitated because the genes are transcriptionally fused to the reporter gene, lacZ, allowing the recovery of regulatory mutants that constitutively express ivi genes in vitro (normally repressing conditions). Our efforts will be focused on those regulatory factors that coordinately regulate ivi genes, thus defining the regulatory components that ensure a multifaceted bacterial response to a multifaceted host challenge. Trans-acting regulatory mutations will be used to characterize the positive and negative ivi regulatory elements that activate ivi transcription in vivo and repress ivi transcription in vitro. These mutants will be screened for those that confer virulence defects in an animal model, suggesting that they, or some of the products they regulate, play an important role in pathogenesis. Cis-acting mutations will be used to define the ivi promoter region with an emphasis on establishing a consensus site for coordinate ivi gene expression. Defining the nature of coordinate regulation provides a means to under stand the sophisticated signal transduction mechanisms that enable bacteria to grow under such different environmental conditions (e.g., laboratory media vs. host tissues). Additionally, the IVET selection will be extended to screening for ivi fusions whose survival is limited to a particular host tissue and thus may contain tissue-specific ivi genes, whose restricted expression will be examined by genetic analysis. These studies have practical applications for both vaccine and antimicrobial development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary The goal of this R21 EBRG project is to develop new optical methods to map high intensity focused ultrasound (HIFU) pressure ?elds. The methods would enable simple, fast, and low-cost in situ HIFU beam measurements, which are needed for quality assurance and safety in the clinic, and to accelerate the pace of research and development of new HIFU applications and technologies. An ideal beam mapping instrument would be low cost, capable of rapidly measuring relevant acoustic parameters of clinical HIFU systems in situ between treatments, and usable by nontechnical experts. Needle hydrophones are currently the gold standard tool for mapping HIFU pressure ?elds, but are poorly suited to the measurement task since they sample only one spatial location at a time, and most can only measure sub- therapeutic pressures. They can be translated in a water tank by a motion stage to produce spatially-resolved pressure maps, but this is a slow and cumbersome measurement that can take several hours to complete. The techniques proposed in this application could meet this clinical need and also provide a fast, ?exible, and spatially- resolved beam mapping instrument that would be invaluable for HIFU research since it would enable rapid val- idation and experimental designs that are currently infeasible, such as mapping pressure ?elds across multiple experimental variables. Standard optical schlieren imaging has a long history in 2D and 3D ultrasound pressure ?eld mapping but has conventionally been applied using sophisticated and expensive high-speed optical setups with limited ?eld-of-view, limited portability and high cost. The methods and devices proposed in this project are instead based on a newer schlieren technique called background oriented schlieren (BOS) imaging, and in their simplest form can be implemented using just a water tank, a tablet PC and a webcam. In essence, BOS trades the sophisticated optical setup for more sophisticated computation, which is a much cheaper commodity. The central innovation in this project is to use BOS imaging to quantitatively map continuous-wave HIFU pressure ?elds in 2D and 3D. The ?rst Aim is to develop portable hardware for BOS imaging and tomography, that can be used with a wide variety of HIFU systems in situ. The second Aim is to develop the mathematical theory underlying the BOS image formation process for HIFU beam mapping, which is different from conventional BOS imaging since the underlying refractive index ?eld is not static. The third Aim is to develop acquisition and reconstruction methods that produce quantitative spatially-resolved pressure ?eld maps, and validate those maps against simulations and optical hydrophone measurements of state-of-the-art HIFU systems. By developing and disseminating BOS hardware, theory, and methods for quantitative 2D and 3D HIFU beam mapping, this development project will lead to fast, simple and robust devices that can be widely adopted and even shipped with each clinical HIFU system for regular quality assurance and exposimetry measurements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Here we will identify nonhuman primate (NHP) neuron types and build an extensive toolbox of vectors for circuit- based neuroscience studies. NHPs share substantial neuroanatomical, genetic, and behavioral homology with humans, and therefore they are indispensable for investigating the neural circuit basis of cognition and devising therapies to treat neurological and psychiatric disorders. Despite the importance of NHPs, we lack the tools to analyze and manipulate complex circuits in the primate brain. This lack severely limits the use of genetically- coded neuroscience tools to examine circuit specific functions and hinders development of targeted gene therapeutics. Current methods for achieving transgenesis in small model species, such as the creation of genetically modified strains, are prohibitively expensive in NHP and not applicable to human disease. AAVs are the leading alternative to germline modification and selective breeding. AAVs infect adult neurons, confer stable transgene expression, and have proven safe in gene therapy clinical trials. AAVs do not have natural cell-types specific properties, but when altered or combined with cell type specific regulatory sequences (enhancers/promoters) they have been able to achieve cell type-specific transgenesis. This has made possible, for example, our previous optogenetic investigation of midbrain dopamine neurons for learning and decision making. However, before AAV-mediated gene delivery can be generalized to circuits across the brain and for multiple behavioral functions, we must create currently lacking vectors and promoters that permit efficient and specific gene delivery to all required cell types. Here, we will combine single-cell RNA-Seq (scRNA-Seq) with high-throughput screening of engineered adeno-associated viruses (AAVs) to create a complete toolbox of viral vectors and promoters enabling minimally invasive monitoring and manipulation of neurons in NHP brain. We have devised a transdisciplinary approach to classify individual neurons according to their gene expression profile and simultaneously screen for adeno-associated virus (AAV) vectors (capsids and regulatory sequences) capable of specific and efficient transgene delivery to classified neurons. First, we will synthesize massive libraries of mutated AAV vectors and synthetic promoters, in which each variant is paired with a unique DNA barcode. We will then scRNA-Seq to capture the transcriptome for each cell and quantify the AAV and promoter- specific barcodes in every cell?s expression profile. Preliminary experiments in Rhesus monkeys have fully validated and demonstrated the promise of this innovative approach. The outcomes of our Specific Aims will include (1) an inventory of cell types in the retina, prefrontal cortex, primary motor cortex, and striatum, (2) cell type-specific AAVs and promoters targeting all defined cell types, (3) AAVs with broad tropisms, (4) a publicly available dataset of transcription profiles for millions of NHP brain cells, (5) an updated and comprehensive Rhesus macaque reference genome, and (6) anatomical, physiological, and functional validation of cell type- specific circuits tools and their function in the NHP brain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Temporal lobe epilepsy (TLE), the most common type of epilepsy, is characterized by destruction of specific hippocampal neurons. Recent evidence indicates that this neuronal damage is the result of excessive stimulation by excitotoxic amino acids (e.g., glutamic acid). Two stages of excitotoxic neuronal damage have been identified: and early phase, occurring in minutes, associated with osmotic imbalance, cellular swelling, and lysis; and a delayed phase, characterized by elevated intracellular calcium. One major homeostatic element which counteracts events underlying both phases of excitotoxicity is the sodium pump, Na,KATPase. Three different mammalian genes encode this family of structurally similar heterodimeric isoenzymes which are responsible for ionic and osmotic equilibrium. Although decreased Na,K-ATPase activity has been recognized in TLE and animal models of epilepsy, few studies have examined the distribution, regulation, or role of the sodium pump isoforms in epileptic hippocampal neuronal death. Our preliminary studies indicate a heterogeneous distribution of each isoform in rat hippocampal neurons. In TLE, we have discovered an inverse relationship between the level of one Na, K-ATPase isoform, and the susceptibility to neuronal death. This proposal outlines studies to explore the hypothesis that the activity of the sodium pump in individual hippocampal neurons is the major determinant of their susceptibility to excitotoxic damage. Using in-situ mRNA hybridization, monoclonal immunohistochemistry, in-vitro autoradiography, and a cytochemical assay, the amount of each isoform in the principal neurons of surgically obtained human hippocampus will be correlated with their susceptibility to epilepsy associated damage. Similar studies will be carried out in rats with seizures induced by graded electrical stimulation. Because the hippocampal somatostatin interneuron appears to be an early excitotoxic target, and thus of possible significance in the pathogenesis of TLE, we will particularly focus on the characteristics of its sodium pump. Finally, using hippocampal neurons in-vitro, we will regulate the activity of the NaKATPase isoforms and determine their role in glutamate-induced cell death. These studies will shed light on the complexity of action of the sodium pump, which is fundamentally critical to normal neuronal function. They will also provide new information about the larger issue of excitotoxic neuronal death, and possibly its prevention, which has now been implicated in the pathophysiological of many human neurodegenerative states, including stroke, Huntington's disease, hypoglycemic brain damage, and Parkinson's disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: The goal of this research is to gain an integrated view of the mechanisms that underlie development an, maturation of the sensory endorgans of the inner ear. Experiments will focus on the auditory and vestibular hair cells of the amphibian, Xenopus laevis. The proposed research will exploit the advantages that Xenopus offers as a classical system for developmental studies. The potassium channels of the inner ear will be a central subject of these investigations, and will be used to test hypotheses about hair cell development and differentiation. A major objective of this research is to establish the pattern of potassium channel gene expression in the developing inner ear. Electrophysiological methods (patch clamp) will be used to study ion channel function in dissociated hair cells. Molecular techniques will be used to clone ear potassium channels (RT-PCR, RACE, recombinant DNA) and will be combined with anatomical methods (in situ hybridization) to examine potassium channel mRNA expression in the ear. The specific aims of this research are to test the following hypotheses: (1) An outward potassium channel homologous to drkl is expressed in the Xenopus inner ear, (2) Xenopus saccular hair cell bundle morphology can be correlated with saccular hair cell electrical membrane properties, (3) The types of ion channels expressed in Xenopus saccular hair cells change as hair cells develop, (4) The properties of ion charmers differ between hair cells of the Xenopus sacculus, amphibian papilla, and basilar papilla. Experimental results are expected to advance fundamental understanding about the genetic basis of hair cell diversity, and about the genetic control of ear development. The long-term goal of this research is to lay the foundation for future studies that will determine how genes expressed in the ear are regulated during development and regeneration. Results of these investigations will provide essential knowledge that can be used to develop effective treatments for hearing loss such as that caused by trauma or genetic disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this Project are two-fold. First, in parallel with Project by Griffin, we will continue to study the effect of survival time and of gene polymorphisms on the manifestation of specific pathological features in traumatic brain injury which mimic the pathology of Alzheimer's disease (AD) in the belief that this will offer valuable insights into the early molecular mechanisms underlying the development of Alzheimer-type pathology. We hypothesis that the genotype of an individual modulates the extent of Abeta deposition and expression of glia-derived cytokines. Seen in the brain post-injury. Having already established that the apolipoprotein E (ApoE) genotype is important in modulating post-traumatic pathology, we will study the molecular basis of this effect in a novel in vivo paradigm of Abeta-induced neurotoxicity. Tissue from a large cohort of head-injury patients with different post-injury survival times and from non-injured control patients will be genotyped with respect to the polymorphisms in genes in the interleukin-1 gene cluster on chromosome 2. Aim 1 will determine quantitatively the changes in expression of the glia-derived inflammatory cytokines IL-1, IL-6, and S100beta throughout the brain after fatal head injury. Aim 2 will assess whether alterations in cytokine immunoreactivity following head injury are spatially and temporally correlated with changes in proteins that are important in the pathogenesis of AD. Aim 3 will determine whether inter-individual variation in the extent of the inflammatory reaction to head injury is associated withy specific polymorphisms in the genes encoding ApoE and IL-1. Aim 4 will investigate the mechanisms by which ApoE4 increases susceptibility to AD using an in vivo model of Abeta-induced neurotoxicity. Results should allow us to determine if genotype affects post-trauma pathology and will provide information on how ApoE4 produces its effects at the molecular level. The effect of genetic makeup on the extent of the inflammatory activity in the brain will be an important factor in predicting long-term sequelae of head injury (i.e., AD) and in targeting any potential therapies for the disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fibroblasts are a major cellular component of the tumor microenvironment and influence cancer cell behavior directly and indirectly through secretion of soluble factors, including growth regulators, angiogenic factors and chemokines. While genetic alterations in breast fibroblasts may exert pro-tumorigenic effects, little is known of the cellular and molecular signals that regulate fibroblast functions in the breast cancer microenvironment. De-regulation of TGF-B signaling significantly contributes to metastatic breast cancer, and may also enhance expression of chemokines in breast fibroblasts, to promote cancer progression. Based on previous studies and preliminary studies, we hypothesize that inflammatory chemokine expression in fibroblasts including CCL2 and CXCL1 mediate epithelial cell and immune cell motility and invasiveness to promote breast cancer progression. The objective of this proposal is determine the functions of chemokines, which are normally suppressed by TGF-B signaling, in fibroblast interactions with tumor cells during breast cancer progression. [unreadable] [unreadable] The following Specific Aims are proposed to address this hypothesis: Specific Aim 1: To determine the mechanisms of CCL2 and CXCL1 inflammatory chemokine expression in mammary fibroblasts as it relates to TGF-B signaling using cell biology, molecular and biochemical approaches. Specific Aim 2: To determine the functional contribution of CCL2 and CXCL1 chemokines expressed by mammary fibroblasts in breast cancer growth, invasion and metastasis, using transplantable and transgenic mouse models of breast cancer, cell biological and biochemical approaches. Specific Aim 3 : To determine the contribution of CCL2 and CXCL1 chemokines expressed by mammary fibroblasts in immune cell trafficking, homing and recruitment in breast cancer progression using transplantable mouse models of cancer, cell biological and biochemical approaches. [unreadable] [unreadable] Specific Aim 1 would be addressed during the mentored phase, while Aims 2 and 3 would be addressed during the mentored and independence phases. This research will address the functions and mechanisms through which inflammatory chemokine expression in fibroblasts contributes to breast cancer. Through these studies, we seek to further understand the functions of stromal cells in the host microenvironment, and how the host microenvironment contributes to tumor progression at the molecular, cellular and in vivo levels. By further understanding the role of the tumor microenvironment in cancer progression, it will be possible to identify new molecular targets for therapy and to develop improved methods for diagnosing and treating metastatic breast cancer. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (provided by investigator): Invasive brain intervention often results in complications and long recovery times. In addition, the delivery of therapeutic agents via the blood supply is often impossible because the Blood-Brain barrier (BBB) protects the brain tissue from foreign molecules. Laboratory experiments have shown that focused ultrasound (FUS) beams can be used for noninvasive interventions. However, the utilization of FUS in the brain has been seriously limited by the difficulty of delivering FUS through the skull bone. The hypothesis of this grant has been that transcranial therapeutic ultrasound exposures can be delivered noninvasively through an intact skull. During our current grant we continued our initial research and developed methods to allow FUS propagation through the skull at large entrance angles thus making whole brain sonications feasible. We have collaborated with industry to develop a prototype device currently in clinical testing. We have further studied the impact of ultrasound exposures on brain tissue and demonstrated chemotherapy delivery across the BBB and an order of magnitude reduction in the required acoustic power for focal tissue destruction when intravascular microbubbles are used. We have also developed computer simulation programs and new array technology and treatment methods that will improve the delivery of FUS energy. Our study plan is to extend our current research and further explore the feasibility of using intravascular microbubbles to enhance the trans-skull sonications. Successful utilization of the bubbles will eliminate skull heating problems and will enable new therapies. Our goals are: First, to modify our prototype phased array ultrasound system to allow acoustic signal feedback, new treatment planning, and advanced sonication methods to be tested for the localization and control of bubble-enhanced treatments. Second, to perform in vivo dual photon microscopy to increase our understanding of ultrasound induced BBB disruption and tissue destruction at the microscopic resolution. Third, to explore sonication methods for localization of bubble-enhanced therapeutic effects. Fourth, to develop and test treatment planning methods for bubble-enhanced therapy that incorporate image-derived information of the blood vessels networks. Finally, to further test the effectiveness of the ultrasound-induced disruption of the BBB for the delivery of chemotherapeutic agents for the treatment of malignant brain tumors. Our vision is that successful trans-skull delivery of FUS could have a major impact on the treatment of many brain disorders. Therefore this research may have a major impact on patient care. PUBLIC HEALTH RELEVANCE: Invasive brain intervention often result in complications and long recovery times. In addition, the delivery of therapeutic agents via the blood supply is often impossible because the Blood-Brain barrier (BBB) protects the brain tissue from foreign molecules. During our current grant we continued our initial research and developed methods to allow focused ultrasound propagation through the skull such that brain sonications are feasible. We have collaborated with industry to develop a prototype device currently in clinical testing. We plan to further develop the ultrasound methods for brain surgery and targeted chemotherapy delivery. Our vision is that successful trans- skull sonications could have a major impact on the treatment of many brain disorders. Therefore this research may have a major impact on patient care. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Research is to be conducted on the visual mechanisms by which the human observer resolves fine details and accurately localizes objects in two and three dimensions. The retinal components of these abilities will be analyzed, using optical and psychophysical techniques, by distinguishing between the limits set by the discrete nature of the mosaic of retinal receptors, by the sensitivity to intensity differences that can be detected in small adjacent retinal locations, and by the interaction between retinal neural entities. Identification of Snellen visual acuity letters and stereoscopic vision require, in addition, the elaboration and relative localization of individual features, and these are functions of the cortex. The interest is in the interaction of separate features and the development of the visual attributes of two-dimensional position and three-dimensional depth. The long-term goal is twofold: (1) to specify rigorously the ultimate performance characteristics of the human visual system and to delimit the range of possible neural circuits that may serve as their substrate, and (2) to gain insight into the optical and neural processes subserving visual acuity, hyperacuity and stereoscopic localization that will aid in the differential diagnosis of patients with reduced acuity and the development of tests of etiological and prognostic value.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY/ABSTRACT Juvenile myelomonocytic leukemia (JMML) is the most common myeloproliferative neoplasm (MPN) in childhood, and tends to occur in very young children less than 4 years of age. JMML is traditionally characterized as being Ras-driven due to mutations in NF1, CBL, KRAS, NRAS, or PTPN11. Traditional cytotoxic chemotherapeutic agents are ineffective in JMML, and the only curative modality is allogeneic hematopoietic stem cell transplantation. Unlike other MPNs, JMML rarely progresses to blast crisis; rather, mortality is due to extramedullary tumor cell expansion leading to organ failure, respiratory failure, bleeding, or infection. Notably, following allogeneic stem cell transplant, 50% of children succumb to leukemia relapse. This relapse rate in JMML is substantially higher than that of individuals who receive allogeneic stem cell transplant for chronic myelogenous leukemia (CML) in chronic phase (approximately 7% leukemia relapse), implicating a strong hematopoietic stem cell (HSC)-independent component of JMML development and progression. We envision two distinct mechanisms that potentially account for a HSC-independent means of JMML relapse after allogeneic HSC transplant. First, the JMML-initiating malignant cells may emerge during embryonic development prior to and independently from HSCs, and persist postnatally as self-replenishing malignant tissue macrophages. Alternatively, regardless of the origin of the JMML cells, the hyperinflammatory nature of JMML may damage the bone marrow microenvironment, prohibiting the expansion of normal donor cells following transplant, permitting residual leukemia cells to outcompete the normal graft, and leading to leukemia relapse. To address these possibilities, we will use the tamoxifen-inducible Cre recombinase system, which will permit yolk sac-restricted expression of the common JMML mutation, Shp2D61Y, to determine if yolk sac-restricted oncogene expression is sufficient for the post-natal development of MPN. Further, we will examine if inhibition of the pro-inflammatory protein, PI3K p110?, improves homing, engraftment, expansion, and myeloid differentiation of WT donor cells into diseased, Shp2D61Y-expressing recipients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A novel anti-HIV activity of CCR6 via APOBEC3G: relevance to CNS infection Abstract We have shown that when defensin hBD2 and chemokine MIP-31 (CCL20) bind to their cellular receptor, CCR6, they induce an intracellular antiviral activity against HIV that is absent in cells that do not express CCR6. While direct inactivation of enveloped viruses including HIV has been implicated as one mechanism of defensins, to our knowledge, HIV inhibition mediated through CCR6 is novel. The mechanism occurs post entry but at or before reverse transcription, and appears to be due to increased expression of the intracellular antiviral protein APOBEC3G. Inhibition is abrogated by pertussis toxin, suggesting the involvement of G1i-dependent signal transduction. CCR6 is expressed on cells that are highly relevant to HIV infection; they include memory T cells, dendritic cells, activated macrophages, microglia and Th17 cells. Defensins that bind to CCR6 are primarily expressed by epithelial cells and astrocytes, while MIP-31 is produced by mucosal epithelial cells, activated peripheral blood mononuclear cells and astrocytes. Since both CCR6 and its ligands are expressed in the central nervous system in cells highly relevant to HIV infection, an HIV suppressive mechanism that targets CCR6 may play a critical role in controlling HIV infection and CNS symptoms. Further, elucidation of the intracellular pathways and the step(s) in the HIV life-cycle limited by CCR6 activation could provide insights into virus-host interactions early in infection and identify new targets for antiviral therapy. We propose to identify APOBEC3G as the effector molecule of the antiviral activity mediated by CCR6 in primary cells; to demonstrate that CCR6- induced signal transduction increases APOBEC3G expression and identify what is the critical specific pathway; to demonstrate the efficacy of CCR6-mediated HIV inhibition on a broad spectrum of primary and non-B viral isolates; and to measure the expression of CCR6, CCR6 ligands, and APOBEC3G in clinical specimens. PUBLIC HEALTH RELEVANCE: The studies proposed here will investigate the mechanism of inhibition of HIV by a cellular receptor called CCR6. These studies are highly relevant to prevention and treatment of HIV infection because they will contribute knowledge that can be used to develop novel anti-HIV drugs that will target CCR6.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The promise of cancer immunotherapy is based upon the exquisite specificity of the immune system, through which a potent machinery can eliminate targeted cells. However, despite some notable examples of success, progress in developing this form of cancer therapy has fallen short of expectations. Major insights explaining the limitations of T cell-based cancer immunotherapies have come from the discovery of inhibitory co-receptors or pathways termed immune checkpoints, which restrain T cell functions in normal physiologic settings as well as in the context of neoplastic disease. Recent evidence suggests that tumors may usurp immunological checkpoint mechanisms to create a barrier against antitumor immune responses - including endogenous responses and those induced by immunotherapies such as cancer vaccines. Animal cancer models demonstrate that blocking the interaction of inhibitory molecules on tumor cells with their co-receptors on tumor-specific T cells can release the brakes on antitumor immunity and cause tumor regression. Thus, checkpoint inhibition, applied alone or in combination with vaccines, represents an important new therapeutic approach for enhancing antitumor immunity. One of the most interesting inhibitory co-receptors is PD-1, that is induced on activated T cells and down-modulates critical functions in both CD4+ (helper) and CD8+ (killer) subsets. The major ligand for PD-1 is B7-H1 (PDL1), a B7 family member normally expressed by several leukocyte subsets upon activation, and aberrantly expressed in many human cancers. These findings, highlighting multiple mechanisms by which PD-1/B7-H1 interactions may inhibit antitumor immunity, have provided a rationale for clinical trials in cancer patients using fully human antibodies blocking PD-1 or B7-H1. Notably, objective tumor regressions were observed in the first phase I trial of PD-1 blockade in patients with advanced treatment-refractory metastatic solid tumors. It is now critically important to better understand the regulation and function of PD-1 and B7-H1, and to discern the effects of PD-1/B7-H1 blockade on antitumor immunity. In current proposal, we will test hypothesis that modulation of B7-H1/PD-1 inhibitory pathway could vastly enhance efficacy of cancer immunotherapy by improving tumor microenvironment and protecting ongoing T cell activity. The current proposal integrates basic and clinical science, and will use animal models and human in vitro systems to achieve the following aims: 1) To define mechanisms regulating B7-H1 expression by tumor cells and other cell types in the tumor microenvironment; 2) To characterize factors influencing PD-1 expression by T cells, particularly in the context of vaccine-induced stimulation; and 3) To characterize immunological mechanisms underlying the clinical effects of B7-H1/PD-1 blockade in cancer therapy. Taken together, results from these studies will enable the rational clinical development of PD-1/B7-H1 blockade, alone or in combinatorial regimens, in cancer therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research outlines a proprietary, potentially patentable fee- for-service method to predict viral leakage, such as HIV, through condoms - - in manufacturers' production lots during or immediately post production. The study would statistically expand results indicating that among 31 brands, some condoms from brands with lowest ASTM/ISO scores leaded HIV; high scorers did not. 100 condoms from each of four lowest scorers will be challenged with high titre bacteriophage phiX-174, (diam.<HCV/HBV). Leakage will be quantitatively assayed with a detection limit of a single phiX-174 particle leaked through the condom. Test condoms will be pre-stressed on a Coital Simulation Instrument to subject them to reproducible conditions akin to those during sexual intercourse. If leakage is adequately confirmed, future work will similarly more fully assess leakage in high and intermediate scorers, correlating all results with individual ASTM/ISO tests values. Goal: a rapid method predicting production lots' leakage potential, through an ASTM/ISO Index of critical score values. The method has potential as a commercially useful step toward lessening the number virally leaky condoms currently on the market in the AIDS/herpes/chlamydia epidemics. Like FDA water leak monitoring, the method would test production lots, not individual condoms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Critical to cryopreservation is that lethal intracellular freezing (IIF) not occur. Its occurrence depends on two major factors. One is the cooling rate. It has to be low enough so that the cells lose nearly all their water osmotically before cooling to the temperature at which IIF becomes possible. Second is the temperature at which IIF occurs. The higher that temperature, the greater the difficulty in avoiding IIF by slow cooling. Our current grant and this renewal proposal deal primarily with the latter. IIF in mouse and Xenopus oocytes and in Arabidopsis protoplasts was found to require the presence of extracellular ice in close contact with the cell membrane. One strong piece of evidence in the mouse oocyte is that IIF occurs at temperatures where about 95 percent of the external medium has frozen. That temperature varies from -14[unreadable]C to -40[unreadable]C depending on the concentration of cryoprotective compounds in the medium. Another factor affecting the IIF temperature may be cell size, for IIF occurs at much higher temperatures in 1 mm Xenopus eggs than it does in <0.1 mm mouse oocytes. The two tools used above were a cryostage that permits us to observe cells during cooling and warming and manifests IIF as \"flashing\", and a differential scanning calorimeter (DSC) that detects IIF as an outburst of heat. In this renewal application we propose to introduce two new instruments. One, in collaboration with the Oak Ridge National Laboratory, is a Scanning and Transmission Electron Microscope (STEM) equipped with a newly designed sample chamber that will permit high resolution images of hydrated cells in aqueous solutions. The other instrument is a directional freezing stage. It permits a cooling rate to be resolved into the thermal gradient (G) and the crystal growth velocity (V). Differences in G and V affect the ice crystal morphology which in turn may affect the temperature of IIF. We will add three new cell types to the study: Yeast and two types of hamster tissue culture cells. These cells are 10-times smaller than mouse oocytes thus permitting us to further explore the role of cell size. Second, cells have to be that small to fit in the STEM liquid sample chamber. Third, IIF has never been observed directly in these cells and we intend to use DSC for this purpose. Other major aims in this renewal proposal are (1) to determine whether the high correlation between observed IIF temperature in mouse oocytes and the frozen fraction holds for other cell types. (2) To determine whether there is a relation between pores in the plasma membrane and the IIF temperature. For this, we will study the freezing of mouse morulae, the 8 to 12 cells of which possess gap junctions and aquaporin pores. Second, we will introduce pores in mouse oocytes and plant protoplasts by exposing them to a toxin from Staphylococcus bacteria. The final goal will be to use the mechanistic data on IIF to devise better methods of avoiding it and thus improve success in the cryopreservation of cells that currently can not be well cryopreserved.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The training and research proposed in this application will be sponsored by Dr. Adams and will take place in both the Arteriosclerosis Research Center and the Comparative Medicine Clinical Research Center. Dr. Wagner is seeking SERCA support to further her development as an independent investigator with special emphasis on cardiovascular health of aging females. The studies will utilize a well characterized primate model, the female cynomolgus monkey, which like women in most Western societies, develops less severe atherosclerotic coronary heart disease (CHD) than its male counterpart. Pathogenetic mechanisms of atherogenesis are difficult to study in women, since atherosclerosis develops slowly over a period of many years. Also, while CHD is the leading cause of morbidity and mortality in American women, clinical events occur relatively infrequently. For these reasons we propose to utilize a primate model to study the effect of sex hormone deficiency, sex hormone replacement and oral contraceptive treatment on the pathogenesis of atherosclerosis. Studies of both human and nonhuman primates have found that only a portion of the beneficial effects of estrogens can be attributed to changes in plasma lipoprotein risk variables. This suggests that estrogens may retard atherogenesis by interacting by either directly at the arterial wall or by modifying plasma components in addition to lipoproteins. The studies proposed will explore molecular and cellular mechanisms by which estrogens and progestins may be acting. The effects of sex steroids on the following parameters will be investigated: 1) functional parameters of early atherogenesis eg., endothelial dysfunction as determined by arterial LDL uptake and degradation, monocyte adhesion to the endothelium, and intimal cell proliferation, 2) whole body and hepatic LSL metabolism, 3) plasma HDL composition and its function in reverse cholesterol transport, 4) expression of cytokines and growth factors by arterial cells, and 5) atherosclerosis-related impairment of vascular responses. If, as suspected, beneficial effects are found, this would provide a rational basis for the prophylactic or therapeutic use of these steroids in CHD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our studies focused on the interrelationships of organelles (endoplasmic reticulum, Golgi apparatus, GERL, lysosomes, peroxisomes, microperoxisomes, mitochondria, cytosolic lipid, coated vesicles, plasma membrane, and microfilaments) to gain a better understanding of organelle functions in normal, pathologic, and malignant cells. To search for differences between normal and malignant cells we used microscopy (light and electron) in combination with enzyme cytochemistry, immunocytochemistry, tracers, freeze cleavage, biochemistry, and molecular biology (e.g., in situ rDNA hybridization). Our key accomplishments have included developing immunocytochemical procedures for localizing preneoplastic antigens, epoxide hydrase, and gamma-glutamyltransferase (gamma-GT) in 2-acetylaminofluorene (AAF)-induced hepatomas; describing organelle translocations in hepatocytes during regeneration; studying hormone effects of functioning islet cells on hepatic ultrastructure; studying the uptake and transport of asialoglycoproteins in hepatocytes; localizing apo-protein B and A-I using immunocytochemistry in rat hepatocytes and absorbtive cells of the intestine; isolating fractions enriched in the Golgi apparatus and GERL from rat liver homogenates and characterizing these fractions biochemically; performing an extensive biochemical, ultrastructural, and enzymatic study of 3T3-L1 cells, including the isolation and characterization of microperoxisomes; describing an extensive lysosomal compartment in Kupffer cells and hepatocytes; developing immunocytochemical procedures for localizing lysosomal and glycoprotein processing enzymes; and localizing antimetabolites in neurons. (E)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mycoplasma pneumoniae is the leading cause of pneumonia in older children and young adults and accounts for 20% of all community-acquired pneumonia. This cell wall-less prokaryote moves by gliding motility, which we contend facilitates colonization of the conducting airways of the respiratory tract. Gliding motility is poorly understood, and no homologs of known motility genes, gliding or otherwise, are found in M. pneumoniae. In the current project period we identified a diverse set of gliding- associated genes, demonstrated conclusively that the terminal organelle alone is the gliding motor, defined the requirement and function of several terminal organelle components in gliding, and generated direct evidence that gliding is required for colonization of mucosal epithelium. The studies proposed here build upon that foundation to define further the role of gliding in pathogenesis and explore the mechanical basis for gliding, encompassing three specific aims. Aim 1 addresses the gliding mechanism, focusing on gliding-associated proteins P65, P41 and P24. The terminal organelle detaches from the cell body with the loss of P41 but retains gliding function. Insertions in the P65 gene result in the dragging of surface adhesin P30 from the terminal organelle to the trailing end, where it detaches to leave a trail behind the gliding cell. In the absence of P41, P24 foci appear to move along the long axis of the cell. In the next project period we will apply biochemical and cell imaging approaches including electron cryotomography to explore further the roles of P24, P41, P65, and other selected proteins in gliding. In Aim 2 we will analyze in detail certain gliding mutants exhibiting a distinctive lawn-like growth as a result of disruption of the genes for the only annotated M. pneumoniae protein phosphatase and cognate ser/thr protein kinase. We will confirm cause and effect for these mutants and explore the impact of loss of protine kinase or phosphatase function on the phosphorylation of terminal organelle proteins HMW1 and HMW2. We will also examine cell behavior of these mutants in detail by microcinematography to establish how lawn-like growth is achieved. In Aim 3 we will use a differentiated normal human bronchial epithelium model and wild-type and gliding-defective mycoplasmas to explore how gliding motility specifically contributes to resistance of mucociliary defenses in the colonization of conducting airways. Mycoplasma pneumoniae is the leading cause of pneumonia in older children and young adults and accounts for 20% of all community-acquired pneumonia. Most infections result in respiratory disease that is chronic and protracted, impacting attendance at school and productivity in the workplace, and permanent lung damage can result. In addition, a growing body of evidence supports a significant, contributing role for M. pneumoniae in onset and recurrence of asthma. The studies proposed here will elucidate the mechanism and role of gliding motility in the colonization of the conducting airways.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY ABSTRACT Focal Adhesion Kinase (FAK) is a major cancer drug target that is overexpressed in multiple tumor types. FAK is a critical regulator of tumor survival, invasion, proliferation, metastasis, and immune evasion. Current FAK inhibitors that target the ATP-binding pocket of the kinase domain do no effectively inhibit FAK in cancer because FAK also functions as a scaffolding protein. The Focal Adhesion Targeting (FAT) domain of FAK is an interesting alternative drug target due to its requirement for FAK localization, activity, and downstream effects. Disruption and mutation of the FAT domain causes significant effects on tumor cell apoptosis, proliferation, invasion, and metastasis. Specifically, the FAT domain interacts with the alpha helical LD2 and LD4 motifs of Paxillin to promote its biological effects. The structure of the FAT-Paxillin complex has been solved by x-ray crystallography however has been challenging to target with small molecules. In this project, we will use hydrocarbon stapled alpha helical peptides that have the advantage of enhanced proteolytic stability, cell permeability, and potent inhibition of the entire protein interaction interface. We have preliminary data of Stapled Peptide 3 showing low micromolar inhibition of FAK-Paxillin binding and NMR/SPR data validating the binding site of the peptide. In the first specific aim, we will perform structure-activity relationships (SAR) on stapled alpha helical peptides for enhanced binding and competitive inhibition. We will perform SAR on stapled peptides by changing hydrocarbon stapling strategy, modifying N- and C-terminal amino acids, and adding alternative amino acids. In addition, we will utilize molecular modeling to optimize peptide-protein contacts, synthesize stapled peptides of homologous peptide sequences, and characterize biophysical/biochemical properties of stapled peptides. In the second specific aim, we will characterize and optimize lead peptides for cellular effects. We will perform robust assays to measure cell permeability of stapled peptides, characterize peptides for protease resistance and effects on membrane lysis, and test peptides in cellular efficacy assays to assess the effects of stapled peptides on cancer cells. In the third specific aim, we will test lead peptides with in vitro DMPK assays and preliminary in vivo efficacy models. We will characterize peptides using plasma binding, metabolic stability, and CYP inhibition/assays. We will also test lead peptides in mouse xenograft models alone and in combination with chemotherapy. In all, these specific aims will be used to discover peptide inhibitors of FAK non-catalytic function that can be the basis for future clinical development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Peptidoglycan recognition proteins (PGRPs) are pattern recognition molecules of the innate immune system that bind and, in certain cases, hydrolyze peptidoglycans (PGNs) of bacterial cell walls (1). They are highly conserved from insects to mammals. PGRPs bind PGNs with high affinity and are important contributors to host defense against bacterial infections. Insects PGRPs, of which there are at least 19, are involved in activating signaling pathways that induce expression of anti-microbial peptides. By contrast, mammalian PGRPs, of which there are four, do not act through host signaling pathways, but are directly bactericidal against both Gram-positive and -negative bacteria (1). Indeed, these PGRPs are a new class of bactericidal proteins that have a different structure and mechanism of action than currently known mammalian antimicrobial peptides. Mammalian PGRP-L is an N-acetylmuramoyl-L-alanine amidase that hydrolyzes PGN. Very recently, it was found that PGRP-L also has a proinflammatory function, unrelated to its amidase activity (2). We previously published structures of human PGRP-Ialpha and -Ibeta in complex with PGN fragments, and have proposed a mechanism for their bactericidal activity (3,4). We now plan to extend our work to PGRP-L, which contains a large N-terminal domain with no sequence homology to other known proteins, and whose structure remains to be established. The PGRP-L structure will provide insights into how three different types of pattern recognition molecules (PGRPs, Nod2 and TLR4) play interdependent roles in inflammatory responses to bacterial pathogens. Recently, We obtained a few small crystals which are too small (~50um) for data collection with our in-house detector. They diffract to a maximum resolution of 7 [unreadable] with unit cell dimensions of", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Multicenter collaboration to reduce the risks inherent in allogeneic hematopoietic cell transplantation (HCT) is essential to dampen the dangers of regimen toxicity, infection, transfusion dependence, graft vs. host disease (GVHD) and malignant relapse. Ongoing efforts to address all these risks have been a focus of Network participation for investigators at the University of Minnesota. Our commitment to the Network has included participation as Steering Committee chair, national PI of four protocols, leading the first four Network publications and committing institutional resources to develop and successfully execute Network trials. Because umbilical cord blood (UCB) transplantation is still limited by delayed or failed engraftment, we initiated studies to augment the homing and hematopoietic recovery of UCB grafts based on preclinical data suggesting that complement fragment C3a can accomplish these goals. An ongoing University of Minnesota phase 1 trial testing C3a priming of one of a pair of UCB units to augment engraftment and enhance predominance of the primed unit provides the clinical background for a proposed multicenter phase 11. We propose a protocol testing whether C3a priming of one UCB unit can increase the likelihood of that unit's predominance as the engrafting hematopoietic source;can augment and accelerate multilineage hematopoietic recovery;and can enhance immune reconstitution thereby lessening risks of infection without excess graft vs. host disease. We offer our continued commitment to the Network's success in developing new protocols, identifying patients suitable for Network trial enrollment and performing protocol specific procedures. We will try to exceed the Network's standards in our commitment to enhance multicenter trial success and advance scientific progress within the Network. Multicenter collaborative trials are essential to formally test new ways to enhance the safety of allotransplantation and improve outcomes for our patients. PUBLIC RELEVANCE: Improvement in the outcomes of allergenic transplantation requires control of early toxicity, risks of infection, transfusion dependence, graft vs. host disease and relapse. Multicenter trials within the BMT CTN have tackled all these areas and at the University of Minnesota we are committed to advancing the development and execution of high-quality Network trials. We propose augmenting the success of umbilical cord blood engraftment by priming one of a pair of UCB units with complement fragment C3a, to extend preliminary University of Minnesota data to a multicenter phase II trial. Improving engraftment and post transplant immune recovery may limit these hazards and augment the success of all transplantation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Many evidence-based treatments (EBTs) for common children's mental health disorders have been developed, but they are rarely delivered in real world care. EBTs must be complemented by evidence-based implementation strategies if they are to achieve their promise. Accordingly, the NIH has prioritized efforts to identify, develop, refine, and test implementation strategies (PAR-10-038). Yet, this research is not likely to be fruitful if it is conducted without a thorough understanding of the service systems in which these strategies and EBTs will (hopefully) be deployed. This mixed methods multiple case study will involve seven organizations that are participating in the control arm of an NIMH funded RCT, Testing an Organizational Implementation Strategy in Children's Mental Health (Glisson, PI). This affords the opportunity to examine implementation as usual (i.e., implementation processes that occur in usual care settings) in order to ensure that newly developed implementation strategies will be acceptable, feasible, sustainable, and scalable in the real world. Semi-structured interviews (with agency and clinical directors) and document review will be used to identify and characterize the implementation strategies used in routine children's mental health care (Aim 1), and to explore how organizational leaders make decisions about which EBTs to implement and how to implement them (Aim 2). Focus groups and a project specific self-administered online survey will be used to determine organizational leaders' and clinicians' perceptions regarding the relative importance, acceptability, feasibility, and appropriateness of implementation strategies (Aim 3). Finally, data from the first three aims will be linked to their corresponding organization's scores on a standardized measure of organizational social context (culture and climate) in order to examine how context influences implementation processes and perceptions about specific implementation strategies (Aim 4). By shedding light on implementation as usual, this study will inform efforts to develop and tailor strategies, propelling the field toward the ideal of evidence-based implementation. It will also benefit the RCT by illuminating strategy patterns in the control group, which may aid in the interpretation of results. The proposed research and training plan will allow the fellowship applicant to receive training in: 1) implementation and mental health services research, 2) organizational behavior and change processes, 3) mixed methods research, and 4) scholarly writing and research dissemination. This training will facilitate the successful completion of the proposed project, and will prepare the applicant for an independent research career dedicated to improving the quality of mental health services for children, youth, and families through the advancement of implementation science and practice. The proposed project is consistent with priorities set forth by the NIH and the Institute of Medicine, and is the first step in a larger research agenda to improve the quality of mental health services for children, youth, and families by developing, refining, and testing implementation strategies. PUBLIC HEALTH RELEVANCE: Mental health problems affect a staggering 20% of children and youth each year. Yet, this vulnerable population continues to receive substandard mental health care, largely because we do not understand how to effectively implement evidence-based treatments (EBTs). This study will address this gap by examining implementation as usual in children's mental health organizations, which will inform efforts to identify, develop, refine, and test implementation strategies that can effectively integrate EBTs into real world care.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Desialylated (i.e. neuraminidase-treated) lymphocytes (DL) induce significantly more cytotoxicity in isolated rabbit hepatocytes than do intact lymphocytes. This phenomenon can be selectively blocked by a desialylated glycoprotein and is attributed to the receptor for desialy- lated glycoproteins (hepatic binding protein, HPB) on the surface of hepatocytes interacting with its specific ligands on the surface of DL. Purified HBP increases the cy totoxicity of DL to target cells which do not bear HBP on their surface presumably by an analogous mechanism. Infused desialylated periperal blood mononuclear cells (PBMC) and DL are selectively taken up by the liver in rabbits. The phenomenon is unaffected by depletion of complement but can be blocked by preincubation of desialylated PBMC with IgG-Fab fragments. These findings suggest that the selective hepatic uptake of desialylated PBMC or DL is not mediated by HBP. They are compatible with desialylated PBMC or DL becoming coated with naturally occurring antibodies in vivo with the result that subsequent recognition of cell bound antibody by Fc receptors on Kupffer cells mediates their selective hepatic uptake.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent advancemens in protein separation, mass spectrometry and bioinformatics have enabled the simultaneous analysis of a large number of proteins in cells and tissues. The interaction of these proteins, especially those that respond to specific challenges, are rapidly identified by ingenious screening techniques such as the phage expression or yeast two hybrid systems. These studies, collectively termed proteomics, have greatly enriched our knowledge of cellular physiology and pathology in normal and diseased tissues. A systematic identification of molecular interfaces between interacting proteins in natural cellular environment represents the next major challenge toward a molecular understanding of the complex and dynamic cellular events. Such a profile of molecular interfaces would also be invaluable as a tool in drug development by targeting crucial molecular interfaces. We are initiating such a proteomics project and focusing on the application of chemical cross-linking approach toward a systematic and global identification of protein-protein interfaces in complex systems such as the contractile machinery of muscle cells, the cytoskeletal filaments and organelles of nonmuscle cells. First, high-resolution gel electrophoresis and fractionation techniques are used to identify and index expressed proteins in skeletal and heart muscle tissues. Second, the neighboring relationship is defined by chemical crosslinking with a wide range of reagents. Third, the molecular interfaces of neighboring proteins are identified by isolating the crosslinked complexes, cleaving the proteins and isolating crosslinked peptides for protein sequencing with conventional chemical methods or by mass spectrometry. Fourth, the peptides at the interfaces will be synthesized and their interaction characterized further by capillary electrophoresis and biosensor techniques. A protein sequence database of molecular interfaces of proteins with known and unknown identities will be established to complement and enhance the utility of genome and proteome databases for basic and clinical research. We are evaluating the feasibility and integration potential of each steps of this multifaceted project. In particular, we have investigated the choice of crosslinking agents and the identification of extent, the stoichiometry of crosslinked complexes to define nearest neighbors. As a first step, contractile machinery, myofibrils from vertebrate skeletal muscle (rabbit psoas) and a novel tubular muscle from Midshipman fish sonic muscle were crosslinked by a zerolength crosslinker. The crosslinked complexes were identified by the gel mobility and Western blot of the complexes. Further efforts are made to purify selected complexes, followed by digestion, purification and sequencing and mass spectrometric analysis. Our work so far has clearly indicated the needs to employee or develop crosslinking agents with spectroscopic and mass tags to facilitate the identification and separation of crosslinked peptides. As a second method, protein interface is identified by screening synthetic peptides. The feasibility is demonstrated by the identification of nebulin/calmodulin interface by capillary electrophoresis of clmodulin with a set of nebulin peptides. The data are being compared with direct crosslinking and mass spectrometry. We conclude that, for our purposes, electrospray mass spectrometry is inferior to laser assisted matrix desorption mass spectrometry. The latter provides simple and interpretable spectra that are sufficiently accurate to calculate the composition of the complexes. - proteomics, protein interface, mass spectrometry, capillary electrophoresis, protein sequencing, crosslinking, genomics", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Humans are able to hear across a wide range of sound levels, both in quiet and in the presence of background noise. The auditory system may achieve this wide dynamic range by turning down its response when there is ongoing sound. Though there is physiological evidence for a feedback loop to the cochlea which adjusts gain in a frequency-specific manner. Temporal effects have been measured in simultaneous masking which would be consistent with a decrease in gain with sound stimulation. It is not clear whether these temporal effects are due to the processing of the feedback system. The proposed research examines this important question in light of more recent evidence about the peripheral auditory system. The experiments are designed so that two measures of gain in the cochlea, input-output functions and frequency selectivity, may be estimated with and without preceding acoustic stimulation designed to activate the efferent system. Listeners with normal hearing and listeners with mild cochlear hearing loss will be tested in the experiments. The hearing-impaired listeners provide an interesting control case in which the gain in some frequency channels has been permanently decreased. The data will be analyzed within a well-established physiologically-based model of the peripheral auditory system, to determine whether temporal effects are consistent with a decrease in gain within the auditory channel which is stimulated. This research is important to our understanding of basic auditory processing in normal hearing, and the effects of cochlear hearing impairment. Listeners with normal hearing may have a type of automatic gain control that works independently in each of the thousands of auditory channels in the cochlea. This gain control could help in the perception of speech in background noise, by improving the signal-to-noise ratio within a channel, and by turning down channels containing only noise. The loss of this mechanism could help explain why listeners with cochlear hearing impairment have particular problems in background noise. This research could suggest novel processing strategies that might be used in hearing aids and cochlear implants to improve perception in background noise.The proposed research examines the hypothesis that listeners with normal hearing may have a type of automatic gain control that works independently in each of the thousands of auditory channels in the cochlea. This gain control could help in the perception of speech in background noise, by improving the signal-to-noise ratio within a channel, and by turning down channels containing only noise. The loss of this mechanism could help explain why listeners with cochlear hearing impairment have particular problems in background noise. This research could suggest novel processing strategies that might be used in hearing aids and cochlear implants to improve perception in background noise.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Administration of manufacture of bulk chemicals and bulk pharmaceuticals for preclinical trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Throughout the years, several practical problems have discouraged drug testing in children. These include the lack of a suitable infrastructure to conduct pediatric pharmacology research; the unforeseeable nature of some clinical responses in immature individuals; the possibility of unanticipated adverse reactions; the threat of effects on growth, development or health, long after the drug's administration, and difficulties predicting dose or concentration-response relationships by extrapolating data obtained in adults or experimental animals. Researchers and clinicians involved in pediatric drug development face daily challenges that include, but are not limited to the following: the need to obtain parental permission and/or the child's assent; ethical issues related to the conduct of non-therapeutic research in children; the lack of technology applicable to children; and the lack of incentives that would encourage pharmaceutical companies to study drugs in neonates, infants and children. There is a significant unmet need for pediatric therapeutic and diagnostic device development. In spite of these challenges, the NICHD is aware of the critical need to establish and maintain an environment in which safe and effective pediatric clinical and device trial activities can quickly be initiated and managed. The NICHD provides direct support for clinical research and clinical research networks that focus on particular diseases or public health issues. One of the goals of NICHD is to fund research to better understand the effects and effectiveness of pharmaceuticals on maternal and child health. Continued use of this Task Order mechanism will ensure that NICHD is able to contract quickly and efficiently to acquire experts with the ability to support studies relating to pediatric drug and device development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Genomics Shared Resource (GSR) is a state-of-the-art genomics facility dedicated to providing the latest genomic research tools at low-cost. The primary goal of the GSR is to facilitate high-impact genomic-based cancer research by Stanford Cancer Center members. This goal is accomplished by providing Cancer Center members with access to cutting-edge technologies and providing the services, expertise, and scientific support necessary for utilizing these genomic tools. Technologies and services currently provided by the GSR include: high-throughput sequencing (Illumina Genome Analyzer II, ABI SOLID, Roche Genome Sequencer FLX, and Helicos HeliScope), microarray services utilizing Affymetrix, Agilent, Illumina, Nimblegen, Stanford, and other platforms, genotyping services utilizing the Affymetrix and Illumina platforms, real-time quantitative PCR. Genomic reagents, such as clones, microarrays, and spike-in controls are also provided. Expert assistance with the use of these technologies is also provided throughout the entire experimental process. Guidance with experimental design, data analysis, data archiving and publication is a major component of the GSR. The resources of the Stanford Functional Genomics Facility (SFGF), Protein and Nucleic Acid Facility (PAN), and Stanford Microarray Database (SMD) have combined to provide the full spectrum of services offered by the Genomics Shared Resource and has a total combined operating budget of $2.8M. John Coiler, PhD (SFGF), is the Director of the shared resource, and Catherine Ball, PhD (SMD) and Michael Eckart, PhD (PAN) co-direct the GSR. Patrick Brown, MD, PhD and Gavin Sherlock, PhD are faculty advisors. There are currently 96 Cancer Center members using the shared resource, representing all program affiliations. Future goals are to increase access to high-throughput sequencing technologies and improve bioinformatics support and infrastructure required to implement the latest generation of genomics tools.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Glutamate excitotoxicity is the presumed mechanism underlying many neurological disorders including acute ischemia and chronic neurodegeneration. Recent work from our laboratories demonstrated that mice lacking the neural isoform of the stress activated protein kinsase SAPK/JNK (JNK3) has remarkable resistance to kainic acid-induced seizures, AP-1 transcriptional activity and apoptosis of hippocampal neurons. These results strongly suggest that the SAPK/JNK signaling pathway is a critical component in the pathogenesis of glutamate excitotoxicity. Based on this hypothesis, the present proposal comprises two sets of closely related in vitro and in vivo experiments. Specific Aim I: Mechanism of JNK -mediated glutamate neurotoxicity ( in vitro studies). Keys issues to be addressed include: (1) the JNK - mediated neurotoxicity through gene regulation and de novo biosynthesis; (2) the regulation of intracellular calcium oscillation through JNK signaling; (3) the interplay between JNK - signaling And oxidative stress, (4) the specificity of activation route of JNK signaling. These potential mechanisms will be tested in primary culture of dissociated neurons using a combination of morphological, physiological and biochemical approaches. Specific Aim II: Neuroprotection of the blockade JNK signaling pathway (in vivo studies) Potential clinical applications of inhibition of the JNK signaling may include: (1) the prevention of ischemic apoptosis; (2) attenuation of chronic glutamate excitotoxicity in amytrophic lateral sclerosis; (3) the prevention of neurodegneration (4) enhanced survival of grafted neurons in transplantation therapy. Preliminary evidence of feasibility has been obtained for each Specific Aim. The potential applications will be tested in experimental models of neuropathology using wildtype and mutant mice lacking the neural-specific JNK3 isoform. In summary, the overall goal of the present proposal is both to understand the mechanism underlying JNK- mediated glutamate neurotoxicity and to test the clinical potential of targeting the JNK signaling pathway for therapeutic intervention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research concerns a comparative approach to diel and seasonal activity patterns in feral Dipodomys species both in the field and in laboratory experiments. Some of the specific research proposed is to measure surface activity of Dipodomys deserti automatically throughout hundreds of nights with the intention of evaluating the effects of moonlight, temperature, of competition with sympatric species, the effects upon foraging patterns of seasonal factors and food availability, and the effect upon the activity patterns of males during the reproductive season. Proposed laboratory experiments involve prey visibility in various lighting conditions simulating twilight, a search for a \"lunar clock\", the effects of food availability upon temporal foraging strategies, and the effects of the presence of a predator upon the timing of activity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "One of the most important mediators of host resistance to a vast number of pathogens is the cytokine Interferon gamma (IFN?). During infection, IFN? is released from specialized immune cells and induces multiple antimicrobial response pathways in most mammalian cell types. An important component of the IFN? response is the induction of cell-autonomous resistance towards bacterial pathogens residing and replicating within vacuoles, including Mycobacterium tuberculosis and Legionella pneumophila. To exert their antimicrobial activities, numerous IFN? -induced host response proteins like Nitric oxide synthase 2 (Nos2), Immunity Related GTPases (Irg) and Guanylate binding proteins (Gbp) translocate to pathogen-containing vacuoles (PCVs). The principles that underlie the ability of the host cell to recognize PCVs and target IFN? -induced host proteins to them are currently not well understood. The goal of this research proposal is to fill this gap in our knowledge by defining the molecular players that direct the localization of IFN? -induced antimicrobial proteins to PCVs. Towards this goal, we are taking two complementary, genomic approaches that aim to identify host factors that either facilitate the translocation of IFN? -induced proteins to a Legionella-containing vacuole (LCV) or translocate to LCVs directly. In Specific Aim 1, we will use gain- and loss-of-function screening approaches to identify host factors required for IFN? -mediated restriction of L. pneumophila replication inside macrophages. Host factors critical for providing resistance to L. pneumophila replication inside IFN? -activated macrophages will be subjected to mechanistic studies to further characterize their role in targeting IFN? - induced proteins to PCVs. In Aim 2, we will use an epitope-tagged kinome expression library to directly identify kinases localizing to PCVs in IFN? -stimulated cells using a high-content imaging approach. This second approach will potentially allow us to identify functionally redundant host factors, as well as capture the dynamic sequence of events involved in the host response. Identification of these targeting pathways is a first and critical step towars understanding how host cells recognize PCVs as 'non-self' and potentially dangerous vesicular structures and, conversely, how host-adapted pathogens can evade recognition. This knowledge will open up avenues for the development of novel therapeutics that aim to boost the inherent ability of the host organism to detect and eliminate intracellular bacterial pathogens. PUBLIC HEALTH RELEVANCE: This proposal aims to deploy new (genomics-based) approaches to identify the key host factors responsible for the delivery of antimicrobial proteins to vacuoles that contain pathogens such as the bacterium Legionella pneumophila. Host factors identified in these studies are likely to play an important role in host resistance to many pathogens, including Mycobacterium tuberculosis and Salmonella enterica, and may serve as drug targets for novel host-based antimicrobial therapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT/PROJECT SUMMARY The research theme of the proposed consortium focuses on the early detection of cancer, deciphering cancer pathogenesis, and the identification of potential biomarkers of Kaposi?s Sarcoma (KS) and ocular surface squamous neoplasia (OSSN), two of the most prevalent AIDS-associated cancers in Zambia. The Zambia AIDS Malignancies Diagnosis and Pathogenesis Program (ZAMDAPP)?s goal, which will link key expertise and resources of the Nebraska Center for Virology at the University of Nebraska-Lincoln, University of Zambia University Teaching Hospital, and Zambia?s Cancer Disease Hospital (CDH), is to develop the CDH cancer research infrastructure in three areas critical to the consortium?s overall research theme: cancer diagnosis, viral oncology, and cancer treatment. An Administrative Core will provide the administrative and programmatic leadership and infrastructure necessary to ensure the consortium functions efficiently and effectively. The Administrative Core will pursue four specific aims: 1) provide the administrative expertise and resources necessary to support the consortium?s Career Enhancement Core, Shared Resources Cores, and research projects in the accomplishment of their goals and specific aims; 2) implement a consortium-wide series of operational processes designed to create a collaborative research environment and build capacity for increased self-sufficiency in Zambian health institutions; 3) provide communications infrastructure to engage key stakeholders and promote the consortium locally and globally; and 4) establish and cultivate a strong working relationship with an External Advisory Group composed of key stakeholders and expertise leaders to connect research efforts with Zambian communities and identify research outcomes that will inform healthy policy changes. The organization of the Administrative Core is designed to be responsive, inclusive, and robust, which is essential to successfully address the broad mission and complex relationships within ZAMDAPP, among partnering and collaborating institutions, and with NIH. This core will focus on meeting the needs of the partnership and collaborators in the current climate of rapidly changing scientific and technological advances, which will result in a sustainable consortium with vibrant research programs that work toward the reduction of cancer in Africa.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Support services for the National Kidney Disease Education Program (NKDEP)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Human papillomaviruses (HPVs) represent some of the most common of the sexually transmitted disease. More than 80 HPV types have been identified and several types have been strongly implicated as causative agents in the onset of cervical vaginal, penile, oral and skin cancers. HPV DNA is present in virtually all cases (93 percent) of cervical cancer and its precursor lesions. Current microbicidal compounds with papillomavirus activity have not been described. In addition, there are not published reports of animal models of genital papillomavirus infections for testing intra-vaginal formulations and co-incidental STD infections. Recently, several in vitro and in vivo papillomavirus infectivity models have been documented that make feasible the screening and testing of microbicidal compounds and formulations for potential anti- papillomavirus activity. The purpose of this renewal proposal is to establish the validity of two in vitro and two in vivo papillomavirus infectivity models for the testing of microbicidal papillomavirus compounds. The in vitro models we propose to use are (I) BPV-1-induced focus formation of mouse C127 cells, and (ii) transient in vitro HPV-11 (and HPV-40) infection of human A431 epithelial cells using an ELISA-based RT-PCR detection of HPV E1 E4 spliced mRNA transcripts. We will use the in vitro models to screen for effective anti-papillomavirus compounds to be later tested in two animal model systems. These include a novel human vaginal xenograft animal model system using infectious HPV-11 and HPV-40 virions, and a rabbit genital papillomavirus model recently developed in our laboratory. During the first 3 years of this Program Project, we have identified several potential papillomavirus microbicidal compounds and have begun to characterize in vivo model of genital papillomavirus infections. These microbicidal agents (neutralizing monoclonal antibodies and alkyl sulfates) represent the first compounds with anti-papillomavirus activity. We will continue their characterizing and expand the testing using formulations delivered intra-vaginally (into human vaginal xenografts and into intact rabbit vaginas) for in vivo efficacy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Recovery from measles is associated with life-long immunity that is reflected by the persistence of antibody. Protection after infection with wt virus or after live vaccine is closely associated with levels of neutralizing and hemagglutination-inhibiting antibody; experiences in immunosuppressed children also suggests that a virus-specific cellular immune response is essential for recovery. The currently licensed live attenuated measles virus vaccine is safe and effective when given at 9-15 months of age, but the vaccine is less effective in infants under the age of 9 months, and urban outbreaks have recently involved many young infants. Although immunization of infants as young as 4-6 months has been successful using a 10-100 fold higher dose of vaccine virus, such immunization has been associated with a poorly understood increase in susceptibility to subsequent infections and has been abandoned as a strategy for improving the vaccine and lowering the age of immunization. An inactivated vaccine was one of the earliest measles vaccines introduced. After 3 doses of alum-precipitated, formalin-inactivated vaccine, neutralizing antibody adequate to protect against wild-type viruses was present for a few months, but protection rapidly waned. In addition, approximately 15% of children immunized with this vaccine who subsequently contracted measles developed a severe form of the disease known as atypical measles. This complication has been ascribed to the low levels of antibody to the fusion protein that were induced by the inactivated vaccine, but this pathogenic mechanism has never been proven. Failure to understand the pathogenesis of atypical measles continues to impede the development of newer vaccines for all paramyxoviruses. The current proposal represents a competitive renewal of an original award granted in 1993 to investigate immune responses to measles and atypical measles in non-human primates. In this proposal the investigators propose follow-up studies and other work to determine the pathogenesis of atypical measles, characterize the immune responses to prototype live and inactivated vaccines, and to continue development of cDNA-based measles vaccines. The specific aims of the proposal are: 1) to characterize the immune responses to formalin-inactivated vaccine and live attenuated vaccines by comparing the specificities and biological properties of anti-measles virus antibodies, T cell effector functions, and the cytokines induced by immunization; 2) to define the nature of the immune responses after challenge with wild-type measles virus in unimmunized monkeys and in monkeys previously immunized with live or formalin-inactivated vaccines; and 3) to determine the nature of the immune response to and the efficacy of H, F an d/or N cDNA vaccines delivered by different routes of inoculation in monkeys.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Research at an early stage of medical education is the best predictor of a future career in research. The College of Physicians & Surgeons of Columbia is especially strong in research focusing on the basic and applied features of diabetes, kidney and endocrinologic diseases. Investigation of the health issues dealing with the mandate of NIDDK requires preparations for the next generation of investigators and this application aims to introduce our medical students to research in these fields mentored by exceptional faculty with proven track record of excellence in research and funding. The curriculum at P &S has changed such that every student is now required to produce a scholarly effort and this proposal will support students performing research either during the summer months between the 1st and 2nd years or later. Students at P& S have traditionally been active in research and this application will provide them with a structured program that includes the research as well as a series of seminars covering topics in modern biomedical research. In addition they will be required to attend a series of lectures in the Responsible Conduct of Research. The Program Director will be assisted in managing the Program by an External and an Internal Advisory Committees. Applying students will be chosen by a Recruitment and Admission Committee and their progress in research will be followed by a Mentoring Committee. Finally, after they complete their research they will be advised by a Career Development committee that will aid them in making important career decisions. Recruitment of under-represented minorities, among mentors as well as students, is strongly supported by an ongoing commitment of the College and by targeted efforts of the Program.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have recently shown that the specific amino acids in the M1 and M2 proteins of influenza A virus are important genetic determinants of filamentous virus morphology. In addition, we have demonstrated that the host cell cytoskeletal complex is an important cellular determinant of filamentous virus formation. The objectives of this proposal expand upon these recent observations and focus on determining the cell biological dynamics associated with filamentous influenza virus entry, and on the pathogenic potential of filamentous influenza virus as it relates to spread of infection and severity of disease. In specific aim 1, experiments are described to address specific questions relating to the mechanisms of filamentous virus attachment and entry. Employing radiolabeled and fluorochrome-labeled viral filaments, we will quantitatively examine whether viral filament entry occurs by partial or complete endocytosis or possibly by a phagocytosis-like mechanism. Membrane fusion assays will be used to determine pH requirements and extent of cell fusion of viral filaments. Live cell video microscopy, confocal and electron microscopy will be used to follow the fate of internalized viral filaments and examine whether budding viral filaments can mediate cell-to-cell spread of infection. In specific aim 2, we will focus on the pathogenic potential of filamentous influenza virus. Using primary human nasopharyngeal organ and epithelial cultures we will determine whether infection is restricted to a subpopulation of epithelial cells and determine whether filamentous virus strains have enhanced ciliopathic and tissue damaging capabilities. Using a fluorochrome-based binding assay, we will assess whether viral filaments can enhance bacterial binding to the mucosal epithelium. Using the mouse and ferret animal models, we will compare in vivo the pathogenic potential of genetically similar variants of influenza A/Udorn virus, which differ only in morphology. Pathogenic criteria that will be assessed include: i) the 50 percent minimal infectious dose, ii) tissue specific viral loads iii) tissue-specific histopathology, and iv) rates of weight loss recovery. In the animal studies, we will examine whether filamentous strains promote a descending spread of infection, from an initial localized infection in the nasal tract to the lower respiratory tract. These studies will set the foundation for establishing further determinants of influenza virus pathogenesis and severity of disease in humans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The biologic changes that occur following menopause have heightened interests in the relationship between declining plasma estrogen levels and health in this aging female population. Specifically, in postmenopausal women, the development of atropic oral epithelial cells, which are easily stripped from the underlying connective tissue, hinders proper oral hygiene and impairs the prescribed utilization of dental prostheses. The broad aims of this research are 1) to understand the actions of estrogens on oral tissues and 2) to elucidate the relationship(s) between extracellular matrix proteins secreted by fibroblasts from young versus senescent gingiva, and the effects of such proteins on normal and aberrant epithelial cell growth. As a model system, human fibroblasts derived from the gingiva of premenopausal, postmenopausal and affected postmenopausal females (i.e., those with gingiva exhibiting a hormone-induced desquamative lesion) will be used to define estrogen action in the gingiva. This proposal will examine estrogen effects on proliferation, glycosaminoglycan synthesis, collagen synthesis, noncollagen protein synthesis and collagen expression in fibroblasts from both young and elderly females. Experiments will be performed to determine if maintenance of estrogen sensitivity is a characteristic of the premenopausal fibroblast secretory phenotype and whether it is lost in fibroblasts of elderly individuals with hormone- induced desquamative lesions. Further experiments will analyze cytosol and nuclear estrophiles as well as estrogen metabolism, to determine the mechanism by which estrogen sensitivity is lost. Finally a study of epithelial growth will be examined by in vitro cultivation of epithelial cells on estrogen-stimulated extracellular matrix proteins secreted by gingival fibroblasts, to determine if gingival epithelial cell proliferation, attachment, and keratin proteins are dependent on extracellular matrices secreted by estrogen-stimulated fibroblasts. Knowledge gained from the proposed research on gingival fibroblast and epithelial cell function in young and elderly females, as ell as a new understanding of how these cells respond in the presence of estrogens, should provide the rational for the development of novel therapeutic approaches in the control of hormone-induced oral and other mucosal desquamations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neurofilament (NF) proteins, though synthesized in neuronal cell bodies, are normally posttranslationally phosphorylated in axons. In some neuro-degenerative disorders (e.g., ALS), however, they are abnormally hyperphosphorylated in cell bodies. We have been studying the factors regulating these topographic pattern of neurofilament (NF) phosphorylation. One kinase, cdk5, identified in our laboratory, specifically phosphorylates KSPXK motifs which constitutes 20% of the total repeats in high molecular weight NF-subunit, NF-H. The remaining 80% repeats are KSPXXXK motifs kinase(s) phosphorylating these motifs are not known. A synthetic KSPXXXK peptide, KSPAEAKSPAEAKS, which repeats 41 times with minor variations at non-KSP residues was used as a substrate to identify the kinase in rat brain that phospshorylates NF- H. On the basis of biochemical, immunochemical, pharmacological and amino acid sequence analysis, the purified kinase appeared to be MEK- activated MAP kinase. To verify this statement, we demonstrated that bacterially expressed MAP kinase phosphorylated expressed rat NF-H in addition to KSPXXXK, KSPXXK and KSPXK peptides. This study suggests that neuronal MAPK can phosphsorylate all KSP repeats in rat NF-H and may be the principal kinase in vivo that phosphorylates serine residues in KSP repeats in neurofilament tail domains. Since cdk5 specifically phosphorylates KSPXK peptides and not KSPXXXK, we investigated the structural differences of these motifs, by analyzing the conformation of four peptides with either KSPXK or KSPXXXK motifs. The KSPXXXK peptide exhibited a CD spectrum indicative of a helical conformation; under the same conditions, however, the KSPXK peptide showed a mainly extended conformation with beta-turns. These differences revealed a structural specificity for the substrate of this kinase. using two dimensional NMR methods and molecular modeling. We are also studying the mechanism of cdk5 activation. Though, cdk5 is similar to other cdc2 kinases, its regulation is completely different from mitotic cdc2 kinases; e.g., cdk5 is active only in postmitotic neurons, its activity is independent of its phosphorylation, nor is its activity regulated by cyclin binding. Instead association of neuron specific proteins, P35 and P67 are responsible for its activation. These regulator proteins share no homology with cyclins. We have begun to locate the activation domains in P35 using various mutations and truncations of P35 in in vitro assays. These studies suggest the potential residues for kinase activation reside in amino acid residues 137-167 and 291-263 of N- and C-terminal tail domains respectively of P35. Further mutational and truncational studies are in progress.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of this project is to identify novel monoclonal antibodies (mAbs) that broadly recognize the HIV-1 envelope glycoprotein (Env) and block infection in vitro to guide vaccine development. This goal will be pursued in local cohorts of HIV-1 infected individuals who control their infections in the absence of anti-retroviral therapy (JAIDS, 50:403-8, 2009). Approximately 13% of these individuals have circulating broadly neutralizing antibodies providing the study population for our studies. A key element of our approach is the development of a new assay to census Env-specific memory B cell clones (BMem) that allows the rapid and direct cloning of full-length molecular clones of the antibodies expressed by these cells (PNAS, 106:3952-7, 2009). This method permits clone identification using native oligomeric HIV-1 envelope glycoproteins (Env) expressed on cell surfaces and by direct neutralization of pseudoviruses. Env-reactive clones are used to produce mAbs that will be evaluated for epitope specificity and neutralization breadth. This information will be used to test the hypothesis that neutralization breadth is determined by monoclonal or pauciclonal responses comprised of one or a very few neutralizing specificities as opposed to a polyclonal response comprised of a mosaic of neutralizing specificities. There are two specific aims. Aim 1- To develop clonal specificity profiles of Env-specific BMem from NVS who have ongoing broadly neutralizing antibody responses- Clonal specificity profiles of anti-Env responses will be determined by limiting dilution analysis, mAb isolation, and epitope mapping to identify broadly neutralizing mAbs. Aim-2- To compare neutralization breadth between plasma antibodies and mAbs representing a full clonal profile of BMem to determine the number of mAbs that must be pooled to reconstruct the neutralization profile of the circulating antibody pool. Neutralization breadth of mAbs will be determined using standardized pseudovirus assays. This data will be used to determine the clonality of an ongoing broadly neutralizing antibody response. This aim will test the hypothesis that neutralization breadth is determined by monoclonal or pauciclonal responses comprised of one or a very few neutralizing specificities as opposed to a polyclonal response comprised of a mosaic of neutralizing specificities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application requests continued support for the long-term objectives of MH 51372, namely to develop and implement high throughput strategies for the detection and fine mapping of mental health-related quantitative trait loci (QTL), for the integration of QTL mapping with functional genomics and for the detection and confirmation of the relevant quantitative trait gene(s) (QTGs). This integrated approach to QTL analysis requires the development and utilization of detailed brain gene expression and sequence datasets. The phenotypes of interest are open-field activity (OFA) and prepulse inhibition (PPI) of the startle response. This renewal application also emphasizes the detection of gene networks associated with OFA and PPI. This competing renewal has 5 specific aims. 1) To fine map in heterogeneous stock (HS) animals (to a resolution of 1 cM or less) high quality QTLs for OFA (2 on Chr 1 at ~ 110 and 175 Mbp) and PPI (Chr 11 at ~ 70 Mbp and Chr 16 at ~ 45 Mbp). 2) To integrate the QTL data with interval relevant gene expression and sequence data. 3) To determine in HS4 animals the gene coexpression networks associated with OFA and PPI. 4) To determine if the modules identified in aim 3 are the same modules that differentiate short-term selective breeding (STSB) lines, selected for High and Low OFA and PPI. 5) To determine if the OFA and PPI modules identified in aims 3 and 4 are also detected in a more genetically diverse mouse population of mice, namely an outbred population derived from the Collaborative Cross. The primary mapping population is a HS formed from the C57BL/6, DBA.2j, BALB/c and LP strains. Mapping in these HS4 will provide QTL resolution of approximately 1-2 cM. As each interval will be completely sequenced, all causative polymorphisms will be detected. In addition to 3' biased oligonucleotide gene expression analysis, we will test the idea that some QTLs may be generated by alternative exon usage. Our ability to sort through the large amount of data generated is made possible by precisely knowing the haplotype of each QTL. The proposed work builds upon data collected during the current grant period and importantly incorporates new genomic technologies which make possible our fully integrated approach. PUBLIC HEALTH RELEVANCE: The proposed work attempts to understand the role(s) of genetic factors in behaviors that are of mental health significance. The eventual goal is to determine which genes and gene networks make some individuals more prone (increased risk) for developing abnormal behaviors such as schizophrenia and anxiety disorders. With this information in hand, it should be possible to develop new therapeutic strategies and interventions. .", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We will continue investigating the intercellular and intracellular signaling pathways that lead to neuronal death (or survival) following the elimination of excitatory synaptic input to the mammalian cochlear nucleus. A major initiative during the current grant period has been to use modern genetic analysis methods to begin assessing the biological mechanisms underlying differential responses of neonatal and adult animals to afferent deprivation (a critical period) of the mouse cochlear nucleus.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is an application written in response to RFA PA-00-004. It describes a systematic 5-year training plan to launch an independent research career in the cognitive and emotional neuroscience of human cocaine addiction through the use of functional magnetic resonance imaging (fMRI), a powerful technique that allows simultaneous, non-invasive measurement of brain activation and cognitive performance. This plan will allow the candidate, a Clinical Psychologist with background in Neuropsychology, to initiate technologically sophisticated research of relevance to treating cocaine addiction. This goal will be achieved through a rigorous training plan, integrating didactic and research components tailored to the candidate's goals and experience. Brookhaven National Laboratory (BNL) provides an ideal training environment: 1) BNL is home to state-of-the-art functional neuroirnaging technologies; and 2) The mentors of this project are world renowned for the application of these cutting-edge technologies to study mechanisms of drug addiction. Under their supervision the candidate will conduct fMRI (4T) activation projects designed to evaluate whether adaptations to the frontolimbic circuit underlie deficits in salience attribution and response inhibition in cocaine addicted subjects, and whether these neuroadaptations and cognitive-behavioral impairments can be used to predict and monitor relapse. In Project 1, subjects receive different levels of Monetary Incentive for correct performance on a go/no-go task. In Project 2, subjects perform a Drug Stroop task, naming the color of cocaine-related words, neutral words, or non-words. Project 3 is a longitudinal study, where subjects undergo Projects 1 & 2 at baseline (2-4 weeks after detoxification) and 6 months follow-up. In all projects, we measure the blood oxygenation level dependent (BOLD) responses of the frontolimbic circuit including the orbitofrontal cortex (OFC) and anterior cingulate gyrus (ACG). We hypothesize: 1) Task specificity: the Monetary Incentive task will primarily activate the OFC while the Drug Stroop task will primarily activate the ACG; 2) Group specificity: the activation of the frontolimbic circuit by the two cognitive-behavioral tasks will differ as a function of drug addiction; and 3) Relapse prediction and monitoring: increased frontolimbic activation and impaired cognitive-behavioral performance at baseline will characterize the cocaine addicted subjects and predict relapse at 6-months follow up. For the subjects who will relapse, abnormalities in these brain-behavior measures will remain the same or get worse during the follow-up period while for the subjects who will remain abstinent, normalization of these measures will be documented.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this research is to investigate the function of a newly discovered mitochondrial progesterone receptor, PR-M. We believe that progesterone acts via this receptor to increase cellular ATP production in multiple tissues to meet the increased metabolic demands of pregnancy. Cardiac function dramatically changes with pregnancy with increased heart rate and contractility requiring increased ATP production. Inadequate cardiac adaptation may contribute to placental insufficiency related to preeclampsia and intrauterine growth restriction. Our aim is to investigate the hypothesis that PR-M increases cellular respiration in the heart. To test this hypothesis we will temporally express human PR-M in the mouse heart using a heavy myosin polypeptide 6 promoter in a Tetracycline-On (Tet-On) system. Tissue and temporal expression of the transgene will be determined by realtime RT-PCR and western blot analysis. Cardiac function in transgenic animals will be evaluated during late gestation and in non- pregnant models of cardiac hypertrophy to mimic volume and pressure overload. Assays will include echocardiography, heart morphometry, microscopic analysis of capillary density, fibrosis and apoptosis and determination of tissue ATP levels. Transgenic overexpression is a valuable tool to analyze the tissue function of PR-M. We believe that enhanced ATP production secondary to PR-M expression will diminish cardiac hypertrophy secondary to volume and pressure overload in the study models. These studies will further develop a novel mechanism in which progesterone modulates cellular respiration via a direct affect on the mitochondria. Additionally, the created PR-M transgenic mouse can be cross-bred with other tissue specific Tet-On strains for future studies of PR-M function in the mammary gland, liver, skeletal muscle, adipose and brain. PUBLIC HEALTH STATEMENT Pregnancy is associated with dramatic changes in a woman's body including a great increase in the workload of the heart. This process requires an increase in ATP which serves as the major fuel source for the cell. Our studies are investigating a new receptor for the reproductive hormone, progesterone, located in the mitochondria of the cell where ATP is produced. We believe that this receptor provides a mechanism whereby progesterone increases cellular ATP formation to meet the many metabolic demands of pregnancy. Dysfunction of this system could contribute to pregnancy related diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of the project is to validate the feasibility of a microwave tomographic (MWT) imaging technology for assessment of functional and pathological conditions of extremity soft tissue. The successful management of a fractured bone involves an understanding of the two major components (boney and soft tissue elements) of any extremity segment. The diagnosis and evaluation of the boney component is obvious to the treating physician by radiographic studies. The accurate assessment of the soft tissue component of the injured extremity segment remains a major deficiency in management of fractures. Consequently there is an important need to develop an effective and rapid method of non-invasive assessing of extremities soft tissue viability. Previous studies (NHLBI grant No HL65657) demonstrate that microwave tomography is feasible for non-invasive assessment of myocardial viability. Our most recent studies suggests that the technology might be feasible for rapid soft tissue functional imaging of extremities Both the imaging capabilities of this technology for assessment of functional conditions of extremities soft tissues within a very short msec range of acquisition cycle and the use of low, safe levels of non-ionizing radiation with cost efficiency have potentials for significant improvement of public healthcare. In this three year study we plan to (i) develop 2D MWT system; (ii) develop an appropriate image algorithm and (iii) assess feasibility of the technology for extremities soft tissue imaging, including sensitivity and resolution of the technology. Relevance of this project to public health. There is important need to develop an effective and rapid method for assessment of conditions of extremities soft tissue during trauma and recovery. The goal of the project is to validate the feasibility of novel imaging modality (Microwave Tomographic Imaging) for non- invasive (non-destructive) imaging of functional conditions of extremities soft tissue. There is an important clinical need to develop an effective, rapid and cost-effective method of non-invasive assessing or imaging of extremities soft tissue viability. The overall goal of the project is to validate the feasibility of a microwave tomographic (MWT) imaging technology for assessment of functional and pathological conditions of extremity soft tissue. The imaging capabilities of this novel technology and the use of low, safe levels of non-ionizing radiation with cost efficiency have potentials for significant improvement of public healthcare. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sounds are encoded by the cochlea in the timing of spikes in the auditory nerve. Auditory nerve fibers converge onto bushy cells in the cochlear nucleus, through synapses called \"endbulbs of Held\". This convergence leads to changes in the timing information passed on by bushy cells to higher auditory centers, which may affect sound localization and processing. Experiments have indicated that the timing of bushy cell spiking is greatly affected by the size of the endbulb synaptic current, which is subject to 2 major influences in vivo. First, endbulb synapses show considerable depression when activated at normal rates. Second, they are subject to a number of neuromodulatory systems. Both these influences can affect the information carried by bushy cells, but neither is well understood. The specific aims of this project are to determine (1) the dynamics and mechanisms of use-dependent changes in the endbulb synaptic current, (2) how the different components of the synaptic current control bushy cell timing, and (3) how neuromodulation changes these relationships. This work will be carried out using patch-clamp recordings of bushy cells in brain slices taken from mice and gerbils. The mechanisms of depression will be considered first, by testing the contributions of presynaptic vesicle depletion and postsynaptic receptor desensitization and saturation. In addition, an unusual form of depression at the endbulb will be examined, which has been proposed to involve reduced presynaptic calcium influx, but has never been directly tested. It will also be determined how depression could be mitigated by both facilitation and the activation of NMDA receptors. In addition, current-and dynamic-clamp studies will test whether delayed release during high levels of activity disrupts the precisely timed responses of bushy cells. Taken together, these studies will provide important information about the mechanisms by which timing information is transformed by convergence of auditory nerve synapses during realistic activity. This work will examine the mechanisms used by cells in the cochlear nucleus to process sound information. It will also provide important insights into the functional role that different receptor types and neuromodulatory systems play in neuronal computation. This work may lead to improvements both in existing cochlear implants and in implants that stimulate the cochlear nucleus directly. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT Neurological and neuropsychiatric illnesses are prevalent and debilitating, affecting more than a quarter of the global population. Learning and memory are implicated in every neurological illness from stroke to Alzheimer?s disease and every neuropsychiatric dysfunction from depression to drug addiction. Understanding the neuroscience of learning and memory, integrating across approaches and methods, is one of the most formidable challenges of the 21st century. While research in these areas has made significant strides in understanding and treating brain disease as well as promoting brain health, progress has been stalled. There are fundamental gaps in our knowledge that stem for lack of integration across approaches and insufficient dialogue among investigators addressing neuroscience questions from different perspectives. Integration across animal and human research as well as basic and clinical research will have substantial impact on facilitating team science, which is essential for successful translation. The 2018 International Conference on Learning and Memory (LEARNMEM2018) covers numerous facets of the neuroscience of learning and memory with an emphasis on integrating across animal and human studies as well as across fundamental and translational science. The 5-day conference that will feature plenary and keynote talks by distinguished neuroscientists, diverse and balanced symposia, short talks, poster sessions, a panel discussion on open science and open publishing, as well as community engagement sessions. Anticipated attendance is ~800 and the conference is open to everyone. AMA PRA Category 1? Continuing Medical Education (CME) credits will be offered to physicians and healthcare professionals. Travel awards for early career scientists will be available, and special emphasis will be placed on multidimensional diversity of contributions across gender, race, seniority, country of origin, and level of analysis. The conference will achieve the following aims (1) Identify knowledge gaps and opportunities for scientists, clinicians, nonprofits, businesses, and community members to share and discuss cutting-edge research in the area of learning and memory; (2) Form collaborative teams among domestic and international investigators, early career and established scientists, animal and human researchers, fundamental and translational scientists, as well as user and developer communities; (3) Disseminate knowledge generated from the meeting through scientific and public venues; and (4) Facilitate engagement between scientists and community members across all levels from schoolchildren to retirees. LEARNMEM2018 will have substantial impact on the field by accelerating the pace of team science and leapfrogging towards a more complete fundamental understanding of learning and memory mechanisms as well as novel therapeutic and preventative approaches to treating brain illness and promoting brain health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The technique of free tissue transfer, has revolutionized many areas of reconstructive surgery, such as limb salvage and reconstruction of the esophagus, where other options have been unacceptable. As free tissue transfer is now being done routinely at institutions where inexperienced staff are unable to adequately monitor viability, the need for a reliable method to monitor the patency of microvascular anastomoses has become increasingly evident. Accurate and objective assessment of blood flow both at the time of surgery and during the post operative period is essential, because if the tissue viability is lost, the consequences can be staggering. Early recognition of postoperative vessel thrombosis is of utmost importance if reexploration is to prove successful. To this end, numerous monitoring techniques have been investigated. Those classified as direct monitors of perfusion have proven to be of little or no value - particularly as free flap monitors. Probes are bulky, very expensive, and require great care in positioning and removal. Indirect monitors such as those which monitor the tissue concentration of a physiological gas such as oxygen, rate ahead of the direct monitors due to advantages of low cost, simplicity, ease of placement and removal, etc. However, these monitors lack the needed specificity of the direct monitors for measuring tissue perfusion. Adding to this specificity problem is the fact that the sensor cannot be calibrated once it is implanted. Our approach is to preserve the simplicity afforded by indirect monitoring devices, while providing the specificity for tissue perfusion measurement, by utilizing a self-calibrating tissue PO2/PCO2 probe, which manipulates the local tissue gas conditions. Tissue PO2 and PCO2 are monitored under alternating depleting and nondepleting conditions. A physilogically based mathematical relationship relates tissue perfusion to the PO2 and PCO2 measurements. This proposal is directed toward the in vivo validation of the method.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have developed a method for preparing a covalent conjugate of proteins with purified gangliosides in the micellar form. The ganglioside is oxidized with periodate, and the protein is attached to it by reductive amination with sodium cyanoborohydride. The ganglioside moiety of the product retains the ability to transfer irreversibly from the micellar form to a lipid bilayer, thereby associating the protein moiety with the lipid bilayer of choice, for example, a liposome or viable cell membrane. We have demonstrated that a ganglioside conjugate with rabbit IgG can associate with human or sheep erythrocytes, since the cells may be hemagglutinated with goat anti-rabbit IgG after incubation with the conjugate. We have also demonstrated that a ganglioside conjugate with monoclonal mouse anti-glycophorin can associate with Sendai virus and confer upon the virus the ability to agglutinate and hemolyze desialylated human erythrocytes. The method will be of considerable importance in cell biology and immunology by allowing one to study: 1) antigen-directed fusion; ii) T-cell, T-cell hybridoma production; iii) delivery of genetic material to cells and the insertion of artificial receptors. We wish to improve the method by examining alternative conjugation methods, by examining the potential of different purified gangliosides as a hydrophobic anchor, and by optimizing conditions for insertion of the conjugates into cells. We will develop a method for detection of ganglioside GD3 in leukemia cells and leukemia cell extracts by production of monoclonal anti-GD3 coated erythrocytes. We also wish to explore the potential usefulness of the method by examining the insertion of conjugates into viable cells. We will characterize the effect of insertion of cell growth and viability, and determine whether conjugates can undergo patching, capping and endocytosis or shedding. We will also examine the usefulness of this technique for antigen-directed fusion in the production of T cell, T cell hybrids. Anti T4 or T8 conjugates will be inserted into BUC cells and fused with human T cells. The resultant hybridomas will be scored for T4 and T8 antigens to establish whether the use of antigen-directed fusion can increase the frequency of specific hybrids.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goals of this Phase II STTR are to evaluate the outcomes of REAL media, an interactive, self- paced, e-learning substance use prevention media literacy curriculum and prepare it for marketing to community organizations, including our partner, 4-H. Substance use increases in frequency and risk through mid-adolescence, yet prevention interventions primarily target early use, are time intensive, and are implemented in a limited number of settings such as schools. Moreover, they often fail to address the media-saturated lives of youth despite research demonstrating the deleterious effects of advertising and entertainment media. This provides a market niche for the proposed project that addresses this curriculum gap through the innovative use of both technology and prevention science. Guided by the new Theory of Active Involvement, REAL media develops critical perspective taking about substance use decisions and confers resistance to pro-drug (e.g., alcohol, cigarettes, e-cigarettes, marijuana, smokeless tobacco) messages through youth analysis of pro-drug media messages combined with interactive media manipulation and active involvement of youth participants in creating their own anti-ATOD prevention messages. These youth-created messages are then entered into an online contest via a social media proliferation strategy (i.e., youth recruit others to view their messages on social media (e.g., Facebook, YouTube) to win the contest) in which messages are diffused to the wider community. The curriculum demonstrated promising results when administered face-to-face during an NIH-funded pilot study, and the Phase I project demonstrated excellent usability and feasibility for online delivery through 4-H clubs. During Phase II, we will finalize the program and conduct a group-randomized clinical trial among 4-H clubs in four states (NJ, OH, MD, and PA). Clubs will be randomly assigned to use the curriculum or continue current practices with the option for delivery at the end of the study. 4-H members (ages 13-15) will complete a pretest, immediate posttest and follow-up posttests at 3 and 9 months to assess effects. Results will guide preparation of REAL media for the market. The flexibility of the brief, online format (four 15-25 minute levels plus a fifth message planning and production level) for youth in individual or group settings makes this ideal for both community and school implementation. Thus, REAL media is well-suited for rapid dissemination through our existing partners, 4-H, D.A.R.E., and Boys and Girls Clubs, as well as other potential community partners (e.g., YM/WCA, Boys and Girl Scouts) and is appropriate for Phase II funding.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To support the development and assessment of two live microbiome-based products.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In an integrated program of laboratory and clinical investigation, we study the molecular biology of the heritable connective tissue disorders osteogenesis imperfecta (OI). Our objective is to elucidate the mechanisms by which the primary gene defect causes skeletal fragility and then apply the knowledge gained from our studies to the treatment of children with these conditions. Structural defects of type I collagen molecule are well known to cause the dominant bone disorder OI. A severe recessive form of OI was first postulated in 1979. We hypothesized that the cause of recessive OI with abnormal collagen biochemistry and normal collagen gene sequence would involve a gene(s) whose products interacted with type I collagen. Ten years ago we identified defects in two components of the collagen prolyl 3-hydroxylation complex, CRTAP and P3H1 (encoded by LEPRE1) as the cause of recessive OI. Our work generated a new paradigm for collagen-related disorders of matrix, in which structural defects in collagen cause dominant OI, while defects in proteins that interact with collagen cause the rare forms of OI. Recessive OI is now a major area of investigation for the BEMB. The phenotypes of types VII and VIII OI are distinct from classical dominant OI, but difficult to distinguish from each other. Biochemically, both groups full overmodification of the helical prolines and lysines by prolyl 4-hydroxylase and lysly hydroxylase, indicating delayed folding of the collagen helix. We showed that mutual stabilization of CRTAP and P3H1 underlies the phenotypic and biochemical similarity of types VII and VIII OI. With collaborators at the Boltzman Osteology Institute, we recently focused on the bone of the non-lethal subset of type VIII OI patients. We demonstrated that there is no redundancy for collagen 3-hydroxylation function, comparing null mutations in bone and skin. Bone histology was similar to type VII OI, although it had the distinctive feature of extremely thin trabeculae and patches of increased osteoid, suggesting mineralization is slower in type VIII than VII OI. BMDD yielded increased mineralization of type VIII bone, as in classical OI and type VIII OI, but the proportion of bone with low mineralization was increased in type VIII bone vs type VII. Type IX OI has a distinctive phenotype without rhizomelia, and distinctive biochemistry. We generated a CyPB KO mouse, which has reduced bone density and strength, but increased brittleness. Only 1-2% 3-hydroxyltion is detected in KO cells, showing the importance of CyPB to complex function. Collagen folds more slowly in the absence of CyPB, but CsA treatment revels the potential existence of another collagen PPIase. CyPB supports LH1 activity and in its absence there is significant reduction of hydroxylation of crosslinking residue K87. The decreased crosslink ratio alters fibril structure and reduces bone strength. With collaborators at the University of North Carolina we showed that CyPB interacted with all LH forms (LH1-3). The effect of CyPB KO in type I collagen of tendon had distinct patterns in the collagen helix versus telopeptide domains. yPB modulates crosslinking by differentially affecting lysine hydroxylation in a site-specific manner. We have extended our work on PPIB function in a collaboration with investigators at Hebrew University in Jerusalem. CyPB was shown to be critical to the folding of presenilin-1, a protein linked to familial Alzheimers Disease. Some substitutions in presenilin-1 make it resistant to folding properly by CyPB. Conversely, the brains of CyPB knock-out mice were shown to have reduced quantities of processed, active presenilin-1. ER-chaperones may thus be targets for the development of counter-neuro degeneration therapies. We delineated a mutation in IFITM5, which encodes the transmembrane protein BRIL, that establishes a connection between types V and VI OI. The BRIL S40L substitution results in minimal expression and secretion of PEDF by mutant FB and osteoblasts. Om contrast to the gain-of-function BRIL mutation that causes type V OI, the BRIL S40 causes decreased mineralization and expression of bone markers. Only type I collagen shows similar expression pattern in both mutations, with decreased expression, secretion and matrix incorporation. Type XIV OI is a moderately severe form of OI which was identified in 2013. It is caused by recessive defects in TMEM38B, which encodes TRIC-B, an ER cation channel. We identified 3 probands with recessive null defects in TRIC-B and studied the function of TRIC-B deficiency in their fibroblasts and osteoblasts. TRIC-B deficiency impaired ER calcium flux, although the calcium channels themselves had normal stability. The impairment in calcium flux resulted in ER-stress along the ATF4 pathway. TRIC-deficiency was demonstrated to be collagen related because it impaired collagen synthesis and assembly at multiple steps. Collagen helical lysine hydroxylation was reduced, although the levels of LH1 protein were increased. Because calcium flux impacts multiple ER chaperones, the collagen was also misfolded and substantially retained in the ER. Further investigations will focus on type XIV osteoblast differentiation. This year we delineated the first X-linked recessive form of OI, made even more exciting by its novel bone mechanism. X-linked OI is moderately severe with pre- and post-natal fractures of ribs and long bones, dysplastic bone with bowing and crumpling. It is caused by missense mutations in MBTPS2, which encodes Site 2 protease (S2P). S2P is a critical component of Regulated Intramembrane Proteolysis (RIP), a process in which S1P and S2P located in the Golgi Membrane sequentially cleave regulatory proteins that are transported from the ER membrane in times of cell stress or sterol metabolite deficiency. The S2P substitutions in X-OI are located in or near the S2P motif critical for metal ion coordination. The levels of S2P transcripts and protein are normal but RIP function on substrates OASIS, ATF6 and SREBP are impaired. At the bone tissue level, hydroxylation of collagen K87 residues in type I collagen is reduced by half, altering collagen crosslinking in bone. The osteoblasts with S2P defects also have a differentiation defect.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Considerable controversy surrounds the question of whether the two different ytpes of asbestos -- amphilboles and chrysotile -- have different carcinogenic potencies. The \"amphibole hypothesis\" holds that chrysotile is less likely to cause lung cancer and mesothelioma than are the amphiboles. The abiolity to clarify this question is limited by a lack of quantitative data with which to estimate separate dose-response curves for the two fiber types. This proposal seeks funds to study lung cancer and mesothelimoma risks in a Sovenian cement asbestos plant in which excellent historical exposure data are available, distinguishing the two broad classes of asbestos fibers. The small Grant Investigator, an occupatinal physician and epidemiology doctoral student, would conduct a detailed historical exposure reconstruction and case control studies of lung cancer and mesothelimoma. The cohort of workers employed in the cement asbestos manufacturing plant of salonit Anhovo, Slovenia will be constructed from existing detailed wage lists which date from the 1940s. Cases will be all incident cases of primary lung cancer or mesothelioma from 1964 to 1994 in those who were hired after 1959 and who worked at least one month between 1964 and 1994. They will be identified using data from the slovenian National Cancer Registry, one of the oldest national registries in Europe. Exposure measurements are available for most exposed jobs begining in 1961. Three different methods of measurement were used: konimeter measuring particles/cm3, a gravimetric method measuring milligrams/m3, and membrane filter measuring fibers/cm3. Regression analyses will be used to calculate operation-specific conversion factors among these methods. Cumulative lifetime exposure to amphilboles and to chrysotile will be estimated sparately for all cases and controls. Logistic regression models will be used to estimate separate exposure-resonse curves for the two fiber types controlling for smoking (smoking history data available on each worker from medical records).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Binge alcoholism is a growing problem worldwide. Young adults (16-24 years of age) are particularly prone to the adverse effects of alcoholic binging, which is often consumed with secondary drugs of abuse including marijuana. Previous research has suggested that alcohol may share pharmacological effects with cannabinoid agonists such as D9-tetrahydrocannabinol (THC), the primary psychoactive substituent of marijuana. Further, previous work has found that these effects may be modulated by alcohol's effects on the endocannabinoid system. Although much of the preclinical work on both alcohol and cannabinoids has been conducted in male rodents, results of sparse studies in female rodents have suggested sex-selective differences, with females often exhibiting enhanced sensitivity to the pharmacological effects of ethanol as well as THC. In the parent grant, our goals are to characterize some of these differences for THC and to investigate potential mechanisms. In this supplement, we propose to incorporate examination of ethanol/ cannabinoid co-abuse into our overall research strategy. As with the parent grant, integrated in vivo and in vitro measures will be used. To take advantage of local expertise on the hepatic effects of alcohol, we have expanded the dual brain and behavior focus present in the parent grant to incorporate research on the effects of alcohol and THC on the liver. In the proposed project, we will investigate sex differences in the acute in vivo pharmacological effects of alcoho and THC co-administration, as well as their rewarding / aversive effects following repeated intermittent co-administration (Aim 1). These behavioral effects will be correlated with in vitro measures of endocannabinoid system functioning in the brain and the liver and with determination of the extent of liver injury induced by co-abuse of both substances (Aim 2). Understanding the sex-selective effects of co-abuse of alcohol and marijuana is particularly critical at this time since the recent loosening of legal restrictions on marijuana use in the U.S. will likely be associated with increased rates of alcohol-marijuana co-abuse in both sexes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application for a Midcareer Investigator Award in Patient Oriented Research (K24) seeks support for a physician nutrition specialist and established investigator in clinical and translational nutrition science. The P.I. is committed to performing patient-oriented research (POR), focused on nutrition, and in mentoring clinical residents, fellows and junior faculty in clinical nutrition research to develop a new cadre of investigators in Clinical Nutrition. This proposal will provide protected time to enable the P.I. to advance his research and mentoring efforts in nutrition-oriented research, and provide additional training and resources that will in turn enhance his ability to mentor physicians and other clinical investigators in nutrition-related studies. The applicant has a unique training background in nutrition and medicine and an excellent record of federal, foundation and industry funding for his investigator-initiated POR and translational research. He has a strong publication record and a history of leadership roles in national nutrition-related societies and service on editorial boards of front-line nutrition journals. At Emory, the P.I. serves in the leadership of the NIH-funded GCRC, K12 Emory Mentored Clinical Research Scholars (EMCRS) and K30 Masters of Science in Clinical Research (MSCR) programs, the Center for Clinical and Molecular Nutrition and the graduate program in Nutrition and Health Science - all of which provide opportunities for mentoring in POR. The P.I. has mentored more than 18 trainees, including a current cadre of 5 physician junior faculty members in Medicine, Surgery, Neurology and Pediatrics, 2 Ph.D. junior faculty clinical investigators, and 3 Ph.D. students who conduct both clinical and translational nutrition research. The candidate's research program includes GCRC-based studies on the efficacy and mechanisms of glutamine in catabolic states, including critical care and sickle cell anemia, on metabolic adaptation in short bowel syndrome and after medical/surgical weight loss, and studies on diet-related redox control. The specific aims of this K24 are: 1) to provide protected time enabling the P.I. to increase his mentoring activities among current and new trainees in POR; 2) to facilitate the P.I.'s development of new research skills in nutritional metabolomics and new redox-related measures within the context of his current clinical research, enabling him to train mentees in these new measures; and 3) to faciliate the career development of the applicant. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary The goal of this project is to focus on the relationship between dental use, overall health, comorbidity, and medical services use. The central hypothesis guiding this study is that dental care use is highly correlated with better-than-average general health and lower levels of comorbidity and utilization of medical services. We will test the hypothesis and conduct this study using secondary data available from the Medicare Beneficiary Survey (MCBS) and Chronic Conditions Warehouse (CCW), Area Health Resources File (AHRF) and the University of Michigan Institute for Social Research (ISR) Health and Retirement Study (HSR). We will also take advantage of a HRS dental specific module co-developed by members of this research team and the survey staff at the ISR. This proposed project builds upon and is an extension of several collaborative studies previously conducted by this research team. Results obtained from this study will have a high impact on the health and welfare of the Medicare eligible community population by describing which co-morbid states are associated with the presence of oral diseases, whether or not preventive treatments are associated with lower levels of those co-morbid states, and whether or not the use of corrective treatments are associated with amelioration of those co-morbid states in comparison to the frequency and severity of those co-morbid states among individuals whose oral diseases are not treated. Specifically we will: Aim 1. Estimate the relationship between utilization of dental care services by elderly persons and co- morbidities over time. The working hypothesis is that elderly persons not using dental care service are more likely to experience specific co-morbidities compared to those using dental services on a regular basis. Aim 2. Estimate the relationship between preventive dental care service use, oral health problems, and co- morbidities over time by elderly persons. The working hypothesis is that elderly persons with regular use of preventive dental care or without oral health care problems are less likely to experience specific co-morbidities over time than elderly persons not regularly using preventive dental care or with oral health care issues. Aim 3. Estimate the relationship between oral disease and co-morbidities over time for elderly persons. The working hypothesis is that elderly persons with oral disease are more likely over time to experience specific co- morbidities compared to elderly persons without oral disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The p53 tumor suppressor and master regulator is central to human DNA repair, damage checkpoints and many aspects of human biology. It controls cell fate in response to many types of internal and environmental stress. Importantly, most cancers are altered for p53 function. We have discovered a greatly expanded universe of p53 targets and human diversity as well as variations in responses to specific stresses. The DNA binding and transactivation of p53 is critical for tumor suppression. There is considerable variation in p53 dependent expression across 100s of targeted genes leading to differences in p53-mediated biological consequences. Much of that is due to target RE sequence. As described below the universe of genes subject to direct p53 control is much larger than originally anticipated based simply on previously established response element (RE) consensus target sequence, consisting of 2 copies of the sequenceRRRCA/TT/AGYYY (R, purine;Y, pyrimidine) separated by a spacer of up to 13 bases. There is considerable flexibility in what constitutes a functional RE. We focus on RE functionality, i.e., the ability of REs to support transactivation by p53. To directly assess transactivation responsiveness of human REs, we developed promoter systems in budding yeast for variable p53 expression (i.e., rheostatable). This has been used to establish the functional evolution of REs across species (discussed below) and has been summarized in our piano model that describes functional variability within a transcriptional network. We have translated many of the findings in yeast to p53 in human cells in culture and ex vivo. Recently, we established functionality of and sites. Also, increases in spacer length of only a few bases in a target RE greatly reduces functionality in terms of p53 transactivation. With Gilbert Schoenfelder (Berlin), we identified a novel mechanism of p53 transcriptional control of genes in studies of a SNP located in the promoter of the Flt1 gene. This C to T SNP results in generation of a perfect p53 -site RE and is present in 5% of the population. Our findings provided the first report of a functional p53 -site. Endogenous p53-dependent induction of FLT-1 mRNA was only observed in cell lines containing the FLT1-T allele in response to several DNA damaging agents. These results established that p53 can differentially stimulate transcription at a polymorphic variant of the FLT1 promoter and placed the VEGF system directly in the p53 stress-response transcriptional network via FLT1. We recently examined transactivation capacity towards a panel of 15 p53 -sites in p53-null human osterosarcoma cells (SaOS2) transiently co-transfected with a WT p53 vector. Some 1/2 site REs supported p53 transactivation to levels similar to a weakly responding full-site RE. This finding greatly expands the potential p53 master regulatory network. We are currently using p53 ChIP-seq approaches to identify targeted genes and networks, investigate crosstalk between regulatory networks especially other stress and DNA response networks. INTERACTION OF p53 AND ER REGULATORY NETWORKS. We identified a -site estrogen receptor RE (ERE) that greatly increased p53 induced transactivation at the FLT1 -site p53 RE, thereby establishing a new dimension to the p53 master regulatory network. Recently, we addressed the generality of synergistic transactivation by p53 and ER acting in cis. The 1/2 site in the FLT1 promoter was replaced with various 1/2 sites as well as canonical weak and strong human p53 REs. p53 transactivation was greatly enhanced by ligand-activated ER acting in cis. Enhanced transactivation extends to several cancer-associated p53 mutants with altered function, suggesting ER-dependent mutant p53 activity for at least some REs and possibilities for reactivation of cancer mutants. We propose a general synergistic relationship between the p53 family and ER master regulators in transactivation of p53 target canonical and noncanonical REs which might be poorly responsive to p53 on their own. We are developing a functional matrix as a tool for genome-wide searches for putative p53 target genes via noncanonical sites and augmenting transcriptional factors. In a limited search to identify motifs containing both strong p53 noncanonical RE sequences (including decamers with a CATG core) and associated ER responsive sequences (FLT1-like motifs), we identified the human RAP80 gene and Toll-like receptors (TLRs) that determine innate immunity. The identification of p53 target REs associated with the TLR genes has led to exciting new findings. Among the 10 human TLRs, nine had canonical and noncanonical p53 REs that may be functional. Using primary human cells obtained in a collaboration with the Clinical Research Unit we examined expression of the entire TLR gene family following exposure to anti-cancer p53 inducing agents. Expression of all TLR genes, TLR1-10, in blood lymphocytes and alveolar macrophages from healthy volunteers is inducible by DNA metabolic stressors. However, there is considerable inter-individual variability. Similarly p53 dependent TLR expression is detected in human cancer cell lines. For some TLRs the p53 control seems to enhance the inflammatory responses, mediated by activation of TLRs in the presence of natural ligands. Furthermore, a polymorphism in the TLR8 promoter provides the first human example of a p53 target RE sequence specifically responsible for endogenous gene induction. CANCER-ASSOCIATED P53 MUTANTS. Nearly all cancers have mutant or reduced expression of the p53 tumor suppressor gene. Using yeast-based and human cell systems, we show that functional p53 mutations can lead to considerable diversity in the spectrum of responses from REs including 1) decrease/loss-of-function;2) subtle changes;3) altered specificity;and 4) super-transactivation all of which lead to variation in biological responses. Most of the functional mutants (8/9) were able to function at 1/2 sites. We extended our analysis of p53 mutants in yeast-based systems to a clinical study of breast cancer patients undergoing neoadjuvant treatment. Several transcriptionally active p53 mutants identified in breast had subtle defects in transcription at REs that can only be revealed at low expression levels achievable in the yeast-based system. It appears that nonfunctional as compared with functional missense mutants are more likely to exhibit stage III progression at diagnosis, high grade cancers and to relapse with cancers at distant sites. P53 NETWORK EVOLUTION. We are investigating evolution of REs in terms of responsiveness to p53. Individual REs exhibited marked differences in potential transactivation as well as widespread turnover of functional REs during p53 network evolution. Only 1/3 of the REs found in humans are predicted to be functionally conserved in rodents. Surprisingly, the p53 responsiveness of the DNA repair/metabolism set of 15 p53 targeted genes in humans has evolved separately from mice suggesting differences in responsiveness to cancer-inducing agents between mice and humans. Importantly, we found functional conservation of weakly responding REs including 1/2 sites. Among validated p53 REs conserved between rodents and humans, one third were comprised of 1/2- or -sites, each with a perfect consensus -site suggesting a selective advantage in retaining weak p53 REs. Importantly, the integration of the TLR gene family into the p53 network also appears unique to primates.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of this research continues to refine and improve the methodology of diffusion tensor imaging (DTI) at very high fields (3T and 7T) for the investigators and collaborators on the P41 RR09784 \"Center for Advanced MR Technology at Stanford\" effort. Preliminary studies from a variety of studies suggest that DTI may predict the cognitive and motor performances of a patient from scans requiring several minutes. Diffusion is measured along at least six non-collinear directions. For each gradient direction, typically four images were acquired and averaged. Two images with no diffusion weighting (b = 0s/mm2) are acquired and a set of Inversion Recovery (IR) images for CSF nulling are acquired with b = 0s/mm2;these images were used to unwarp the diffusion weighted images, which resulted in a more robust unwarping than using the non-IR b=0 images. The development of SENSE and more robust parallel imaging sequences and post-processing methods has accelerated the use of high fields for diffusion studies. Issues regarding the opimal protocol and set of sequence parameters needed for various diffusion studies are under investigation, which includes more rapid protocols for the assessment of stroke and more high resolution protocols for diffusion tensor imaging of white matter tracts. The effort is being translated from 3T to newer field field studies at 7T.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "OBJECTIVE: To determine whether behavioral and hormonal responses to stress in marmosets are altered by social and reproductive status. RESULTS In human neuropsychiatric disorders, chronic alterations in baseline levels of the stress hormone cortisol are often associated with altered hormonal responses to stress. I have been investigating whether chronic suppression of baseline cortisol levels in socially subordinate, reproductively suppressed female marmosets are similarly associated with altered hormonal or behavioral responses to psychological stressors. Results indicate that psychological stressors, including restraint and exposure to novelty, cause dramatic elevations in circulating levels of both cortisol and the pituitary hormone adrenocorticotrophic hormone (ACTH), as expected, and that the magnitude of the endocrine stress response increases with increasing stressor intensity. Preliminary analyses, however, do not support the hypothesis that individual differences in baseline hormone levels, associated with differences in reproductive and social status, are accompanied by differences in the hormonal response to stress. If confirmed, these results woud indicate that the neuroendocrine mechanisms mediating the endocrine stress-response, unlike those controlling baseline cortisol levels, are not highly responsive to social and reproductive factors. FUTURE DIRECTIONS In the coming months, I will complete the analysis of the hormonal and behavioral stress esponses to either confirm or disprove the hypothesis that suppression of baseline plasma cortisol in socially subordinate female marmosets is associated with altered endocrine and behavioral responses to stressors. KEY WORDS dominance, subordination, ovarian cycle, adrenal cortex, adrenocorticotrophic hormone, glucocorticoids. stress", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "That early postnatal experience may have long-term repercussions has been demonstrated for visual, auditory, tactile and chemical sensory systems. Multiple studies employing abnormal stimulation of these systems shortly after birth revealed significant alterations in their functionality in the adult individuals. These studies also pointed to three types of most-frequently observed early experience-induced alterations in the sensory circuitry which are likely to underlie the aberrations in the functionality of sensory systems in animals subjected to abnormal early experience; (i) alterations in connectivity of the neuronal processes, detectable as changes in the density and distribution of projecting sensory fibers; (ii) alterations in neurochemical organization of the circuitry, detectable as changes in the density and distribution of specific neurotransmitter and receptor-expressing cells as well as modifications in the levels of expression of neurotransmitters and receptors in the relevant centers of there central nervous system; and (iii) alterations in receptive field size and/or stimulus-response properties of neurons within the sensory circuit, detectable as changes in cells electrophysiological responses to appropriate sensory stimulation. Nociceptive circuitry undergoes significant postnatal maturation, and thus should also be vulnerable to modulations by early pain-associated experience. Indeed, recent studies demonstrated that exposure to pain-associated events early in life could induce long-term alterations in responses to pain by the affected individuals. Furthermore, experiments conducted in newborn rats have demonstrated that even a single local inflammation of a handpaw lasting for approximately 24 hours can result in significant changes in future withdrawal responses to noxious stimulation in the affected animals. However, the specific parameters of the \"window(s) of vulnerability\" of the developing nociceptive circuitry to such short- lasting inflammation, as well as long-term changes in nociceptive circuitry underlying the altered nociceptive behaviors, have not been well investigated. It is unknown how global are the long term-effects produced by local inflammatory insult. Therefore, in this project: I. We will characterize the early postnatal sensitive period(s) of nociceptive circuitry to modulation by short-lasting inflammation; II. We will test the hypothesis that short-lasting early local inflammation is capable of inducing long-term alterations in morphological, biochemical and electrophysiological properties of the peripheral and spinal nociceptive circuitry serving the affected peripheral region (this specific aim will be performed in conjunction with Project 3, which focuses on the ability of early local inflammation to affect ion channels and electrophysiological properties in cells of the dorsal root ganglia); and III. Together with Projects 1 and 4, we will test the proposition that early local inflammatory insults may have a global long-term effect on nocifensive responses of the organism and that this global effect may be associated with alterations in the descending modulatory circuitry.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this proposal is the characterization of a locus that confers resistance to mammary and intestinal tumor development. These studies will provide insight into factors that control the complex process of tumor development. The identification of genes that affect tumor development will aid in our ability to design prevention or treatment strategies. Mouse models provide an excellent system with which to identify genes that affect cancer development. ApcMin/+ mice are predisposed to develop intestinal and mammary tumors. Gtrosa26 mice carry a retroviral gene-trap insertion of LacZ-neoR on chromosome 6. These mice express the fusion protein ubiquitously and are thus a useful tool in chimeric analyses. We have found that when ApcMin/+ mice also carry the Gtrosa26 insertion, they are resistant to mammary tumor development and the intestinal tumors are reduced in size, but not number. The first aim of this proposal is to test two hypotheses for the molecular mechanism of resistance to tumor development in mice carrying the Gtrosa26 insertion. One is that the resistance is a function of the high levels of expression of the beta-galactosidase-neomycin resistance fusion protein that is encoded by the insertion. The second hypothesis is that the disruption of the expression of two non-coding transcripts at the insertion site is the cause of the resistance. In the second aim, we propose experiments that explore the mode of action of the insertion. Specifically we will test for an effect of the insertion in other mammary tumor models. This will test whether the effect is a general effect on tumor development or is specific to the Apc pathway of tumor development. Second, we will test whether the Gtrosa26 insertion acts in a tissue autonomous manner. This information will be vital in the understanding of how the insertion exerts its effects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objective of this research effort is to identify mechanisms which may be responsible for or contribute to differences in disease severity among patients with bronchial asthma. The research effort considers both immunologic mechanisms. A lymphocyte beta-adrenergic receptor defect has been demonstrated to be present among some patients with bronchial asthma. The magnitude of the receptor abnormality, as determined by specific dihydroalprenolol binding assays, appears related to disease severity and degree of airway obstruction. Drug consumption and documentation was noted on all patients studied. A significant relationship was shown between quantitation of airway reactivity by methacholine challenge and disease severity score. Higher disease severity scores also correlated with urinary norephinephrine and dopamine excretion. The results suggest that a major determinant of disease severity among patients with bronchial asthma is differences in nonspecific airway hyperreactivity among individuals. Abnormalities of beta-adrenergic receptors in conjunction with alpha-adrenergic hyperresponsiveness contribute to disease severity. The importance of adrenergic factors in disease severity may have important clinical and therapeutic implications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primate visual cortex uniquely possesses a regular array of metabolically active, cytochrome oxidase (C.O.)-rich zones (blots or puffs) in the supragranular layers with distinct physiological properties, particularly those related to color processing. Our previous studies indicate that unilateral retinal impulse blockage in the adult severely affects the most metabolically active neurons and induces synaptic reorganization within the puffs. These findings suggest that the mature visual cortex is not static but, rather, responds dynamically to altered functional demands. Besides changes in puffs, our preliminary light microscopic analysis of surrounding C.O.-poor interpuff regions indicate compensatory increases in C.O. levels within zones related to the non-treated eye. Such dynamic changes in the adult are of obvious clinical and functional importance, consequently our initial ultrastructural and quantitative analyses of the puffs will be extended to the interpuff regions. Selective vulnerability of the most metabolically active neurons deserves further investigation, since it appears to implicate a specific neurotransmitter type, GABA. Whether all puff neurons with intense C.O. activity are GABAergic, or whether GABAergic neurons encompass a wide range of oxidative metabolic capacities, will be examined by means of combined C.O. histo- or immunohisto-chemistry and GABA-immunohistochemistry on the same histological section. To directly address the relationship of C.O. levels to physiological activity and visual processing, single neurons will be recorded extracellularly from C.O.-rich and poor zones in normal cortex and during periods of physiological modification: 1) following monocular retinal impulse blockage in the adult and 2) during the critical period of postnatal development, when the innately determined cortical organization undergoes further physiological maturation. To understand the anatomical basis of these changes during development and specifically to explore the dynamics of maturational plasticity, we will examine structural reorganization in developing visual cortical neurons following retinal blockade. In summary, our approach is to combine histochemical, immunohistochemical, ultrastructural, and physiological observations to yield an integrated understanding of metabolic adjustments to altered functional demands within developing and mature neurons of the primary visual cortex.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chromatin structure is critical to the regulation of eukaryotic gene expression. The basic unit of chromatin, the nucleosome, regulates transcription by limiting the accessibility of transcriptional factors to specific DNA sequences within promoters. A better biochemical understanding of how eukaryotic genes are regulated will require reconstituted transcription in the context of genomic DNA packaged with properly organized nucleosomes. Our lab recently used the latest in DNA sequencing technology to show that the in vivo pattern of nucleosome positioning can be recapitulated in vitro across the S. cerevisiae genome by chromatin remodelers. Here we propose to develop a genome- wide immobilized template assay that will allow us to ask fundamental questions regarding nucleosome dynamics, transcription factor occupancy, and RNA transcription on a genome-wide scale in the context of proper chromatin structure. With this assay aspects of gene expression regulation will be investigated in regards to transcription factor binding and the transcriptional requirements of histone modifications. These studies are intended to shed light on two fundamental questions in gene regulation: Can transcription factors intrinsically access their binding sites in native chromatin, or is the cooperation of remodelers needed? Does the physiological pattern of active histone modifications arise before or after transcription occurs? The development of this assay will lead to a technique in which this in vitro system will be programmed to contain specific aberrant chromatin states as are seen in disease states in order to determine how these changes in chromatin structure lead to the development of cancer, along with cardiovascular, neurodegenerative, and autoimmune diseases. PUBLIC HEALTH RELEVANCE: Chromatin structure is critical to the regulation of eukaryotic gene expression, and alterations of chromatin structure are known to cause aberrant regulation of gene expression that can lead to cancer. Here we are proposing to develop a biochemical assay in yeast that will later be used to study the aberrant regulation of gene expression caused by specific alterations of chromatin structure to understand how it leads to the development of cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This Unit investigates the mechanism of action of excitatory amino acids as synaptic transmitters and neuromodulators in the vertebrate CNS, utilizing cell culture and electrophysiological techniques. Substantial progress has been made in developing a fast perfusion system for applying drugs and ions to nerve cells, and this is now used routinely. The divalent cations, zinc and cadmium, have two major effects on hippocampal neurons: (1) block of excitatory responses to NMDA receptors (Zinc Kd = 13 micromolars); (2) an increase in due to block of postsynaptic inhibitory GABA receptors (Zinc Kd = 11 micromolars), and zinc suppression of a transient potassium current, which normally slows repetitive firing. Zinc block of NMDA responses is reduced on raising the extracellular calcium concentration suggesting competition between zinc and calcium or screening of the zinc binding site by calcium. Low concentrations of zinc (50 micromolars) also potentiate responses to kainate, quisqualate, and response to glutamate at non-NMDA receptors. Fast application of excitatory amino acids produces three patterns of response: fast (tau = 20 ms) desensitization of quisqualate receptors: slow (tau = 200 ms) desensitization of NMDA receptors, sustained activation of kainate receptors. Concanavalin-A, which binds to glycoproteins, reduces desensitization at quisqualate but not NMDA receptors, and does not alter responses to kainate. Low concentrations of L- glutamate @ 1 micromolar) were found to depress excitatory synaptic transmission via activation of a novel presynaptic receptor: L-AP4 mimics this effect. Neither agonist produces a substantial postsynaptic response in glycine free medium. Culture medium is conditioned by substances secreted into the extracellular space, including L-glutamate, which tonically inhibits synaptic transmission; on the other hand neuronal survival in F-12 medium reflects activity of a glial sink for neurotoxic amino acids, which rapidly reduces the L-glutamate concentration from 100 less than 10 micromolars.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Methamphetamine is one of the most growing drugs of abuse, causing neurological impairments, and associated with behaviors that favor exposure to HIV, a virus that penetrates the CNS leading to neuroAIDS. Meth users infected with HIV usually present a more severe form of neuroAIDS, due to effects on neurons. IT is known that free radicals participate in the enhancement of brain pathology, but the cell types as well as pathways affected by Meth that lead to free-radical mediated stress. The hypothesis behind this proposal is that Meth interacts with cell types other than neurons will be explored, particularly regarding its potential to induce and/or modulate reactive oxygen and nitrogen intermediates. We will investigate the action of methamphetamine (Meth) on oxidative metabolism, directly on macrophages, and on other cell types in vitro, as well as in vivo, in conditions of co-morbid factor interaction. For that we will examine the action of Meth on cell lines, by investigating how the drug affects the intensity and quality of the oxidative burst, and searching for genes within ROI and RNI pathways that exhibit transcriptional pattern changes upon Meth exposure. Such transcriptional changes will be validated by looking for similar changes in cells obtained from Meth-treated animals infected with SIV, which have been previously characterized for CNS disease parameters. The results from this project will offer important clues on the participation of Meth in pathology mediated by oxidative stress by acting on ROI and RNI induction pathways, not only in the Central Nervous System, but also in peripheral organs. PUBLIC HEALTH RELEVANCE: We will investigate the action of methamphetamine (Meth) on oxidative metabolism, directly on macrophages, and on other cell types in vitro, as well as in vivo in conditions of co-morbid factor interaction. The hypothesis behind this proposal is that Meth affects the intensity and quality of the oxidative burst by acting directly on macrophages and/or glial cells, in addition to its action on neurons, contributing to the development of pathology mediated by oxidative stress, not only in the Central Nervous System, but also in peripheral organs. Initially, we will focus on the effect of meth on various human and mouse macrophage (THP1, RAW264.7), microglia (MG5, EOC 2), astrocyte (C8-D1A, CCF-STTG1), and neuronal (SH-SY5Y, SK- N-MC) cell lines, regarding the production of O2_, H2O2, and other reactive oxygen intermediates (ROI), as well as Reactive Nitrogen Intermediates (RNI) using various chromogenic methods. Mitochondria activity upon interaction with meth will be also addressed on cells stimulated with Meth. Following a characterization of the different cell lines regarding production of free radicals and mitochondrial changes in reaction to Meth, we will choose good responders to investigate transcriptional changes on molecules within ROI and RNI pathways, caused by action of Meth. For that we will use multiplex technology, as well as enzymatic checkpoint inhibition strategies. These approaches will help dissect the molecular basis of respiratory burst induction by Meth, and will distinguish whether Meth interferes over mitochodria, peroxisome, or both. Following the in vitro approach using cell lines, we intend to validate findings in the context of pathology. We will use cryopreserved cells and tissues from monkeys infected with SIV (as a model for HIV infection, a common co-morbid condition) subjected to a Meth treatment schedule. We will identify transcriptionally altered oxidative pathways on cryopreserved brain and peripheral tissues derived from SIV-infected Meth-treated monkeys by qRT-PCR. This brings relevance to pathology in humans. The present proposal will identify the basis of oxidative stress in the brain and other organs in Meth users. Meth is one of the most growing street drugs of abuse, exposing users to HIV infection, and potentially facilitating the penetration of virus across the blood brain barrier (BBB) into the brain. In this proposal we will dissect mechanisms by which Meth can induce and modulate free radical production, causing an aggravation on the severity of damage associated with virus presence in the brain tissue, and consequently aggravate AIDS-associated CNS dysfunctions. The results obtained in this proposal will allow the generation of data on direct effects of the drug on cells, relative susceptibility of different cell types to direct actions, and on pathways, aiming a future R01 application on modulation of oxidative stress in Meth abuse in vivo. These will lead to the development of important therapeutic tools for rehabilitating individuals, especially patients presenting co-morbid factors, such as HIV infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this project is to understand the systems-level neural mechanisms by which general anesthetics suppress consciousness and allow its return during emergence in the human brain. Our fundamental hypothesis is that consciousness emerges from brain function as a network phenomenon, and that general anesthetics suppress consciousness by disrupting the communication across large-scale neuronal networks that support information integration in the brain. Our previous findings suggest that anesthetics alter the functional connectivity of thalamocortical systems, particularly in the nonspecific thalamocortical division, as well as in other intrinsic cortical cognitive networks. Here, we build on our decade-long developments in blood-oxygen level-dependent (BOLD) functional MRI (fMRI) and resting-state functional connectivity MRI (R- fcMRI) methods with high spatial and temporal resolution applied to test specific hypotheses about the anesthetic modulation of cognitive functioning, network organization, reorganization, and information integration during wakefulness and at graded levels of suppressed consciousness. In Specific Aim 1, we will employ concurrent behavioral and BOLD fMRI assessment of semantic discrimination to test the hypotheses that deepening anesthesia with propofol will suppress conscious awareness by diminishing integrative functional networks of the brain in a graded, top-down manner, suppressing the most complex systems first and the simplest systems last and that these changes can be characterized by the anesthetics' effect on the behavioral response and neural activity to a series of tasks that depend on different levels of information integration. In Specific Aim 2, we will test the hypotheses that propofol confers differential changes in resting- state (baseline) functional connectivity, modularity and network integration in thalamocortical and cortical intrinsic networks, particularly those involved with attention, executive control, and salience, vs. others, such as the default mode and sensory networks. Finally, in Specific Aim 3, we will test the hypotheses that anesthetic- induced loss and subsequent return of consciousness are mediated in part by different neural mechanisms that show prior state dependency, as reflected by various brain network interaction measures and that the restoration of consciousness from anesthesia requires additional neural resources over those required for the maintenance of consciousness, reflecting the reconfiguration capability of the brain as a self-organizing system for resource management and functional resilience. The proposed work should advance our understanding of the neural mechanisms of anesthesia with respect to its effect on human consciousness at an integrative level. The work should reveal the order in which cognitive functions are lost during sedation and anesthesia and the degree of residual cognitive functions based on direct, noninvasive detection of neural events. It should illuminate how anesthetics alter resting-state intrinsic brain networks and how the latter may reconfigure to cope with anesthetic challenge. Novel quantitative approaches to assess residual cognitive functions by fMRI network analysis may eventually be extendable to neurological patients with disordered consciousness. In a wider context, the findings should facilitate our understanding of the scientific basis of human consciousness, including its aspects for sensory awareness and voluntary action.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose to continue the study of glutathione synthesis and metabolism in the lens in which we have already made considerable progress. The maintenance of a high level of reduced glutathione is recognized as essential in the prevention of certain types of cataract formation and progression. Utilizing procedures already established in this laboratory for organ lens culture, enzyme purification, assay, and kinetic studies, we plan 1) to investigate certain key enzymes in glutathione synthesis and metabolism, especially in relation to aging and safe use of pharmaceuticals. These enzymes include coenzyme A-acetyl transferase, glutathione S-transferase and the mercapturic acid pathway, and the first three enzymes in the transformation of methionine to cysteine. 2) To determine the amount of free and bound glutathione in situ in both aging and cataractous lenses by means of NMR spectroscopy; and the concentrations of ADP and substrates of glutathione synthesis in the aging human lens by HPLC or the amino acid analyzer. 3) To establish the roles of glutathione peroxidase and catalase in the prevention of oxidative damage in the organ cultured lens. 4) To study the effect of both reduced and oxidized glutathione on Na-K-ATPase. 5) To examine the possible role of a glutathione analogue, arising from homocysteine, in patients suffering from homocystinuria. These studies will further expand our knowledge of glutathione and its role in the maintenance of a clear lens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research project consists of experiments on the induction of rhabdomyosarcomas in rats. The two stage model of tumor induction is used. The initiating agent is a subcarcinogenic dose (0.4 mg) of nickel subsulfide (Ni3S2) injected into the rectus abdominis muscle; the promoting treatment consists of three slight compressions made at weekly intervals, of the area infiltrated with the carcinogen. It is known that a single muscle compression induces promptly proliferation of mononucleated cells located at a short distance from the site of compression. Cell division is considered to be vital importance for neoplastic transformation. The use of the two stage method makes it possible to schedule tissue removal in concomitance with cellular mitotic activity. Study with the electron microscope will lead to establish whether the cells in divisions are cells that have cleaved from injured multinucleated muscle fibers or are satellite cells. Information will also be gained on the role of the dividing cells in tumor formation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application is being submitted to continue the University of Pennsylvania's role as a main institution in the Eastern Cooperative Oncology Group beyond its 37 years of continuous particpation. The overarching objectives are to continue to serve as a scientific and administrative resource for the Group, to enter patients onto ECOG protocols, and to coordinate and support a large affiliate network. In the latter role, this application will support the diffusion of knowledge of clinical trials, assist in the proper conduct of clinical trials, and make these trials accessible to the broadest possible population of patients with cancer. Although the University of Pennsylvania has had an outstanding track record in all aspects of cooperative group activities, including accrual, administrative and scientific input and data quality, the specific aims of the proposal for the next six years are to: 1. continue to enhance accrual in all disease site and modalities, beyong our current particpation, particularly in breast and lung cancers. 2. increase physician, nurse, and data management participation in Group activities, including protocol development. 3. attract new affilaites through the Abramson Cancer Center network, allowing for enhanced availability for clinical trials participation in the larger community The relevance of this project to the ultimate cure of cancer is self-evident. Cooperative groups are the main force outside of the pharmaceutical industry dedicated to the definitve tesing of new treatments, studying not just new drug development but also non-drug treatment, such as radiation therapy and surgery, for which there would be nooter funding. These cooperative group activities, and the specific involvement of the University of Pennsylvania, represent the chief way in which the ultimate model of multimodal, collaborative patient care can be performed. The main ECOG grant describes in detail the very specific way in which clinical trials have led to RELEVANCE: Malignancies remain an enormous burden for individuals in the United States and for society as a whole. Without public support of clinical trials, both by financial support as well as participation in clinical trials, progress will not be made. If left in private hands, large areas of cancer treatment will go unexplored, denying patients the possibility of progress.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract Endosomes, the membrane compartments inside living cells, are increasingly recognized as discrete ?hubs? that regulate the network of cell signaling circuits in space and time. Physical phenomena like the clustering of proteins on endosome membranes and the active transport of endosomes are hypothesized to play a key role in these regulatory mechanisms. Unfortunately however, the direct evidence needed to support this level of mechanistic understanding of endosome signaling is lacking. Over the last four years, the research program of my lab has focused on exploring the wealth of physical phenomena involved in the endocytic process. We have uncovered new physical mechanisms of endocytosis in immune cells, but the questions we pose are of general relevance to many kinds of cells. We aim to test the general hypothesis that endosomes are a specialized platform for the spatiotemporal regulation of cellular signal transduction. This proposal highlights two of our developing project areas that are designed to test this hypothesis by identifying the biophysical mechanisms of endosome signaling regulation. Both are enabled by our established biophysical tools that allow us to manipulate and analyze the signaling activities and dynamics of endosomes in living cells. One research direction focuses on the mechanisms of signaling crosstalk on endosome membranes. Our ultimate goal for this research direction is to identify mechanisms by which physical interactions between endosomal receptors lead to their signaling crosstalk. By developing a novel approach that physically manipulates interactions between receptors on endosome membranes, we will establish the quantitative relationship between receptor clusters on endosomes, their signaling crosstalk, and the end-point cell response. The other research project addresses the functional roles of endosome trafficking in signaling. Our ultimate goal for this second research direction is to determine mechanisms under which the transport and subcellular location of endosomes regulate their signaling functions. By developing a particle reporter system that will allow us to magnetically control trafficking of single endosomes and simultaneously detect their signaling activities, we will reveal direct connection between the dynamical, mechanical and biochemical activities of individual endosomes. The proposed research directions are enabled by the novel integration of nanomaterial engineering, quantitative physical measurements, and advanced optical techniques, with live cell experiments. In the long term, we will expand our research scope from endosome signaling in immune cells to that in other cell types. Our ongoing and future research directions share the overarching goal of establishing a quantitative understanding of endosome signaling in living cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This training grant proposal is entitled The cellular and molecular foundations of biomedical sciences. It is a renewal of a successful program started six years ago and seeks to train graduate students in the broad fundamentals of biomedical sciences to provide them with the tools to understand important problems in modern biology and health. This is accomplished initially through a two semester Core course covering Genetics, Genomics and Bioinformatics, Eukaryotic molecular biology, Molecular biophysics, Protein structure and function, and Cell biology. Students follow this with electives in specific fields of their interest. This interdepartmental program is based in the Department of Biological Sciences, but has been expanded to include established faculty from Columbia's College of Physicians and Surgeons. Students undertake rotations in up to three laboratories and can do their thesis work in any of the 47 faculty in the program, roughly half of which are on the Medical School campus. This has provided expanded opportunities for training of the students in outstanding laboratories and has fostered collaboration between faculty and students at Columbia's Medical School and main campuses. The faculty members work in a broad range of fields including structural biology, molecular biology, genetics, bioinformatics and systems biology, developmental biology and neurobiology. Experimental systems are also broad including bacteria, yeast, drosophila, C. elegans, frogs and mice. Human tissue culture systems are also utilized and most systems consider problems relevant to human health and physiology. Twelve graduate student traineeships are requested per year to support six students for two years each, usually in their third and fourth years. This will fund 19% of our current training grant eligible student pool of 63. The best students in our program will be chosen to be funded by this grant. This will be determined by their performance in the Core and other courses, recommendations from their rotation and thesis sponsors, and a short research proposal reviewed by the grant's steering committee. The qualifications of the faculty and the research facilities at Columbia are outstanding and provides a superb training environment for the students. RELEVANCE: The training of the next generation of biomedical scientists is essential to continued excellence of research in the U.S. and advances in understanding, controlling and curing human disease. Laboratories in the program address specific basic problems in biomedical sciences, many with direct applications to human disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Flow Cytometry Project Summary/Abstract The primary purpose of the Flow Cytometry Module is to provide Principal Investigators with a centralized service for cell phenotyping, sorting (FACS) to obtain viable populations of cells according to specific cellular markers, and biomarker detection. Such a service makes expensive equipment for fluorescent activated cell sorting and analysis available to researchers. In addition, the module helps develop experimental procedures and assists with analysis as well as the interpretation of results. Finally, the module provides full time technical support for the maintenance and calibration of the equipment, a centralized internet access to data and a database for techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Reduction of racial and ethnic disparities in cancer incidence, stage of presentation and outcome requires resilient strategies that link medically underserved communities into the continuum of care. Community- based participatory research has the potential to generate such strategies, through processes of collaborative planning, experimentation and discovery. The proposed Queens Library Health Link program is designed to test the efficacy of a comprehensive participatory research approach to reduce disparities by improving use of cancer prevention, screening and treatments interventions. In this project, public library branches provide a base for neighborhood organizing, education, data gathering, planning and implementation of local health promotion experiments and dissemination of results. We will work with 42 of the 63 branch libraries in Queens, each serving its own large, diverse segment of the Queens community. Although we will follow the same process consultation protocol, every community will undertake unique health promotion activities most suited to local priorities, cultures, preferences and opportunities. This project is designed to address the following aims: (1) To determine whether community-based participatory action research (PAR) leads to more effective cancer prevention and control in underserved communities compared to standard, \"top-down\" programs; (2) To examine how communities build upon and sustain locally-developed initiatives for cancer prevention and control; (3) To compare whether cancer prevention and control strategies developed through PAR are more widely accepted and more efficacious than standard strategies when they are transferred to new communities. This project will be carried out in two phases. In phase 1, we compare health behavioral outcomes of community-tailored strategies to address cancer needs with top-down alternatives. In phase 2, we examine the sustainability and dissemination of tailored strategies. Queens neighborhoods will be randomized to serve as settings for program development or dissemination. Data will be gathered from multiple sources to examine the participatory processes as well as behavioral outcomes at the individual, library and community levels. Core organizations involved in this project comprise a four-way partnership including the Queens Borough Public Library System, the Queens Cancer Center, the American Cancer Society of Queens, and Memorial Sloan-Kettering Cancer Center.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this research is to define the relative roles of intra-and extra-mitochondrial genetic systems in determining cellular and mitochondrial phenotype. Major emphasis will be placed on analyzing the regulation of mitochondrial biogenesis. We propose to apply both genetic and biochemical techniques to this end. In this project attention will be focussed on the biosynthesis of heme and its relationships to the formation of cytochromes. In addition, a comprehensive study will be made of the role of copper in regulation of heme formation and cytochrome aa3 biogenesis. Three projects are proposed. In the first, the mechanism by which delta-aminolevulinate (Alv) synthase is transported into mitochondria will be studied. Evidence for specific membrane receptors and pilot sequences on cytoplasmic Alv synthase will be sought employing specific mutants. In the second, a mutant which over-produces cytochromes will be characterized to determine whether the primary defect involves alteration in heme production or in a general regulator of apocytochrome formation. The third project will examine the mutual interactions between copper metabolism, heme biosynthesis and cytochrome formation. This study will include a characterization of copper transport, copper proteins and the regulation of copper metabolism as it pertains to mitochondrial biogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Development of an effective therapy against post-exposure Anthrax. Anthrax is an infectious disease caused by the bacterium Bacillus anthracis. This rod shaped bacterium infects humans through the respiratory system, skin, or digestive tract. Dependent upon the entry route into the human body, Anthrax can be highly lethal. Although cutaneous Anthrax is rarely lethal, inhalation Anthrax is dangerous and usually fatal. Upon inhalation, the Anthrax spores adhere to the alveolar macrophages and germinate. Bacteria migrate to the lymph node, in which they rapidly multiply and excrete a tripartite exotoxin comprised of protective antigen (PA), lethal factor (LF, a Zn2+ dependant metalloproteinase) and calmodulin-activated edema factor adenylate cyclase (EF). The combined actions of these proteins constitute the Anthrax toxin which induces cell death particularly in macrophages by the action of LF. Once in the cytosol, LF is able to cleave several members of the MAPKK family near their N-terminus preventing interaction with, and phosphorylation of, downstream MAPKs, thereby inhibiting one or more signaling pathways. High liter of the bacteria and concomitant high level of LF in the blood stream will lead to the death of the host organism. Therefore, the developments of treatments that include a combination of antibiotics with orally active, potent, safe and selective LF inhibitors should provide an efficacious countermeasure to post-exposure Anthrax. Our three specific aims are: (I) To increase, through iterative optimizations selectivity, potency and drug-likeness of selected LF inhibitors. (II) To perform in vitro and in cell toxicity studies to refine hits and identify drug-like inhibitor(s) of LF for subsequent in vivo studies. (Ill) To perform in vivo efficacy evaluations and necessary formulation, PK and toxicity studies. Relevance Inhalation Anthrax is the most deadly form of this disease. After infection, Anthrax spores are engulfed by alveolar macrophages where germinate to generate the bacteria, which spread through the lymph nodes to the bloodstream, eventually leading to systemic fatal shock presumably due to the action of the secreted Anthrax toxin. LF (lethal factor) is a key component of the secreted toxin and plays a major role in inducing apoptosis (cell-death) of macrophages by cleaving the specific human cell signaling proteins. Our plan is to develop potent, safe, selective and orally active compounds that inhibit LF, which should prevent macrophage cell-death and promote termination of the disease. Such inhibitors will be tested as single agents and in combination with antibiotics, in order to develop the most efficacious post-exposure therapy that should facilitate and assure the recovery of patients who harbor the disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Summary: Despite their high prevalence, the molecular basis for common genitourinary (GU) congenital defects is poorly understood. We have identified and validated gene lesions at the KCTD13/16p11.2 locus as responsible for lower GU tract abnormalities, particularly cryptorchidism and hypospadias. KCTD13 encodes a substrate-specific adapter of a BCR (BTB-CUL3-RBX1) E3 ubiquitin-protein ligase complex, which regulates the actin cytoskeleton and cell migration via ubiquitination and degradation of RHOA. Notably, RHOA has been implicated in the maintenance of Sertoli-germ cell junctions to promote gametogenesis and gonads development. E3 ubiquitin ligases also regulate the androgen receptor (AR), which is key to male genitalia development and differentiation. We found that KCTD13 is robustly expressed in the GU tract and have recently shown that in comparison to normal controls, gene copy number variants are remarkably common in patients with GU birth defects. Moreover, we obtained in vitro and in vivo evidence that KCTD13 loss affects AR levels in testis and penis. We also observed that haploinsufficient and Kctd13 deficient mice had significantly higher incidence of cryptorchidism and decreased size of testes, seminal vesicles and penis size in conjunction with spermatogenic defects, causing severe subfertility. Further, Kctd13 null mice revealed a significant decrease in masculinization factor SOX9 levels and concomitant upregulation of the feminization factor RHOA. While our collective data suggest a key role of KCTD13 in male GU development, the mechanisms of how this molecule impinges upon the AR and/or SOX9 axes remain a major gap of knowledge. We hypothesize that gene dosage changes in KCTD13 alter the signaling of the masculinization axes, leading to abnormal GU tract development, defective gonad formation, undervirilization, and subfertility. In line with our reasoning, we will test two plausible independent mechanisms by which KCTD13 mediates lower GU tract development and differentiation. First, we hypothesize and test that gene dosage changes in KCTD13 affect GU tract development by affecting AR degradation, subcellular localization and downstream gene targets (AIM 1). Second, we hypothesize and test that KCTD13 affects the expression of masculinization factor SOX9 directly by modulating SOX9 ubiquitination or indirectly by regulating RHOA degradation, which in turn leads to defects in testis and penile development and differentiation (AIM 2). Finally, we identify and characterize the mutations in KCTD13- CUL3 pathway that are critical in GU development such that they could be used to generate a diagnostic kit for patients with disorders of sexual development (AIM3). Completion of the studies in this proposal will advance our understanding of the molecular mechanisms that underlie common GU birth defects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 34 million people are infected with human immunodeficiency type-1 (HIV-1) world-wide. More than two million new infections occur annually (1,2). These statistics could be reduced if at-risk individuals were provided antiretrovirl drugs for HIV pre-exposure prophylaxis (PrEP). We propose to develop a combination nano-microbicide containing cellulose acetate phthalate (CAP) as a HIV-1 entry inhibitor with dolutegravir (DTG), a HIV-1 integrase strand transfer inhibitor, for highly effective PrEP. Our preliminary data show that novel CAP-DTG combination nano-microbicide reduce cytotoxicity of DTG to cells. In vitro HIV prophylaxis using HIV indicator TZM-bl cells show that CAP-DTG combination nano-microbicide offers significantly higher protection from HIV-1 infection as compared to DTG solution and CAP nanoparticles indicating the importance of combination nano-microbicide. This proposal will extend our preliminary experiments and evaluate CAP-DTG nano-microbicide in thermosentive gel and DTG solution based conventional gel for cytotoxicity, intra-cellular delivery of DTG, in vitro HIV PrEP, and vaginal pharmacokinetics. In order to reach this goal, Specific Aim 1 focuses on development of DTG solution based conventional gel (DTG-gel), optimization and characterization of CAP-DTG combination nanomicrobicide for DTG loading, and incorporation of CAP-DTG combination nanomicrobicide into a thermosensitive gel (CAP-DTG-NPs-Gel) for vaginal delivery. Chemical stability of DTG in conventional gel and CAP-DTG-NPs-Gel will also be assessed. Specific Aim 2 employs various cell lines and primary human cells to evaluate DTG-gel and CAP- NPs-DTG-Gel for cytotoxicity and intra-cellular delivery of DTG. The ability of DTG-gel and CAP-NPs-DTG-Gel to offer protection from HIV-1 infection will be tested in TZM-bl cells, PBMCs, and MDMs. We will evaluate vaginal pharmacokinetics of DTG-gel and CAP-DTG-NPs-Gel using female NOD SCID gamma (NSG) mice for up to 7 days. Drug dissemination will be determined using HPLC analysis to measure DTG levels in dissected tissue. We will evaluate tolerability of DTG-gel and CAP-DTG-NPs-Gel after repeated administration for 7 days by analyzing inflammatory cytokines in cervicovaginal fluid and immunohistochemical analysis of NSG tissue. As a laboratory within an undergraduate biology department with collaborators in attached professional schools, we are uniquely situated to expose undergraduates to state-of-the-art nanoparticle research. This project will incorporate education of undergraduate students in our investigation of the utility of combination CAP and DTG nano-microbicide in a thermosensitive gel delivery system as cost-effect, highly efficacious modality for HIV pre-exposure prophylaxis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study will compare the efficacy and safety of two doses of a new inhaled corticosteroid, fluticasone proprionate (FP) usinf one of two delivery systems. Asthma is a potentially debilitating disease in children, likely accounting.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neuroendocrine (NE) tumors such as carcinoid, islet cell tumors, and medullary thyroid cancer are the second most common cause of isolated hepatic metastases. These tumors often cause debilitating symptoms due to excessive hormonal secretion. Besides surgery, there are limited curative and palliative treatments available to patients with metastatic NE tumors, emphasizing the need for development of other forms of therapy. In this proposal, we will present data illustrating that GSK3B inhibition may be a viable strategy for the treatment of NE tumors. Furthermore, we will show that the potent GSK3B inhibitor, lithium chloride, suppresses NE tumor proliferation and hormone production. Thus, we propose to conduct a Phase II clinical trial to determine if lithium is a potential therapy for patients with NE tumors. The primary objective in this aim is to evaluate the response rate for patients with NE tumors treated with Lithium. The secondary endpoints are: 1) the progression-free and overall survival of patients with NE tumors treated with Lithium; 2) the effect of Lithium on tumor markers specific for NE tumors; 3) the toxicity and tolerability of Lithium in this patient population; 4) the quality of life of patients with NE tumors treated with Lithium and 5) the expression of GSK3B signaling pathway members in the NE tumor samples from patients enrolled in this clinical study before and during lithium therapy. In summary, these studies are part of our long-term goal of designing and developing novel methods to treat and palliate patients with unresectable, metastatic NE tumors. Neuroendocrine (NE) tumors such as carcinoid, islet cell tumors, and medullary thyroid cancer are the second most common cause of isolated hepatic metastases. These tumors often cause debilitating symptoms due to excessive hormonal secretion. Besides surgery, there are limited curative and palliative treatments available to patients with metastatic NE tumors, emphasizing the need for development of other forms of therapy. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "TheVARR&DCenterforNeurorestorationandNeurotechnology(CfNN)attheProvidenceVAMedical Center(PVAMC)unifiesadistinguishedteamofscientists,engineersandcliniciansfromtheVA,Brown University,andaffiliatedhospitalstodevelopandtranslateneurotechnologiesanddevice-basedtherapies thatrestorefunctionforVeteranswithdisordersaffectingthenervoussystem.CfNN?sthreeFocusAreasare eachdedicatedtoperformingtherigorous,peer-reviewedsciencethatleadstonewdiagnosticand therapeuticstrategiesforVeteranswithspinalcordinjury,stroke,amyotrophiclateralsclerosis(ALS), epilepsy,limbloss,post-traumaticstressdisorder(PTSD),depression,chronicpain,andothernervous systemdiseasesorinjuries.CfNN?sthreeFocusAreasare:(1)RestoringCommunicationandMobility(2) RestoringAffectiveandCognitiveHealth,and(3)RestoringLimbFunction.TheFocusAreas?interdisciplinary researcheffortsaresupportedbythreeCoresprovidingessentialinfrastructuresupportandspecialized services:(A)NeuromodulationandNeuroimaging(B)Recording,Decoding,andComputational Neuroscience,and(C)Assessment,OutcomeMeasurement,andImplementation.CfNNresearchisfurther facilitatedbyadedicatedadministrativestaffwhosupportscientificproductivity,coordinatethesynergistic relationshipsbetweenCfNNanditsacademicandclinicalaffiliates,maintainclosecontactwithlocaland nationalVAofficials,andoptimizeourservicetoVeterans.CfNN?sresearchstrategymaximizesinteractions viacomplementaryexpertiseandresourcesamongitsFocusAreasandCores.Thisinturnsupportsahigh- impactresearchcommunityfocusedonapplicationofmedicaldevicestoneurologichealth,bothwithinthe VAandinthewiderregionalandnationalbrainsciencecommunity.Throughthisrenewal,overthenextfive yearsCfNNwillleverageandexpanditscollaborativeresearch,bothwithintheProvidenceVAMCand togetherwithotherVACenters,towardrehabilitationgoalsimportanttothelargenumberofaffected Veteransandtheirfamilies.Theseinclude:restoringarmandlegmovementforVeteranswithstroke,spinal cordinjury,oramputation;?improvingcommunicationforVeteranswithALS;?improvingtheunderstanding, earlydetection,andmanagementofseizures,and;?moresuccessfullytreatingVeteranswithPTSD, depression,obsessive-compulsivedisorder,orotherseverecognitiveoraffectivedisorders.CfNNalsoaims todevelopandvalidatetheoutcomesmeasuresneededforeffectiveclinicaltrialsofnewrestorative neurotechnologiesfortheseconditions.Inadditiontotheresearchofitscoreinvestigators,CfNNisdesigned tofacilitatethetrainingandresearchcareersofjuniorresearchersandtorecruitestablishedcliniciansand scientiststofocustheirresearcheffortsonimprovingrehabilitationforVeteranswithnervoussystemdisease orinjury.CfNNiscommittedtoprovidingseedfundingforpilotstudiestoenableaffiliatedinvestigatorsto launchpromisingresearchandcollectpreliminarydatatoinformandstrengthenlargerresearchprojects. ThroughitscloseassociationwithBrownUniversity,CfNNprovideseducationalopportunitiesto undergraduates,graduatestudents,neuropsychologyinterns,andpostdoctoralresearcherstolearnmore abouttheVA,inspiringthemtoshapetheircareerstowardnotonlygainingabetterunderstandingof fundamentalhumanneuroscience,buttowardusingthatknowledgetohelpdesign,test,anddeploynovel device-basedneurotherapeuticstosupportVeteransHealth.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The specific goals of the research study proposed for this fellowship are to examine interpersonal obsessive-compulsive disorder (OCD) and to assess how these processes can be modified in order to improve patients' treatment outcomes, while simultaneously providing invaluable training in the administration of a family treatment. The investigator will design and implement a brief family intervention for families with the aim of reducing family members' accommodation of patients' OCD symptoms. The intervention will take place during two 90-minute sessions and will encompass psychoeducation regarding OCD and anxiety, as well as modeling and role-play of appropriate responses to requests for accommodation. Half of the family members in the study will be randomized to receive the intervention, and half will be randomized to the no-intervention condition. Measures of family accommodation will be taken at 5 timepoints throughout the study, including a 1-month follow-up after patients have completed treatment. The investigator will also assess levels of depression and anxiety in family members, as well as family members' critical or rejecting attitudes toward the patient. The study will examine: 1) whether participation in the intervention is associated with greater change in family accommodation compared to a no-intervention condition, 2) whether family members' participation in the intervention is associated with greater symptom reductions in patients, 3) whether patients' treatment outcomes are mediated by changes in the level family accommodation, 4) the nature of the relationship between family functioning and accommodation and the effect of the intervention on measures of family functioning, and 5) whether improvements in accommodation and patients' symptoms are maintained at follow-up assessment. The broader aim of this study is to begin determining specific parameters for an effective family intervention, including targets for treatment and appropriate treatment length. In light of the growing acknowledgement of important underlying similarities across anxiety and mood disorders (Barlow, 2002; Brown, 2007; Brown & Barlow, 2009), this deeper understanding of the effective components of family interventions is likely to have significant implications across a range of different diagnoses. The lifetime prevalence of anxiety and mood disorders in America are estimated to be 29% and 21% of the population, respectively (Kessler et al., 2005), and empirically-supported interventions which address patients' broader interpersonal contexts have the potential to bolster the success rates of our current treatments, reducing dropout and relapse rates.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goals of the currently funded competing renewal grant P50 DC000242-21 are the following: 1) To determine the boundaries of selection criteria for the acoustic and electrical speech processing implant. 2) Refine and expand how electrophysiologic measures of auditory function can be used to understand how the auditory system responds to electrical stimulation. 3) Investigate factors that influence music perception and enjoyment of Cl recipients. 4) Describe and explain individual differences in perceptual, linguistic, and social outcomes, of individuals receiving cochlear implants in childhood. The purpose of this supplement is to complement the above goals of the currently funded P50 grant by studying ways to: improve and determine benefits of binaural cochlear implants using more diverse and realistic soundfield hearing tests. Specifically, our goals are to improve: (1) bilateral signal processing; (2) bilateral fitting; and (3) the evaluation of bilateral and unilateral devices. Several experiments are planned and will determine if: (1) moderately directional microphones are superior to highly directional microphones (the latter might exclude important binaural cues); (2) fixed directional microphones are superior to adaptive directional microphones; (3) linear amplitude coding is superior to amplitude compression; (4) synchronous amplitude compression across devices can preserve binaural cues; (5) the use of a \"conditioning\" stimulus improves binaural hearing; (6) electrophysiological measures that match electrodes across ears result in improved bilateral performance; and (7) bilateral fittings are superior to unilateral fittings in patients who are experienced unilateral listeners. We use several new tests, including ones of distance perception, movement direction, speech recognition with localization cues, and speech recognition while performing a visual thinking task. We believe our proposal is innovative in the comprehensive set of new tests we have designed, in our exploration of new bilateral fitting and signal processing approaches, and our comprehensive evaluation of bilateral and experienced unilateral implant users. The impact of these experiments is very high, in that we hope to be able to improve hearing in challenging everyday conditions and will have direct relevance for the larger population of hearing-aid users. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of these studies is definition of the role of cyclic nucleotides as possible regulators of mammalian gastric secretion, with particular emphasis on the source and activity of gastric cyclic nucleotides and the \"second messenger system\" of cyclic nucleotides as mediators of gastric acid secretion in response to gastrointestinal hormones, other hormones, cholinergic release, adrenergic blockade, fasting, refeeding, H2 receptor antagonism, hyperchlorhydria and achlorhydria. We plan to determine whether cyclic nucleotide responses in the intact canine stomach are representative of events occurring exclusively in parietal cells by measuring intra- and extracellular cyclic nucleotide concentrations after gastric secretory stimulation by histamine or pentagastrin. In order to resolve the problem of measurement from parietal cells exclusively, we will attempt to study responses in dogs using the antral (acid-secreting) and fundic (acid-secreting) mucosa to secretagogue stimulation. We plan also to measure the responses to a variety of stimulatory and inhibitory agents with measurements in vitro of the key enzymes involved, including cyclase, phosphodiesterase and protein kinase in addition to gastric cyclic nucleotide activities. We also plan to further develop an isolated mammalian parietal cell preparation which would permit direct metabolic study of the parietal cells and resolve the problem of structural localization of presumed key enzymes inth cyclic nucleotide system in the analytical approach of acid generation mechanisms. We plan to further define the localization of cyclic nucleotide activity by developing immunofluorescence techniques which will selectively localize cyclic AMP and cyclic GMP to the parietal cells and indicate their response to secretagogue stimulation. We plan to determine the source of cyclic nucleotide activity in gastric juice by utilizing double isotopic techniques and inert markers to calculate both plasma and gastric cyclic nucleotide content in order to determine whether simple plasma clearance or active secretion from the mucosa accounts for the presence of cyclic nucleotides in canine gastric secretions. BIBLIOGRAPHIC REFERENCE: Schwartzel, E.H., Jr., Bachman, S., and Levine, R.A.: Cyclic nucleotide activity in gastrointestinal tissues and fluids. Fed. Proc. 35:583, 1976 (abstract).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ATAD2 is an important co-activator of the estrogen and androgen receptors. ATAD2 is known to be up- regulated in multiple different types of cancer including breast, lung, gastric, endometrial, renal, and prostate. Up-regulation of ATAD2 is often correlated with poor patient outcomes, and can be used as prognostic marker. Furthermore, silencing the expression of ATAD2 through RNA interference inhibits cell proliferation and promotes apoptosis in ovarian carcinoma, and inhibits migration and invasion of hepatocellular carcinoma and colorectal cancer cells. ATAD2B, is a poorly studied paralog of the ATAD2 gene, and although ATAD2 and ATAD2B are highly conserved, there is little known about the function of ATAD2B or its role in oncogenesis. Both the ATAD2/B proteins contain two conserved domains: an AAA ATPase domain and a bromodomain. The overall objective of the proposed research is to determine how di-acetyllysine recognition by the ATAD2/B bromodomains regulates the cellular function of these proteins. This proposal aims to: (1) characterize how cross-talk between histone modifications modulate acetyllysine recognition by the ATAD2/B bromodomains; (2) outline the molecular mechanism(s) of di-acetylated histone recognition by the ATAD2/B bromodomains; (3) determine the functional significance of di-acetyllysine recognition by the ATAD2/B bromodomains. A unique combination of in vitro biochemical, biophysical, and structural biology studies on the ATAD2/B bromodomains will be coupled with in vivo functional genomic investigations using a breast cancer progression model to characterize the biological roles of the ATAD2/B bromodomains. We will evaluate the impact of neighboring histone modifications on histone H4 tail recognition using peptide array assays in combination with isothermal titration calorimetry (ITC) and nuclear magnetic resonance (NMR) chemical shift perturbation techniques. We will determine the structural features of ATAD2/B bromodomains required for recognition of di-acetylated histone tail ligands using NMR and/or X-ray crystallography. To characterize the binding mode of the ATAD2/B bromodomains with their histone ligands we will carry out analytical ultracentrifugation, size-exclusion chromatography, ITC, and NMR T1/T2 relaxation experiments. Site-directed mutagenesis coupled with NMR and ITC will be used to measure the effects on ligand binding, and identify differences in the binding pockets of the ATAD2/B bromodomains. We will compare the genome-wide associations of ATAD2/B with histone H4 acetylation patterns in a breast cancer progression model to determine the impact of ATAD2/B on breast cancer cell phenotypes using ChIP-seq and RNA-seq, followed by cellular migration and invasion assays. Our multi-faceted approach will correlate specific histone modifications with ATAD2/B binding and action, which will allow us to connect histone H4 acetylation marks to bromodomain function in cancer cell proliferation. Overall, our integrated biochemical, biophysical, structural biology and functional genomics approach will reveal the biological roles of ATAD2/B and facilitate the discovery of novel drug targets to help overcome cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sustaining adherence to antiretroviral therapy (ART) is central to HIV management. For children in resource- limited settings, no well-validated measures of adherence exist. A reliable, valid measurement tool would provide more efficient, effective HIV care for this vulnerable population. My long-term goal is to work as a clinician-scientist who develops instruments to improve pediatric HIV care systems In resource-limited settings. In the near term, I want to focus on a reliable, valid, cost-effective way to measure children's ART adherence because this is central to the challenge of providing long-term HIV management. The objective of this application is to develop and test a reliable, valid instrument to measure pediatric ART adherence for children ages 0 to 14 years in western Kenya. The rationale for developing a validated adherence measure is to gain the ability to measure adherence accurately, test interventions to improve adherence, and assess if therapies optimize clinical outcomes. This work will be done within a long-standing US-Kenya partnership, the Academic Model for Providing Access to Healthcare (AMPATH). AMPATH cares for over 80,000 adult and pediatric HIV-infected patients in western Kenya, including over 2,800 children on ART. I plan to accomplish the research objective of this application by pursuing the following three specific aims: Aim 1: Develop a reliable, valid comprehensive pediatric ART adherence measurement questionnaire (CAMP - Comprehensive ART Measure for Pediatrics);Aim 2: Develop a reliable, valid, short-form version of the pediatric ART adherence measurement tool (SF-CAMP) for use as an adherence screening measure in busy clinical care environments;Aim S: Evaluate the field-readiness, implementation feasibility, and clinical utility of CAMP and SF-CAMP within the AMPATH HIV clinical care system in western Kenya. I also propose a course of training, study, and mentorship in the design and testing of health behavior measurement tools for International settings, the use of psychometric analyses to develop and validate clinical tools, and evaluation of pediatric HIV care. The combination of research work and my career development plan will enable me to become an independent clinician-scientist who can improve pediatric HIV care provision. PUBLIC HEALTH RELEVANCE: The contribution of this study is expected to be a reliable, valid instrument to measure pediatric ART adherence within a resource-limited setting. The proposed research is significant because we must measure adherence accurately to provide long-term clinical management for HIV-infected children and to find effective interventions to prevent viral resistance and poor outcomes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Purpose of this grant application is to perform studies aimed at understanding the structure, structure-function relationships, and molecular pathology of the platelet membrane receptor, glycoprotein (GP) Ib-IX complex, GP Ib-IX is essential for normal platelet adhesion to and activation on thrombogenic surfaces. It exerts its function by interacting with the adhesive molecule, von Willebrand factor (vWF). and with the platelet agonist, thrombin. Deficiency or defective function of GP Ib-IX complex leads to the congenital bleeding disorder known as Bernard-Soulier syndrome. The interaction of platelets with vWF through the GP Ib-IX receptor has also been shown to be relevant for the development of pathological platelet thrombi responsible for vascular occlusion. Therefore, studies on the structure and function of GP Ib-IX are relevant for understanding the pathogenesis of the thrombotic complications of atherosclerosis, as well as for unraveling the physiological mechanisms of platelet function. We propose to use an integrated approach based on the use of proteolytic fragments, synthetic peptides, monoclonal antibodies, and expressed mutant molecules to identify the amino acid residues responsible for vWF and thrombin binding to GP Ib-IX. The amino terminal domain of the alpha-chain of GP Ib (the 45 kDa domain),previously shown to contain the vWF- and thrombin-binding sites, will be subjected to limited proteolysis to generate smaller fragments retaining binding function. Synthetic peptides will be constructed to represent the sequence of functionally relevant epitopes of GP Ib-Alpha. The identification of amino acid residues related to the binding function of GP Ib-IX will represent the basis for selective mutagenesis experiments that will be used to obtain final confirmation of the chemical structure of functional domains. Moreover, the molecular pathology of different forms of Bernard-Soulier syndrome, including a newly described variant, will be explored to establish the bases of the genetic abnormalities responsible for this diseases, and also to obtain information on the functionally relevant structures of the GP Ib-IX complex from the study of naturally occurring mutants with defective binding activity. These studies will provide concepts and reagents to probe the functional relevance of the GP Ib-IX complex in normal hemostasis and pathological thrombosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A unique method for measuring filtration in single isolated glomeruli has been developed. This technique permits study of the ultrafiltration characteristics of glomeruli from any species. Values from the ultrafiltration coefficient of normal glomeruli of rats, rabbits and dogs have been established. An increase in glomerular size and ultrafiltration in the developing rabbit has been documented. Glomerular function in several forms of acute renal failure has been studied. Further studies will be directed toward developing a more precise description of the changes in capillary volume which occur during filtration in vitro and further defining the relation between glomerular size and cell morphology and filtration. In addition, glomerular function in several physiologic states, eg. intravascular loading and depletion will be studied, and the in vitro effects of several vasoactive substances will be investigated. These studies will enhance our understanding of normal glomerular function and of the mechanisms which regulate glomerular function in normal and disease states.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Aging is the single most important risk factor in human disease in developed countries, and it is thought that an understanding of aging mechanisms would prompt design of rational therapies against age-related diseases. A corollary is that a pharmacological agent that slows aging itself is likely to be effective against a wide spectrum of diseases. To discover what classes of chemical compounds slow aging, we will screen a large and diverse chemical library for extension in the lifespan of the nematode C. elegans. We have developed a number of automated screening methods that exploit C. elegans handling technology. We will also screen a library of compounds generally recognized as safe (GRAS) for extended lifespan to bias our search towards compounds that will prove useful in mammals. We will establish whether the compounds extend lifespan by acting on known pathways and determine their impact on general metabolism and stress resistance. We expect some compounds will prolong lifespan by novel mechanisms in which case we will identify the target proteins. Overall, we aim to identify a range of compounds that slow aging;these compounds will be leads for the development of therapeutics against a range of age-related diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The central objective of this proposal is to determine the regulatory mechanisms controlling the accumulation of specific proteins during mouse central nervous system development. 1) The accumulation of the tissue specific enzyme glutamic acid decarboxylase will be examined to determine if the increase in the quantity of the protein is due to a change in the rate of synthesis, the rate of degradation, or a modulation of both parameters. 2) The tissue specific S-100 protein is actually a class of related proteins; we will complete experiments to determine whether each of the mouse S-100 proteins is a direct product of mRNA translation or whether they result from an interconversion process. 3) Mouse CNS mutants showing the appearance of abnormalities at 10 to 25 days postnatally will be surveyed for quantitative and qualitative alterations in soluble proteins. The mutant spastic which does have alterations in brain amino acid pool levels will be examined in detail to determine if this is a result of a change in the activity of an enzyme. If this is found, we will determine whether this change is due to the synthesis of an altered protein or the production of an abnormal amount of the enzyme.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "EXCEED THE SPACE PROVIDED. Scientific research conducted in a Culture of Mentoring is fortified vastly with respect to both qualityand productivity. This culture, which is being actively cultivated at the Oklahoma Medical Research Foundation, has played a vital role in the generation of numerous innovative and fundamentallyimportant biomedical research breakthroughs credited to this Institution. Recognizing the important role of this culture in strengthening the research infrastructureat the OMRF and connected research environments, we describe in this proposal a mechanism by which we seek to formalizeand stratify the mentoring process at OMRF, with the goal of creating a self-sustaining system designed to maximize both individual and collective research potential. Within the context of this mission we seek to: 1) integrate and diversify individual and Program research efforts with the aim of achieving specific research objectives in the areas of immunology and neurobiology, and 2) strengthen individual and collective research efforts by reinforcing selected Core Research Facilities. The cumulative impact of these aims is the cultivation of a strong, supportive, and thoroughly progressive research environment at OMRF, which functionsboth directly and indirectly to reinforce and energize biomedical research here and throughout the state of Oklahoma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Interparental conflict is a common early environmental stressor with potential long-term consequences for emotional development and risk for psychopathology. Current research findings suggest that interparental conflict is associated with behavioral and physiological indices of emotion regulation during infancy (Crockenberg, Leerkes, & Lekka, 2007; Moore, 2010). Further, differences in biological systems associated with conflict during infancy have been identified as moderators of risk for school-aged children exposed to conflict (El-Sheikh et al., 2009). However, researchers have not yet explicated how exposure to conflict during infancy may get under the skin to affect development of key neural systems. In the proposed study, we will utilize a neuroimaging technique, resting state functional connectivity magnetic resonance imaging, to examine differences in functional neural networks between infants from high and low conflict families. The cortical and limbic (cortico-limbic) level of the biological stress response system and a set of functionally connected regions known to be more activated during rest states, the default network (Raichle et al., 2001), will constitute the focus of this study due to evidence linking the development of these networks both to early adversity and to risk for psychopathology. This work will build on the applicant's ongoing study of two groups of 6- to 12-month- olds screened for high and low interparental conflict. Within this study, fMRI scans are conducted with infants during natural sleep. The purpose of the proposed investigation is to examine whether, among infants from high conflict families, regions of the cortico-limbic network involved in emotional reactivity are less functionally connected to regions important for regulatory processes. We will also investigate whether functional connectivity between regions of the default network previously shown to be less established in preterm infants (Smyser et al., 2010) will be less strong in infants from high conflict families. The current study represents a step towards examining common early environmental stressors in terms of potential consequences for neural networks that have been implicated in mental health disorders. Eventually, such work may facilitate the identification of targets for prevention and intervention and provide a basis for program evaluation in terms of reducing risk at a biological level. The proposed fellowship training covers stress neurobiology in the context of environmental risk, fMRI methods with challenging populations, and rs-fcMRI analyses. Training also focuses on the collaborative process that plays a central role in translational research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The gram negative bacterium Neisseria gonorrhoea is the causative agent of gonorrhea in man. Many basic questions concerning gonococcal pathogenicity remain unanswered due to the lack of an adequate animal model for this disease. The research outlined is designed to examine genetic mechanisms by which the gonococcus regulates the expression of a major surface structure, the pilus. Gonococcal pilin is a member of a family of pilins that share amino acid sequence homology. The pilin genes from N. gonorrhoea, Pseudomonas aeruginosa, Bacteroides nodous, and Moraxella bovis all show significant DNA sequence homology within their structural genes, and also show DNA sequence homology to the sigma factor binding sequence present in the 5' untranscribed regions of nitrogen fixation and nitrogen utilization genes. In E. coli, expression of a pilin beta-galactosidase fusion is stimulated 40- fold by expression of a cloned Klebsiella pneumonia nifA gene. Pilin expression has been found to be regulated by two gene products from pilA and pilB. The relationship between pilA and pilB regulation and the sigma factor binding site will be investigated utilizing gene fusions and site directed mutagenesis. The gene encoding the sigma factor will be isolated and the effect of mutations in this activity assayed. The possibility of a regulatory system related to nifA, but distinct from pilA and pilB will also be investigated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Several investigators have demonstrated cytoplasmic particles from human leukemia cells with characteristics of oncornavirus. We have successfully induced the production of oncornavirus-like particles from cultures of human leukemia cells. The presence of similar particles has been demonstrated in patients suspected to have \"premalignant\" conditions, two of which subsequently developed leukemia and lymphoma respectively. Based on this work, the broad objectives of this proposal are (1) to determine the biological significance of oncornavirus in human leukemia and preleukemia, and (2) to determine the nature and activity of particles from human leukemia cells. Bone marrow cells from preleukemic, immune deficient, and renal transplant patients will be studied following in vitro culture for the presence of oncornavirus-like particles. Human leukemia cells will be grown in vitro. Proteins from these particles will be studied and compared to known oncogenic agents. Human leukemia cells will be cocultivated with human and possibly non-human cell lines in an attempt to induce a chronically infected cell line. These studies will provide data relevant to the occurrence of oncornavirus-like particles in various disease states. The establishment of long term productive cultures of human oncornavirus-like particles and/or the demonstration of related proteins in easily accessible mammalian oncogenic agents will provide material for further biological survey by biochemical and immunological techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a report on progress achieved during the third year of the program. The aims have been to establish a program of social science research and training in the study of drug use. Research has focused particularly on drug behavior among youth, cross-cultural studies of drug use within the U.S. and in other countries, the evaluation of drug treatment programs, and the methodologies of drug research. During this period, drawing on computerized sets of data from drug studies at Columbia, a number of cross-study analyses--as well as new research projects--have been undertaken in these areas of interest. Other activities during this period include the expansion of drug research training facilities at Columbia and the continuation of a Columbia University Seminar on Drugs and Society. Goals for the coming year include conducting additional cross-study analyses of data already collected as well as new research in the U.S. and abroad in the areas of interest. Also planned for the coming year is a conference on the contributions of social science research to policy-making in the drug field.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project has been expanded to encompass two related research studies. The first study has analyzed data derived from a randomized clinical trial of diagnostic ultrasound use during pregnancy conducted by the team of Norwegian investigators in Trondheim, Norway. The purpose of the analysis is to examine fetal growth patterns using longitudinal measurements throughout pregnancy of: (1) symphyseal-fundal heights; (2) weight gain at each prenatal visit; (3) serial biparietal and abdominal diameter measurements from ultrasound; and (4) maternal hemoglobin level. Regression models have been fit to the serial measurements for each mother. The coefficients of the regressions have been analyzed in relation to various indicators of birth size such as weight, crown-heel length, ponderal index, and birth weight-for-gestational age percentile. Using an analysis of covariance procedure, additional factors (e.g., cigarette smoking, alcohol intake, low maternal prepregnancy weight, etc.) will be tested for significance in modifying intrauterine growth patterns. In addition to the study described above, a prospective study to determine risk factors for intrauterine growth retardation, or small-for-gestational age birth, was begun in 1984 through the research contract mechanism with both the University of Alabama in Birmingham and University of Trondheim, Norway (in collaboration with the Universities of Bergen and Uppsala). The study protocol includes recruitment of pregnant women before 17 weeks gestation. Those enrolled in the study will be carefully monitored throughout the remainder of their pregnancy. Symmetric and asymmetric forms of intrauterine growth retardation will be assessed prenatally and at delivery. Infants born to the study mothers will have follow-up exams during the first year of life to assess catch-up growth and attainment of early developmental milestones.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The bi-directional translocation of lipids from one side of a biological membrane to the other is termed flip-flop. Lipid flip-flop across the endoplasmic reticulum (ER) membrane is required for protein N-glycosylation and GPI-anchoring. These protein modifications are essential in eukaryotes; for example, their genetic abrogation causes embryonic lethality in mammals and renders yeast unviable. Lipid flip-flop across the ER is also required for membrane biogenesis: phospholipids that are synthesized on the cytoplasmic face of the ER must be translocated to the opposite face to enable the membrane bilayer to grow uniformly. The demand for lipid flip-flop at the ER is likely to be exceptionally high when the ER membrane expands and glycoprotein secretion increases; this occurs, for example, during the differentiation of B-lymphocytes to antibody-secreting plasma B cells. Unassisted flip-flop is extremely slow because of the energy barrier to taking the polar lipid head group through the hydrophobic interior of the membrane, yet lipids flip-flop rapidly across the ER membrane on a time-scale of seconds. This is because the ER possesses specific transport proteins (flippases) that accelerate lipid flipping to a physiologically sufficient rate. Lipid flipping in the ER occurs by an ATP-independent mechanism in which the flippases facilitate 'downhill' transport of lipids; this distinguishes ER flippases from other translocators, typically found in the eukaryotic plasma membrane, that couple ATP hydrolysis to concentrative 'uphill' transport of lipids. We estimate that there are as many as six different ER lipid flippases but none of these have been identified at the molecular level. We developed biochemical reconstitution systems that recapitulate the activity of three of the flippases required for ER membrane bilayer expansion and protein glycosylation. These flippases specifically translocate glycerophospholipids, oligosaccharide diphosphate dolichols and mannose-phosphate dolichol. Our aim is to identify these physiologically important translocators with the long-term goal of understanding their mechanism of action. We propose to do this via a two-pronged approach involving protein purification and mass spectrometry on the one hand, and screening of systematic collections of yeast ER membrane proteins on the other. Our purification efforts will be aided by the use of novel affinity matrices. We will also use partially purified flippase preparations to continue our efforts to define the specificity of these proteins. Our published work and preliminary data put us in an excellent position to accomplish these aims. PUBLIC HEALTH RELEVANCE: Flipping of lipids from one side of a biological membrane to the other is necessary for membrane expansion during cell growth, as well as for the biosynthesis of molecules that play critical roles in human and microbial physiology. These molecules include glycoproteins such as the neural cell adhesion molecule, GPI-anchored proteins such as acetylcholinesterase, glycolipids such as the receptor for cholera toxin, components of the cell walls of bacteria and yeast, and the O-antigen of E. coli lipopolysaccharide. We are interested in identifying the transport proteins that catalyze lipid flipping in yeast and mammals and understanding how they work.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Microbial transcriptional dynamics have been known to be highly dynamic and heterogeneous between cells, seen from microscopy based and targeted sequencing approaches. Such heterogeneity can be an asset or a liability; from a fitness perspective, transcriptional heterogeneity is a prerequisite for survival under changing environments; however, the modern scourge of antibiotic resistance may be ascribed to such heterogeneities. In either case, there is an enormous need to characterize the transcriptional dynamics at the resolution of individual microbial cell. However traditional approaches relying on one or a few selected reporter genes are inadequate for this challenge. Recent technical advances now allow us to use RNA-Seq to profile single mammalian cells. Massive and early barcoding followed by pooling or manipulation by microfluidics have increased the scale to tens of thousands of cells. However, these technologies have thus far failed to translate to single microbial cells due to (1) difficulty in single microbial cell lysis, especially those with thick cell wall; (2) difficulty to capture and barcode relatively sparse microbial mRNAs, especially when lacking polyA tails (in bacteria); and (3) large population size and complexity of microbial population that require orders of magnitude more cells be sampled in an experiment. We will leverage droplet microfluidics, develop physical, chemical and enzymatic lysis methods, and investigate novel molecular biology and sequencing techniques to develop a single-cell microbial genomics pipeline to (1) Isolate and (2) lyse single microbial cells; (3) capture and barcode the mRNA of single microbial cells; and (4) process 104-105 cells per sample with hundreds of distinct transcripts per cell. Barcoded RNA will then be pooled and sequenced at high depth. These tools will be modular and have broad applicability beyond RNA-Seq, including single cell epigenomics and proteomics. New physical lysis modes investigated will include MEMS, laser ablation, acoustic waves, and plasmon resonance. The proposed project will significantly advance current technologies which are limited in throughput or the number of RNA molecules measured. Currently, there is no successful strategy for single microbial cell RNA- Seq at scale. Our strategy will enable cost-effective and generalized single-cell RNA-Seq in microbes at massive throughput. Novel, hybrid microfluidic devices containing silicon, nanomaterials and elastomeric components will be developed for single microbial cell lysis, barcoding and library prep.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Six studies of the adult mentally retarded living and working in community facilities will test the influence of opportunity to interact; similarity choice for cognitive level and individual differences in sociability on enduring friendship choice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lyme disease (LD) is a tick-borne, multi-system, infectious disorder caused by the extracellular spirochetal bacterium Borrelia burgdorferi (Bb). Since Bb lacks exotoxins or known bacterial secretory system, it is widely believed that the inflammatory manifestations of the disease result from the host's innate and co-evolving adaptive immune responses to the bacterium. Monocytes and macrophages are considered to be critical cellular elements of the innate immune response to the spirochete. For more than a decade, Bb-mediated cell activation was thought to occur chiefly as a result of the interactions of the spirochete's abundant outer membrane-associated lipoproteins with CD14 and Toll-like receptors (TLR) 1/2 on the surface of these cells. We now have extensive evidence that phagocytosis of live spirochetes by human monocytes and murine macrophages generates a more intense and far broader inflammatory response than can be attributed to lipoprotein-mediated, cell surface TLR1/2 activation. Phagocytosis of intact Bb also induced transcription of interferon-2 (IFN-2) and type I interferon-stimulated genes (ISGs), independently of TLR2. Recent evidence from the investigator's laboratory provides substantial evidence the TLR2-independent signaling events elicited by Bb in human monocytes are MyD88-dependent and occur via TLR8. On the basis of our collective findings we now propose a new model of Bb-induced monocyte activation, which emphasizes the importance of phagocytosis and the cooperative role of TLR2 and TLR8 signaling. In this model, binding of Bb to the monocyte/macrophage cell surface, through a yet to be characterized phagocytic receptor, is followed by a broad sequence of immune signaling events which mechanistically can only be integrated following internalization of the bacterium and formation of the phagolysosome. The validation of the phagosomal signaling model, the centerpiece of our research strategy, will significantly enhance our understanding for how the bacterium triggers the inflammatory processes that under actual disease conditions cause tissue damage and/or that promote bacterial clearance. To accomplish our goals and examine mechanistic aspects of the proposed model, we have formulated the following Specific aims. In Aim 1 we will use a ex vivo stimulation technique to characterize in detail the mechanisms by which phagocytosis of Bb elicits activation and TLR1/2 and TLR8 dependent inflammatory signals in human monocytes. In Aim 2, we will examine key elements of the phagosomal signaling model in Bb-infected human and murine macrophages. The use a large repertoire of available knockout mice will allow us to define mechanistically how Bb is sensed and triggers immune responses in these cells. In Aim 3 we will characterize MyD88 dependent and independent responses to Borrelia burgdorferi both in vitro and in vivo using human monocytes and macrophages obtained from children and adults with known congenital deficiencies in components of the MyD88 signaling pathway (IRAK-4 and MyD88 deficient). PUBLIC HEALTH RELEVANCE: Lyme disease (LD) is a tick-borne infectious disorder caused by the spirochetal bacteria Borrelia burgdorferi (Bb), which has continued to increase in endemic areas and has spread geographically, paralleling the distribution of its primary vector, Ixodes ricinus complex, and the explosive growth in the white-tailed deer population. To study the inflammatory responses to the LD spirochete, the investigator will use a powerful stimulation model that allows a very complete characterization for how the bacterium activates human immune cells and in parallel experiments takes advantage of the large repertoire of available knockout mice to study these responses. Using this combined translational-experimental approach, in this application the investigator will validate key elements of a proposed new model of spirochetal recognition, where the phagosome is a central platform for recognition of diverse borrelial ligands and which involves a cooperative interaction between TLR2 and TLR8 in pro- and anti-inflammatory cytokine responses, and TLR8 in IRF-7 mediated induction of IFN-2.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The L-type Ca2+ channel provides the initial triggering influx of Ca2+ (I/Ca) for excitation-contraction coupling in cardiac muscle. This channel is an extensively regulated protein complex composed of three subunits: alpha1, beta and alpha2-delta. Stimulation of neurohormonal receptors such as endothelin (ET), angiotensin II, and alpha1-adrenergic receptors result in the production of the second messenger diacylglycerol (AG), which can activate protein kinase C (PKC). The effects of activation of PKC on the contractility of the heart and on the L-type Ca2+ channel are controversial. The objectives of this proposal are to clarify the effect of neurohormonal activation of PKC on native cardiac L-type Ca2+ channel are controversial. The objectives of this proposal are to clarify the effect of neurohormonal activation of PKC on native cardiac L-type Ca2+ channels and determine the molecular mechanisms underlying this regulation. Preliminary data using the perforated whole-cell patch clamp technique demonstrate a potent increase in Ica in response to ET and photorelease of a DAG analogue. The hypothesis that PKCepsilon directly stimulates cardiac L-type Ca2+ channel activity by phosphorylation of the alpha1C subunit will be tested. Experiments will utilize isolated rat ventricular myocytes and cultured embryonic stem cell- derived cardiomyocytes, which will be studied using a series of complementary cellular electrophysiology, biochemical, and molecular biology approaches. We propose to address the cardiac myocytes in response to ET and photorelease of caged diC8; 2) Determine the PKC isoform specificity by modulation of cardiac L-type Ca2+ channels; 3) Characterize the phosphorylation of the alpha1C subunit by PKC and determine the functional consequences. Overall, these results will provide a new molecular framework to understand the regulation of L-type Ca2+ channel by the CAG/PKC system. Activation of PKC has been implicated in myocardial hypertrophy and heart failure as well as in ischemic heart disease. Understanding the detailed regulation of the L-type Ca2+ channel by PKC may provide insights and potential novel therapeutic strategies for these disease processes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The eventual long term scientific goal of this research proposal is to achieve an understanding of the molecular mechanisms which are important for vesicle transport and digestive enzyme secretion leading to invasive activities of prostatic tumor cells. We are searching for ways to destroy tumors more effectively by identifying critical cellular targets (ATPases) and more specifically studying drugs which inhibit the ATPases critical for cell motility events related to tumor invasion. Estramustine, a synthetic product of nor nitrogen mustard and estradiol represents a class of anti-tumor drugs we have worked with whose effectiveness in inhibiting a specific class of ATPases, vesicle transport, enzyme secretion and invasive activities of prostatic tumor cells will be assessed. We have found that a novel microtubule associated cytoplasmic ATPase, termed kinesin powers secretory vesicle streaming along microtubules isolated from DU 145 cells. The motion is inhibited in vivo and in vitro with micromolar levels of estramustine (Piazza and Stearns, Nature, in preparation), and agents (vanadate) which are known to inhibit dynein and kinesin ATPase activity. Because of these preliminary findings, in depth biochemical studies of estramustine and kinesin are planned. Specifically, these include, (a) purification and characterization of prostatic cell kinesin from a variety of tumor cell lines; (b) immunofluorescent and immunogold localization studies of kinesin(s) and assessment of the relevance of such localizations to secretory vesicle motility along microtubules, (c) drug combination studies with estramustine and several analogues in an attempt to establish if ATPase activity and vesicle motility are inhibited and more importantly, to determine if drug combination studies inhibit collagenase secretion by invasive DU 145 cells or primary tumor cell lines established from human prostate biopsies. The results from these studies should establish if different tumor cell lines contain kinesin or novel kinesin(s) important for critical cellular functions during tumor invasion. The effect of estramustine dosages to cell performance will be determined with respect to its affinity for kinesin and inhibitory effects on kinesin ATPase activity and related motility functions in tumor invasion including vesicle transport and secretion.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have established the University of Massachusetts Center for Clinical and Translational Science (UMCCTS) which includes the five UMass campuses. Operationally, the Center is located at the University's Worcester campus and managed by the School of Medicine. The Center serves as the \"academic home\" for clinical and translational scientists across all UMass campuses. The CTSA program will build clinical and translational research upon a foundation of world class basic science as evidenced by the award of the Nobel Prize in Physiology or Medicine to Craig Mello in 2006 and the Albert Lasker Award to Victor Ambros in 2008. The UMCCTS aims are to: 1) Accelerate early phase translational (Tl) studies by recruiting outstanding T1 research leaders;establishing innovative research core facilities;and developing new therapies, devices, and interventions based upon UMass discoveries;2) Integrate unique networks of clinical research and healthcare delivery in Central New England and Massachusetts at-large to build and expand T2 (clinical effectiveness research) and T3 (implementation and dissemination research) capacity by recruiting outstanding T2 and T3 research leaders, and by developing innovative T2/T3 core facilities;and enhancing patient and community outreach and participation in clinical/population-based research;3) The UMCCTS will work collaboratively with all campuses and schools ofthe UMass system in developing programs, curricula, and faculty support systems that promote careers in clinical and translational research. Governor Deval Patrick and the Massachusetts Legislature recently enacted legislation committing $1 billion in state investment to be directed to the Life Sciences in the Commonwealth. With $90 million in seed funding, the University has established the UMass Advanced Therapeutics Cluster (UMATC) which includes the RNA Therapeutics Institute, the Center for Stem Cell Biology and Regenerative Medicine, and the Gene Therapy Center. The UMATC joins the Massachusetts Biologic Laboratories, Meyers Primary Care Institute, and Commonwealth Medicine, four innovative components that are rarely found in academic centers, to speed bench to bedside to community discoveries in clinical and translational research. RELEVANCE (See instructions): The University of Massachusetts Center for Clinical and Translational Science (UMCCTS) will transform the conduct of clinical and translational research by providing an \"academic home\" for all university clinical investigators. The UMCCTS will provide robust pilot grant programs, comprehensive education and career development programs, and innovative core facilities to support and nurture clinical investigators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Genomic imprinting is a mammalian-specific phenomenon whereby the expression of a subset of genes is dependent on parental origin of the allele. Monoallelic expression is generally correlated with differential methylation of the parental alleles. Differences in methylation can be inherited as a primary imprinting mark from the gametes or acquired secondarily as a post-fertilization event. The significance of establishing the appropriate methylation pattern and achieving imprinted expression is illustrated by the fact that in the absence of parent of origin-specific expression, developmental defects ensue. Human diseases such as Prader-Willi, Angelman and Beckwith-Wiedemann syndromes result from the inappropriate expression of imprinted genes. The objective of this research is to enhance the understanding of how imprinted expression is achieved via an analysis of methylation acquisition at imprinted loci. The first goal is to conduct a temporal analysis of the acquisition of methylation during male gametogenesis at the paternally-methylated Gtl2 and Rasgrf1 loci. Second, the role of DNA methyltransferase 3L in establishing methylation at imprinted loci during male gametogenesis will be examined. Finally, the post-fertilization establishment of methylation at the Gtl2 promoter and its relationship to monoallelic expression will be investigated. Bisulfite mutagenesis and DNA sequencing will be the primary methodology utilized to examine methylation in these analyses. This research will advance the field of imprinting by determining whether the establishment of imprinting marks is globally or independently controlled. Furthermore, it will test the hypothesis that DNA methyltransferase 3L is the enzyme responsible for methylation establishment in the male germline. In addition, this project will enhance undergraduate education by providing students the opportunity to conduct independent research in molecular genetics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Current therapy for Rheumatiod arthritis comprises non-steroidal anti-inflammatory drugs (NSAIDs) in early stages of the disease, ultimately given way to oral steroids and the disease-modifying anti-rheumatic drugs (DMARDs) as the disease progressively worsens. Methotrexate (MTX) has become the DMARD of choice of many rheumatologists because of its faster mode of action and better record of prolonged use. However, despite the use of high doses of MTX, many patients only experience partial relief of symptoms and still have features of active disease. Experimental evidence implicates TNF as an important mediator in the pathobiology of RA and underscores TNF as an appropriate therapeutic target. Chimeric A2 (cA2) is a monoclonal antibody that binds with high affinity and specificity to human TNF and neutralizes its biologic activity. The current trial is designed to show in a larger patient population (n=300) that cA2 given for up to one year will provide a rapid and sustained reduciton of clinical signs and symptoms of RA in patients who have active RA despite MTX therapy. Four cA2 treatment groups will be evaluated and compared to a placebo group. The primary endpoint in this study will be a reduction in the clinical signs and symptoms of RA at 30 weeks following the onset of treatment. Continuing reduction in signs and symptoms, as well as disability, joint damage, disease remission and quality of life will be evaluated at 1 year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is well recongnized that ketone bodies are excellent fuels for energy production in developing rat brain. Our on-going studies have shown that ketone bodies are also good precursors for lipid synthesis in developing brain. Since hypothyroidism and maple-syrup urine disease, an inborn error in the branched-chain amino acids are known to cause retardation of mental development and function, the effects of these two conditions on the metabolism of ketone bodies in developing rat brain will be investigated. Specifically, the effects of hypothyroidism on the following parameters will be evaluated: (1) the development of ketone body-utilizing enzymes and of several key mitochondrial enzymes, (2) the distribution of these enzymes in the free-mitochondrial and synaptosomal fractions, and (3) the synthesis in vitro of cerebral lipids from ketone bodies and glucose. Hypothyroidism will be induced in new-born rat pups by injecting 131I. In an experimental model for maple-syrup urine disease, the effects of leucine and alpha-ketoisocapruate on the metabolism both in vivo and in vitro of ketone bodies and glucose by developing rat brain will be studied.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Objectives: The aim of the present study is to increase our understanding of the development of self-concepts in children and adolescents and to add to our knowledge of the social factors that influence self-image formation. Over the years psychiatry, psycholgy, and sociology have each contributed to knowledge of the structure of the self-concept, its place in the personality system, its bearing on cognitive, affective, and behavioral processes, and its determination by interpersonal interaction, culture, and location in the social structure. Our main purpose is to add to this accumulated body of knowledge by focusing on certain important issues especially in need of further research. First, we hope to learn more about self-concept change and development. Speculation on the biological, psychological, and social factors giving rise to self-concept change is ample, but systematic data are rare. A second aim is to subject certain products of clinical experience and psychoanalytic insight--in particular, the \"idealized image\"--to systematic data analysis and to learn how location in the social structure may affect its formation. A third objective is to examine certain important but generally neglected global dimensions of the self-concept; the stability of the self-concept is of particular interest. A fourth aim is to learn more about how membership in certain groups, statuses, or social categories are associated with various self- concept components. Although some literature on minority group status and on sex is available, many important questions remain unanswered or, if answered, are inadequately documented. Fifth, although it is contended that the maintenance and enhancement of the self are of prime motivational importance (Lecky, 1945; McDougall, 1932; Snygg and Combs, 1949; Scwartz and Stryker, 1971; Allport, 1955), hard knowledge about the concrete manifestations of these mechanisms is still inadequate. Our special objective is to examine further the selectivity mechanism; currently, speculative observations (Rosenberg, 1967) outrun systematic data. Finally, our aim is to shed light on certain other important but neglected topics, such as ego-extensions, identification, and self- values, or those having special practical import, such as academic achievement.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Relapse to alcoholism remains a vexing clinical and national health problem. Efforts to match alcohol dependent patients to specific treatments based on their clinical characteristics have produced mixed results. Pharmacogenetics (the study of genetic influences on therapeutic response to drugs) offers a powerful new tool to match specific elements of an individual patient's complex genetic blueprint with targeted pharmacotherapies to which that individual may optimally respond. The purpose of this proposed research is to apply pharmacogenetic techniques to predict which alcohol dependent patients will respond favorably to a trial of a selective serotonin re-uptake inhibitor (SSRI) for the prevention of alcoholism relapse. Our central hypothesis is that genetic differences affecting serotonin transporter function will influence an alcohol dependent individual's treatment response to the SSRI, citalopram. To test this hypothesis, we will perform a 14-week, randomized, double blind, parallel group comparison of citalopram and placebo in treatment seeking outpatients who meet DSM-IV criteria for alcohol dependence. All subjects will receive a single Motivational Interview and 9 brief sessions of a manual-guided Compliance Enhancement Therapy designed to promote treatment adherence and enhance motivation to quit or cut down on drinking. Post-treatment follow-up assessments will be conducted at 4, 12 and 24 weeks. Subjects' DNA will be genotyped to determine allelic variants in the promoter region of the serotonin transporter gene that have been found to markedly affect serotonin re- uptake and influence treatment responsiveness to SSRIs. We predict that individuals who carry two long variant alleles (1/1 homozygotes) of this polymorphism will exhibit a significant reduction in drinking days in response to citalopram compared with patients homozygous for the short variant allele (s/s homozygotes). To our knowledge, this will be the first study conducted in alcohol dependent patients to test whether pharmacogenetic differences in the function of the serotonin transporter (the site of action of these medications) influence the treatment response to a SSRI in nondepressed women and men. The study is designed to maximize the likelihood of finding treatment efficacy for citalopram over placebo by excluding subjects with severe alcohol dependence and marked impulsive traits in which SSRIs have not been found to be effective, controlling the exposure to the concomitant psychosocial intervention to minimize a psychotherapy ceiling effect, and by controlling the potential moderating effects of sex and cigarette smoking. The successful completion of this single center study may lead to future multicenter trials in more heterogeneous populations, and to studies using serotonin receptor subtype-specific medications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Acute thromboses of arteries are usually a medical emergency. The long-term objective of this project is to develop a pulsed fluidjet system for intravascular use to hydraulically remove clots and debris from occluded arteries via small caliber angiographic catheters. We propose a pulsed fluid irrigation system combined with aspiration through the lumen of the catheter. A specialized pump, catheter, and control system will be developed. The pump will provide variable pressure, flow, and pulse pattern. The attenation of the pressure pulse in the catheter will be analyzed to optimize the jet. The controls will allow the physician to select various operating parameters for the cutting jet. The goal of Phase I is establish the feasibility of such a system through laboratory testing of catheters and determination of cutting parameters on tissue samples. During Phase II, the system will be fabricated and refined through in vitro and in vivo animal testing. The resulting system for clearing thrombi will have immediate commercial application and could improve the health care for millions of patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY/ABSTRACT Chemoresistant metastatic disease presents the most serious threat to cancer patients despite the increased arsenal of targeted therapeutic options available to clinicians. While intense study into late-stage cancer progression has revealed a number of mechanisms that contribute to chemoresistance, little is known about the intracellular signaling mechanisms that desensitize cells to cytotoxic chemotherapy. In an effort to address this knowledge gap, we recently performed a large-scale RNA-interference (RNAi) screen intended to comprehensively identify critical kinases and phosphatases in the human genome that alter or modify tumor cell sensitivity to chemotherapeutic agents. In this RNAi screen, we identified a novel phosphatase, MK-STYX, which potently suppressed the response of tumor cells to a wide variety of chemotherapeutic drugs. Our central hypothesis is that MK-STYX specifically controls mitochondrial function by regulating phosphorylation of the machinery required for ATP synthesis, and thereby serves an essential role in the induction of chemotherapeutic-induced cell death. The objective of this project is to determine how MK-STYX regulates cellular ATP levels, and thus modulates intrinsic apoptosis. We propose the following specific aims to address this hypothesis and to understand its significance in the context of metastatic colorectal carcinoma: (1) Identify the catalytic mechanism of MK-STYX in the mitochondria; (2) Identify the mechanism whereby MK-STYX regulates chemoresistance; (3) Establish the role of MK-STYX in colorectal cancer progression and chemoresistance. Consistent with our central hypothesis, we have shown that loss of MK-STYX increases ATP production. Therefore, we predict that the elevation in cellular ATP due to loss of MK-STYX is sufficient to inhibit apoptosome formation and entry into apoptosis. We have shown that MK-STYX interacts with two additional mitochondrial proteins and we will mechanistically determine the mitochondrial function and the molecular consequences of each of these interactions. We have also shown that loss of MK-STYX expression correlates with colorectal cancer progression. To determine whether loss of MK-STYX mediates chemoresistance in vivo, we will test the efficacy of standard chemotherapies on a colorectal xenograft model using cell lines that demonstrate variable expression of MK-STYX, or have been manipulated to decrease endogenous MK-STYX levels. We will also determine the prognostic significance of MK-STYX protein levels in a cohort of patients with colorectal cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The vast majority of current human vaccines function by eliciting protective antibody responses. Neutralizing antibodies are an important component of protective immunity against a wide range of viral infections in humans and in animal models. T cell help to B cells is a fundamental aspect of adaptive immunity to viruses and the generation of immunological memory. Follicular helper CD4 T cells (Tfh) are the specialized providers of help to B cells. Tfh cells depend on expression of the transcription factor Bcl6. Tfh cells are important for the formation of germinal centers. Once germinal centers are formed, Tfh cells are needed to maintain them and to regulate germinal center B cell differentiation into plasma cells and memory B cells. There is compelling evidence that Tfh cells are limiting for the magnitude of germinal centers and hence limiting for the germinal center derived high affinity antibodies, memory B cells, and long-lived plasma cells that are the basis of long term humoral immunity. Therefore, there is substantial potential for an understanding of Tfh cells to facilitate better antiviral immune responses and vaccine-elicited humoral immunity. Despite these recent advances, our understanding of Tfh cells is still in the early stages, and the transcription factor (TF) pathways that control Tfh differentiation and define Tfli functions remain poorly understood. In Project 1, we will identify, characterize, stratify, and interconnect TFs that control Tfh differentiation and function to an acute viral infection, while also determining important TF regulators of Th1 differentiation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this proposal we are dissecting immunologic responses in the human respiratory tract that are part of the host's normal defense appartus. Airway derived proteins and phagocytic cells comprise the immune components of these responses and are retrieved from the lungs by bronchoalveolar lavage (BAL) during fiberoptic bronchoscopy. Immunoglobulin G is a particularly important element and we are concentrating on unraveling its role: initiating lung parenchymal inflammation, enhancing alveolar macrophage phagocytosis through opsonic antibody, and causing immune complex formation. Important to this analysis of IgG was to accurately quantitate it and other immunoglobulin and protein constituents of the airway milieu and to characterize the 4 gamma heavy chain subclasses within the IgG population. Lung washings (and blood) from patients with a variety of diffuse interstitial lung disease will provide a contrast with normal values. During the past year of this project, we have made substantial progress in 1) characterizing the relative amounts of immunoglobulins 2) developing better methods to isolate IgG antibody from lung fluids, 3) identifying inflammatory mediators secreted by alveolar macrophages and in BAL fluid and 4) adapting new methods for measuring Ig secreting plasma cells and respective gamma subclasses in BAL. In the next year, we plan to quantitatively measure IgG subclasses in BAL from smokers, nonsmokers and patients with diffuse interstitial lung diseases. In addition we will pursue the function of IgG opsonins derived from patients with cystic fibrosis and will investigate the regulation of chemotactic factors secreted by alveolar macrophages.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of the proposed research is to develop new types of organic molecules that can bind metal ions, forming complexes that are soluble in organic solvents and membranes. In these new molecules, pyridine and other heterocycles are oriented toward the metal-binding cavity, cleft or channel by fusion of rings, forming the most highly preorganized ligands known. Eight large-ring or helical complexing agents are proposed for study. Six are torands in which a large ring is formed by complete fusion of rings. Four of these torands have already been synthesized. Studies of the first torand will be extended to include complexes with metals throughout the periodic table. Particular emphasis will be placed on complexes of heavy metals and lanthanides do may result in applications, such as metal sensors, luminescent markers and NMR imaging agents. A new torand synthesis will be applied to formation of \"tubular\" discotic mesophases (liquid crystals). Two new \"expanded\" or \"mixed-heterocycle\" torands are proposed and their complexation of large metal ions and transition metal clusters will be examined. Two helical ligands of different sizes are proposed for study and their binding of alkali metals will be examined. These helical ligands and the tubular liquid crystals are potential approaches to synthetic ion channels that could be used to model biomembrane channels or to produce permselective membranes. Double-helical or even triple-helical polynuclear complexes may be formed by these novel ligands. Analogues of these complexes might be developed to interact specifically with helical biomolecules, such as proteins and nucleotides. All complexes will be studied spectroscopically and crystallographically to improve fundamental understanding of the interactions of metals with biomolecules. The new complexing agents will be tested as ionophores in ion-selective electrodes for analysis of metals in biological fluids. The polynuclear transition metal complexes formed by expanded torands may be useful as metalloenzyme models and as polyfunctional or multiple-electron catalysts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (adapted from application abstract): Dr. Diamond received his Ph.D. in 1992 (Cell/Dev Biology) and M.D. degree from Harvard University, where he worked with Timothy Springer, who describes him as `an extraordinary level of graduate student that one may see only several times in a lifetime.` The candidate has nineteen publications, eight of these as first author, the first four research done in college and the next fifteen from his Ph.D. work in immunology, focusing on the role of CD11b/CD18 on leukocytes. After graduating from the MD/Ph.D. program he initially began a post-doc to continue his study of integrins in the Drosophila genetic system, but then decided to continue clinical training in order to be able to combine clinical work with basic investigation. Having now completed a residency and in the midst of an infectious diseases fellowship, he requests funding to obtain new training in viral pathogenesis and molecular epidemiology so that he can apply his skills in a system that is new to him. Dr. Diamond is at the very highest level of prior research training and experience that can be considered appropriate for the K08 award. The career development plan is outlined for five years, with courses in epidemiology and biostatistics during phase I, followed by the period of independent research in phase II. The proposed research addresses the role of interferon in infection by dengue virus (DV), both to investigate how IFN modulates DV infection, and how DV alters the IFN response. The candidate proposes to use this award as a vehicle for learning a new area of research that will allow him to add virology and molecular pathogenesis to his knowledge base and to apply his prior research and clinical skills in a novel system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Thin filament-associated actin-binding proteins control both actomyosin-based muscle contraction and cytoskeletal formation. To elucidate the mechanisms required for muscle thin filaments to function, it is crucial to determine the structural interactions of the regulatory proteins involved. As a means of achieving our objective to understand the physiology of cardiac, skeletal and smooth muscle control systems, we will examine the architecture of muscle thin filaments at a fundamental structural level and characterize the changing interactions of thin filament-linked proteins that regulate muscle activity. We will use state-of- the-art electron microscopy and electron tomography coupled with image analysis and 3D reconstruction to establish the macromolecular structure of actin-binding proteins on thin filament actin. Using these techniques: (1) We aim to determine the structural basis of troponin-tropomyosin regulation of cardiac and skeletal muscle activity by analyzing interactions of tropomyosin and troponin on thin filaments, which are governed by Ca2+binding to troponin and myosin-crossbridge binding on actin. To accomplish this goal, (A) we will test our newly proposed atomic model for troponin-tropomyosin localization on thin filaments by generating single particle and electron tomographic reconstructions;(B) we will test the hypothesis that mobile domains of troponin-I latch onto actin to constrain tropomyosin in the inhibitory \"blocking\" state characteristic of relaxed muscle;(C) we will test both the hypothesis that tropomyosin assumes the contours of the F-actin helix as a relatively stiff coiled coiled-coil and the alternative view that tropomyosin is flexible. (2) We will test the hypothesis that mutant cardiac troponin and different tropomyosin variants perturb muscle regulation by causing an imbalance in tropomyosin's position that alters the regulatory state of thin filaments. (3) We will assess the regulatory role of thin filament-linked caldesmon and calponin in defining tropomyosin position in vascular and visceral muscle. (4) We will determine the structure of nebulin bound to actin to complete our map of thin filaments. In each study, reconstructions fitted to the atomic resolution maps of F-actin will demarcate molecular contacts of binding proteins with actin at near atomic resolution (\"hybrid crystallography\"). Lay summary: Studies on troponin-tropomyosin regulated filaments, with particular attention devoted to normal and mutant proteins derived from cardiac muscle, will lead to an elucidation of the molecular regulatory mechanisms governing cardiac contraction, which is essential for tracing cardiovascular disease processes. Studies on smooth muscle filaments will aid in understanding the fine-tuning of smooth muscle contraction thus revealing key controls for vascular tone and pulmonary airway resistance, determinants in, e.g., hypertension and asthma. Actin filaments and associated proteins are major participates in diverse cellular systems, underscoring the broad significance of the proposed work. Our goal is to elucidate the control mechanisms that regulate cardiovascular and skeletal muscle activity. We will examine structural changes at a molecular level that are orchestrated by regulatory proteins and which control muscle shortening and force production. Understanding the underlying molecular physiology governing contraction and relaxation in heart muscle and blood vessels is key to deciphering cardiovascular disease processes, controlling blood pressure and identifying novel targets for drug development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Applicant's Abstract) Despite advancing cure rates in childhood acute lymphoblastic leukemia (ALL), 25-30% of all patients eventually succumb to their disease. The long-term objective of the proposed research is to improve clinical outcome in this subset of patients. By early identification of patients who are likely to relapse, it should be possible to instigate potentially curative therapy in a more timely manner, thus boosting the proportion of long term survivors. This goal will be pursued through three interrelated projects. In the first, overexpression of WT1 and BCL-2 genes will be assessed as markers of minimal residual disease (MRD) in childhood ALL patients. The underlying hypothesis is that these two indicators are more widely associated with leukemia than current markers, and will significantly expand capabilities for prospective identification of high risk patients. Specific Aim 2 seeks to expand results obtained during the previous period of support, suggesting that immunologic monitoring of MRD has clinical utility in the assessment of childhood ALL patients. Immunologic findings in sequential bone marrow samples from patients with B- and T-lineage ALL will be compared with event free survival, as well as presenting clinical and biologic risk features, to establish the independent predictive strength of this assay. The data will also provide opportunities for cross comparisons with results of WT1 and BCL-2 screening in Specific Aim 1. Based on encouraging preliminary results, studies in Specific Aim 3 seek to assess the clinical utility of MRD investigations using peripheral blood instead of bone marrow. Success in this endeavor will radically improve remission studies in patients with ALL, by overcoming the practical and ethical constraints posed by sequential bone marrow aspirations in children. The clinical significance of MRD has been in doubt because of the lack of prospective studies in a large group of uniformly treated patients. The studies proposed in this application should meet that need and demonstrate the feasibility of clinical management decisions based on MRD detection in children with ALL.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The inflammatory bowel diseases (IBD) are chronic inflammatory disorders of the intestine with autoimmune characteristics. Although the pathogenesis of these disorders is poorly understood, many of their features point to a dysregulated T cell response as a major, if not dominant, etiologic factor. An understanding of the inductive events that regulate development of T cells of the gut-associated lymphoid tissues (GALT) under normal and inflammatory conditions will be required to better understand the evolution of these diseases and mechanisms by which they can be managed. Recent studies indicate that commitment to different cytokine/effector phenotypes by naive T cells occurs during the initial antigenic stimulation and is controlled by APC-associated molecules and cytokines. We propose that the abnormal T cell responses in IBD may follow from aberrant phenotype differentiation in the gut. A major focus will therefore be to define where antigen presentation to naive T cells occurs in the normal and inflamed GALT and how expression of costimulator molecules and cytokines in these sites contribute to induction of alternative T cell differentiation pathways. The experimental approach will be to use an alpha-beta-TCR transgenic (Tg)mouse with specificity for ovalbumin (OVA), as a model to allow control of antigen exposure to naive Tg T cells in vitro and in vivo. Novel in situ and immunohistochemical analyses will be used to determine single-cell T cell responses. The first aim is to examine the sites of OVA antigen presentation in the normal and inflamed gut and to determine the cytokine and proliferative responses of naive T cells in these sites. The evolution of distinct T cell phenotypes in the absence of the critical regulatory cytokine IL-4 will be examined by breeding the OVA TCR transgenes into mice carrying homozygous null mutations for IL-4. The second aim is to explore the hypothesis that tolerance to enteral antigens is initiated by antigen presentation on costimulator-deficient enterocytes. Initial studies will examine the APC function of isolated enterocytes for responses of OVA-TCR naive T cells and Th1 and Th2 clones. We will then examine the effects of costimulator expression on these cells in vivo and in vitro, by generating mice in which a B7 transgene is expressed on the small intestinal epithelium under control of the intestinal fatty acid binding protein (I- FABP) promoter. This transgene will be bred into the OVA-TCR Tg mouse and oral tolerance to OVA will be examined. The third aim will be to characterize the phenotypes of proinflammatory and protective GALT T cells divided on the basis of expression of the CD45RB isoform. By using T isolated from OVA-TCR Tg mice, we will be able to control exposure of the separate populations to antigen after adoptive transfer to SCID mice. This will allow us to dissect the earliest antigenic responses of these two populations during disease progression or prevention. In parallel studies, T cell clones of distinct cytokine phenotypes will be derived from OVA TCR Tg mice in vitro and will be examined for their capacity to produce or prevent inflammatory enterocolitis upon exposure to OVA antigen after transfer into SCID hosts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The a9[unreadable]1 integrin is highly expressed in airway smooth muscle. Mice we have generated lacking this integrin only in smooth muscle cells have marked in vivo airway hyperresponsiveness and lung slices from these mice have increased airway narrowing. We have previously shown that the a9 subunit cytoplasmic domain directly binds the enzyme spermine/spermidine acetyltransferase (SSAT), the rate limiting step in catabolism of higher order polyamines, and that this association and polyamine catabolism are important modulators of a9[unreadable]1 function. Pharmacologic stabilization of SSAT also augments a9[unreadable]1-dependent prevention of airway smooth muscle contraction. In the current application we will systematically evaluate the effects of a9[unreadable]1 on responses of airway smooth muscle to multiple contractile agonists and to isoproterenol-induced relaxation using 4 parallel experimental systems (in vivo AHR, tracheal ring contraction, airway narrowing in lung slices and shortening of airway smooth muscle cells). We will utilize a variety of mutant and chimeric constructs of a9 and SSAT, in vivo and in vitro studies with SSAT knockout mice, catalytically inactive SSAT mutants and genetic and pharmacologic inhibitors to thoroughly examine what contribution interaction of a9 with SSAT makes to this response. Force generation in smooth muscle depends on calcium-dependent actin-myosin cross-bridging and parallel actin polymerization, and we will systematically evaluate the effects of a9[unreadable]1 on each of these pathways. Because the two major cytosolic effects of higher order polyamines are prevention of potassium efflux through Kir channels and activation of the lipid kinase, PIP5K1?, we will pay special attention to the roles of Kir channels and PIP5K1? in this process. The proposed studies will test the overall hypothesis that ligated a9[unreadable]1 normally serves as a brake on airway narrowing by concentrating SSAT, catabolizing polyamines and thus inhibiting potassium efflux and/or PIP2 production, resulting in reduced calcium oscillations, decreased actin-myosin cross-bridging and/or impaired actin polymerization. Abnormalities in this pathway, either acquired or genetic, could contribute to diseases such as asthma that are characterized by enhanced airway narrowing. PUBLIC HEALTH RELEVANCE: Project Exaggerated airway narrowing is a central feature of asthma. We have found that mice lacking a specific protein (an integrin) only in smooth muscle have exaggerated airway narrowing. In this proposal we will determine how this integrin normally prevents airway narrowing, a process that could be perturbed in diseases such as asthma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal seeks support for the training of Dr. Supriya Saha, a medical oncologist at the Massachusetts General Hospital (MGH) Cancer Center, toward his goal of becoming an independent physician-scientist with a focused clinical practice in hepatobiliary cancers and a research laboratory dedicated to the study of cholangiocarcinoma. Dr. Saha will be mentored by Dr. Nabeel Bardeesy, a leader in mouse models of gastrointestinal cancer, and Dr. Andrew Zhu, a clinical expert in hepatobiliary cancers. Intrahepatic Cholangiocarcinoma (ICC) has an average survival of < 1 year and has been increasing in incidence for several decades. With no effective targeted therapies or methods for prevention or early detection currently available, there is an urgent need for a greater focus on basic and translational ICC research. Recent studies have identified missense mutations in the metabolic enzymes isocitrate dehydrogenase 1 or 2 (IDH1/2) as the most common genetic alterations in ICC. Mutant IDH (IDH*) has been proposed to contribute to tumorigenesis through an intriguing mechanism whereby the IDH* enzyme produces an `oncometabolite', 2-hydroxglutarate (2HG), that inhibits a family of enzymes requiring alpha-ketoglutarate as a co-factor. Unfortunately, the lack of animal models of IDH*-driven ICC or human cell lines with endogenous IDH mutations has hindered elucidation of the oncogenic mechanisms and of effective therapies against IDH* disease. Over the last 3 years, Dr. Saha has worked in Dr. Bardeesy's laboratory to overcome these hurdles by defining the mechanism by which IDH* promotes ICC and generating the first genetically engineered mouse model of IDH* ICC. Simultaneously, Dr. Saha designed and implemented a clinical protocol to obtain fresh patient samples to generate a rich panel of human ICC cell lines and patient-derived xenografts. In collaboration with Dr. Cyril Benes, these tools were subjected to a high-throughput drug screen to identify a class of kinase inhibitors with potent and specific efficacy against IDH* ICC. During the award period, Dr. Saha will complete studies required for the effective translation of his work into an early phase clinical trial. This includes evaluating the se of targeted kinase inhibition in an autochthonous mouse model of IDH* ICC, using proteomic approaches to identify the specific targets and mechanism of action of kinase inhibition in ICC and exploring the potential mechanisms of resistance that may develop in this disease. This work will benefit from the breadth of expertise at MGH, which houses leading research groups in cancer genetics, molecular biology, signal transduction and cancer therapeutics, and was the first institution to identify IDH mutations in ICC. Given his extensive background in laboratory research, promising preliminary results and the superb mentorship he will receive, Dr. Saha anticipates applying for independent funding within the third year of this award.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Na,K-ATPase is the plasma membrane enzyme that actively transports Na + and K+ ions against their electrochemical gradients, with the hydrolysis of ATP. It is thought to be the single largest consumer of energy in the central nervous system, accounting for 30-40% of ATP hydrolysis. Three different isozymes of the Na,K-ATPase are expressed in the CNS, and in some instances are rather strikingly localized within different cell types. Their individual properties and physiological roles are not yet well known. Available evidence suggests that the isozymes may differ in their affinities for ATP and cardiac glycosides, and in their susceptibility to regulation by unidentified intracellular factors. In addition, ischemic injury has been reported to cause a selective and rapid inactivation of one of two isozymes in the heart. The hypothesis of this proposal is that the Na,K-ATPase isozymes in the CNS are expressed in different cell types, where their affinity for ATP or susceptibility to regulation or degradation may either exacerbate or ameliorate ischemic injury and edema. The properties of the Na,K-ATPases may play a role in explaining the different vulnerability to ischemia of different classes of neurons. The proposal has several objectives: to determine which cell types express which isozymes; to investigate their functional properties and predict which isozymes will remain active in marginally perfused tissue; and to investigate their inactivation secondary to a rise in intracellular Ca2+. We will utilize immunocytochemistry with specific monoclonal antibodies to determine the cellular and subcellular distribution of the Na,K-ATPases in the rat CNS. We will separate the isozymes, and determine their individual affinities for ATP, Na and K+, as well as investigate the possible use of GTP as a substrate. Finally we will investigate the mechanism of modification or inactivation of each isozyme by Ca 2+-dependent proteolysis and by calnaktin-mediated Ca 2+ inhibition. This basic research should lay the groundwork for a rigorous analysis of ion movements, energy consumption, and their role in cell death and edema in ischemic brain, and contribute to a detailed understanding of ion pump function.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During the forthcoming year the following main topics will be studied: (1) The effects of electrical acupuncture conditioning on single unit activity evoked in the spinal nucleus of the trigeminal by stimulation of the infraorbital nerve. (2) The effects of drugs which block presynaptic effects on primary afferents on the conditioning effects of electrical acupuncture stimulation. (3) Efforts to determine at a microscopic level which peripheral structure or structures are activated when the demostrated conditioning effects are obtained.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Human polymorphonuclear leukocytes (PMNs or neutrophils) are essential to the innate immune response against invading microorganisms. In contrast to the acquired immune response, which is dependent on previous interaction with specific bacteria, the ability of PMNs to kill microorganisms is immediate and non-specific. Inasmuch as PMNs produce highly toxic microbicidal components, moderation of infection-induced inflammation is critical for limiting host tissue destruction. This moderation is especially important given that PMNs are the predominant immune cell in most bacterial infections. A key aspect of our research investigates how PMNs ingest and kill bacteria, and elucidates post-phagocytosis sequelae such as apoptosis, processes crucial for the resolution phase of inflammation. Thus, one of our research objectives is to elucidate molecular processes in human PMNs that facilitate resolution of infection. To that end, we used genomics methodologies to establish a global model of host cell-pathogen interaction that provides fundamental insight into the resolution of infection in humans. A second focus of research in my laboratory investigates how bacterial pathogens such as Staphylococcus aureus and Streptococcus pyogenes (group A Streptococcus or GAS) evade human innate host defense to cause disease. Although most bacteria are killed readily by PMNs, some human pathogens have evolved mechanisms to inhibit phagocytosis and death resulting from exposure to ROS and microbicidal products. For example, strains of S. aureus which produce Panton-Valentine leukocidin cause lethal necrotizing pneumonia in non-immunocomprimised individuals, the molecular basis for which is unknown. We hypothesize staphylococcal pathogenesis includes evasion of PMN killing and undetermined host-susceptibility factors. GAS successfully evades PMN phagocytosis and killing to cause human infections such as pharyngitis and necrotizing fasciitis (flesh-eating syndrome). To date, our studies include identification of genes and proteins used by S. aureus and GAS to evade destruction by human neutrophils, hence contributing to virulence, survival and pathogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Alcohol consumption and HIV infection are frequently co-existent pathologies. Muscle wasting is a common feature of both conditions. The alterations in immune responses resulting from chronic alcohol consumption have been hypothesized to enhance the transmission and acquisition of HIV or the progression from HIV infection to acquired immunodeficiency syndrome (AIDS). Based on available information, it is possible to speculate that these are either related to the direct effects of alcohol on the immune system, or are secondary to the impact of alcohol consumption on the nutritional state of the individual. Excess alcohol consumption is associated with a approximately 50% incidence of skeletal muscle myopathy. Alcohol consumption impairs the nutritional state of the individual either as a result of decreased food consumption or as a result of decreased absorption. affecting micronutrients which in turn have been shown to modulate circulating and tissue levels of growth factors. Hence the effects of alcohol consumption on muscle wasting appear to be multifactorial. Alcohol-induced myopathy appears to be predominantly the result of decreases in muscle protein synthesis, and is characterized by decreased weight, protein, RNA and DNA contents in skeletal muscle. The general hypothesis of the present proposal is that alcohol consumption accelerates and worsens the muscle wasting associated with HIV infection, leading to increased morbidity and mortality. Infection of Rhesus monkeys with simian immunodeficiency virus (SIV) has been established as an excellent model system for studying the pathogenesis of HIV-like infection. The disease is characterized by diarrhea, weight loss, lymphopenia, thrombocytopenia, and lymphadenopathy/lymphoid hyperplasia progressing to immunosuppression with marked reduction in CD4+ cells and in the CD4+/CD8+ cell ratio, and opportunistic infections. The aim of the present proposal is to characterize the time-course and relative contribution of alterations in muscle protein synthesis and proteolysis to the progression of muscle wasting associated with chronic alcohol consumption and SIV infection individually and combined. These studies will allow for the longitudinal investigation of the progression of the alterations in muscle metabolism beginning with a healthy, non-infected animal, throughout the acute infectious period and throughout progression to full blown AIDS. These will provide the preliminary data for a more mechanistic approach to the study of the etiology of alcohol-induced muscle wasting and its impact on a chronic infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our lab is interested in understanding the fundamentals of centrosome biogenesis. We aim to uncovering the mechanisms that control the centriole duplication and centrosome maturation cycles. Through our own genome-wide RNAi screen and from other published screens, we now have a large number of candidate genes that play critical roles in these two cycles. What is lacking in the field is a true understanding of these proteins functions. Our lab uses a variety of biochemical and cell biological methods, including high resolution live cell imaging, to perform structure/function studies on known and candidate centrosomal proteins. One such protein we are currently investigating is the human Pericentrin ortholog, termed Pericentrin-Like-Protein in Drosophila. Our in-vivo structure/function analysis of PLP has revealed many new functional aspects of this protein, which are most likely conserved from Drosophila to Humans. Similar experiments are being carried out on a group of proteins that have been shown to preferentially localize to either the mother or the daughter centriole. In support of this project, we have generated many molecular tools and transgenic animals that will be used to perform standard cell biological experimentation in combination with genetic loss-of-function analysis using RNAi in cell culture and allelic analysis in the animal.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Human serum enzyme assays for clinical diagnosis usually make use of measurements of catalytic activity. In many situations, this may not be ideal; e.g., isoenzymes reflecting different damaged tissues may be indistinguishable or inactive enzymes may be released. Radioimmunoassay offers a very sensitive and specific way in which to measure enzyme concentration and to distinguish isoenzymes. Creatine kinase was chosen as a model for this approach because of its clinical importance and its simple isoenzyme distribution. The enzymes from human skeletal muscle (MM) and brain (BB) have been purified, used as immunogens to raise antibodies, and labeled wth 125I. Radioimmunoassays for each of these enzymes were developed and were found to correlate well with standard enzyme assays in clinical use. An attempt is now being made to develop a specific assay for the hybrid MB species.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 10-20% of adults experience chronic abdominal symptoms (abdominal pain/discomfort and associated bowel changes [constipation and/or diarrhea] compatible with a diagnosis of irritable bowel syndrome (IBS). In the US as well as other western countries, women seek health care services disproportionately to men. IBS is defined as chronic abdominal pain/discomfort accompanied by altered bowel pattern (diarrhea, constipation or both [mixed]). The public health impact of IBS in the US is enormous with direct and indirect costs totaling approximately $6 billion/year. Studies suggest an interplay between increased gastrointestinal (GI) permeability ('leaky gut'), abnormalities in the composition of the GI microbiome (defined as bacteria, their genomes and interaction with the host), altered immune responses, autonomic dysfunction (high sympathetic tone), and psychosocial distress lead to the symptoms and the subsequent functional impact of IBS. Our long term goal is to delineate the contribution of pathobiology and psychosocial distress to better inform patient management e.g., diet versus cognitive behavioral approaches). We propose to compare GI microbiota, intestinal permeability, cytokines, and GI and psychological distress symptoms in menstruating women with IBS and without IBS. We also will derive important information about women with IBS with no evidence of abnormal GI markers. The current proposal will also allow us to define microbiome composition and pathophysiological features tied to symptoms, and likely etiology. Thus, the first specific aim is to compare GI microbiome, permeability and cytokines in women (18-45 yr. of age) with IBS (n=100) vs. Healthy Controls (HC) (n=50) without IBS. Based on pediatric data, we hypothesize that among women with IBS, those with a GI microbiome enriched with Proteobacteria/Enterobacteriaceae, increased GI permeability or increased cytokine levels will have greater abdominal pain symptoms versus those without this Proteobacteria/Enterobacteriaceae enriched microbiota composition, normal permeability, or lower cytokine levels. The second aim will be to compare abdominal pain, other IBS symptoms and psychosocial distress symptoms in those IBS subjects with abnormal versus normal GI biomarkers. In addition we will explore patterns of associations among GI biomarkers, and of GI biomarkers with abdominal pain and other symptoms. The goal is to identify possible IBS subgroups based on biomarkers. This research is innovative because we will use biomarkers (e.g., GI microbiome analyses, permeability, and serum markers applied novelty to characterize pathobiologically what heretofore has been primarily a phenotypically and arbitrarily defined condition. We will integrate psychosocial factors (e.g., anxiety, somatization, co-morbid functional conditions) that our preliminary data suggest are likely to predominate in some women with IBS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is submitted to help organize and conduct the 1989 Gordon Research Conferences on Calcium Phosphates, to be held July 24 to 28 at Salve Regina College, Newport, Rhode Island. This meeting is the ninth in a continuous series and allows the interaction of approximately 135 international scientists from universities, industry, and government who study chemistry, biology, physics, dentistry, medicine, biomaterials, mineralogy, oceanography, geology, soil science, environmental science, and related fields and share a common interest in calcium phosphate research. The general theme of the 1989 Conference is \"Calcium Phosphate Research - Linking the Basics to Applications,\" and topics planned for discussion include (1) the nucleation and growth of calcium phosphates; (2) interactions between calcium phosphate minerals and organic matrices in biological systems; (3) remineralization of tooth enamel and root; (4) physicochemical mechanism of dental caries; (5) synthesis, fabrication and properties of new calcium phosphate biomaterials; (6) interactions between calcium phosphate implants and tissue; (7) roles of calcium phosphates in diseases in which mineralized tissues are destructed or pathological calcified deposits are formed; (8) calcium phosphates in industrial and environmental processes; and (9) new experimental techniques in calcium phosphates research. The strength of this Conferences lies in the fact that scientists with highly diverse backgrounds bring their expertise together to exchange scientific information, and generate new approaches for research solutions for problems in all areas of calcium phosphate studies. The meeting thereby contributes to the scientific cooperation and understanding between laboratories both within and outside this country.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies of the biology of mucosal HIV-1 transmission are limited because the circumstances of viral exposure usually cannot be observed and can never by controlled. Nonhuman primate models are limited because massive titers of SIV are required to establish infection after mucosal challenge and sexual transmission between macaques has not be observed despite frequent sexual contact and the presence of infectious virus in genital secretions. Serological data from the mangabey colony at Yerkes were analyzed during this study period. This analysis indicated that the risk of seroconversion began at the age when mounting behavior is initiated and rates of seroconversion are higher in female mangabeys compared with males. A series of experiments were planned to test the hypothesis that SIV is sexually transmitted among mangabeys and to determine if sooty mangabeys could be a useful model for studies of viral selection during sexual transmission and for the evaluation of topical virucides for the prevention of viral transmission. These studies may greatly facilitate vaccine design and the development of topical agents for the prevention of sexual transmission of lentiviruses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Summary: The purpose of the research is to study the cellular and molecular mechanisms of adaptation to stress with emphasis on the regulation of the various components of the hypothalamic pituitary adrenal (HPA) axis. This includes the expression of hypothalamic corticotropin releasing hormone (CRH) and vasopressin (VP), pituitary CRH and V1b VP receptors, and adrenal steroidogenesis. At the hypothalamic level, in vivo and in vitro studies suggest that cAMP can mediate positive and negative regulation of CRH transcription. Activation of CRH expression by cAMP is markedly dependent on downstream elements in the CRH gene. Stimulation of cAMP production by forskolin increased luciferase activity driven by the CRH-promoter. This was caused by increases in transcription, shown by nuclear run-on-assay, as well as increases in mRNA stability and translation, shown by experiments using tetracyclin inducible constructs. Addition of the 3 prime untranslated region downstream of luciferase in a CRH promoter driven reporter construct markedly potentiated foskolin-stimulated CRH luciferase activity due to a potentiation of cAMP-stimulated mRNA translation. Concerning negative regulation, in vivo studies showed that stress induces expression of the CREM isoform, inducible cAMP early repressor (ICER) in the hypothalamic paraventricular nucleus (PVN). Studies using the recently characterized cell lines (4B and H32) revealed that transfection of ICER markedly inhibit cAMP-induced stimulation of CRH promoter activity after cotransfection with a luciferase reporter gene driven by a CRH promoter fragment. Western blot analysis of nuclear extracts of cells incubated with forskolin revealed time dependent increases in ICER. This paralleled formation of ICER-CRH CRE complexes in the electromobility gel shift assay (EMSA) suggesting that endogenous levels of ICER can interact with the CRH CRE. EMSA using hypothalamic nuclear extracts of control and stressed rats and CRH CRE radiolabeled oligonuclotides showed several shifted bands, of which the two lower bands were supershifted with CREM antibody. Extension of studies on immune-neuroendocrine interactions showed that alterations in fluid homeostasis secondary to endotoxemia are due primarily to renal insensitivity to VP rather than defective hypothalamic VP secretion. These studies revealed marked and sustained decreases in VP V2 receptors and aquaporin 2 in kidney medulla following LPS induced endotoxemia, and that these alterations lead to impaired capacity to concentrate urine. Transcriptional and post-transcriptional mechanisms regulating the number of CRH and VP receptors in the pituitary play an important role in the control of HPA axis activity. An essential element regulating V1b receptor transcription is a GAGA box located in the proximal promoter, which binds a protein complex found in pituitary nuclear extracts. Stress causes transient increases in GAGA binding activity an effect possible mediated by VP. Experiments using the hypothalamic cell line, H32, which expresses endogenous VP receptors showed that VP caused a rapid increase in GAGA binding activity followed by a decline. This effect of VP paralleled changes in ERK phosphorylation and it was mimicked by epidermal growth factor (EGF) and blocked by inhibitors of the EGF receptor and MAPK pathway. While the early increase was blocked by intracellular calcium chelators, the declining phase was prevented by protein kinase C inhibition. Studies using receptor subtype specific analogs or different cell lines transfected with either V1a or V1b receptors showed that the pathways involved in transactivation of the MAPK pathway by VP are cell specific and independent of the V1 receptor subtype. This data suggest that VP released into the pituitary portal circulation during stress activates V1b receptor transcription through transactivation of the EGF receptor. At the translational level, studies showed that the 5 prime untranslated region (5?UTR) of the V1b receptor mRNA contains elements, which can inhibit or stimulate translation. While the presence of upstream open reading frames in the 5 prime untranslated region play a role maintaining low translational activity in basal conditions, an internal ribosome entry site (IRES) can initiate translation independently of cap. Stress conditions which upregulate V1b receptors stimulate IRES activity through protein kinase C and PI3 kinase dependent pathways, providing a mechanism for rapid stimulation of V1b receptor translation to meet physiological requirements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "SUMMARY/ABSTRACT Reverse-phase protein arrays (RPPAs) offer a powerful functional proteomic approach to investigate molecular mechanisms and response to therapy in cancer. MD Anderson Cancer Center has been a leader in the implementation of this antibody-based technology that can assess many protein markers across large numbers of samples in a cost-effective, sensitive and high-throughput manner. The platform currently assesses ~300 protein markers, covering all major signaling pathways and most drug targets. Its utility was demonstrated through its selection as the sole platform for characterizing >10,000 patient samples through The Cancer Genome Atlas (TCGA); and recently it has been designated as one of two NCI Genome Characterization Centers, and will characterize up to ~10,000 samples from ongoing NCI initiatives and other consortium projects. For TCGA project, the applicants built The Cancer Proteome Atlas (TCPA), a web platform for visualizing and analyzing RPPA data, which has a community of >5,000 users worldwide. The long-term goal is to promote the ability of functional proteomics to impact cancer research and the development of relevant therapeutic strategies. The current objective is to expand the scope of TCPA by adding new functionalities and datasets, and to enhance and improve its existing analytic capabilities. Working relationships have been formed to link TCPA with other widely used bioinformatic resources (e.g., cBio, UCSC Genome Browsers, Firehose and Synpase) and other ITCR projects. An experienced, multidisciplinary team has been assembled to pursue four specific aims: Aim #1. Develop an open source, all-in-one software package for processing RPPA data. This effort will standardize each informatic step for RPPA data generation including experimental design, quality control, and data normalization. The resultant program will be exported to other RPPA facilities. Aim #2. Expand and enhance our existing web platform for the analysis of patient-cohort RPPA data. The web platform will cover other patient cohorts, incorporate other types of molecular/clinical data, and provide pathway/network-based analytics. Aim #3. Build a user-friendly, interactive, open web platform for the analysis of cell line RPPA data. This effort will collect and compile RPPA data of >1,500 cell lines, and develop a web platform parallel to Aim #2. Aim #4. Promote TCPA and active interaction with the user community. This effort will provide documentation, hands-on workshops, and bug fixes, and build web APIs for interaction with other tools. The expected outcome is the first, dedicated bioinformatic resource that fully integrates RPPA data generation, analysis and user feedback, allowing for fluent exploration and analysis of high-quality proteomic data in a rich context. The project is important because it will greatly enhance the quality and reproducibility of RPPA data from important consortium projects; substantially reduce barriers biomedical researchers face in mining complex functional proteomic data; serve as a hub for integrating proteomic data into other widely used bioinformatic resources; and directly facilitate development of protein markers for precision cancer medicine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Transcription factors in the nuclear factor ?B (NF-?B) family are evolutionarily conserved master regulators of immune and inflammatory responses. They are activated in response to ligation of many receptors including T-cell receptors, B-cell receptors, members of the tumor necrosis factor (TNF) receptor superfamily and the Toll-like receptor/interleukin-1 receptor (TLR/IL-1R) superfamily. The I?B kinase (IKK), comprising IKKa and IKK, is at the heart of NF-?B activation and mediates two NF-?B activation pathways. The canonical NF-?B pathway is triggered by microbial and viral infections and pro-inflammatory cytokines and is dependent on IKK phosphorylation and activation. The alternative pathway is triggered by certain members of the TNF cytokine family and selectively activates IKKa. Activated IKK phosphorylates I?Bs, leading to their polyubiquitination and subsequent degradation by the proteasome. The freed NF-?B dimers translocate to the nucleus to mediate transcription. Because of its importance in NF-?B activation, IKK, especially IKK, has become a potential therapeutic target for many human diseases. The regulatory protein NEMO (also known as IKK? or FIP-3) interacts with IKKa and/or IKK to form the IKKa, IKK or IKKa/ holo-complex. The intact IKK holo-complex is approximately 700-900kD in molecular mass containing multiple copies of IKK2 and NEMO. IKKa and IKK both contain the following conserved recognizable domains: a kinase domain (KD), a leucine zipper domain (LZ), a helix loop helix domain (HLH) and a C-terminal NEMO-binding domain (NBD). NEMO contains an N-terminal kinase-binding domain (KBD), a minimal oligomerization domain (MOD) that is also the ubiquitin binding domain (UBD) and a C-terminal zinc finger domain (ZF). IKK and NF-?B signaling has attracted tremendous attention with more than 30,000 papers published on the subject. Despite the biological importance, not a single successful structure determination has been reported on IKK, an indication on the difficulty of the project. To elucidate the molecular basis of IKK function and to assist the discovery of IKK inhibitors, we propose a series of structural and functional studies on IKK, in particular, IKK and its regulatory protein NEMO. Public Health Relevance: The I?B kinase (IKK) is at the heart of NF-?B activation and a potential therapeutic target for many human diseases. The proposal seeks structural studies of IKK, which will enhance our understanding on the molecular basis of IKK function. In addition, the proposed studies will provide a structural basis for discovery and optimization of IKK inhibitors in the treatment of inflammatory diseases and cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Progress in healthcare research depends heavily on demonstrations of efficacy and safety via clinical trials. Bayesian methods offer a valuable alternative mode of analysis for clinical trials. By comparison with standard frequentist methods, Bayesian methods provide more interpretable outputs, use all the available evidence, and lend themselves to the more complex analyses demanded by complex healthcare research. Unfortunately, few biostatisticians have received training in Bayesian methods. We propose to produce user-friendly software to enable Bayesian analysis of data from clinical trials. This software will implement the complete functionality of Spiegelhalter s BART software, but in a much easier-to-use package. We will develop an associated web-based tutorial. In Phase II we will develop the software into a full-featured Bayesian analysis toolkit for clinical trialists. PROPOSED COMMERCIAL APPLICATION Clinical trials represent a 10 billion dollar industry. Statistical software for this industry is itself a significant business. We sell iBART both to drug company trialists and to federally sponsored clinical researchers. We expect iBART s tutorial and the Bayesian approach in general to appeal especially to the many trialists without specialized statistical qualifications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal will investigate several interrelated immunologic aspects of the basic host-parasite relationship in the mouse model of schistosomiasis mansoni. This important, wide-spread worm infection is prevalent in much of Africa, the Caribbean and South America. The long term objective of this research is to understand sufficiently the balance between host and parasite to allow manipulation of the system in favor of the host. These studies should improve the understanding of cytokine/lymphokine involvement in the production and immunoregulation of the primary pathogenic mechanism in murine schistosomiasis the schistosome egg-induced granuloma (Specific Aim #1). Specific Aim #2 seeks to define major cross-reactive, immunoregulatory idiotypes which are expressed on antibodies specific for schistosome soluble egg antigens (SEA) and elucidate their role in granuloma modulation at the antibody and T lymphocyte levels. Examination of the potential role (s) of gamma/delta (gammadelta+) T cell receptor- bearing T lymphocytes at different stages and in certain anatomical locations during schistosomiasis (Specific Aim #3) should yield useful information about the function of these newly studied cells, and may clarify previously unstudied aspects of host responsiveness in schistosomiasis in the dermis, spleen, liver and intestine. Specific Aim #4 states, and will explore the validity of, a new hypothesis regarding differential T cell responses which result in eosinophilopoiesis in schistosomiasis. The answers to questions posed in these specific aims should provide information that will contribute to our fundamental immunologic knowledge as it applies to schistosomiasis. When germane, data and concepts from these studies bay be useful in understanding other medically important conditions, such as chronic infections, autoimmune diseases, cancer and transplantation. These settings parallel schistosomiasis in the sense that in each the body's immune system must \"learn\" to deal with chronic antigenic exposure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cerebral palsy (CP) is the most prevalent physical disability originating in childhood of which spasticity and weakness are primary clinical signs. Spasticity management in this population has changed dramatically in the past two decades, first with the introduction of selective dorsal rhizotomy (SDR) and most recently with the introduction of intrathecal baclofen (ITB) pump implantation. ITB is efficacious for persons with spasticity of spinal origin; however, the clinical results in CP, while generally positive, are less well-established, reduction in spasticity is typically less marked, and functional gains are less impressive or even equivocal. Reported positive effects of ITB include a relaxation in spasms and spasticity which may reduce associated discomfort and improve ease of movement. A major unresolved question is whether muscle weakness is a direct effect of ITB or whether only an apparent weakness occurs due to elimination of spasticity, as seen after SDR. Exacerbation of weakness could prove problematic in these patients who are already significantly weak. The Specific Aims of this project are to:(1) quantify the changes in voluntary torque production, spasticity and selective control as a result of ITB; and (2) determine the interplay of these clinical changes on functional motor outcomes so as to improve clinical application of this therapy in CP. The following hypotheses will be tested: a) ITB will have a negative effect on isometric and eccentric peak torque production of eight (8) major lower extremity and two (2) representative upper extremity muscle groups in a dose-dependent manner; b) Isolated control of muscles opposing spastic agonists during movement tasks and gait will be conversely improved in a dose-dependent manner; c) Functional gains, including changes in gait temporal-spatial parameters and the Global Function Scale of the PODCI, will not be dose-related. Alternatively, these will depend on the individual's underlying motor capabilities, the amount of change in spasticity and strength produced by ITB, and how these changes interact to alter functional performance. Our long term goal is to improve spasticity management in CP by more precise patient selection and dosage adjustment, and greater consideration of adjunct therapies such as strength training post-operatively.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "KS is the most common neoplasm occurring in patients with AIDS. Kaposi's sarcoma-associated herpesvirus (KSHV) is universally present in Kaposi's sarcoma tissue and in a rare subset of non-Hodgkin' lymphomas termed primary effusion lymphomas (PEL). A causal role for KSHV in the pathogenesis of KS has been suggested by epidemiological data, and is supported by the presence of KSHV gene products, which can alter the cellular activation state, activate cell cycling, and inhibit apoptosis. HIV infection greatly increases the risk of KS development in KSHV-seropositive individuals. In a PEL cell culture-based model, HIV replication stimulates lytic phase replication and transmission of KSHV. The major goal of this research proposal is to determine the mechanism underlying the stimulation of KSHV replication by HIV. A second goal is to define the potential influence of KSHV gene products upon replication of HIV in the PEL cell model. To achieve these goals, the HIV gene product(s) required for stimulation of KSHV replication will first be determined. Proviruses with knockout mutations in key gene products will be analyzed in the HIV-PEL model for their ability to induce lytic KSHV replication. The role of HIV-1 Tat, Env, Vpr, and Nef in altering the replication of KSHV in the absence of other HIV gene products will be examined by expression of these gene products or controls within cells harboring KSHV in latent phase. Although HIV- and KSHV co-infected cells undergo lytic phase replication, a soluble factor released from HIV-infected PEL cells is also capable o stimulating KSHV lytic replication. The identity of this soluble factor will be pursued through measurement o released cellular cytokines, measurement of released HIV and KSHV gene products, and by biochemical purification techniques. HIV replication may act to induce lytic replication through the activation of an. immediate-early KSHV gene, KSHV Rta. The role of HIV infection in inducing transcriptional activation of KSHV Rta and of additional KSHV transcripts will be assessed in the HIV-infected PEL-cell model. The mechanism of HIV spread in two PEL cell lines will next be investigated, with an emphasis on identifying KSHV gene products, which facilitate transmission and replication of HIV. Finally, the effect of HIV infection upon KSHV lytic replication will be investigated in a model of primary monocyte-derived macrophage infected with KSHV and HIV. These studies have relevance to the pathogenesis of AIDS-KS, and may provide insights into interactions between HIV and KSHV, which occur in co-infected individuals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In persons with congenital nystagmus (CN), the incessant to and fro oscillations of the eyes produce continuous motion and smear of the retinal image. However, unlike many patients in whom nystagmus is acquired, individuals with CN rarely report the visual world to be moving. A robust extraretinal signal has been shown to exist for the eye movements in CN, which neurologically \"cancels\" much of the retinal image motion. One aim of the proposed research is to document how operation of this \"cancellation\" process depends upon the foveation periods of the CN waveform - brief intervals of relatively low eye velocity during which a target of regard is imaged at or near the fovea. A related aim is to compare the temporal characteristics of the \"cancellation\" of CN to that for smooth eye movements in normal subjects, to determine whether the \"cancellation\" mechanism is altered in CN. In addition to perceiving a stationary world, persons with CN typically don't perceive visual targets to be smeared. A second aim of this research is to quantify the extent to which perception of image smear is reduced in subjects with CN and evaluate the contributions of three potential mechanisms: l) masking of smeared retinal images by a clearer image that is available during the foveation periods in the CN waveform, 2) suppression of image smear by the extraretinal signals for CN, and 3) neural sharpening of the moving retinal image by \"deblurring\", as has been reported to occur in normal vision. The third aim of this research is to document the effects of the retinal image motion in CN on two basic visual functions. First, sensitivity to changes in target orientation will be measured in subjects with CN and compared to the variability of torsional eye position, during both the entire CN waveform and just the foveation periods. Second, thresholds for perceiving stereoscopic depth will be measured and related to parameters of CN eye movements and to visual acuity. The stereothresholds of normal subjects for targets that move to simulate the retinal image motion in CN will document how stereopsis is affected by the CN retinal image motion per se. These results will also clarify how normal subjects maintain fine stereopsis, despite substantial retinal image motion and vergence errors that accompany voluntary head movements. Over all, the results of the proposed research will define how the potentially debilitating symptoms of oscillopsia and perceived image smear are prevented in subjects with CN, and how the characteristics of CN affect visual orientation sensitivity and stereopsis. A more complete picture than currently available of the visual capabilities and limitations of individuals with CN will result.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To understand how the several mesenchymal cell types in the adult mouse mammary gland interact with mammary epithelium to regulate its development. We expect to mimic normal mammary morphogenesis and terminal differentiation, in monolayer cultures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "MINOS will develop new methods and techniques to test the hypothesis that changes in conformation and/or assembly determine biological outcomes. The staff and scientists of the SIBYLS beamline provide comprehensive expertise in the targeted areas of nucleic acid binding proteins. High Throughput (HT) Small Angle X-ray Scattering (SAXS), Macromolecular Crystallography (MX), and hybrid computational methods. MINOS builds upon our results developing and employing SAXS to define accurate conformations and assemblies in solution in combination with PSI high-resolution crystal structures for detail. Biological information involves changes in shape as well as active site chemistry. SAXS provides robust analyses of shape and conformational change in solution whereas crystallography provides precise information on structural chemistry. Leveraging our existing SAXS and molecular biology expertise, we will innovate new methods and technologies to integrate and advance PSI and community characterizations of key human proteins and their complexes (with partner proteins, DNA, and RNA). MINOS will work closely with PSI centers and individual researchers to identify promising targets and constructs, optimize solution conditions, and provide solution conformation and assembly results that complement PSI high resolution crystal structures. MINOS will provide new methods, tools, and strategies to characterize key human and higher eukaryotes proteins and their complexes for structural biology and medicine, which have been challenging for current PSI and community efforts. Technical goals include identifying and optimizing SAXS data collection strategies for human proteins and their complexes in concert with high resolution structural studies within the PSI centers, rescuing stalled protein targets, and developing hybrid methods and techniques for easing the bottlenecks that currently place real limits on overall PSI productivity. The Specific Aims will endeavor to 1) develop and apply innovative HT SAXS methods to solve solution structures of PSI:Biology defined targets, and 2) use solution scattering technologies to link PSI and community structures to biology. Structures determined by PSI and community collaborations will direct SAXS experiments, test functional implications from SAXS structures, and provide critical details for defining conformational trajectories in solution. Collectively the proposed Aims provide a clear path to leverage PSI and research community strengths and technologies for imaging human and higher eukaryote proteins and their complexes with major impacts on biological understanding.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the last year, upgrades were performed on the spinning disk confocal system in the Laboratory of Cellular and Molecular Biology Microscopy Core. 27 researchers used the resources of the Laboratory of Cellular and Molecular Biology Microscopy Core. While most of the researchers come from the Laboratory of Cellular and Molecular Biology, the Core has been used by scientists from the Laboratory of Immune Cell Biology, the Section on Cellular and Developmental Biology at the National Institute of Child Health and Human Development, the Genetic Disease Research Branch and The University of Maryland Department of Physics. Almost all of the Principal Investigators in the Laboratory of Cellular and Molecular Biology have projects that involve the Core facility. Dr. Lawrence Samelson uses Core resources for the project Biochemical Basis of T Cell Activation. Dr. Carole Parent's projects, Signaling Events Regulating Chemotaxis and Chemotactic Signals Regulating Human Neutrophil and Breast Metastatic Migration use Core instruments. Dr. Paul Randazzo has made extensive use of the Core for the projects Regulation of focal adhesions and Turnover of invadopodia. Dr. Ying Zhang uses Core instruments for the project Molecular Mechanisms of TGF-beta Signaling Pathway. The Core has been involved with the project Cbl Proteins as Regulators of Tyrosine Kinase Signaling from Dr. Stanley Lipkowitz, who is now Chief of the Women's Malignancies Branch. In addition, the Core facility has been used by personnel working with Principal Investigators from other groups including work with Dr. Jonathan Ashwell on the role of ZAP-70 in T cell activation. This research usually involves the use of a Leica SP8 Laser Scanning Confocal Microscope or a PerkinElmer UltraView Spinning Disk Confocal Microscope, with some usage of our Total Internal Reflection Fluorescence (TIRF) microscope. Most of the users view immunofluorescent staining on fixed samples with the Leica LSCM while the spinning disk confocal is generally used for live cell imaging. We routinely use our TIRF microscope for PhotoActivation Localization Microscopy (PALM) and direct Stochastic Optical Reconstruction Microscopy (dSTORM). These high resolution techniques allow us to determine the location of single proteins clustered in signaling complexes in T cells with an localization error of around 20 nmfor PALM and 5 nm for dSTORM. We can now perform two color PALM imaging and multiplexed 3-D dSTORM imaging.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Optimizing HIV Protease Inhibitor Safety and Efficacy via Intracellular Targeting Abstract The AIDS epidemic is a global crisis with 31 million people worldwide who are living with HIV. According to the World Health Organization's 2010 revision of antiretroviral treatment therapy guidelines, creating less toxic therapies is a top priority to reduce adverse effects and improve compliance with therapeutic regimen. HIV protease inhibitors are a mainstay of highly active antiretroviral therapy (HAART) with annual sales of over $2.5 billion. All marketed HIV protease inhibitors (PI's) are substrates for cytochrome P450 and are co- administered with ritonavir, a pharmacokinetic booster. Boosting maintains therapeutic levels of compound to suppress viral replication and avoid incidence of viral mutation. Ritonavir is a strong inhibitor of the cytochrome P4503A4 isoform with a of Ki 5-70 nM which helps reduce P450 interactions of the co- administered PI. Eliminating P450 interactions of PI's would also eliminate the need for ritonavir. Toxicities attributed to ritonavir used alone or in combination with other HIV protease inhibitors (PI's) include hepatotoxicity, carotid artery thickening, hypercholesterolemia, hyperglycemia, and lipodystrophy. The elimination of P450 interactions for PI's is a high-priority research focus at several pharmaceutical companies, including Merck, Sequoia Pharmaceuticals, and Concert Pharmaceuticals. The elimination of ritonavir has been shown to reduce toxicities associated with HAART in short-term studies, and would increase compliance with therapeutic regimens. The specific aims of this proposal are: Aim 1. Design and synthesize a novel library of PI fragments linked to FKBP ligands. Aim 2. Screen and select library compounds for the ability to inhibit HIV protease employing an enzymatic assay and evaluate in vitro pk/pd. Aim 3. Re-screen the best PI candidates resulting from Aim 2 for microsomal stability, potency in cell infectivity assays, and p-glycoprotein assays to select candidates for further preclinical evaluation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "RTI International has proposed innovative methodological and applied research entitled Geospatial and Time Series Analysis of Food Prices and Obesity: Evidence from Sugar-Sweetened Beverage Prices to enhance scientific capabilities for conducting obesity policy research. This project is motivated by the research community's need for high-quality community-level measures of food prices and by the currently active debate among academic researchers, policy analysts, and lawmakers on the efficacy of large sugar- sweetened beverage (SSB) taxes. The proposed spatial economics research will determine the feasibility of constructing high-quality community-level food price measures using optical scanner data on supermarket sales and household purchase records collected by The Nielsen Company. The utility of the price measures in informing obesity policy will be validated by matching these measures with geocoded National Health Interview Survey (NHIS) data and examining the causal effect of SSB prices on body mass index (BMI). Price endogeneity will be controlled by using instrumental variables that are correlated with the costs of SSB and non-SSB supply but uncorrelated with SSB and non-SSB demand. The proposed research will directly benefit obesity policy researchers and policy makers through (1) the development of new food price microdata and (2) the enhanced understanding of the prospect of leveraging economic incentives such as junk food taxes and/or healthy food subsidies to reverse the obesity epidemic. Upon completion, the price microdata will be made available to the research community. The proposed 2-year project has two aims: Aim 1: Gain insights regarding the feasibility of developing high-quality community-level measures of food prices based on existing scanner data on supermarket sales and household purchases and matching these price measures with geocoded health survey data. Aim 2: Validate the utility of the food price microdata in an econometric model relating obesity to SSB prices and measures of the built environment. Use the results to shed light on whether large SSB taxes are likely to affect health outcomes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "TMD is a highly prevalent (i.e., up to 15-20% among women of reproductive age) chronic pain syndrome with profound consequences including disability, psychosocial dysfunction, and reduced quality of life. While psychosocial interventions have shown tremendous promise in reducing TMD symptomatology, standard cognitive-behavioral treatments for pain are costly and time-consuming to administer. As a consequence, significant interest has been generated in the development of interventions that minimize the logistical burdens on healthcare providers and patients (e.g., \"minimal-contact\", \"self-help\", and \"brief treatments\"). While these treatments are widely used in the context of certain conditions (e.g., arthritis), their application in TMD has been minimal. Moreover, none of the few existing studies of psychosocial interventions for TMD make explicit use of social support, a powerful intervention with benefits for many chronic conditions. Thus, the principal goals of the present proposal are as follows: (1) to develop a minimal contact/home-based psychological intervention based on cognitive-behavioral principles for pain management that also makes explicit use of social support, and (2) to test the efficacy of this intervention, using an appropriate control group, in reducing pain and improving functioning. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cell differentiation, reprogramming and malignant transformation are major events characterized by remarkable changes in the epigenome and involve remodeling of DNA methylation patterns. In cancer tissues, DNA methylation patterns are drastically different from those in normal tissues. Two major events are observed, (i) global DNA hypomethylation in cancer affecting predominantly repetitive DNA sequences, and (ii) gene-specific hypermethylation of CpG islands affecting hundreds of genes. The mechanisms how these cancer-associated DNA methylation patterns arise are largely unknown. In 2009, it was reported that a sixth DNA base, 5-hydroxymethylcytosine, is present in substantial amounts in certain mammalian cell types. 5-hydroxymethylcytosine (5hmC) is created from 5-methylcytosine (5mC) by enzymatic oxidation carried out by the TET family of proteins. One model proposes that 5hmC is an intermediate in DNA demethylation. Our hypothesis is that defects in the 5mC oxidation pathway are responsible for altered DNA methylation patterns in human tumors. We have established methodology for precise quantification and genome-wide mapping of 5mC and 5hmC. Our goal is to determine the level and the genomic distribution of 5hmC in normal human tissues and in malignant tumors. These data will be compared directly with the distribution of 5mC in the same tissues. We will focus primarily on two tumor types: (1) human grade II/III astrocytomas, because these tumors frequently contain mutations in isocitrate dehydrogenases (IDH1 or IDH2), an enzymatic activity potentially impacting on the 5mC oxidation pathway; and (2) myelodysplastic syndrome (MDS), because this malignancy often is characterized by mutations in one of the TET genes, TET2. The third Aim will focus on functional studies of TET and TET-associated proteins and their aberrations in cancer. PUBLIC HEALTH RELEVANCE: DNA cytosine-5 methylation patterns in cancer are aberrant and are characterized by frequent hypermethylation of CpG islands. It is unknown how these changes in tumors are initiated. Recently, it has been shown that 5-methylcytosine can be oxidized enzymatically to 5-hydroxymethylcytosine. In this application, we propose that changes in 5-hydroxymethylcytosine patterns are hallmarks of malignant transformation and are related to the aberrant DNA cytosine methylation patterns seen in tumors. We will analyze this novel epigenetic mark as well as 5-methylcytosine in normal and malignant tissues. We will also investigate if and how mutations affecting the 5- methylcytosine oxidation pathway have an effect on genomic methylation patterns in human tumors. Functional studies of 5-methylcytosine oxidases (TET proteins) and their associated factors will support these studies so that a comprehensive picture of the importance of this pathway in human tumorigenesis can be obtained.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fish are known conduits of metal exposure to humans and wildlife. Forty-eight states currently issue public advisories limiting fish consumption to reduce human exposure to Hg and other toxins. Yet, there is much unexplained variation in metal levels in fish from different lakes even within the same region. Project 7 studies mechanisms driving lake-to-lake variation in burdens of top priority metals (Hg, As, Cd, Zn, Pb) in aquatic organisms. We focus on the trophic transfer !of metals (i.e., movement of metal from water through the food chain to fish) in a multiple stressor context (e.g. multiple metals, pH, DOC, nutrients, low food) across a gradient of lake types. We hypothesize that much variation in fish metal burdens is driven by fundamental differences in the food web structure among lake types and in the ability of particular taxa to accumulate, magnify or dilute metals. We will predict this variation based on environmental properties that vary across lakes, such as lake productivity, adjacent land use, dominance of \"key conduit\" species, use of \"conduit\" habitats for foraging, timing and nature of metal inputs, and the toxicological responses of the conduit organisms. Our research across 60+ lakes identifies three aspects of food web structure that increase trophic transfer of metal to fish: (1) larger bodied, lower complexity zooplankton food webs, (2) dominance of key conduit zooplankton taxa (e.g., Daphnid), and (3) lower algal biomass and zooplankton abundance. We also plan to test for additional influences of factors including lake productivity, use of the littoral zone for feeding, presence of other environmental stressors. We further seek to strengthen the scientific basis for lake-specific management and public health warnings and plan to develop Daphnia as a model organism for toxicogenomic analysis to understand gene-environment interactions underlying effects of multiple metal stressors in aquatic ecosystems. This proposal has five specific aims. Aim 1 characterizes metal trophic transfer pathways in the field and tests whether the transfer to fish diminishes!in eutrophic or urbanized lakes. Aim 2 determines strength and consistency of specific taxa and specific lake habitats (littoral Vs. pelagic) as conduits of metals to fish. Aim 3 links temporal patterning in deposition of multiple metals to watersheds and inputs to lakes, and determines the source of key depositional events using multielement emission source fingerprinting, a new stable Hg isotope technique, and meteorological trajectory analysis. Aim 4 quantifies combined toxic effects of multiple environmental stressors associated with metal burdens in situ on Daphnia. Aim 5 characterizes genomic response to metals among natural populations of Daphnia, in order to further the development of Daphnia as a general toxicogenomic model species for assessing metal exposure and effects in natural field populations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Catecholamine neurotransmitters have been implicated in the pathophysiology of several neurological and psychiatric disorders such as schizophrenia, depressive illness and Parkinson's disease. The enzyme which catalyzes the first and rate limiting step in the biosynthetic pathway of catecholamine neurotransmitters is tyrosine hydroxylase (TH) and its gene expression is limited to catecholaminergic neurons. In this proposal, human neuroblastoma cell lines which are either adrenergic (TH-expressing) or cholinergic (TH-nonexpressing) will serve as the model system. The molecular events and factors governing this differential expression of the TH gene will be investigated. In order to gain a deeper understanding of gene regulation in the nervous system, it is of critical importance to define which nuclear proteins (trans-acting elements) specifically bind to the promoter DNA sequences (cis-acting elements). Characterization of the molecular interactions between trans-acting and cis-acting elements should lead to the successful isolation of the gene(s) for the relevant trans-acting factor(s). As an extension of these studies, tissue-specific expression will further be investigated in transgenic mouse models. The results from these studies may provide insights not only into the regulation of catecholamines in general but also into the etiology of some human brain disorders that exhibit altered catecholamine expression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aim of the proposed study is the further delineation of B cell differentiation and characterization of B cell triggering in mice. The basic tools in these studies will be antisera specific for the two B cell differentiation markers Lyb3 and Ia.W39 which we have defined previously. Both these antigens are expressed selectively on a functionally defined B lymphocyte subset that is absent in adult mutant mice carrying the xid gene and in newborn normal mice. Since we have previously shown that Lyb3, an isogenic B cell marker, is a receptor for triggering signals, we will now analyze the nature of these signals by studying whether Lyb3 has binding capacity for T cell replacing factor(s). The role of Ia.W39, coded for by a gene in the I-A region of the H-2 complex, as an effector molecule in cell interactions will be tested. In particular, we will examine whether Ia.W39 is essential for optimal presentation of antigens, which are under immune response gene control mapping in the I-A region. Since the membrane expression of both antigens is controlled by a gene on the X-chromosome, there might be a structural or organizational relationship between the two molecules: We will analyze their chronologic appearance during ontogeny. Further we will attempt to in vivo and/or in vitro modulate their expression in order to test whether there is a mutual interdependence in the mechanism of surface expression. We will also study whether a selective loss of receptor/effector function (see above) is seen in these manipulated B cells. We will try to produce hybridomas secreting monoclonal anti-Lyb3 and anti-Ia.W39 antibodies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to demonstrate that lead CounterAct compounds atenolol (AT) and levetiracetam (LV) given after organophosphate (OP) pesticide and nerve agent induced status epilepticus (SE) are a safe and effective treatment to reduce OP SE mortality and morbidity, including behavioral abnormalities, cognitive impairment, acquired epilepsy (AE) and mossy fiber sprouting. SE is a major medical emergency seen with exposure to OPs from chemical threats from terrorist attacks or from exposure by accident or natural disasters. Advances have been made to treat the seizures associated with SE and the cholinergic crisis from OP exposure, but at present there are no therapies available to prevent mortality and the long term morbidities associated with OP SE. Our research has made a major advance in understanding how OP SE causes mortality. Our PR indicate that cardiac irritability in the first 7 days after OP SE is the major cause of mortality and this can be reduced by treatment with AT and LV. We made a breakthrough in our preliminary results (PR) indicating that AT plus LV may reduce mortality by greater than 70% and also significantly reduce morbidity by greater than 50%, including behavioral abnormalities, cognitive impairments and the development of AE. This PR also suggests that AT and LV can reduce cardiac irritability and cardiac and neuronal damage after OP SE. This study will use the OP pesticide paraoxon (POX), the OP nerve agent surrogate diisopropyl- fluorophosphate (DFP) and the nerve agent sarin to induce SE in rats. Our laboratory is ideally suited to conduct these studies and has developed the necessary skills to carry out the following specific aims: Aim 1: Determine whether AT and LV can reduce mortality following POX induced SE and conduct pharmacokinetic and pharmacodynamic analyses for CounterACT lead compounds AT and LV when administered intra- muscularly and orally. Aim 2: Determine whether AT and LV can reduce mortality following DFP and sarin induced SE and evaluate the acute and chronic effects of intramuscular injections on injection site musculature. Aim 3: Evaluate whether AT and LV can reduce cardiac irritability and cardiac pathological changes following POX, DFP and sarin SE. Aim 4: Determine whether AT plus LV can reduce the development of depression-like symptoms and provide neuroprotection following POX, DFP and sarin SE. Aim 5. Evaluate whether AT plus LV can reduce the development of cognitive impairment, the development of AE and mossy fiber sprouting following POX, DFP and sarin SE. The PR demonstrate the feasibility of these studies and underscore the potential significance of conducting this research. AT and LV have been used for many years clinically to treat hypertension and seizures, respectively, and thus their use in humans has been well established. If these preliminary findings are documented in this study, we will conduct a pre-IND meeting with the FDA for ultimately getting an IND for the use of AT and LV by intramuscular administration as an effective treatment for reducing mortality and morbidity from OP SE.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a response to RFA # RFA-AG-09-002. The major goals of the proposal are to: (1) Identify biomarkers that are associated with progression from normal cognitive status to mild cognitive impairment (MCI) or dementia, with a particular focus on the dementia of Alzheimer's disease (AD) - these biomarkers may potentially include measures based on cognitive testing, magnetic resonance imaging (MRI), blood, or cerebrospinal fluid (CSF); (2) Determine which cross-sectional or longitudinal biomarker measures (taken alone or in combination) are the best predictors of progression from normal cognition to varying levels of cognitive dysfunction (i.e., MCI or AD); (3) Create a publicly accessible data base containing the clinical, cognitive, imaging, blood and CSF data - raw imaging data and samples of blood and CSF will also be available to investigators in the field, as appropriate. A team of investigators has been assembled at the study site with substantial experience in the clinical evaluation of older individuals, as well as expertise in the analysis of the biomarkers in question. Several external advisory groups will be assembled in order to provide guidance concerning the analysis of the data (particularly the CSF and blood samples, which are a non-renewable resource). In order to accomplish these goals we will: (1) complete a comprehensive clinical evaluation on as many prior participants in BIOCARD as possible in order to determine their current clinical and cognitive status - this will represent an approximate 10 year follow-up of the cohort; (2) initiate annual, longitudinal, clinical and cognitive evaluations on as many of these individuals as possible in order to complete a 15 year follow-up of the cohort by the end of the funding period; (3) complete analyses of the previously collected MRI scans, CSF samples and blood, using state-of-art techniques; (4) complete analyses of the relationship of the previously collected data to the current status of the subjects; and (5) provide these data to the scientific community through a publicly accessible database.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this study is to characterize the neurophysiologic mechanisms underlying the age-related decline in human visual cognitive function. This research will delineate specific neurophysiologic measures of subcortical functions that contribute to the age-related decline in cognitive function. It is hypothesized that the impairment of the sub, cortical function of alertness contributes to the age-related decrement in visual cognitive function and is related to right hemisphere dysfunction. Alertness is experimentally modulated in a double blind placebo-controlled drug study using a CNS depressant (diphenhydramine) and CNS stimulant (methylphenidate). Alertness is operationally defined using computerized EEG frequency analysis; slow later\" al eye movements; blink rate; sympathetic nervous system activity (skin blood flow based, on laser Doppler velocimetry); parasympathetic nervous system activity (heart rate, variability). Visual cognitive function is assessed by performance on serial and parallel search tasks and a directed attention task. Measures of cognitive performance include: reaction time; accuracy; variability of reaction time. The hypothesis will be tested with 120 subjects, 60 in each of 2 age groups, 25-35 years and 65-75 years. Subjects will participate on 5 separate mornings. On each morning they will be given either a placebo, 0.5 mg/Kg diphenhydramine, 1 mg/Kg diphenhydramine: 0.1 mg/Kg methylphenidate or 0.2 mg/Kg methylphenidate. They will perform the cognitive tasks before and after administration of each drug condition. The research impacts clinical management of patients with cognitive deficits. Clinicians will be provided with measurement tools to document alertness deficits in cognitively impaired patients with subcortical lesions secondary to aging, focal lesions, neurodegenerative diseases and medications. If deficits in alertness can be measured and contribute to cognitive decline in the elderly, then strategies to increase alertness could be offered by clinicians. This may be particularly helpful in problem areas for the elderly such as driving and ambulation, where intermittent lapses in alertness or attention may produce significant injury. The normative data for cognitive function does not presently account for the subject's level of alertness. Since the level of alertness may account for a large amount of the variability in cognitive function, especially in the elderly and neurologically impaired population, the definition of a measurable state of alertness will improve the sensitivity and specificity of all clinical cognitive testing.GRANT=R35AG09014 The aim of this LEAD application is to test the value of examining cellular and molecular abnormalities in extra-neural tissues from patients with Alzheimer's disease (DAT). It will test whether or not reported abnormalities relate to the presence of -the clinical syndrome of DAT or certain of its subgroups (familial vs sporadic, early vs.late onset, with vs without Parkinsonism, myoclonus, depression, or early aphasia). DAT patients and disease and intact controls of comparable age and sex will all receive detailed examination including neuropsychological testing; follow-up where possible will be to autopsy. Skin cell cultures including biopsy will be meticulously standardized to ensure that DAT and control cells are studied under identical conditions including identical biological age in culture. Parameters measured in the cultures will include two related to the materials which accumulate in DAT brain: amyloid precursor protein, and materials which react with antibodies to paired helical filaments (PHF). (Recent studies by the PI and co-workers indicate that skin cells accumulate anti-PHF reactive materials when grown under specified conditions, much more in DAT cells than in controls). Other parameters to be measured have been reported abnormal in DAT calls in at least two laboratories: isoproterenol-stimulated cyclic AMP synthesis, cellular calcium homeostasis, and [U-14C]glutamine oxidation. Data will be stored in a relational data base (SIR-software) and relations among clinical and laboratory findings analyzed in detail (SAS statistical software). Dr Ronald Black, an assistant professor of Neurology, will develop methods to quantitate anti-PHF reactive materials, compare the amounts of these materials in soluble and insoluble fractions of affected and unaffected areas of DAT and control brains, and then compare their amounts in cultured DAT and control cells. He will gain expertise in clinical as well as laboratory research in dementias by participating actively in the clinical evaluations. One pilot study will examine a possible increase in anti-APP reactive materials in DAT granulocytes, and a second possible abnormalities in phosphokinase activities in cultured DAT skin cells. Future pilots will also study other potential markers. The proposed investigations will extend the PI's ongoing mechanistic studies of abnormalities in cultured DAT cells. They will test directly whether or not the abnormalities studied relate closely to the clinical syndrome of DAT or to DAT subgroups.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Several aspects of obligate intracellular parasitism are being studied in vitro using cell culture and electro-optical systems developed with the program. Future research will be directed at (1) obtaining a clearer understanding of the mechanism of penetration of vertebrate cells by Trypanosoma cruzi trypomastigates and (2) developing in vitro systems for the continuous cultivation of malaria merozoites.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Comprehensive nutritional analysis and food management for metabolic kitchens in research settings require a different set of functions than those supplied by the general software created for nutritionists. These research nutritionists need to be able to create diets with a specific nutrient composition. The solution of this class of problem requires linear programming techniques that meet nutritional researcher's needs and are not available with existing programs. The objective of Phase II is to develop ProNutra, a research dietitian software tool for metabolic research studies, which was specified in Phase I. ProNutra contains several other innovative features, such as the ability to: automatically modify a study's diet schedule to meet the diet requirements of each of the subjects, create kitchen management reports that specifically handle the needs of a research metabolic kitchen, and store and export study, subject, and diet data to third-party statistical software packages with complete user control over the data selection, format, and order. These innovations will result in new methodologies that will: increase accuracy and efficiency of diet design and preparation, provide standardization in research methods among metabolic wards, facilitate the use of valid nutrition research methodologies, and improve the ability to monitor subject response. PROPOSED COMMERCIAL APPLICATION: The completed product in Phase III will be marketed to the general nutritionists in the industry but especially those at the clinical Research Centers, the USDA's Human Nutrition Research Centers, military, industry, hospitals, teaching institutions, and private practice. A modified version with multimedia extensions will be created for home recipe management.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cigarette smoking remains the single most preventable cause or morbidity and mortality in the United States, accounting for approximately 440,000 deaths per year. It is thus essential that researchers continue to investigate the factors relevant to the treatment of nicotine dependence. Drug-related expectancies represent the consequences that an individual expects from the use (or non-use) of any given substance. With respect to smoking, research has focused on smokers'expectancies for the use of cigarettes. These expectancies, as measured by the standard smoking expectancy questionnaires, are robust predictors of smoking motivation and behavior. However, no prior study has examined smokers'expectancies for the non-use of cigarettes. That is, no previous study has investigated the consequences that smokers expect when they quit smoking (that is, abstinence-related expectancies). However, these expectancies likely have considerable significance to the treatment of nicotine dependence. The primary goal of the current study is to examine smokers'expectancies for abstinence via the development of an abstinence-related expectancies questionnaire: the Smoking Abstinence Questionnaire (SAQ). An initial pool of SAQ items will be developed via reference to the literature, focus groups with current smokers, and expert panel review. A draft version of the SAQ will be administered to 500 current smokers and refined with the use of established psychometric procedures, including factor analysis. The SAQ's relationship to smoking-relevant variables will be examined, including its relationship to nicotine dependence, withdrawal, motivation to quit, abstinence self-efficacy, negative affect, and smoking expectancies. We hypothesize that the SAQ will have significant relationships with each of these variables, thereby providing evidence for its construct validity. The proposed study will provide valuable information regarding the process of quitting and important treatment-related process variables. Moreover, the development of the SAQ will allow for continued, systematic, research of the role of abstinence-related expectancies in the quitting process. Finally, this investigation may help inform smoking treatment. Thus, the current study will advance the development of nicotine dependence treatments. Relevance: No prior study has investigated the consequences that smokers anticipate when they quit smoking, although these expectancies likely have significance in the treatment of nicotine dependence. The primary goal of the current study is the development of an abstinence-related expectancies questionnaire. Thus, the current study will advance the development of nicotine dependence treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mesenchymal stem/progenitor cells (MSCs) hold considerable potential for a wide range of tissue regeneration therapies. While their differentiation capacity has been demonstrated extensively, mechanisms that control their plasticity remain poorly understood. Observations by our laboratories and others have shown that proliferating MSCs express lineage-associated molecules prior to induction of differentiation. This has led to a suggested model of differentiation where commitment to a specific cell type results from the combined effects of increased expression along the induced pathway and repression of genes related to other lineages. MicroRNAs are endogenously expressed, small RNAs that may regulate this process. By acting as transcriptional repressors, they have recently emerged as regulators of cellular differentiation in cancer and stem cells. In this proposal we will investigate the role of specific microRNAs and their gene targets on MSC differentiation. We have performed microRNA profiling on MSCs undergoing osteogenic and chondrogenic differentiation and identified miR-130b, miR-432 and miR-559 as differentially expressed miRNAs among both lineages. Functional studies in MSCs suggest multilineage regulation by miR-130b; its overexpression enhanced the osteoblast phenotype and repressed smooth muscle differentiation. Differential gene expression analysis in miR-130b transfected MSCs has identified several mRNA targets that could serve as control points for osteo- and myogenic differentiation. Building on these findings, Aim 1 of this proposal will identify direct targets of miR-130b and assess their role in osteo-, chondro- and myogenic differentiation. The same approach will be followed to assess the functional effects and gene targets of miR-432 and -559. In Specific Aim 2, we will investigate whether the functional effects of these microRNAs are replicated in vivo. MicroRNA overexpression and inhibition experiments will be performed by systemic delivery of lentiviral vectors encoding mimics and hairpin inhibitors to developing mouse embryos. At specific developmental stages embryonic tissues will be evaluated for bone, cartilage and smooth muscle formation. Successful completion of this study will 1) determine how miRNA-mediated gene silencing in mesenchymal cells alters commitment to osteo- chondro- and myogenic lineages and 2) identify potential regulatory roles for these specific microRNAs during musculoskeletal development. This should improve our current understanding of mesenchymal stem cell differentiation and aid future cell-based therapies for musculoskeletal repair.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During the past two decades there has been a dramatic increase in the use of drugs by pregnant women. While the target for drugs administered during pregnancy is the mother, the fetus often becomes an unwanted recipient. Unfortunately, use of drugs during pregnancy can cause structural malformations in the fetus. We propose that many commonly used drugs also can produce more subtle biochemical, physiological and behavioral teratogenic defects in the fetus. The effects of these drug teratogens may not be apparent at birth or even in childhood, but may lie dormant or \"latent\" until years later. We propose that many drugs may interfere, in utero, with the normal differentiation of the liver and brain, resulting in permanent defects in drug metabolism, disposition and action in the adult. By administering commonly used drugs (i.e., barbiturates, benzodiazepines, analgesics, etc.) to pregnant and lactating rodents we plan to study in their adult offspring the delayed teratogenic effects of the drugs on in vitro hepatic microsomal drug metabolism and drug receptor levels in the brain and to correlate these biochemical findings with n vivo measures of drug action.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose that nutrition intervention and physical activity will decrease the amount weight gained, the amount of protein consumed, dyslipidenias, hypertension, and hyperinsulinemia and affect the incidence of progressive fibrosing and atherosclerotic lesions in the renal allograft.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a multi-center, open-label, safety and efficacy study to assess the use of rhFIX in the treatment and prevention of bleeding in severe and moderate hemophilia B patients who have previously received blood products. It is comprised of three segments: (1) a baseline PK segment, (2) a treatment segment, and (3) a surgical Segment (if appicable). Patients will undergo screening during a 30-day period before the first dose of rhFIX is administered in the Baseline PK Segment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ras family GTPases are critical components of cell regulatory systems that control proliferation, differentiation, and cell survival. Inappropriate regulation of these systems directly contributes to initiation and progression of human cancer. This proposal is directed at increasing our understanding of the composition, organization, and function of cell regulatory networks engaged by Ras family GTPases. Our focus is on the dominant effector pathways that mediate oncogenic Ras-induced cell transformation. Our specific aims are 1) revealing the molecular basis of the contribution of Ral GTPases to support of human tumor cell proliferation and survival; 2) assessing the role of two candidate Ras effectors in limiting cellular responses to mitogenic signals; and 3) defining the contribution of scaffolding proteins to the generation of signal fidelity on the ERK1/2 MAP kinase cascade. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Yersinia enterocolitica is a Gram-negative pathogen responsible for a range of clinical syndromes but is primarily associated with gastrointestinal disorders. A number of important paradigms of pathogenesis have emerged from the studies of Y. enterocolitica and Y. pseudotuberculosis. In addition these enteropathogens have served as important models of bacterial invasion, a process primarily encoded by inv. Virulence properties such as invasion can be characterized at the molecular level due to the ease of manipulation of these bacteria in the laboratory and the existence of an excellent murine model of infection for dissecting the host-pathogen interaction. In studies to further our knowledge of inv and its role in virulence we identified a gene, rovA, that regulates expression of inv both in the laboratory and during infection. Subsequent studies demonstrated that RovA acts as a DNA binding protein and promotes inv expression by displacing a repressor complex from the inv promoter. The rovA mutant was less virulent than either the wild type strain or the inv mutant. The rovA virulence defect is characterized by reduced systemic dissemination and an increased LD50 after oral infection. Infection using the i.p. route abrogates the rovA defect, suggesting RovA is required for events occurring either in/from the intestine/colon or in the Peyer's patch. Because the rovA mutant virulence defect was more significant than that of an inv mutant alone, this suggested RovA regulates additional virulence determinants. However, RovA did not appear to regulate the expression of previously identified virulence determinants. Using whole genome microarray analysis we identified 64 genes potentially regulated by RovA, suggesting the regulon may be quite large. The long-term goals are (i) to understand how these genes are regulated by RovA and how that is coordinated with expression of other virulence factors, and (ii) to determine which RovA regulated genes (rrg) contribute to virulence. Specifically we propose the following: (Aim 1) What is the role of the RovA regulon during infection? Recently our understanding of how enteric pathogens interact with the intestinal mucosa and spread systemically has changed significantly but questions still remain. In addition, questions regarding how these bacteria spread from host-to-host are more tractable. Given the known virulence defects of the rovA mutant, we hypothesize that the RovA regulon will play a role in these host-pathogen interactions and feel that a more detailed understanding of both wild type and the rovA mutant with respect to these aspects of infection is warranted. (Aim 2) Which rrgs are important for virulence and how do they contribute to individual RovA-associated phenotypes? (Aim 3) Are all rrgs regulated in the same way as inv? By knowing when and where these gene products are expressed combined with information regarding the phenotype of mutations in these genes we may be able to gain a better understanding of their function. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE: Gastrointestinal disease is a significant cause of morbidity and mortality in infants and of morbidity in adults. Our long-range goal is to gain a full understanding of the Yersinia enterocolitica virulence factors and their contribution to the disease process at the molecular level. This will expand our understanding and ability to intervene therapeutically not only with Yersinia but with many other gastrointestinal pathogens as well. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed Drug Education Proect seeks to lower the number of youth in the Model Neighborhood Area who become drug dependent by reducing their predisposition toward drug use. Specific program objectives include extention of education and training efforts towards broader coverage of the MNA, strengthening of the training program and periodic review, evaluation and refinement of program methods and activities. Program activities include: an educational component which features value clarification and decision making exercises, and which is devised for neighborhood groups, skill-training centers and schools; a training component enabling particpants to develop drug education projects within neighborhoods; an evaluation component which aims at providing pre- and post program analyses, sophisticated attitude probes and feedback mechanisms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "As the applicant notes, herbal medicines are part of folk medicine in developing and developed countries and have been used extensively to reduce pain and discomfort. Unfortunately, very little controlled research has been conducted to determine whether these agents actually produce analgesia, their possible sites and mechanisms of action, and their efficacy and potency relative to known classes of analgesic agents. The major hypothesis of this proposal is that purified components of herbal medicines have potential as analgesic and antihyperalgesic agents and their efficacy and potency can be evaluated using animal models of inflammatory persistent pain. The first aim is to characterize the efficacy of systemic administration of active components of some herbal medicines in a rat model of inflammatory pain and hyperalgesia. The investigators will use their well-characterized model if inflammation to examine the effects of these active components on pain and hyperalgesia that persists for hours or days. This model will also distinguish between analgesic and antihyperalgesic activity. The second aim is to investigate the possible sites of action of agents that have analgesic efficacy by administering these agents to the site of inflammation or to localized sites in the central nervous system. In this instance the investigators will be examining whether these agents have actions in the periphery at the zone of injury, or in the control nervous system at sites of hyperexcitability and sensitization. The third aim will be to compare the efficacy and potency of active components of herbal medicines with known antihyperalgesic and analgesic agents. The principal investigator and his collaborators will compare the relative potency of active components of herbal medicines to known opioids, excitatory amino acid antagonists, neurokinin receptor antagonists and nonsteroidal antiinflammatory agents. The investigators will also administer herbal-derived agents in combination with specific antagonists of analgesia (e.g., naloxone) to reveal possible mechanisms of action.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We are investigating the mechanisms by which the virus, Q-beta, subverts the host's protein synthetic machinery into performing functions in viral RNA replication. Specifically, do the protein synthesis elongation factors Tu and Ts perform functions in Q-beta replicase related to those they perform in protein biosynthesis? We are also studying the structure and mode of action of Q-beta replicase, as well as its mechanism of template specificity. We are building Q-beta replicases with mutant elongation factors and with host polypeptides from distantly-related procaryotes. Other projects include a study of subunit relationships of RNA polymerase and a study of the roles played by Tu and Ts in protein synthesis and in other host processes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pre- and postnatal exposure to opioids can profoundly affect CNS development and function. Since the fastest growing population of opiate (heroin) abusers are young women of childbearing years, there is a public health interest in understanding how opioid signaling affects CNS development. Opioids given experimentally in vivo or to slice cultures during pre- or perinatal periods can alter survival and proliferation of neurons and astroglia, cause permanent changes in CNS structure and adversely affect learning and memory. Opiate exposure also reduces neurogenesis in adult hippocampus by 40 percent. Opioid effects are complex. Depending on the cell type or receptor (mu, delta or kappa) targeted, opioids can be toxic or protective and can have distinct effects on cell maturation. The survival of oligodendrocytes (OLs) and formation of myelin is critical for CNS function. Although opiate abuse can result in myelin pathology, essentially nothing is known about opioid effects on OLs either during neonatal or perinatal periods or in the adult. Our work has defined the existence of opioid signaling pathways in cultured OLs by showing that: (a) OLs express mu- and kappa-opioid receptors in a temporally specific pattern; (b) OLs have physiologic responses (survival, proliferation, myelin production) to selective manipulation of receptors; (c) OLs synthesize, process and probably secrete 2 classes of endogenous opioids (dynorphins, enkephalins). The central goals of this proposal are to determine the spatiotemporal expression patterns of opioid receptors and peptides within the developing and mature CNS, and to determine the role that opioids play in modulating the survival and function of OLs. Functional studies center on the role of dynorphin peptides since our findings show dynorphin mediates effects on the survival of OLs and neurons. A secondary goal is to determine whether manipulation of opioid signaling pathways can promote OL survival and myelination in clinical conditions with myelin pathology. Proposed studies use complementary in vivo and in vitro approaches employing mice deficient in opioid receptors and dynorphin to: (1) Identify spatiotemporal patterns of opioid receptor expression on OLs in the CNS; (2) Identify spatiotemporal patterns of dynorphin expression on OLs in the CNS and determine if dynorphin peptides are secreted; (3) Test the hypothesis that signaling through kappa-opioid receptors promotes OL survival and activates the PI3-kinase/Aktl pathway; (4) Test the hypothesis that some dynorphin peptides have glutamatergic effects on OLs. Techniques used include cell culture, immunostaining, in situ hybridization, immunoblot, confocal microscopy and adenoviral transfection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this application is to study the role played by van Willebrand factor (VWF) in initiating platelet deposition at sites of vascular injury, particularly in areas of the circulation characterized by rapid blood flow. The goal is to elucidate the mechanisms of adhesive interactions and cellular responses that are relevant to the arrest of bleeding from wounded tissues but also to the development of common and serious diseases caused by arterial thrombosis, such as myocardial infarction and stroke. The first aim is to define the biomechanical characteristics of the bonds formed between the VWF A1 domain and the platelet ilycoprotein (GP) Ib-alpha in relation to specific structural aspects of the A1 domain. The second aim is to define the mechanisms through which soluble VWF multimers and membrane tethers contribute to stabilizing platelet adhesion mediated by GP Ib-alpha. The third aim is based on structural evidence that alpha-thrombin may stabilize the VWF A1 domain-GP Ib-alpha interaction, and may thus play an unexpected role in modulating the initial adhesion of platelets to a thrombogenic surface through a mechanism that is independent of platelet activation. The proposed studies will clarify the biological significance and mechanism of this novel athrombin function. The fourth aim is to characterize the signaling events related to the interaction between the VWF A1 domain and GP Ib-alpha, and obtain a more definitive definition of the mechanisms through which the interaction contributes to platelet activation. The fifth aim is to define how VWF binds to extracellular matrix components, in particular to define the potential biological significance of the interactions with collagen type VI and the oligosaccharides that are present in proteoglycans. Extensive interactions with all the other projects in the program and with a core A will facilitate the development of these aims. The results of the proposed studies will have an impact on public health by providing novel mechanistic information on processes that are central to normal hemostasis and pathological arterial thrombosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of the proposed research is to investigate the functions of the placental members of the prolactin/growth hormone family in the mouse. Since prolactin has a broad spectrum of activities in animal physiology, these related placental hormones are likely to be important regulators of the physiological changes that occur during pregnancy. Four placental members of the prolactin/growth hormone family have been identified in the mouse. We hypothesize that the interactions of these hormones with their receptors are important in regulating maternal physiology, fetal development, and the successful completion of gestation in the mouse. We propose to prevent the synthesis of one of these placental hormones (placental lactogen II) by generating a mouse strain in which the gene encoding this protein has been disrupted. Since this protein (as well as placental lactogen I) binds to the same receptor that recognizes prolactin, we will investigate the locations, timing, and levels of prolactin receptor expression in the developing mouse fetus. We will also generate mice that have a mutated gene for the prolactin receptor; this will test if the interaction of this receptor with these two placental hormones is essential for normal fetal development. If mice that lack the prolactin receptor are able to develop, then these mice would be of value in investigating the requirement for this receptor in such processes as growth and metabolism, reproduction, osmotic balance, lung development, and immune response in the adult. Two of the placental members of the prolactin/growth hormone family in the mouse do not bind to either the prolactin receptor or the growth hormone receptor, and their physiological roles and targets have not been defined. Our recent data indicate that physiological concentrations of these two proteins regulate angiogenesis: one of these proteins (proliferin) stimulates angiogenesis, while the other (proliferin-related protein) inhibits this process. We therefore hypothesize that these proteins play important roles in initiating and limiting placental angiogenesis and that they bind to specific receptors present on capillary endothelial cells. The actions of these two placental hormones in regulating angiogenesis will be further characterized, and the receptors through which these hormones act will be identified. In addition, we hypothesize that the expression of proliferin may occur in tumors, with this hormone then acting as a paracrine angiogenesis factor that promotes tumor growth and metastasis. By introducing an expression construct for proliferin into tumor cells, we will determine if we can increase the tumorigenicity of these cells. Similarly, the effect of expression of proliferin-related protein will be tested as a possible means of inhibiting angiogenesis, and therefore the growth, of tumors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To assess the effects of intrathecal baclofen on persons with generalized dystonia secondary to cerebral palsy or traumatic brain injury. The research design has two phases. First an open label trial of intrathecal baclofen is infused by an external pump. If the patient responds to the trial, they may continue to a second phase: a randomized, double-blind trial of baclofen versus placebo after programmable pump implant. Outcome measures for this study are based on our dystonia scale and functional assessments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Informatics/Data Management Core will be responsible for providing computer-based data management tools that facilitate the storage, retrieval, and analysis of the data generated in this research. The system must be accessible to each laboratory participating in the project, and should work as a communications gateway and server. It should have power and memory to quickly process large amounts of data for analytical and simulating statistical analysis. The system will build on the prototype database of FCCC and that being developed by the Human Genome Initiative for management of pedigree data and laboratory reagents and results and will have an analysis component to perform linkage segregation and risk analysis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary goal of this proposal is to systematically examine the psychological and physiological responses of narcotic addicts to stimuli previously associated with drug-taking behavior. Drug addicts and non-addicted control subjects are shown stimuli reminiscent of drug-taking behavior in the form of a videotape (VT) and an assortment of drug paraphernalia. Prior to the stimulus presentation, measures of mood state and craving are taken. During the presentation, physiological measures of heart rate, skin resistance, skin temperature and respiration are monitored along with subjective level of craving. Subsequent to the VT, the psychological measures are taken and compared to the pre-stimulus scores. The same measures are also taken before, during and after viewing control films, including a non-drug-related, emotionally neutral film and non-drug-related stress film as controls for general arousal and stress factors. Goals of our study include an assessment of the stability of drug addicts' response systems. Initial results indicate that a longer period of time might be necessary for habituation to the experimental set-up, as compared with controls. In subsequent studies we will attempt to answer the following questions: (1) Are the subjects' responses specific to drug-related stimuli? (2) Are the responses limited to addicts, i.e., those with conditioning experiences - or at least, is the response topography different from those control groups? (3) Are the responses greater in subjects who have been addicted longer - thus having a longer conditioning history? and (4) Are the conditioned responses a function of drive or current levels of opiate use? This project is an attempt to expand on prior studies by the applicants demonstrating psychological and physiological conditioned responses to a VT. The present design should accomplish this in a systematic manner. The use of appropriate control subjects, precise presentation of stimuli and additional physiological measures will allow us to draw firm conclusions for our results.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "GOAL: To decrease the morbidity and mortality of cancer in Wisconsin. OBJECTIVES: 1. Planning and Legislation. To develop a comprehensive plan for control of cancer in Wisconsin and a strategy for its enactment into formal legislation. 2. Epidemiology and Evaluation. To further develop and maintain a strong WCCC Biometry component to aid in the planning, implementation and evaluation of Wisconsin cancer control programs. 3. Public Information and Eduction. To develop and promote needed programs to inform the public about cancer. 4. Professional Information and Education. To conduct necessary and evaluable programs to inform and educate health professionals about cancer. 5. Prevention and Early Detection. To promote optimal practices in cancer prevention and early detection. 6. Diagnosis and Treatment. To organize regional, cooperative, site-oriented programs, networks and task forces dedicated to improving the diagnosis and treatment of cancer. 7. Rehabilitation and Continuing Care. By demonstration, to inform the public and professional communities in Wisconsin about effective rehabilitation and continuing care possibilities and practices for cancer patients. 8. New Projects Development. To catalyze, encourage and support the development of new ideas in cancer control.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary: 518 human protein kinases modulate the activity of -30% of all proteins, and collectively control almost all complex pathways and decisions of a cell. Despite tremendous experimental analysis on some kinases, we know little of the detailed function of most members of this uniquely important family. We propose to use genome sequences to tap hundreds of millions of years of evolutionary experimentation, in order to clarify the link between kinase sequence and biological function. We will use our extensive knowledge from the discovery of the human and mouse kinomes to predict all kinase orthologs in up to 40 vertebrate genomes. We will then map the evolutionary constraints on every residue of every kinase, and predict domains, motifs, phosphorylation sites, and other functional regions of proteins, extending the kinome catalog to unprecedented resolution and generating a wealth of hypotheses for experimental testing. Finally, we will apply this knowledge to predict the functional impact of both SNPs and somatic mutations in cancer, providing a valuable preview of the proposed cancer genome atlas. By using kinases as a model family to explore the predictive power of comparative genomics, we will develop well-validated tools and parameters which can then be applied to any human gene or gene family. Relevance: Protein kinases key controllers of cell function, are one of the most important gene families in disease and development of new drug therapies. By exploring how human kinases vary from those of other vertebrates, we can predict whether any human sequence change can predispose to disease or alter drug response, and can distinguish cancer-driving mutations in tumors from background mutations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research aims to study mechanisms of synaptic function with emphasis on the origins and locations of individual synaptic membrane polypeptides. By reacting epsilon-amino groups of lysines which are exposed on membrane surfaces with sodium boro (H3) hydride in the presence of pyridoxal phosphate it is possible to selectively label exposed peptide areas of intact synaptosomal membranes. Combining this technique with purification and subsequent gel electrophoresis of synaptosomal sub-fractions from chick brain allows classification of membrane polypeptides into four groups: those occurring entirely within the membrane, those exposed on both the internal and external faces, and those exposed only at one surface, either external or internal. The nature of the peptides around the exposed lysine residues will be characterized by labeling polypeptides eluted from the gels with I125, cleaving with trypsin or cyanogen bromide, and performing autoradiographic peptide mapping on the resulting peptide mixtures. Whether or not synaptic vesicles fuse with the external membrane will be investigated by comparing labeled gels and peptides of these two fractions. Distances between polypeptides in the membrane will be determined with bifunctional reagents with various bridge lengths between the functional groups. When basic parameters are defined synaptosomes prepared from synapses in differing states of activity will be examined. Alterations of membrane conformation and subsequent protein exposure may reflect neural activity. Synaptosomal membranes will be prepared from electrically stimulated brain slices and chick optic lobes that are either denervated (by enucleation) or receive minimal input (by eyelid suture). Comparing such preparations with normal synaptosomal membranes may reveal a relation between synaptic membrane structure and its functional use. Such data will have important implications for theories both of normal brain function and of the etiology of neurological and psychiatric illness.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "P. aeruginosa (PA) frequently infects immunocompromised individuals with HIV-1, cancer, and cystic fibrosis (CF). Since PA is increasingly resistant to antibiotics, its infection often leads to either severe states or chronic situations with a persistent inflammatory response. Better understanding of host-pathogen interaction may suggest a more effective approach to combating this pathogen. MCP-1 is a major chemokine secreted by alveolar epithelial cells type II (AECII). Recent research has illustrated an immune role of AECII in PA infection, but the underlying mechanism remains unidentified. Our long-term goal is to understand the mechanism of host immunity and develop new strategies for controlling respiratory infections. The objective of this application is to elucidate the immune function of AECII, in particular through their secretion of cytokines and activation of AM. Our central hypothesis is that AECII can secrete cytokines (MCP-1) to enhance AM's anti-bacterial immunity through a lipid raft- mediated mechanism. We have formulated this hypothesis based on our recent findings that both AECII and AM participate in innate immunity against PA. We further found that membrane lipid rafts may be instrumental for regulating cytokine secretion. Using our primary cell model, we have discovered an immune role of AECII in enhancing AM's immunity using a conditioned AECII medium. Our data also suggest that AECII play a critical role in PA infection by secreting MCP-1 and recruiting the classically activated macrophages (CAM). The rationale is that elucidating how AECII enhance AM immunity will indicate a potential strategy to bolster immunity against PA. Our laboratory is ideally suited for this research, having the relevant expertise in isolation and culture of AECII a well as in lung infection models. We propose the following three specific aims: Specific Aim 1: Define the immune role of AECII cells in secreting cytokines during PA infection. We will identify the source of MCP-1 using in situ hybridization with AECII marker SPC. We will also use primary AECII culture to show MCP-1 as a dominant cytokine. Furthermore, AM and AECII from MCP-1-/- mice will be examined for their decreased immune function against PA infection. We will determine the ability of AECII in recruiting the classically activated macrophages (CAM). Specific Aim 2: Evaluate how lipid rafts regulate MCP-1 secretion in AECII. We will study the underlying mechanism for MCP-1 secretion and hopefully identify the involvement of ceramide-rich membrane microdomains. Acid shingomyelinase will be blocked by siRNA and chemical inhibitors for analyzing sphingolipid hydrolysis during PA early infection. Specific Aim 3: Assess the potential of super-AECII over-expressing MCP-1 in enhancing anti-PA capacity of human AM. We will create super-AECII using retroviral vectors to secrete high levels of MCP-1 and test their host defense in PA infection. We will also demonstrate that human AM can be activated by AECII and that this translational research may imply the clinical value of the immune AECII. This research will be performed by graduate and undergraduate students. Our efforts are expected to substantially advance understanding of this previously unrecognized immune function of AECII in activating AM, and may provide insights into mechanisms of cytokine secretion, with indications in development of novel therapeutics for treating this infection. PUBLIC HEALTH RELEVANCE: P. aeruginosa (PA) is a bacterium that causes severe infections, particularly in immunodeficient individuals who are suffering tuberculosis, cancer, AIDS, severe burns, and cystic fibrosis. Because PA is increasingly resistant to antibiotics, its infection usually leads to a chronic state of persistent inflammatory response. We have made the surprising discovery that MCP-1, a versatile cytokine from alveolar epithelial cells, regulates host defense and inflammatory response in PA infection. We have also noted that lipid rafts may be important for regulating cytokine production. Through secretion of MCP-1, the alveolar epithelial cells may recruit a particular subset of macrophages (i.e., classically activated macrophages) to promptly respond to infection. Studying the immune role of alveolar epithelial cells may provide new insights into the development of novel treatment for PA infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of this project is to develop strategies to improve the treatment of breast cancer. The aromatase inhibitors we pioneered in the early phases of this grant are now proving to be of value in the clinic. Although AIs appear to be more effective than tamoxifen, some patients may acquire resistance or have de novo resistance to AIs. The unique preclinical model we have developed to study the effect of aromatase inhibitors (AIs) has provided accurate predictions of clinical outcome. We propose to use this model to study mechanisms of tumor resistance. We have also developed unique tumors and cell lines that are resistant to letrozole and other AIs for use in these investigations. In the proposed studies, we will focus our efforts on several significant observations made during the current period. We plan to determine the mechanisms of resistance to aromatase inhibitors and how these might be reversed so that response to well tolerated AI treatment can be restored. The Specific Aims of the proposal are: 1) to investigate the functional the role of HER2/MAPK in estrogen receptor regulation and development of resistance to AIs. We will explore whether other inhibitors of HER2, such as lapatinib, pertuzumab and HKI-272 also reverse resistance to letrozole; 2) to investigate whether down-regulation of the estrogen receptor (ER) by fulvestrant prevents crosstalk with tyrosine kinase receptors (TKR) in AI resistance. We will determine whether combining an AI and estrogen down regulator is more effective than complete estrogen suppression in controlling tumor growth. Based on mechanisms identified, the optimal efficacy of these agents in combination or sequential strategies will be determined; 3) to investigate mechanisms involved in reversing resistance of tumors to letrozole and other AIs following withdrawal of AI treatment; 4) to investigate whether histone deacetylase (HDAC) inhibitors will convert ER negative cells to hormone responsive cells that are sensitive to HDACI + AI treatment and reduce the cancer progenitor cells or side population in these and AI resistant cells and tumors. Our studies should provide information with which to plan new strategies to treat breast cancer patients. PUBLIC HEALTH RELEVANCE: This competitive renewal application is to continue studies to understand the mechanisms involved in resistance of breast cancers to aromatase inhibitor treatment. Having identified mechanisms of resistance to aromatase inhibitors, we will then apply this information to develop strategies to reverse resistance and restore sensitivity to aromatase inhibitor treatment. These strategies will be tested in our unique model to determine their anti-tumor efficacy. The results of these studies could improve treatment for breast cancer patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Summary We know very little about the mechanism of INO80, how it disrupts nucleosomes and the factors governing its activity. We will take detailed ?snapshots? of INO80 during nucleosome remodeling to find how INO80 and nucleosomes are moved during remodeling. A series of orthogonal approaches will be used to arrest INO80 remodeling at distinct stages and examine conformational changes in the core nucleosome and INO80. We will build on our recent observations of the motor domain being engaged at the H2A-H2B interface and persistently displacing DNA from this surface to find why displacement occurs, the factors that control displacement and whether this displacement weakens the interactions of H2A or H2A.Z dimers with the rest of the histone octamer or otherwise disrupts the nucleosome structure. Based on the proximity of Arp5 to nucleosomal DNA, we will test the premise of Arp5 as the ?gatekeeper? regulating DNA traversing through the center of nucleosomes with wild type INO80 and mutant Arp5 in which either its histone or nucleosome binding regions have been deleted or mutated. We will also test whether the Arp8 module regulates Arp5 interactions with the acidic pocket of nucleosomes or nucleosomal DNA and if communication between these two domains is mediated by the Ino80 catalytic subunit. INO80 will be arrested at different stages in remodeling by limiting DNA translocations to specified distances, arresting with non-hydrolyzable ATP analogs, limiting linker DNA length and mutation of Arp8 and Arp5. We will probe the role of DNA sequence in INO80 remodeling because we observed coupling of ATPase activity to nucleosome movement being dramatically affected by the DNA sequence of the core nucleosome. We will find as suggested in these experiments if INO80 interactions and conformation varies depending on the DNA sequence bound by nucleosomes. In order to better examine the importance of DNA sequence in a ?native? context, we will use yeast chromatin reconstituted with recombinant histones and simultaneously examine the differences of INO80 binding and remodeling with many thousands of nucleosomes, each with a different DNA sequence. We will use our expertise of mapping protein-DNA interactions in these genomic assays to sort with high precision the interactions of the INO80 subunits along with nucleosome movement, composition and structural features at ~bp resolution to provide a detailed analysis of each of these nucleosomes in a time resolved manner when remodeled. This approach will provide more insights into the DNA sequence specificity of INO80 and if there are ?hot spots? for mobilizing/ destabilizing nucleosomes or exchanging H2A.Z in the yeast genome that doesn?t require additional factors. To confirm if INO80 behaves the same in vivo as in our in vitro assays, we will transfer several of these approaches to yeast cells so that we can measure chromatin dynamics in vivo with the same resolution as in vitro. We will compare how mutations in Arp5 and Arp8 change nucleosome dynamics in vivo, the importance of genomic position, and other factors for INO80 remodeling not present in our yeast reconstituted chromatin.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Diarrhea is the most commonly reported illness in travelers from industrialized nations to the developing world. It is estimated that of the 7.6 million U.S. residents traveling to these regions annually, 30-70% contract Enterotoxigenic E. coli (ETEC) related diarrhea [1-3]. The objective of the studies proposed here is the commercial development of TravelGAMTM, an Anti-ETEC hyperimmune Milk Immunoglobulin, intended for oral prophylaxis against diarrhea caused by ETEC. In Phase I, the clinical importance of including colonization factor antigen I (CFA/I) in the bovine vaccine was determined. During that period, ImmuCell gathered preliminary efficacy data for TravelGAM in a human challenge study. In Phase II the objectives are: (1) Evaluation of improved methods of in vitro CFA expression for use in CFA screening by utilization of cfaD constructs too up-regulate pilus expression in toxin-positive diarrheal ETEC isolates which, in their native state, do not express immuno- detectable CFAs in vitro; (2)Manufacture of TravelGAMTM containing an expanded range of anti-colonization factor antigen (CFA)s) activity suitable for prospective field testing; (3) Scale-up and cGMP manufacture of enteric coated formulation of TravelGAM; (4) evaluation of the efficacy of formulated TravelGAM in a prospective field study in collaboration with the US Navel Medical Research Unit No. 3 in Cairo, Egypt under ImmuCell's Investigational New Drug Application BB-6049. In Phase III, we hope to build upon the prospective study proposed herein by conducting a pivotal effectiveness study in collaboration with a corporate marketing partner. PROPOSED COMMERCIAL APPLICATION: ImmuCell envisions manufacturing a purified-antibody product based on immune bovine milk whey, for oral administration to patients suffering rom Travelers' diarrhea caused by ETEC. Of the 16 million people who travel from industrialized to developing countries each year 6 million are Americans. Best estimates predict that at least one third, or 2.7 trillion individuals from the U.S. will experience Travelers' diarrhea per year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The potential of proteins to form structural complexes is a most important property which determines their function in biological systems. A rheological method to resolve and characterize this potential is proposed. Dominant intermolecular interactions will be identified in concentrated solutions of globular proteins from the concentration dependence of their viscosity. Three hydrodynamic parameters are defined to analyze the factors affecting the viscosity of proteins; the interaction factor (delta) determines the maximum interaction radius, the constraint number (Nc) measures the lower limit of this interaction radius, and the compressibility factor (Fc) evaluates the capacity of proteins to compress their radius of interaction under increasing concentration. These hydrodynamic parameters depend on the geometry of molecules or aggregates and the nature of protein-protein interactions. These parameters have been evaluated for common geometrical shapes and aggregates to serve as a basis to identify dominant interactions in concentrated solutions of globular proteins. Dynamic viscoelastic properties of these protein solutions will be used to characterize the elasticity (energy storage capacity) of these interactions and to establish a basis for their later resolution from complex systems. Using a model globular protein, the predictive capability of current structural models will be evaluated. In addition, the properties of globular proteins at higher concentrations will be related to hydrodynamic properties in dilute solutions, based on their potential to self-associate and/or interact electrostatically. The proposed research represents a pioneering effort to resolve dominant interactions in concentrated solutions of globular proteins with steady and dynamic viscoelastic measurements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Scavenger receptors (SR) are cell surface proteins that bind chemically modified lipoproteins and exhibit broad ligand binding specificities. We have identified three classes of vertebrate and invertebrate SRs: class A (SR-A), class B (SR-B) and class C (SR-C). They participate in or influence lipoprotein metabolism, development, host defense (innate immunity, protection against septic shock and viral infection), possibly asbestosis, recognition and clearance of damaged (apoptotic) cells and macromolecules, red blood cell maturation, female fertility and atherosclerosis/coronary heart disease (CHD). Many of their functions are directly related to health and disease and are consequences of their broad ligand binding specificities. One of these, SR-BI, is a physiologically relevant HDL receptor that controls the levels and fates of plasma HDL cholesterol, including delivery to the liver and steroidogenic tissues. SR-BI mediates selective uptake of HDL cholesterol, a poorly understood mechanism distinct from classic lipoprotein endocytic uptake and cholesterol effiux. The overall goals of this proposal are 1) to elucidate the biochemical and structural bases for the high affinity, broad ligand binding specificities of these receptors by determining how their ligand binding domains (e.g., collagenous and alpha-helical coiled-coil domains of SR-AI/II) recognize diverse arrays of structurally distinct ligands, 2) to provide additional insights into the novel molecular mechanism underlying selective lipid uptake and cholesterol efflux, and 3) to provide both experimental tools and a biochemical framework with which to assess further the functions of these unusual receptors. The work will rely on the generation and functional analysis of mutant receptors generated using standard and novel methods. Detailed characterization of the structures and distinctive binding properties of mammalian and invertebrate scavenger receptors will provide important tools for the analysis of scavenger receptor function and will probably suggest new approaches for the treatment and prevention of at least some of the related diseases (e.g., atherosclerosis, infectious disease, female infertility). The proposed work may lead to methods for predicting which physiologically relevant molecules are receptor ligands; this would provide additional avenues for exploring receptor function and, possibly, the design of pharmacologic reagents. In addition, clarification of the molecular bases of the broad bind specificities of scavenger receptors may provide insight into other biological systems in which broad binding specificity is important, e.g., multidrug resistance. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Annually, more than 300,000 upper extremity injuries in the U.S. require operative treatment for repair of tendons injured in their midsubstance or at their insertion sites. These injuries lead to an estimated loss of 4 million workdays. Recent improvements in the treatment of flexor tendon midsubstance injuries have been driven by advances in the scientific understanding of repair and rehabilitation variables. By contrast, there have been no significant changes in the treatment of flexor tendon insertion-site injuries in several decades, in part because there have been few scientific investigations to support such changes. As a result, many patients have a poor clinical outcome after repair of the flexor tendon insertion site, as evidenced by decreased range of motion and loss of grip strength. Our long-term objective is to identify repair and rehabilitation techniques that will consistently produce excellent clinical function for immediate and delayed treatment of flexor tendon insertion-site injuries. In this project, we will apply a canine model of flexor tendon insertion-site injury and repair to investigate several clinically relevant variables that have not been previously addressed: 1) suture technique for reattachment of tendon to bone, 2) increased tendon force and excursion applied during rehabilitation, 3) time interval from injury to repair, and 4) growth factor enhancement of tendon-bone healing. Our primary hypothesis is that the stiffness and strength of the repair site are improved by application of increased tendon force during early, passive motion rehabilitation. In addition, we hypothesize that healing of the tendon-bone repair site can be accelerated by delivery, at the time of repair, of targeted gene products that enhance expression of growth factors that are important to early tissue healing. These include: basic fibroblast growth factor (bFGF), platelet- derived growth factor (PDGF) and vascular endothelial growth factor (VEGF). A multidisciplinary approach will be utilized for evaluation of the experimental variables, with biomechanical, histological and biochemical outcomes analyzed. The final determination of practical benefit will be based on biomechanical assessment of digital range of motion and repair-site stiffness and strength during the critical first 6 weeks of healing. Identification of improved repair and rehabilitation techniques for insertion-site injuries will be an initial step toward modernizing clinical treatment and improving patient outcomes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-range objectives of this laboratory are to determine the mechanisms by which enzymes catalyze reactions, and in a more general way to determine the nature of biological interactions at the molecular level. To these we have now added an attempt to gain some understanding of the energetics of the catalytic process and to learn the relationship between the energetics of individual steps and the chemical events occurring in those steps. For some years we have focused our investigations on the glutamic dehydrogenase system which offers a wide variety of phenomena with which to pursue such studies. Having already blocked out the order of occurrence of the nine major complexes formed along the reaction time course, we now turn to the study of the detailed chemical events involved in the interconversions of those complexes. In this proposal we focus our attention on two critical areas of the reaction sequence: 1) What is the role of the carbonyl group of Alpha-ketoglutarate, and at precisely which point does the required water molecule enter the reaction course? This aim will be explored through the use of an analog lacking the carbonyl group and by a variety of 0 1 8 studies. 2) What are the events and complexes preceding and accompanying the hydride transfer step? This will be explored by a study of very pertinent model reactions and by parallel studies on the early transient steps. This latter study will include cryoenzymology and alternative kinetic approach to the study of transients which may have some application to the study of enzymes in general. Finally, we will attempt to relate a recently discovered temperature-dependent poised equilibrium between two isomeric forms of the free enzyme to changes in hydration and (or) to the large negative DeltaCp's observed in many of the complexes of pyridine nucleotide dehydrogenases. These initial experimental probes are a prelude to the definition of a serious effort in this area on our part to determine whether, as Jencks has suggested, part of the substrate binding energy can be channeled into driving a later catalytic step, and if so, how it is accomplished.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term goal of this research is to understand the functional basis for the expression of apolipoprotein (apo) E in peripheral tissues. The proposal consists of 4 aims that are focused on activities of apoE that influence cellular cholesterol metabolism in adrenal cells and activities that are anti-atherogenic within the vascular wall. Experiments in Aim 1 will test the hypothesis that low levels of systemic apoE act within the vascular wall to block an early step in atherosclerotic lesion development. Aim 2 will explicitly test the hypothesis that low levels of apoE provide protection against atherogenesis initiated by apoB48 lipoproteins but do not protect against atherogenesis due to apoB100 lipoproteins. Aim 3 will test the hypothesis that adrenal gland apoE expression increases adrenal cholesteryl ester storage and diminishes steroid production. Aim 4 will test the physiological importance of a newly discovered apoE-stimulated pathway for the selective uptake of low density lipoprotein cholesteryl ester.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is a delicate balance between food intake and metabolic expenditures. Small mismatches lead to quite large changes in body energy content (weight). It seems likely, both on ecological and empirical grounds, that exercise constitutes an important regulator of appetite and/or body weight. The proposed experiments will examine the temporal relationship between food intake and voluntary exercise in golden hamsters and rats. It has been reported that hamsters increase their food intake during exercise, and we shall determine whether that increase reflects meal size or frequency changes. We shall examine the blood metabolites (glucose, fatty acids, ketones) associated with exercise and post-exercise periods, in relation to meals. We shall relate these findings to the failure of hamsters to increase their food intake on a variety of restricted food access schedules. Hamsters exhibit nocturnal activity bouts of several hours duration, and hence are an interesting model in which to study the relationships between metabolic expenditure and food intake. Rats are known to eat in excess of their metabolic needs during their active (night) phase, yet hamsters eat about the same by day and night and are active only at night. It is thus likely that the circadian lipolysis-lipogenesis cycles are different in these two species, and we hope to document this.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The review of our CDP plan was positive and noted that the potential candidates described were very strong. However, our reliance on a funding source for senior level post-doctoral fellows was considered not consistent with the SPORE guidelines. We have therefore revised the entire CDP to more cleariy provide research support for advanced fellows, junior faculty and established investigators who wish to develop or refocus their careers on translational breast cancer research. CDP support for research fellows through the Thomsen Family Fellowship will be used only when it is for support of research fellows transitioning to independent faculty appointments. In this submission, we have clarified our goals and our adherence to the SPORE guidelines. Our proposed plan describes in detail how promising individuals with an interest in, and expressed commitment to, translational breast cancer research will be selected. It further addresses how the CDP leadership will seek out and recruit qualified women and minorities for participation in the program. More specifically, the CDP Leadership will use program funds to recruit and nurture qualified investigators at all levels. CDP leaders will provide a system for mentoring such individuals in a broad range of disciplines and create a framework in which investigators can gain exposure to, and possibly training in, aspects of translational breast cancer research outside their areas of expertise. Ensuring that innovative ideas can develop into promising translational studies in breast cancer research, and that women, minorities and developing faculty who can make key contributions to translational breast cancer research at the FHCRC and UW, are strongly recruited and retained are also key goals ofthe current plan. In a response to the comment: Although the commitment to diversity is not questioned, a paragraph stating the gender/minority composition ofthe research programs at FHCRC/UW was not included, we have added two tables to section C.2 to address this oversight. The first presents the ethnic breakdown of faculty, postdoctoral candidates and pre-doctoral students at FHCRC as of December 31, 2008. The second provides enrollment figures by ethnicity for the UW School of Medicine (UWSOM) in 2007.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Office of National AIDS Policy (2000) estimates that half of new HIV infections occur in youth under the age of 25. Sexual risk behaviors and substance use appear to be highly prevalent among HIV positive youth. The few studies of HIV positive youth suggest adherence rates that are dismally inadequate to manage the disease. Adherence is a multi-dimensional construct that includes taking medication, keeping appointments, and general health behaviors. Poor adherence increases transmission risk resulting from elevated viral loads. This triad of risk behaviors in HIV-infected youth (drug use, sexual behaviors, and health behaviors) must be targeted in intervention studies. We plan to pilot an empirically validated risk reduction intervention, Motivational Enhancement Therapy (MET), that can be easily disseminated and integrated into existing HIV clinics and community-based organizations addressing the needs of HIV positive youth. The proposed study is a randomized clinical trial with a wait-list control examining the utility of MET. This pilot study will use a sample of 60, ages 16-24, 30 of whom will receive MET immediately after baseline and 30 of whom will begin MET after the 9-month post-test. A repeated measures design will be used for the proposed study with a multiple baseline for the control group to examine level and trend effects. Youth will complete an initial data collection sessions (baseline), and then 30 youth will be assigned to the treatment condition. These youth will complete a three month post-test designed to coincide with treatment completion. Subsequent post-tests occur at nine months and 15 months after baseline data collection (6 and 12 months after treatment completion). At that point, the wait-list group will begin treatment. The waitlist control will receive standard care during the initial treatment period. For those in care, all four sites offer comprehensive, multidisciplinary care including social work and case management services. Those youth in the control condition who are not in care will be given referral for medical care. Youth in the control condition w ill participate in the three month and nine month post-test for the multiple baseline design. They will than enter treatment and receive additional post-tests at 12 months (post-treatment), 18 months (6months after treatment completion) and 24 (12 months after treatment completion). If successful, this intervention will provide immediate assistance to vulnerable population to prevent the spread of HIV and to minimize its negative physical and psychological effects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The adrenergic and cholinergic components of the autonomic nervous system serve an important modulatory role in the cardiovascular system. Norepinephrine is the primary physiologic agonist in the adrenergic arm of the autonomic system. In the heart, norepinephrine is known to activate three types of receptors: alpha 1, alpha 2, and beta 1. The increase in cell calcium by norepinephrine both augments action potential triggered sarcoplasmic reticulum (SR) calcium release to cause an enhanced contractility and increases the likelihood for arrhythmogenic diastolic SR calcium release, seen in myocytes as spontaneous contractile waves. While the positive inotropic effect of alpha1-adrenergic agonists on myocardial contractility is thought to be mediated via an increase in cell inositol 1,4,5-tris phosphate (IP3), alpha1-adrenergic agonists also increase 1,2-diacylglycerol which activates protein kinase C. We examined the relative potency of alpha and beta mechanisms and the effect of phorbol ester, an activator of protein kinase C, on these neurotransmitter effects in single adult rat myocytes. Contractility was measured as the velocity of shortening during stimulation at 1 Hz. Waves were measured in a 30 sec window following 2 min of stimulation. In absence of drugs average velocity of shortening was 70 plus/minus 32 um/sec (x-SEM, n=6) and no waves occurred. Norepinephrine (1x10-5M) increased velocity of shortening to 300 plus/minus 70% control (n=6), and caused 3.6 plus/minus 1.55 waves to occur (n=6). Beta (norepinephrine plus prazocin (1x10-5M) had a similar effect: velocity of shortening increased to 310 plus/minus 93% control and 1.8 plus/minus 0.95 waves occurred (n=6). In contrast alpha (norepinephrine plus propranolol (1x10-6M) increased velocity of shortening by 37 plus/minus 28% (n=6) and no waves occurred. Thus, the increased contractility and enhanced probability for spontaneous diastolic calcium release to occur in response to neurotransmitter release in situ are essentially beta rather than alpha in nature. In Quin2 loaded myocytes, norepinephrine and beta decreased resting calcium and increased the rate of calcium uptake with KCl depolarization compared to control. Alpha had no effect on resting calcium and decreased the rate of calcium uptake with KCl depolarization.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A Pre-Clinical Model of TBI Heterogeneity. The majority of traumatic brain injuries are considered mild. Mild TBI (mTBI) is difficult to diagnose, and despite recent progress in public awareness and clinical management, mTBI (or concussion) is a persistent problem in sports, the military, and in the general population. In order to further understand injury mechanisms, identify more sensitive diagnostic tools, and begin to delve into the apparent risk for neurodegeneration, there is a need to improve pre-clinical laboratory studies. This is no easy task given patient heterogeneity and the physiological complexity of TBI. Despite decades of basic research, successful translation of potential treatments from animal to patients is extremely poor. Clinical population heterogeneity is not captured in pre-clinical studies, gravely limiting the ability to validate animal models as reliabl research surrogates. The long-term goal of this research is to improve lab-to-clinic translation using a bi- directional systems approach. The overall goal of this R21 Research Proposal is to reflect the heterogeneity of the mTBI patient population in a pre-clinical animal study and use informatics-based analysis to determine features that contribute to the injury response. The central hypothesis is that systematic institution of animal heterogeneity will result in more reproducible and robust pre- clinical TBI studies and the emergence of clinically relevant risk factors. The experimental aims are: 1) Develop a pre-clinical experimental design for mTBI that selects patient-relevant variables (gender, age, genetic variety, previous mTBI, and chronic stress) and applies them to a heterogeneous rat population using stratified randomization; and, 2) Assess acute neurological response, balance, working memory, and biomarker signature acutely following mTBI using a heterogeneous rat population. Informatics tools will be used in place of traditional multivariate statistics to extract common features of the injury response, classify them, and build predictive knowledge models. Several imbedded hypotheses will be tested to examine the response to mTBI as a function of animal sex, age, strain, previous mTBI, and chronic stress. It is expected that unanticipated relationships among the data will emerge as a result of the robust experimental design and knowledge-based model. The experimental platform presented here is novel and, if successful, can be used as a template for other studies. Addressing patient heterogeneity and clinically relevant acute outcome measures is highly significant and will increase understanding of the complexity of mTBI.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The results indicate distinct differences with maturation upon the neuronal-glial reaction of the hypoglossal nucleus to axonal injury in rats. Up to some 10 days postnatal, the neuronal membrane engulfs the presynaptic terminals in contact with its membrane and the proliferative glial phase is delayed to 1-3 weeks postoperative. In the mature neurone (21 days postnatal) the microglia proliferate earlier and lift the presynaptic terminals off the neuronal somata.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this protocol is to determine the safety of porcine fetal neuronal cell implantation into the putamen and caudate of patients with Parkinson's diseasue, under two different methods of manipulating the human immune system to prevent rejection of the transplanted tissue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Now that we have lowered the death rate of patients with gram negative bacteremia and septic shock by treatment with human antiserum to core lipopolysaccharide (LPS), we propose to extend these successful results by pursuing 3 specific aims: 1. To determine if J5 antiserum is an effective in preventing bacteremia and shock as it is in treating it. In these studies the antiserum would be given as prophylaxis to patients predisposed to bacteremia. In the first trial on prophylaxis with J5 antiserum we cut sharply the incidence of febrile attacks in neutropenic patients. We now propose to extent the trial by giving J5 antiserum to enough subjects to allow an evaluation of its efficacy against bacteremia as well. 2. To produce an effective, safe, human gamma globulin preparation for intravenous use in the treatment and prevention of gram negative bacteremia. Our goal during the period of this project is an IgM preparation that is free of non-specific anticomplementary activity but retains its ability to prevent and treat lethal bacteremia. After the effectiveness and safety of this preparation has been demonstrated in experimental animals, a new proposal will be submitted for a clinical trial of its safety and efficacy. 3. To produce in vitro human monoclonal antibody against core LPS and to test its ability to protect against experimental gram negative bacteremia and endotoxemia.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Interstitial cystitis (IC) is a sterile bladder condition of unknown etiology characterized by increased urinary urgency and frequency as well as suprapubic pain. Two important features of the disorder that must be explained are that 1) IC affects predominantly females, and 2) IC patients exhibit enhanced sensitivity to bladder distension. In addition, clinical observation suggests that symptoms of IC are exacerbated perimenstrually. The research proposed in this exploratory/developmental (R21) grant application is based on the overriding hypothesis that these clinical features can, at least in part, be explained by a generalized alteration in central nervous system nociceptive processing in IC, which is influenced by the hormonal fluctuations that accompany the female menstrual cycle. Based on this hypohesis, two predictions regarding IC will be investigated: first, that IC is associated with a generalized increase in pain sensitivity, which includes enhanced response to somatic as well as visceral stimuli; and second, that both clinical symptoms and responses to experimentally-evoked pain will be influenced by the menstrual cycles of the IC patients. In order to examine these predictions, the responses of IC and healthy, female controls to three, clinically-relevant laboratory pain induction procedures will be evaluated: 1) temporal summation of thermal pain, 2) ischemic arm pain, and 3) visceral pain produced by fluid distension of the urinary bladder. In addition, clinical symptoms as well as responses to the same three experimental pain procedures will be assessed aross the menstrual cycle in both IC patients and controls. It is anticipated that: 1) IC patients will demonstrate greater sensitivity to both somatic and visceral pain stimuli relative to controls, 2) clinical symptoms will be greater among IC patients during the luteal versus the follicular phase of the menstrual cycle, while menstrual cycle effects among controls will be minimal, and 3) experimental pain sensitivity for both groups will be greater during the luteal versus the follicular phase; however menstrual cycle effects will be greater for IC patients than controls. The results of this research will provide important and novel information regarding pain sensitivity and menstrual cycle effects in IC and will establish a foundation of knowledge to support more extensive investigations of hormonal influences on nociceptive processing in interstitial cystitis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This R21 application seeks support for an investigation of the roles of marital status, social support, and self- efficacy in the effectiveness of diabetes management in a diverse population of middle-aged and older adults in the U.S. The proposed research will use structural equation modeling to analyze data collected in 2002 and 2004 from core interviews of the nationally representative Health and Retirement Study (HRS), plus data from the 2003 supplemental diabetes-specific mail survey to examine these relationships by gender, age group, and major race-ethnicity categories. The significance of the proposed research lies in the skyrocketing rates of type 2 diabetes in the U.S. over the past few decades in all segments of the population, but primarily in middle aged and older adults. Concerns about obesity and physical inactivity and their links to the onset, complications, and societal costs of diabetes have led to public health mandates at the national level-such as those included in Healthy People 2010-aimed at reversing those trends. Although prevention of diabetes is certainly an important goal, more effective management for individuals already diagnosed with diabetes is also critical for reducing the risks of future complications. Currently, less than one in eight of all adults with diabetes follow the guidelines for good management;and older adults follow them at even lower rates. Many past studies showing that self-efficacy and social relationships are key factors in diabetes-related behavioral and health outcomes are limited by their use of non-representative data sets, cross-sectional analyses, and inadequate controls on other factors that may also be important to consider. This application's goal is to examine how social relationships (and particularly marriage) and self-efficacy influence adherence to diabetes-related lifestyle behaviors and health outcomes in order to develop and test interventions that provide more effective supports for adults living with diabetes. In particular, we will: (1) examine marital status as a predictor of behavioral (diet and exercise) and health outcomes (glycemic control and diabetes-related quality of life) in middle-aged and older adults living with diabetes;(2) investigate whether self-efficacy and diabetes support mediate the link between marital status and behavioral and health outcomes of diabetes;and (3) discern whether associations among marital status, diabetes support, self-efficacy, and behavioral and health outcomes are moderated by gender, age group, or race-ethnicity. Understanding more clearly the role of marriage and self-efficacy, and particularly if and how diabetes management differs between middle-aged and older married and unmarried adults, may inform the development of more effective interventions for adults living with diabetes. This knowledge may also be all the more important given recent studies showing spouses may be at increased risk for developing diabetes themselves because of the marital context and shared environment. PUBLIC HEALTH RELEVANCE Concerns about obesity and physical inactivity and their links to diabetes have led to public health mandates- such as those included in Healthy People 2010-aimed at reversing those trends. Although prevention of diabetes is an important goal, more effective management for individuals already diagnosed with diabetes is also critical for reducing the risks of future complications. Currently, less than one in eight adults with diabetes follow the guidelines for good management;and older adults follow them at even lower rates. Understanding how social relationships (and particularly marriage) and self-efficacy influence good diabetes management in middle-aged and older adults living with diabetes may be key to developing and testing interventions that provide more effective supports, and all the more important to married couples because spouses may be at higher risk for developing diabetes themselves if their partner has diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "! Project Summary Exposure to particulate matter (PM) air pollution is the fifth leading cause of premature disease and death on the planet and the number one environmental risk factor for the global burden of disease (posing a greater danger than all other environmental risk factors combined) according to the World Health Organization. Despite the growing need (and demand), the state-of-the-art for assessing personal exposure to PM is based on decades old technology that is inefficient, burdensome, and expensive. The physical burden posed by these monitors (noise, visual aesthetic, and weight) make them difficult to wear. The cost of these monitors also prevents air pollution exposure monitoring at scales relevant to epidemiologic research and occupational hazard surveillance. This Phase II proposal will develop and commercialize a novel, lightweight, and inexpensive personal sampling technology based upon ultrasonic piezoelectric pumping modules. The ultrasonic personal aerosol sampler (UPAS) has the potential to gain a wide share of the air pollution monitoring market. AST proposes developing this sampler into a commercial prototype that is inexpensive (<$200 bill-of-material cost at scale), provides both time-integrated (filter-based) and real-time measurements, is accurate (flow control and PM sampling efficiency within 4% of reference), compact, lightweight (<200g), provides better subject fitment, has no annual maintenance requirements, and virtually silent in operation. Furthermore, the technology will be versatile ? able to collect PM2.5 (along with other relevant size fractions such as PM4 and PM10, respirable, and inhalable PM) for sample durations of up to 24 hrs. Taken together, these innovative aspects suggest that the UPAS will be highly competitive with potential for rapid and substantial market penetration. Three aims are proposed: (1) Integrate a novel, real-time PM sensor into the UPAS hardware/firmware; (2) Develop a suite of different plug-and-play size-selective inlets to make the UPAS more versatile and optimize the UPAS for weight, power, performance, and usability; (3) Validate performance of the prototype through laboratory and field testing. !", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Clinical Tools, Inc., in collaboration with the UPMC Health System, will create and evaluate internet-based education for physicians, residents, and medical students interested In the topic of opiate addiction. Our goal is to increase accurate recognition and treatment of opiate addiction through education of physicians and physicians-in-training. The courses will use clinical scenarios and multimedia presentations to provide a low cost distance learning opportunity that is available 2~hours a day, seven days a week. In Phase I we will create design specifications for a suite of courses related to opiate addiction based on the results from the 1997 NIH Consensus Development Conference on Effective Medical Treatment of Heroin Addiction. We will demonstrate our ability to create internet-based courses by developing a single course which is an overview of opiate addiction. We will evaluate the effect of the course on physicians' knowledge, attitude, behavior and satisfaction using assessment forms developed in Phase I. In Phase II we will create and evaluate the full suite of courses. Phase III work will involve modifying and expanding the courses to teach a full range of health professionals. PROPOSED COMMERCIAL APPLICATIONS: Quality continuing education is essential for health care providers. Providers and their organizations will request easy to use, inexpensive and engaging courses that help them detect opiate addiction and provide cost- efficient, effective treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronic smoking is a national public health problem. Associative-based interventions (cue-exposure therapy) can be effective at reducing smoking and decreasing relapse rates, indicating that acquired associations with nicotine contribute to chronic smoking and relapse. The long-term goal of this research program is to elucidate these associative processes and their role in chronic nicotine use. Paviovian conditioned associations between a conditioned stimulus (CS) and an unconditioned stimulus (US) is one such process. In contrast to research treating nicotine as a US, little work has explored the possibility that the pharmacological effects of nicotine might serve as a CS and acquire additional excitatory properties. The present application will eliminate this deficit using a rat model. In this model, nicotine (CS) is reliably paired with the appetitive effects of sucrose (US). An association is evidenced when nicotine differentially evokes anticipatory food-seeking behavior. Specific Aim 1 will test whether changing the nature of the nicotine CS alters development or expression of conditioned responding; alterations include changes in nicotine dose and time between nicotine administration and testing. Aim 2 will test whether an associative learning or a state-dependent learning process is responsible for differential control of food seeking by nicotine. Aim 3 will assess the ability of several ligands (ABT-418, bupropion, nornicotine, cytisine, epibatidine) to prompt nicotine-like conditioned responding (stimulus substitution). Differential substitution patterns will provide insight into the neurobiological processes mediating the CS effects of nicotine. Aim 4 will assess whether extinction with a ligand that shares stimulus properties with nicotine will lead to the development of a competing association that transfers to a nicotine CS. This transfer of extinction (associative substitution) would be evidenced if rats trained with a nicotine CS, but receive extinction with another drug, show loss of conditioned responding when retested with the unextinguished nicotine CS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This case-control epidemiological study is designed to ascertain whether there are any associations, either positive or negative, between use of exogeneous estrogenic hormones for therapy during or following menopause or for oral contraception during reproductive years, and cancer of the corpus uteri, cancer of the breast and cancer of the ovary. Previous estrogen use in women in the age group 45-74 admitted to several large Connecticut hospitals with a first diagnosis of any of these cancers will be compared to estrogen use in women in the same age group admitted to the surgical services of the same hospitals for other conditions. Information on estrogen use and other relevant variables will be obtained by means of a standardized structured questionnaire administered by trained interviewers; this will be supplemented by information provided by the patients' physicians when necessary. Diagnosis of the cancers will be confirmed by a pathologist, and particular attention will be focused on specific histological types. The search for associations will include attempts to determine whether specific estrogenic compounds are involved, how long they must be used before an effect is seen, and whether any subgroups of women are more likely to be affected than others. This study will thus involve collaboration of epidemiologists, pathologists, biometricians, and surgeons.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "E. coli p-aminobenzoate synthetase and anthranilate synthetase are structurally and genetically related enzymes that catalyze similar, but slightly different, reactions. Each enzyme is composed of two dissimilar subunits, one responsible for glutamine amidotransferase (GAT) activity, and the other responsible for the aromatization of chorismate. The difference in the products is the disposition of amino- and carboxy- substituents on a benzene ring. The subunits of PABS are encoded by pabA and pabB and the subunits of anthranilate synthetase are encoded by trpE and trpG. The nucleotide and amino acid sequences of these four genes have been determined, and clearly indicate divergence from a common ancestor. Similarity is approximately 30% between the large subunits and 45% between the small subunits of the enzymes. The GAT subunits perform identical functions, and differ only in the specific interactions with the appropriate large subunit. The nucleotide sequence of a GAT subunit that functions in both anthranilate and PABA synthesis in A. calcoaceticus has also been determined, and is very similar to the E. coli pabA sequence. The experiments in this proposal focus on the determination of specific differences between the pab and trp genes that are responsible for divergence of function. The experimental approaches include construction of hybrid genes by exploiting in vivo recombination and in vitro DNA replication techniques, in vitro inter- and intrageneric subunit exchange, in vitro mutagenesis, and analysis of second site reversions. The results of the proposed studies will include information on the ways in which genes evolve from a common ancestor after a gene duplication event. The extent of total nucleotide and amino acid sequence divergence has been documented, and my aim is to determine the fraction of the divergence that is responsible for alteration of function. We will also gain insight into a mechanism in which biochemical specificity has been maintained while function has diverged. The approach and methodology are unique in that a functional determination of the consequences of sequence divergence will accompany a descriptive approach on the extent of sequence divergence.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this project is to develop a peptide sequencing procedure based in part on novel reactions first noted in this laboratory. By this procedure peptides generated by enzymatic and/or chemical cleavages will then be converted by the novel reactions to volatile components separable by gas chromatography. The novel reactions are catalyzed by pyridine - 2 -carboxaldehyde Schiff base(s). Gas chromatography/mass spectrometry provide the information requisite for sequence assignments. A dedicated computer and software facilitate data work up and sequence assignment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Over half of all deaths in non-ischemic heart failure (HF) are due to ventricular arrhythmias, primarily ventricular tachycardia (VT) degenerating to ventricular fibrillation (VF). In non-ischemic HF, VT can initiate by non- reentrant (focal) mechanisms likely due to delayed after depolarizations (DADs). The heterogeneous HF substate may then allow focal activity to convert to reentrant VT/VF. Thus, lethal arrhythmias in HF likely require a focal triggr for initiation and a vulnerable substrate that promotes reentry. HF electrophysiological remodeling can promote DADs and cause pro-arrhythmic changes to the substrate. -adrenergic receptor (-AR) stimulation can exacerbate DADs and further perturb the abnormal ionic currents and Ca2+ transients (CaT) found in HF. Moreover, cardiac sympathetic nerve remodeling in HF may lead to localized -AR stimulation and spatially heterogeneous effects. Electrophysiological and sympathetic remodeling has been individually linked to arrhythmia in HF. However, the interplay between local -AR stimulation and altered HF electrophysiology in arrhythmogenesis has not been explored. The overall objective of this proposal is to systematically determine the role of local -AR stimulation in producing the trigger and substrate for ventricular arrhythmias and how electrophysiological remodeling in non-ischemic HF exacerbates these effects. Our over-arching hypothesis is that local -AR stimulation causes 1) spatiotemporal synchronization of DADs across many cells to provide focal triggers; and 2) local electrophysiological heterogeneity to produce the substrate for reentrant VT/VF. We further hypothesize that electrophysiological remodeling in HF exacerbates the pro-arrhythmic effects of local -AR stimulation, both in generating triggers and in modifying the substrate. To address these hypotheses, dual optical mapping of Vm and Ca2+ will be performed on isolated rabbit hearts while administering local norepinephrine (NE). Aim 1 will focus on the mechanisms by which local -AR stimulation triggers focal arrhythmia in healthy rabbit hearts. We will then systematically test the effects of HF- associated electrophysiological remodeling on the propensity to focal activity by pharmacologically mimicking key HF phenotypes. Aim 2 focuses on the role of local -AR stimulation in contributing to the substrate for reentry via dispersion o action potential and CaT properties and development of alternans. We will also test the effects of key HF-associated electrophysiological mechanisms in contributing to reentry during local -AR stimulation. In Aim 3, we will determine the arrhythmogenic role of localized -AR stimulation in a rabbit model of non-ischemic HF by applying exogenous as well as invoking endogenous sympathetic stimulation followed by a quantitative assessment of neurochemistry in HF. We will then assess the therapeutic potential of reversing key HF phenotype(s) with newly proposed anti-arrhythmic strategies. Overall, the results of this project will define the firt mechanistic link between sympathetic dysfunction and ventricular arrhythmias in non-ischemic HF and will greatly advance our long-term goal of predicting and preventing sudden cardiac death in HF. PUBLIC HEALTH RELEVANCE: Heart failure predisposes patients to deadly cardiac arhythmias (disturbances of the normal heart rhythm). Increased activity of the cardiac sympathetic nerves is known to be associated with arrhythmias. But how exactly sympathetic nerve activity leads to arrhythmias and the interaction between nerve activity and other changes that occur in the heart during heart failure are unknown. The goal of this project is to determine the role of local sympathetic nerve activity in leading to arrhythmias in heart failure. The result of this study will shed new light on proper therapeutic treatments for heart failure patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Pre-exposure prophylaxis (PrEP), formulated as a once-daily pill, represents an effective form of biomedical HIV prevention for men who have sex with men (MSM), but its use among African American MSM remains suboptimal and low relative to White MSM. African American MSM are more likely to report suboptimal adherence and subsequently seroconvert while using PrEP. As such, the use of long-acting injectable (LAI) antiretroviral drug formulations as PrEP may be an attractive alternative for MSM interested in biomedical HIV prevention but who may have difficulties with daily pill-taking. However, little is known about what factors may influence willingness to use LAI-PrEP among African American MSM. The proposed research will leverage existing data from the N2 Study, a population-based longitudinal cohort of young HIV-negative African American MSM residing on the South Side of Chicago (n = 350). Using detailed information on the social and sexual networks of young African American MSM, we will assess whether one?s willingness to use LAI-PrEP is impacted by their position within their networks. Using a complex systems approach known as agent-based modeling, we will simulate HIV transmission in the dynamic networks of young African American MSM, allowing us to assess the potential effects of a network-based intervention in improving LAI-PrEP use and reducing HIV incidence. Identifying the social dynamics associated with future use of LAI-PrEP may make a significant impact on the future success of this prevention method and the speed at which an effective LAI-PrEP formulation is taken up among young African American MSM. In achieving our aims, the findings from the proposed research may be used to optimize the uptake of LAI-PrEP through innovative network-based interventions, potentially maximizing its ability to make a sustained impact on the HIV epidemic in the United States.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Perhaps few questions in development have been so intractable as the means by which different cell fates are produced in response to a gradient of signals. These analogue-to-digital switches underlie the determination of cell fates in many organisms. T Lymphocyte development may be one of the best systems to understand this general problem because the critical signals are given at a stage where biochemical, genetic and cell biologic methods can all be brought to play. In lymphocytes, weak or transient signals are thought to produce positive selection (differentiation and proliferation) of T cells capable of reacting to self-MHC on thymic stromal cells. On the other hand, strong signals produced by self-antigen lead to death of cells responding to self-antigen. The pro-apoptotic protein Bim is required for negative selection but is not necessary for positive selection. Conversely, we have recently found that calcineurin is essential in T cells for positive selection, but dispensable for negative selection. Surprisingly calcineurin specifically controls the activation of ERK but not other MAP kinases or IkB, suggesting a revision of the accepted signaling pathways of thymocyte selection. These observations set the stage for a biochemical march from Bim and calcineurin to the molecule(s) that divert signals from positive to negative selection with increasing signal intensity. Current studies support several possible mechanisms by which signals of different intensity could control selection. To avoid the difficulties encountered with forward analysis of biochemical pathways we will work backward from Bim and calcineurin to define the biochemical pathways that control their activity in CD4+, CD8+ thymocytes. Our goal in these studies will be to define the lowest common mediator necessary for activation of both Bim and calcineurin and hence positive and negative selection. We will then determine the mechanism by which this molecule is induced to channel high intensity signals to Bim and low intensity signals to calcineurin. We will also define the processes downstream of calcineurin that mediate positive selection including the mechanism of NFATc nuclear import and export, the set of genes that are dependent on calcineurin activity in positive selection and how these genes give rise to a population of immunologically competent peripheral lymphocytes. Defining these mechanisms should lead to a more complete understanding of immune defense and provide useful information for development of new therapies. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application requests partial funding for the Reproductive Tract Biology Gordon Conference to be held July 2-7, 2000. Please note that due to the scientific content of the meeting we request the National Institute of Child Health and Human Development as the primary sponsor of the meeting. We also request joint sponsorship by the Office of Women's Health, the National Cancer Institute, National Institute of Diabetes, Digestive and Kidney Diseases, the National Institute for Environmental Health Sciences, and the National Institute of General Medical Sciences. The number of requested sponsors reflects the unusual nature of this conference which focuses on the biology and the pathology of the organs and organ systems of the male and female reproductive tracts, rather than on specific molecules or processes. Therefore, this meeting attracts participants from a wide variety of fields and backgrounds ranging from basic science to clinical practice. Accordingly, the principal goal of this conference is to stimulate cross- disciplinary exchange and integration of information concerning the reproductive tract. Three specific aims will allow us to accomplish this goal: First, the scientific content of the meeting will be at the highest level. To achieve this standard of excellence, we are inviting speakers who are internationally recognized experts at the forefront of their fields. There is an interesting blend of established investigators who have been leaders in their respective areas for many years and equally renowned researchers in other fields whose phenotypic analyses of transgenic mice have uncovered unexpected and novel actions of these molecules in the reproductive tract. Second, there is a strong emphasis on research that is highly relevant to important clinical questions and has potential for translation into medical practice. For example, one entire session focuses on mechanisms of hormonal signaling in the context of the mammary gland with a special emphasis on aberrations that lead to breast cancer. Third, there is a strong emphasis on the inclusion of students and junior colleagues because the future of any field depends on the next generation of investigators. In summary, we have planned an exciting meeting that is in accord with the rich traditions of the Reproductive Tract Biology Gordon Conference, which has been in existence for nearly 30 years. We envision that the new site (Connecticut College, New London Connecticut) and the intent to meet on a permanent basis with the Mammalian Gametogenesis and Embryogenesis Conference will enhance the quality of this meeting, which has consistently been rated extremely high by the attendees.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Inhibitors of the enzyme sarco/endoplasmic reticulum Ca2+ATPase (SERCA) are valuable research tools for the study of the enzyme's role in physiological processes and they also have the potential of being developed into new anti- cancer agents. A series of 2,5-disubstitued hydroquinones will be synthesized and their ability to inhibit SERCA will be assessed in bioassays. Based on the results, computational techniques such as structure-activity relationship modeling and ligand docking will be used to develop models capable of predicting the activities of yet untested, hydroquinone-based compounds. With the aid of these models, compound libraries will be screened virtually for novel SERCA inhibitors that then will be obtained and tested in bioassays. PUBLIC HEALTH RELEVANCE: The research described in this application is aimed at the development of novel, hydroquinone-based inhibitors of the enzyme sarco/endoplasmic reticulum Ca2+ ATPase (SERCA). As documented by an impressively large number of publications on the naturally occurring inhibitor thapsigargin, SERCA inhibitors are of tremendous value for the study of the enzyme's role in physiological processes. In addition, it has been shown that SERCA inhibitors can be developed into prodrugs for the treatment of prostate cancer. Since hydroquinone-based compounds are structurally simple and quite different from other SERCA inhibitors, their synthesis from inexpensive starting materials is relatively straightforward. Thus, they have the potential of being developed into valuable alternatives to currently available agents with unique physicochemical and pharmacodynamic properties.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent work in this laboratory has shown that a variety of cancer cells secrete nerve growth factor. In particular, mouse L cells secrete large quantities of the factor in culture. Further, the chemical properties of L-cell NGF are different from any form of NGF that has so far been studied. Our study is: (1)\\to grow large quantities of L cells; (2)\\to isolate and to completely purify the NGF secreted by these cells; (3)\\to determine the molecular properties and structure of this protein; and (4)\\to ascertain whether this naturally occurring form of NGF has biological actions that have heretofore not been recognized, both within the nervous system as well as outside of it. Further work will completely purify human nerve growth factor from serum to characterize it chemically and to develop specific immunoassays for it. Such assays then will permit the specific measurement of NGF in man. More recent studies have shown that NGF can significantly promote the healing of experimentally induced wounds in animals, that it can substitute specifically for the first component of human complement, and that it is chemotactic for leukocytes. These findings indicate that NGF may play an important role in systems other than neural tissue. The chemical and biological meaning of this new information is under study at present. (J)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The incidence of obesity worldwide continues to escalate with the spread of the Western diet and with it there has been a corresponding increase in cardiovascular disease, diabetes, end-stage renal disease and other obesity-related disorders. Of all the causes for obesity, the predominant one seems to be choice - the choice to eat more and exercise less. One of the choices that has been linked with obesity has been the selection of a high fat, calorically dense diet. Despite much research on the link between fat intake and obesity, relatively little is known about the chemosensory mechanisms that underlie the taste of fat and how these mechanisms might contribute to dietary fat intake. Our previous research has identified differences in how the taste systems in obesity-prone and -resistant rodents respond to fatty acids, a cue for dietary fat, and how this was correlated to differences in gene expression and receptor function. Moreover, we have demonstrated that these chemosensory mechanisms are modulated by diet and the development of obesity. The revised proposal focuses on pursuing a more focused, single remaining specific aim that seek to explore how the gustatory response to fat is modulated by dietary experience. There is new and emerging data that argues to the plasticity of the peripheral taste system, yet our understanding of how the system changes in relation to experience is poor at best. Specifically, we will use an approach including a multidisciplinary approach to answer the following question: 1. Is the fatty acid transduction pathway modulated by diet? Our multidisciplinary approach analyzing genes through behavior will be used to test the hypothesis that fat receptor expression in taste cells is altered during high fat feeding in a manner that results in an increased responsiveness to fatty acids. Following high fat dietary regimens, molecular, cell-based and behavioral assays will be performed to measure changes in four primary receptive elements in the fatty acid signaling pathway, including CD36, GPR 120, GPR 84 and fatty acid-sensitive DRK channels. Our extensive preliminary data generated in the previous period would argue that these are the primary components of the pathway that underlies the ability of the gustatory system to recognize and respond to free fatty acids.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There has been little systematic study of the function of tissues within the oral cavity during aging, either describing normal processes of alterations resulting from specific diseases and therapeutic procedures. The purpose of this project is to focus on 3 oral health problem areas for the elderly (salivary secretion, oral motor function and cervical caries) and examine the status of certain biological factors which would likely influence the course of such problems. Major effort has been directed at evaluating electrolyte secretions from the stimulated parotid glands (reflecting ion fluxes in various gland components) and assessing several oral motor functions (postural, masticatory, speech, swallowing).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposal is to investigate the use of MYXV as potential oncolytic virotherapy agent against pancreatic cancer in preclinical animal models. The proposal focuses on MYXV, and is based on our previously published reports and supporting preliminary studies. MYXV is a rabbit-specific poxvirus with oncolytic activity against many types of human cancer models in vivo, including brain tumors, melanoma and rhabdoid tumors. In addition, we reported recently that MYXV is able to infect and kill pancreatic cancer cells in vitro. Our preliminary studies provided in the main body of the proposal show that MYXV exhibits potent oncolytic activity in both immunodeficient and immunocompetent animal models of pancreatic cancer. Based on these results, we seek to evaluate MYXV in combination with current chemotherapy regimens, especially gemcitabine, for pancreatic cancer. We hypothesize that MYXV will have potent oncolytic activity against pancreatic cancer in vivo and may be combined and/or engineered to enhance current chemotherapy treatments. Wildtype MYXV and recombinant MYXV \"armed\" with chemosensitizing gene(s) will be evaluated as single agent therapies and in combination with current chemotherapy drugs approved for the treatment of pancreatic cancer. For the purposes of this grant, we propose to: 1) Evaluate the efficacy of wildtype MYXV oncolysis in murine models of pancreatic carcinoma. MYXV will be evaluated: a) as a single agent therapy compared to standard chemotherapies, b) in combination with gemcitabine and/or erlotinib, and as c) second line treatment therapy for chemotherapy resistant tumors. These experiments involve the use of the most common first line chemotherapies for pancreatic cancer and will therefore evaluate MYXV in the context of a clinically relevant scenario. Immunodeficient and immunocompetent murine models of pancreatic cancer will be established intraperitoneally (IP) and virus will be administered locally by the IP route. Tumor burden and survival curves will be compared between treatment groups in the presence or absence of gemcitabine and in gemcitabine refractory tumors to determine if MYXV virotherapy under the three regimes mentioned above results in an enhancement of therapeutic benefits as measured by tumor burden and survival. 2) Generate recombinant MYXV armed with chemosensitizing genes that will enhance gemcitabine-based chemotherapy. Recombinant \"armed\" MYXVs that express deoxycytidine kinase (CDK) or the human equilibrative nucleoside transporter 1(hENT-1) will be engineered. These viruses will be characterized in vivo to determine if the expression of the transgenes enhances gemcitabine-based oncolysis of pancreatic cancer cells in the two models described above in Specific Aim 1. Tumor burden and survival curves will be compared between armed and wildtype MYXVs treated groups to determine if the use of an armed MYXV provides a therapeutic advantage over wildtype MYXV by enhancing gemcitabine-based chemotherapy in vivo. This proposal will be the first study to evaluate the oncolytic potential of MYXV in preclinical animal models of pancreatic cancer as single agent therapy and in combination with chemotherapy drugs, as well as the characterization of armed MYXV capable of sensitizing cells to gemcitabine chemotherapy. Thus, if successful, these pilot experiments will identify MYXV as an effective oncolytic virus that can be further developed as a novel, safe virotherapy for the treatment of pancreatic cancer. In particular, if the results are as positive as we anticipate, and given the excellent safety profile of MYXV, we will pursue the production of clinical grade stocks of MYXV and the filing for an investigational new drug application (IND) at the completion of the proposed study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract The Department of Occupational and Environmental Health (OEH) at the University of Oklahoma Health Sciences Center (OUHSC) Hudson College of Public Health provides graduate education leading to the Master of Science (MS) degree in Industrial Hygiene and Environmental Health Sciences (IH/EHS) and the Doctor of Philosophy (PhD) degree in Occupational and Environmental Health. The MS in IH/EHS at OUHSC is one of only four ABET-accredited masters level industrial hygiene programs in the south central United States. The goal of the MS in IH/EHS is to prepare professional practitioners to apply scientific knowledge to the anticipation, recognition, evaluation, and control of environmental hazards or stresses affecting human health. The MS curriculum, which emphasizes both quantitative skills and effective communication, consists of 48 semester hours of coursework, including masters thesis research and a field practice experience, and can be completed in 22-24 months of full-time study. The goal of the PhD degree in OEH is to prepare graduates to impart and/or add to knowledge in occupational health through careers in academia or research. The PhD curriculum includes advanced didactic coursework in environmental sciences, epidemiology, biostatistics, and research methods related to occupational health, and a doctoral dissertation within the broad field of occupational and environmental health. The specific aims of the proposed renewal of the NIOSH training project grant (TPG) are: (1) to attract highly qualified and motivated students, including individuals with cultural or linguistic fluency related to underserved populations, into the industrial hygiene profession; (2) to recruit students with diverse technical backgrounds who are interested in entering the industrial hygiene field; and (3) to recruit and train very promising research-oriented students in industrial hygiene research. TPG funds will be used to support six traineeship slots per year, to be filled by full-time masters-level trainees and up to 2 full-time doctoral-level trainees. Trainees in the MS program may be supported on the TPG for up to 24 months. Trainees in the PhD program may be supported for up to 5 years. Oklahoma colleges have relatively high American Indian enrollments reflecting the state's unique ethnic mix, presenting an excellent pool of well qualified prospective trainees from under-represented minorities. Recruitment for the MS program will also target individuals with work experience in the health professions, engineering, chemistry, and environmental science who wish to make the transition into industrial hygiene practice. Recruitment for the PhD will target highly qualified individuals with BS or MS degrees in science or engineering that provide a solid foundation for industrial hygiene research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although epidermal growth factor (EGF) stimulates the growth of various cells in vitro, the biological role of this polypeptide is not known. Evidence that EGF may function as an autocrine or paracrine factor is suggested from previous studies demonstrating submaxillary gland preproEGF mRNA in other organs. We have shown that primary cultures of mouse uterine epithelial cells possess specific binding sites (Kd approximately 1.8 nM) for 125I-EGF and approximately 50 x 10-3 receptors/cell. EGF stimulated proliferation of the uterine cells in vitro, whereas other known growth factors did not. Immunolocalization of EGF in pronase-treated sections of the mouse uterus revealed staining at the luminal aspect of epithelial cells. Localization required protease treatment and the material reactive with anti-EGF antiserum was apparently not estrogen-dependent since this pattern of staining was readily observed in uteri from animals ovariectomized for over two weeks. Protease-treatment of sections was also required for localization of EGF in epithelial cells of the mouse kidney and mammary gland (mid-pregnant). We have found that kidney EGF occurs predominantly as the apparent precursor form (140 kDa) bound to cell membranes. Thus, it is likely that the observed pattern of EGF localization in uterus represents a membrane-bound form of the precursor. Hybridization with a 32-P-labelled cDNA probe for submaxillary gland preproEGF mRNA occurred with samples of uterine mRNA, although at much lower levels than that of kidney and submaxillary gland. The level of preproEGF mRNA increased in immature mouse uteri following treatment with estrogen. Northern blot analysis of uterine A+ mRNA from estrogen-treated mice revealed a single band of 4.9 kilobases, equivalent to that of submaxillary gland preproEGF mRNA. Studies are in progress to further understand the synthesis sorting and processing of the EGF precursor in different organs and the potential for various hormones to regulate these events.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Residential histories collected during a national survey of oral health of U.S. schoolchildren conducted in 1986-1987 were used to establish two groups: those children who always lived on a public water supply in a fluoridated community (N=8,165) and those who never lived in a fluoridated community (N=8,233). Comparisons of the mean levels of Decayed, Missing and Filled Surfaces were made by age, sex and region of the country. Mean DMFS for children with lifelong exposure to water fluoridation was 2.8 compared to 3.4 mean DMFS for children who had never lived in areas with fluoridated water. Mean DMFS for mesial-distal surfaces was about one-third higher in children without water fluoridation. Both groups of children reported high use of supplemental and/or topical fluorides. Regional differences between groups varied greatly from 61% difference in Region VII to 6% difference in Region III. These findings were presented at a workshop \"Mechanisms of Fluoride\" and published in the Journal of Dental Research, Sp. Iss. Feb. 1990. Mean dfs for deciduous teeth was computed for those children ages 5-9 who had a lifelong history of community water fluoridation or no history of water fluoridation. The mean dfs was lower in all age groups for children on fluoridated community water supplies, with a combined difference of 23% fewer dfs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is well known that the aged brain undergoes a series of subtle but progressive alterations that eventually lead to altered neuronal function and, ultimately, altered behavior of the individual. Together with the well-described age related decline in cognitive performance, it has also been found that motor function undergoes a steady and progressive decline in most aged humans. The midbrain dopaminergic neurons, which are important for regulation of motor activity, appear to be especially vulnerable to age-related factors, leading to a specific increase in neurodegenerative disorders afflicting these neurons, such as Parkinson's disease (PD) with age. This program project grant is focused on understanding why midbrain dopamine neurons are selectively vulnerable to environmental and endogenous agents, leading to lost function and structure during aging. We will work along an overall \"dual-hit\" hypothesis, proposing that it is a combined endogenous inability to cope with oxidative stress and external factors making it worse that leads to a selective degeneration of these neurons in some individuals but not others during aging. Every project is devoted to the understanding of age-related deterioration in the function and morphology of the midbrain dopaminergic system, with a focus on motor impairment following specific intrinsic and extrinsic damaging events. To this end, all primary projects in the program (Project 0001-0005) will provide a distinct level of analysis of three specific determinants of motor impairment, namely: growth factor reduction (intrinsic, genetic alteration), endotoxin exposure (prenatal extrinsic factor, LPS), and methamphetamine exposure (extrinsic factor to young adult mice). The Animal Core will provide all subjects to guarantee consistency between each project, and thereby facilitate the capacity to unite discoveries emerging from different research approaches into a coherent understanding of the neurobiological basis of factors that might affect the impact of aging in this transmitter system. The subjects will be examined at many levels of analysis, including cellular (Projects 0001 - 0004), tissue culture (Project 0005) and behavioral (Project 0001 and 2). Project 0003 will integrate the different treatments (GDNF/BDNF partial deletion, LPS treatment, and methamphetamine) from the perspective of neuroinflammation, and Project 0004 will examine the same three factors from the perspective of in-depth analysis of neurotransmission itself. Finally, Project 0005 will examine intrinsic versus extrinsic determinants of the alterations observed by isolating the brain regions (substantia nigra and striatum) in slice cultures or transplants. This thematic integration around an animal model (GDNF/BDNF knockouts) with external factors will focus multiple dimensions of research activity on a single problem (age-related decreased motor function) with the goal of providing a relatively complete description of the neurological basis of age-related dopamine neuron cell dysfunction leading to motor impairment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Renal osteodystrophy (ROD) is a multifactorial and pervasive disorder of bone remodeling in chronic kidney disease (CKD). The prevalence CKD in the United States is estimated at 15.1% in 2011. With the aging population, the number of patients affected by ROD, therefore, is expected to increase. Despite the widespread use of therapies, such as phosphate binders, calcium, and vitamin D, bone-fracture rates and mortality risk following fractures are markedly greater in patients on dialysis, compared with the general population. Furthermore, the effectiveness of treating dialysis patients with conventional osteoporosis drugs is not established. The high cost and numerous side effects of drug therapies have spurred the search for alternative treatment options for ROD. It has recently been shown that low-magnitude mechanical stimulation (LMMS) is osteogenic. Although many theories exist, the mechanism of action of LMMS is not well established. A more precise understanding of how vibrations work is needed to optimize the efficacy of LMMS treatment and increase its appeal as a viable non-pharmacological solution to the problem of weak bone. One major hurdle in monitoring efficacy of treatment to LMMS in particular and other interventions in general is the lack of non-invasive tools to study the anabolic response at micro-structural levels in humans. All LMMS trials conducted to date use BMD as the outcome variable, an approach which has many limitations. In high-turnover ROD, for example, increased trabecular bone volume may offset cortical bone loss, resulting in normal or increased areal-BMD despite poor bone strength. The proposed project focuses on overcoming a critical barrier towards understanding the mechanism of action of LMMS therapy in humans by developing a non-invasive high-resolution imaging based biomechanics technique to study the dependence of local-strain on bone formation at the microstructural level in response to LMMS intervention in dialysis patients. If the aims of this project are achieved, scientific knowledge of LMMS treatment in humans and technical capabilities to monitor bone's anabolic response to treatment will be improved. Most importantly, the successful completion of the aims has the potential to change the current paradigm of managing ROD by adapting LMMS as an attractive and preventive intervention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Clostridium difficile recently has been linked to antibiotic-associated enterocolitis and pseudomembranous colitis which are severe and sometimes fatal diarrheal syndromes. This toxin-producing, strictly anaerobic bacterium was first described in 1935, but has only been recognized as a clinically significant pathogen within the last 8 or 9 years. Evidence exists that the pathogenic effect of this organism is mediated by two toxins and that adherence of C. difficile to the host may possibly play a role in pathogenicity. Conclusive studies on the genetic determinant(s) responsible for the expression of C. difficile toxins have not been reported. However, plasmids and bacteriophage mediated production of toxin and other extracellular enzymes have been previously demonstrated in other clostridial species such as Clostridium perfringens, Clostridium tetani, Clostridium botulinum and Clostridium novyi. Because of the previously reported linkage of toxin genes to extrachromosomal DNA and the documented occurrence of adhesive or invasive genes being coded for by plasmids in other bacteria, the high molecular weight extrachromosomal DNA of C. difficile will be characterized. Preliminary work in our laboratory has demonstrated a large extrachromosomal DNA element in 16 out of 16 strains of C. difficile tested. Also bacteriophage particals were identified with electron microscopy to be associated with C. difficile. The major objectives of this two year pilot study is to determine if the large extrachromosomal DNA observed is plasmid or phage in nature and to determine if this DNA codes for assayable determinants responsible for pathogenicity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "B3. Molecular Genetics and Epigenetics The Molecular Genetics and Epigenetics component of the IHSFC, formerly called Facility Core C: Microarray and Molecular Biology, has been transformed to address the expressed current and future needs of Center members for instrumentation and expertise support in the areas of gene expression, epigenetics and functional genomics. The goal of this activity is to provide to Center members access to cutting edge technologies in the analysis of gene expression and modulation through access to instrumentation, instruction, assistance in methods and training in the analysis of data. We have added the epigenetics analysis to this iteration of the grant, replacing resources that had been previously directed towards proteomics. This decision was made because the Proteomics Core at Johns Hopkins School of Medicine is (http://www.hopkinsmedicine.org/msf/) now widely available for Center members and we choose not to duplicate these efforts. The objectives of this component of the IHSFC are to: provide state-of-the-art analysis of gene expression and analysis using the Affymetrix platform for mouse (genome 430A 2.0 array), rat (expression 230 2.0) and human (genome U133 Plus 2.0 array) gene chips that comprise the global gene profiling for these respective species; provide infrastructure so that an investigator can understand the global gene expression changes modified by environmental exposures that are manifested through epigenetic changes; integrate with the Bioinformatics/Biostatistics core for using Bioconductor and other tools for the analysis of genetic and epigenetic changes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract There is significant evidence that anesthetics which act at the GABAA receptor alter immune function and that multiple immune cell types express functional GABAA receptors. However, the mechanism of this immune modulation is unknown. In this proposal, we present exciting preliminary data demonstrating that mice lacking expression of the GABAA receptor ?4 subunit (gabra4), a subunit expressed in immune cells including CD4+ lymphocytes, have significantly greater lung inflammation after sensitization with house dust mite antigen, a common allergic asthma model. This heightened inflammation results in greater in vivo airway reactivity. We also demonstrate that gabra4 knockout CD4+ cells express increased inflammatory cytokines compared to wild type CD4+ cells when stimulated via the T cell receptor in vitro, suggesting the hyper-inflammatory phenotype of the gabra4 knockout mouse may be CD4+ cell-mediated. We hypothesize that GABAA receptor currents, which are depolarizing in CD4+ cells, alter calcium signaling processes key in lymphocyte activation and function, specifically calcium oscillations and store-operated calcium entry. We propose to demonstrate that ?4 subunit containing-GABAA receptors mediate currents in CD4+ cells which are augmented by GABAA receptor ligands, including XHE-III-74A, a novel ?4 subunit selective positive allosteric modulator developed by a collaborator. We will also determine if the CD4+ cell is key in producing the gabra4 KO mouse?s hyper-inflammatory phenotype by performing adoptive T cell transfer experiments. Furthermore, we propose to determine if altered GABAAR signaling affects CD4+ cell calcium dynamics (oscillations and store operated calcium entry) using a novel in situ calcium imaging technique that utilizes precision-cut lung slice technology. This offers the key advantage of maintaining the native environment of the inflamed lung. Finally, we will determine if XHE-III-74A, an ?4 subunit- selective GABAA receptor activator that does not cross the blood brain barrier, will limit house dust mite antigen induced lung inflammation and airway hyper-responsiveness when administered chronically during the sensitization process. Although the studies proposed here focus on lung inflammation, the implications of this work may be relevant to inflammatory processes of many types. Given the widespread use of GABAergic medications, this research promises to be of high clinical significance. My department and Columbia University provide an ideal environment to complete these studies, as all necessary support is already in place. In addition, my proven mentoring committee provides a perfect blend of skills to help guide the science, including expertise in ion channel physiology, lung biology, immunology, and calcium signaling. A comprehensive career development plan has been established, including coursework and additional training. Taken as a whole, this K08 proposal outlines a robust pathway to scientific independence and the foundation of a successful and sustained career as a physician-scientist.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Solid Phase Sciences Corporation (SPSC) proposes to produce a library of substituted acridine and pyridine based heterocycles. The library will consist of 5-10 milligrams each of a total of 384 compounds. SPSC will accomplish this by utilizing it's novel high-loading support and commercially available solvent delivery robot. Specifically, libraries will be made off of this high-loading support to separate the desired products from undesired starting material. A liquid handling robot will be used to add reagents and solvents. The goal of the Phase I effort will be to integrate the use of this high-loading support in solid phase synthesis with automated liquid handling into one automated system to produce a library composed of a series of acridine and pyridine based heterocyclic compounds for general screening.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Pseudomonas aeruginosa is a versatile bacterial pathogen that causes life-threatening acute and chronic infections in diverse patient populations. Complicating treatment is the ability of P. aeruginosa to resist the majority of antimicrobial therapies. It is therefore critical to identify virulence properties that can be targeted for the development of novel therapeutics. Several studies demonstrate the importance of iron homeostasis for P. aeruginosa pathogenesis. Recent work from our lab shows that the P. aeruginosa prrF chromosomal locus, which encodes the iron-responsive PrrF1 and PrrF2 small regulatory RNAs (sRNAs), is required for acute lung infection in mice. The PrrF1 and PrrF2 sRNAs contribute to iron homeostasis by repressing the expression of iron-utilizing pathways when this nutrient is limiting. This ?iron sparing response? further impacts diverse virulence properties, including quorum sensing and biofilm formation. The prrF locus produces a distinct sRNA (PrrH) that is regulated by heme, an abundant source of iron in the human body, thus linking iron and heme homeostasis pathways of P. aeruginosa. While our studies have established the broad impact of this locus on P. aeruginosa physiology and virulence, the mechanisms by which the individual sRNAs transcribed from this locus mediate gene expression and pathogenesis remain unknown. Based on our preliminary and published studies, we hypothesize that the PrrF and PrrH sRNAs play critical yet distinct roles in regulating P. aeruginosa iron homeostasis and virulence. We will test our hypothesis by 1) identifying PrrF target mRNAs responsible for virulence attenuation of the ?prrF1,2 mutant; 2) determining the genetic basis of heme regulated expression via the PrrH sRNA; and 3) defining the mechanisms by which PrrF and PrrH regulate gene expression. These studies will define the specific mechanisms by which the prrF-transcribed sRNAs mediate iron homeostasis and virulence of P. aeruginosa.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal aims to understand the changes in visual processing that occur when subjects shift between alert and non-alert waking states. While great advances have been made in understanding central mechanisms of visual perception of alert, attentive subjects, there is little understanding of cortical processes that come into play when alertness wanes. The awake, non-alert state is not equivalent to anesthesia, or to sleep states. When non-alert, we are capable of perception, but our perceptual capacities differ. It is commonly believed that accidents happen when we are not alert, but the extent to which early thalamic or visual cortical mechanisms may be responsible for this (as opposed to higher cognitive processes) is an open question. This proposal relies on a unique model system that is very well-suited to address this question: the awake rabbit, an animal who's inner mental life transparently and frequently shifts between alert and nonalert EEG-defined states, and who's stable eyes and diffident nature make it an ideal subject for these experiments. The proposed research will examine how changes in the brain state of awake subjects influence the multiple, sequential stages of information processing that occur within the visual thalamocortical, intracortical, and cortical output networks. The experiments will compare state dependent changes in the visual response properties of excitatory and inhibitory neurons at the input layer and within several output systems of the cortex and will investigate the underlying mechanisms leading to these changes, at the subthreshold and spiking level. This work will lead to a better understanding of cortical mechanisms of visual processing in a dynamic, awake brain. From a health perspective, these studies will have an important impact on our understanding of how alertness/vigilance deficits can impact visual perception and performance, and will provide the basis for future clinical studies of human mental health and behavioral disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our primary aim is to study the morphological, biochemical and immunological properties of the mouse mammary tumor virus (MuMTV). The viral rosettes, consisting of viral membrane fragments with attached projections, and viral projections alone, are two structural components which have been isolated and their protein composition studied. Work is in progress on the isolation of viral cores and ribonucleoprotein complexes, which will be analysed by electron microscopy and polyacrylamide gel electrophoresis in order to identify the various proteins of the virus with its architecture. Purification of the various viral proteins, including reverse transcriptase, is an integral part of our program. The MuMTV-reverse transcriptase and two of the five major viral polypeptides have been purified. Within the next six months we hope to have enough purified enzyme from RIII milk borne MuMTV for antibody production. This antisera will be used to study the degree of relatedness between the 4 known variants of MuMTV and other oncornaviruses. The techniques for the isolation of MuMTV-polypeptides are being standardised. The purified viral proteines will be used for the production of monspecific antisera in order to determine the level of expression and distribution of several MuMTV antigens in various mouse tissues by radioimmunoassay.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of this research is to gain a better understanding of the physiological mechanisms underlying the hormonal control of behavior. This specific project is the initial step in determining if catecholaminergic neurotransmitters are involved in mediating the effects of hormones on behavior. The first goal of this project is to establish and validate methodology for delivering small amounts of drugs to into the ventricular circulation. Once the methodology is established, two studies will be run. They will serve both to validate the methodology and determine the effects of lowering catecholamine function on behavior. The first will examine the behavioral effects of decreasing norepinephrine levels primarily in telencephalic brain areas. The second will examine the effects of a more generalized depletion of both norepinephrine and dopamine. In both studies, norepinephrine and dopamine depletion in brain areas known to be involved in controlling the behaviors of interest will be monitored by high performance liquid chromatography with electrochemical detection. This will allow examination of the relationship between catecholamine depletion in specific brain areas and alterations in normal behavior patterns. This work will serve as the foundation for a more extensive investigation of the involvement of the catecholamines in modulating hormone-sensitive behaviors. It is obvious that catecholamine dysfunction is involved in a number of serious mental health problems. This research seeks to clarify how catecholamines modulate normal behavior. The neuropharmacological studies will demonstrate how different brain areas respond to drug treatment and the behavioral consequences of neurochemical changes in specific areas.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. SCIENCE RESEARCH CORE The Arkansas INBRE proposes to continue its Science Research Core, which will comprise services offered by three scientific research facilities at two lead institutions. These core facilities will provide investigators throughout the state with access to the instrumentation and expertise needed for three key areas of modern biotechnology[unreadable]proteomics, digital microscopy, and DNA damage and toxicology. These three areas were chosen because the technical expertise and, especially, the instrumentation of each facility are not commonly available, even in an average, well-funded research laboratory, particularly not at four-year PUIs. These three facilities were especially chosen to address the specific needs of the 13 Arkansas INBRE Project Leaders (see Table 11). With access to these technologies for specific studies, researchers can then pursue and extend research projects in their own laboratories. The use of these facilities should help researchers obtain necessary preliminary data to apply for extramural research funding. Arkansas INBRE support for the Science Research Core and its three facilities is primarily intended to guarantee access to these facilities by PUI investigators (both faculty and students). This support is not sufficient to completely sustain these facilities, which derive additional, more substantial support from other grant funding sources and from the institutions at which they are located (UAMS and UAF). The three facilities, described in detail below, have the goal of providing expertise and equipment for the following purposes: + Proteomics: To enable researchers to study protein structure and function using state-of-the-art instrumentation; + Digital Microscopy: To allow researchers to locate macromolecules within cells and tissues, thereby better assessing their function in normal and diseased states;and + DNA Damage and Toxicology: To allow researchers to incorporate DNA damage and toxicity analyses (as in cell injury and death) into their studies of normal and diseased cells and tissues. Each Science Research Core facility will also be committed to educating and training Arkansas INBRE faculty and students at both partner and non-partner PUIs, in sophisticated technologies specific to each facility. Hence, all facilities will provide workshops or demonstration sessions relevant to the instrumentation and expertise available. These offerings will be organized by leaders of all Science Research Core facilities and publicized by the Administrative Core statewide. The Science Research Core will also provide access to other Core Facilities located at the lead institutions, for example, DNA Sequencing and Flow Cytometry, for all researchers within the Arkansas INBRE. In addition, through the INBRE consortium, the Arkansas INBRE will network with other INBREs to obtain access to facilities that may not currently be available in Arkansas (see http://louisville.edu/research/genomics/content/microarray_services.shtml and letter of support from the directors of the Kentucky INBRE and the University of Louisville Microarray Facility, Louisville, KY). The Science Research Core is also committed to assisting both student and faculty participants in the Summer Mentored Research Fellowship Program (see Outreach Core), enabling summer fellows to become familiar with technologies that facilitate modern biomedical research and, ultimately, to participate more fully in the INBRE program. As evidence of this commitment, faculty investigators at non-partner PUIs have been using the current INBRE facilities (see below under each facility). Workshops offered during the summer introduce researchers from across the state to available, state-of-the-art technologies. All facilities within the Science Research Core will interact as needed with the Bioinformatics Core to provide for the needs of the Arkansas INBRE partner PUIs (see Objectives, Bioinformatics Core). At present, one of the most daunting aspects of biomedical research concerns data management and bioinformatics tools for analysis. For proteomics research, data collection for protein structure determination requires new information technologies (protein informatics) to analyze and interpret the data. Faculty associated with the Bioinformatics Core will be available to provide the requisite expertise for data analysis, including more conventional statistical analysis of all types of data and analysis of data generated by microarray or proteomics techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Inflammation or peripheral nerve injury increases neuropeptide Y (NPY) expression in pain-relevant areas of the nervous system, such as large primary afferents and lamina l/ll of the dorsal horn of the spinal cord. However, the contribution of endogenous NPY to chronic pain is unclear. This NRSA proposes to test the overall hypothesis that NPY tonically inhibits nociceptive signaling in the spinal cord and brain following inflammation or nerve injury. Specific Aim #1 will test the hypothesis that genetic knockdown of NPY increases allodynia and/or hyperalgesia associated with nerve injury and inflammation. Aim #1 experiments utilize a conditional knockout mouse (Npytet) that contains a doxycycline (Dox)-regulated cassette (tTA) upstream of Npy exons to evaluate the contribution of NPY to the development (Aim 1 A) and maintenance (Aim 1B) of chronic pain, and correlate these changes with NPY expression using immunohistochemistry and western blot (Aim 1C). While Aim #1 establishes a contribution of NPY inhibition to behavioral manifestations of chronic pain, Specific Aim #2 uses direct microinjection techniques to investigate the site of action of NPY. NPY Y1 and NPY Y2 receptors decorate lamina l/ll of the dorsal horn. I propose to target these receptors with intrathecal injection of selective Y1 or Y2 receptor antagonists (Aim 2A). Previous studies indicate that inflammation up-regulates the expression of NPY and Y1 in the arcuate nucleus of the hypothalamus (ARC), and that microinjection of NPY into the ARC attenuated behavioral signs of heat hypersensitivity. In Aim 2B, I propose to microinject Y1 or Y2 antagonists into the ARC, and predict that this will exacerbate allodynia and hyperalgesia. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The ability to predict the response of normal tissues to a given course of radiation might allow a clinician to anticipate and avoid undue toxicity. Recent studies have demonstrated that the intrinsic radiosensitivity of skin fibroblasts from individual patients is predictive of the severity of acute and late radiation reactions experienced by these patients. While radiosensitivity assays are cumbersome, newer techniques which measure DNA double strand breaks (dsb) or the level of p53 induction (a central molecule involved in radiation-induced apoptosis) following irradiation may prove to be useful as predictive assays. The primary goal of this project will be to assess whether these assays performed on skin fibroblasts obtained form patients treated with radiation are predictive for acute or late skin reactions. Early passage fibroblast cell lines will be established from 26 breast cancer patients exhibiting severe skin reactions and from case-matched controls treated on a prospective radiation fractionation trial at the Royal Marsden Hospital. DNA dsb will be measured with pulse-field gel electrophoresis or single- cell gel electrophoresis. The levels of p53 and WAF1 (a downstream effector of p53) induction will be measured by various methods including Western blotting, enzyme linked immunosorbent assay, electrophoretic mobility shift assay and Northern blotting. We hypothesize that the level of DNA dsb induction following in vitro irradiation of skin fibroblasts should correlate with p53 induction, and that higher levels of p53 and DNA dsb induciton may predict for more severe acute or late normal tissue reactions in patients. The ability to accurately predict the severity of radiation reactions can potentially aid the clinician in planning a course of radiotherapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The University of Maryland requests support to develop a broad- based inter-departmental curriculum and program in Preventive Cardiology. This will be targeted to medical students and a wide range of house officers and fellow in training. A concerted effort to reach both junior and senior faculty will also be made. The program will build on a unique alliance between the Division of Cardiology and the Department of Epidemiology and Preventive Medicine. Led by the above, the Departments of Family Medicine, Pediatrics, Internal Medicine and the Office of Medical Education have prominent roles, with support from basic science departments as well. Objectives include: A) Development of a Center for Preventive Cardiology to foster education, patient care and research; B) Development of an integrated, core curriculum aimed at teaching, reinforcing and verifying knowledge, attitudes, and clinical skills with respect to prevention of cardiovascular disease; C) Training in \"Health Risk Assessment\" for medical students, house staff, fellows and faculty; D) Development of a Faculty Educator Development Core ensuring teaching by role model throughout the program; E) Development of a weekly conference series in preventive cardiology ; F) Preparation of a regular column in the Maryland Medical Journal, \"Perspectives in Preventive Cardiology,\" and a preventive cardiology campus newsletter; G) Development of teaching materials and evaluation instruments; H) A program for the applicant's own educational development in preventive cardiology, epidemiology and medical education. Plans for perpetuation of the program beyond the proposed funding period are also presented.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this study is to examine how the daily light-dark cycle in the environment affects the timing of hormone release, the timing of the daily cycle of body temperature, the sleep-wake cycle, and inner body functions which ordinarily vary with the time of day.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We will continue our investigation of the assembly of the bacterial cell wall. The main emphasis will be on the study of teichoic acid biosynthesis. We have discovered a new carrier for cell wall assembly, a lipid teichoic acid carrier, and are in the process of determining its structure. We believe this is an important key to the assembly of the bacterial cell wall by membrane bound enzymes. We will examine the structural relationship between the E. coli UDP-glucose 4'-epimerase and TDP-glucose oxidoreductase. These closely related enzymes and their structure have implications for the evolution of related enzymes to adapt to specific function. We are examining membrane recognition by embryonic cells. We will try to elucidate the membrane components involved in specific cellular recognition in embryonic cell aggregation and ultimately apply the same techniques to recognition problems involved in contact inhibition in normal and malignant cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The chief objective of this investigation is to clarify the mechanism of action of a simple flavoprotein oxidase, putrescine oxidase, as part of a general research interest in the mechanisms of flavoprotein catalyzed reactions. To this end, the kinetic component of the mechanism of putrescine oxidase will be studied utilizing standard steady-state and transient state (rapid-scan stopped-flow) kinetic techniques. The mode of cofactor binding to apoprotein will be explored using circular dichroism, fluorescence, and absorbance techniques together with studies of the effects of group-specific reagents on binding of the FAD. Effort will also be directed toward the design, construction and use of various types of specialized on-line instruments to be used in biochemical research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Project Director, Dr Rathod, will rely on the extensive infrastructure of his home institution, The University of Washington, for normal management (eg interactions with NIH and US institutions subcontractors). However, given the complexity ofthe project particularly in South Asia, the PD will emply a experienced, professional, part time Project Manager (50% effort) for the first two years ofthe Project. The Manager will go to 100% time commitment as the Project scales up in South Asia and the US in years 3-7. The South Asian part of project management will be handled through a new and innovative arrangement All South Asian contracts, and their administration, will flow through Dr Narayansamy's unique organization that has a team of business managers, scientific project managers, and technology support staff. Their scrupulous track record, and their familiarity with business and political norms in South Asia, will allow the main ICEMR scientific team to focus on the research and training aspects ofthe project.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Study of protein expression and protein-protein interaction is of central importance to understanding oncogenesis and cancer progression. Robust protein capture arrays will greatly facilitate cancer proteomics research. There is an acute need in designing novel types of affinity binders that are easy to express, smaller in size, highly stable, and extremely economical to screen and manufacture. This proposal describes a novel engineered protein scaffold for rapidly generating antibody-like molecules by using molecular selection processes. If the proposed project progresses to phase II/III, it will offer great commercialization opportunity in areas of proteomics research, molecular diagnostics and assay development for drug screening. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE: This project is related to development of a type of protein capture array for cancer research and diagnostics. The focus of the proposed work is to create a novel class of affinity reagents to accelerate deciphering gene function in the post-genomic era, with broad implications in improving human health. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "New molecular information from blood is being used to improve patient outcomes. Assays for established biomarkers have recently advanced (so-called 'high sensitivity', 'variants'and 'ultra', for examples), allowing physicians to use this additional information to provide better care. The enhanced patient outcomes result from more detailed, accurate and precise diagnostic and risk stratification, resulting in more timely and useful care (such as pharmaceuticals, treatments options, and timing of treatments) and avoiding unnecessary and costly actions. The objective of this proposal is to enable a new capability to isolate and concentrate biomarkers from blood in small volumes (200 microliters), at high sensitivity, over short periods of time, and to monitor multiple markers. We initially will focus on biomarkers for myocardial infarction and stroke. Upon completion of the entire project-which this proposal enables-enhanced information will be provided to physicians, allowing for accurate and fast diagnostics and treatments providing better patient outcomes-saving lives and money. To accomplish the proposal objective, a pair of microfluidic techniques will allow processing of small samples of blood to remove unwanted materials and isolate and concentrate the target biomarkers. These two techniques enable the objective because they keep the sample volume minimal and remove unwanted materials that could degrade detection, while quickly isolating and concentrating target species. Physically, this is made possible by exploit a unique combination of dielectrophoretic, flow and electrophoretic forces combined into the two techniques (gradient dielectrophoresis and electrophoretic capture) pioneered in the PI's laboratory. Gradient dielectrophoresis will remove cells and debris (and perhaps concentrate the targets) and electrophoretic capture will isolate and concentrate individual biomarkers away from possible interfering species. The four target biomarkers identified for proof of principle to enable the larger project are: two cardiac markers- myoglobin and cardiac troponin I, (cTnI), a stroke marker-neuron specific enolase (NSE), and an inflammatory marker-tumor necrosis factor-alpha (TNF1). These targets will provide a reasonable test for the success of the strategy and techniques.. PUBLIC HEALTH RELEVANCE: Developing an ability to isolate and concentrate biomarkers from blood using gradient dielectrophoresis and electrophoretic capture improving detection limits by at least two orders of magnitude while keeping the sample volume very modest (200 microliters).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In vitro fertilization (IVF) provides a unique opportunity to isolate and observe certain human reproductive processes, and thus potentially to differentiate among possible mechanisms of reproductive toxicity. In this clinical protocol, the woman undergoes pharmacologic hyperstimulation of her ovaries, and multiple ova are surgically extracted and fertilized in a dish, usually with her husbands sperm. Thus, the quality of each ovum is assessed, rates of fertilization and cleavage can be studied, early development of the embryos can be observed, and finally implantation into the uterus can be assessed, after the transfer of a known number of embryos. In collaboration with staff at UNC School of Medicine, we undertook a study to relate exposures to clinical outcomes in IVF. Although the patients are themselves infertile, this clinical protocol allows us potentially to demonstrate effects of certain exposures, such as caffeine, on specific reproductive processes. We have now completed the analysis of number of ova harvested, and fertilization and cleavage of the ova and the findings are being written up for publication. - infertility, in vitro fertilization, reproductive toxicants - Human Subjects & Human Subjects: Interview, Questionaires, or Surveys Only", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project focuses on the development of a coronavirus vaccine candidate for severe acute respiratory syndrome (SARS).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Description: The overall goal of the OSUEHS Centers COEP is to increase the public's ability to understand and to make informed decisions on issues relevant to the role of environmental factors in human health and disease. The COEP has eighteen current or past partnerships and collaborations with other OSU programs. The Center's COEP works actively with the Science and Math Investigative Learning Experiences (SMILE) program. These programs jointly received an NIEHS dissemination grant in 1996 to bring environmental health science education to teachers, students and community in rural Oregon. The program has reached an increasing number of students, teachers and community members since 1996. The majority of the population affected by this program consists of Native Americans (25 percent) and Hispanics (46 percent). Each year this program focuses on a different environmental health science theme, and as a portion of the program each summer teachers come to OSU to be trained in an environmental health science subject area. Of the four themes presented yearly since 1996, two were directly related to a research focus of the Center (1996-Chemicals in the Home Environment and 1998-Water Quality and Human Health). The \"Lunch with a Scientist\" portion of this program also involved Center investigators. Family Science Nights are also offered as a portion of this program through the COEP each fall. The final component of this program is the High School Challenge Weekend. Center members have served as environmental experts in this program by having students work in cooperative groups to develop solutions to environmental problems. Through the OSUEHS Center/SMILE Program, educational programs have been provided to over 1225 elementary, middle, and high school students, 125 teachers and 2000 community members. Rural communities (775 individuals) have received informal science education through family science nights. Center members are actively involved in programs such as Adventures in Learning, American Women in Science, and Science Education Partnerships. Teacher training components are currently being integrated into the program. Teacher workshops are being offered in the curriculum, Exploring Environmental Issues: Focus on Risk. Sixty-five teachers in 1999 have been trained in this curriculum, with future workshops planned in 2000. Teacher training in the ToxRAP program also will be a major focus of the Center through support from the Society of Toxicology. The OSUEHS Center sponsors a yearly course, Your Health and Chemical Risks. This course and environmental health science articles in the monthly newspaper reaches over 3,500 community members each month.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application for a comprehensive Training Program in Brain Tumor Biology at UAB offers the rigorous interdisciplinary education required for the development of independent research scientists in this field. The Training Program is based in the Departments of Cell Biology, Pathology, and Surgery. The 16 faculty members of the Training Program have been selected based on their complementary research expertise in basic and translational brain tumor research, dynamic research programs, and training records. Research projects include: studies of the molecular aspects of brain tumor migration/invasion, angiogenesis, cell surface receptor-initiated signaling, the regulation of apoptosis, and novel viral approaches to therapy. The exposure of the trainees to the full spectrum of intellectual and experimental approaches to this disease will be integrated formally through a new graduate level course in Brain Tumor Biology, a weekly Brain Tumor Research Conference, a monthly Brain Tumor Research and Treatment Seminar Series, and an annual UAB Brain Tumor Biology Scientific Retreat. The Training Program will be administered by Dr. Etty Benveniste (Director), Dr. Candece Gladson (Co-Director), and Dr. Jim Markert (Co-Director), and by an Executive Committee (Drs. Benos, Gillespie, Roth and Sontheimer). This group of faculty will be responsible for development of individual training plans, integration of research and didactic components, oversight of formal requirements, recruitment, competitive review of formal applications, and oversight of the trainees'progress. An additional strength of this Training Program is the close association with the UAB Brain Tumor SPORE Grant (PI: Dr. Yancey Gillespie), and the UAB Comprehensive Cancer Center Neuro-Oncology Program. Funding is requested for both advanced pre-doctoral students and postdoctoral trainees. Thus, this Training Program in Brain Tumor Biology will prepare future investigators in this important and clinically relevant area of research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The cytochrome P-450 dependent monooxygenase system plays a pivotal role in both the detoxification and bioactivation of drugs, environmental contaminants, and other potential chemical toxicants. The balance between detoxification and activation is largely dependent on the relative amounts and activities of different isozymes of cytochrome P-450. The long-term objective of the research proposed in this application is to design irreversible inhibitors of specific isozymes of cytochrome P-450. Such isozyme-specific inhibitors could be used in vivo either 1) diagnostically to assess the role of the various cytochromes in mediating or protecting against chemical toxicity or 2) therapeutically to redirect the metabolism of xenobiotics from potentially harmful to innocuous pathways. This proposal will focus on the mechanism, isozyme specificity, and structural requirements of the suicide inactivation of rat liver cytochrome P-450 by chloramphenicol. As the only known suicide substrate of cytochrome P-450 which acts by virtue of the modification of the protein rather than the heme moiety, chloramphenicol or one of its analogs should prove a unique tool for studying and modulating the various functions of the enzyme. Emphasis will first be placed on elucidation of the mechanism by which a single isozyme of rat liver cytochrome P-450 is inactivated in vivo upon covalent modification of specific amino acid residues in the protein by metabolites of chloramphenicol. Subsequent studies will focus on determining the specificity of chloramphenicol as an inhibitor of different isozymes of rat liver cytochrome P-450 and on elucidating which structural features of the chloramphenicol molecule are responsible for its effectiveness and specificity as a suicide substrate of cytochrome P-450. These studies should provide the rational basis for the design of isozyme-specific inhibitors for modulating monooxygenase function in vivo.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this program since its inception has been to define the pathobiological response of the mammalian respiratory system to the inhalation of ambient concentrations of oxidant air pollutants. The focus of this renewal application will be on mechanisms of environmentally induced asthma in young children, using the model of environmental asthma in infant rhesus monkeys that the investigators have developed through the support of this program. Using this model over the previous five years of funding, the investigators have made a number of startling discoveries regarding the effect of chronic ozone exposure on lung development and growth during infancy, including: stunting of airway growth, postnatal loss of airway generations, impaired establishment of the fibroblast growth factor (FGF)-2 ternary signaling complex by basal cells, the failure of epithelial surfaces to innervate, impaired central nervous control, enhancement of the allergic response, airway hyperreactivity, disrupted alveolarization, and airway remodeling. The analytical framework in which all of the studies proposed for this renewal will be conducted is the epithelial/mesenchymal trophic unit (EMTU), whose cellular components establish trophic interactions via an extracellular signaling complex modulated by the basement membrane zone (BMZ). The overall hypothesis is that environmental exposure to oxidant air pollutants promotes the development of allergic asthma in the developing lungs of young children and exacerbates its severity by: 1) disrupting the homeostasis within the EMTU and 2) fundamentally compromising the establishment and differentiation of the trophic interactions that promote normal airway growth and development. These changes result from the superimposition of continual cycles of acute injury, inflammation, and repair on the immune response to allergen exposure. Each of the four projects within the program will focus on different components of the EMTU: Project 1, epithelial and mesenchymal cells (fibroblasts, smooth muscle) and the BMZ; Project 2, mucosal immune system; Project 3, innervation and neural control; and Project 4, vasculature.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Connexin proteins form gap junction channels that mediate direct intercellular molecular communication crucial in development, physiology and response to trauma/inflammation. Defects in connexins cause human pathologies. Elucidation of the molecular mechanisms that regulate connexin channel function is essential for understanding their roles in human physiology and pathophysiology, and to identify targets for translational and basic science studies. The long-term goal of this project is to understand the cytosolic interdomain interactions that control the gating of connexin channels. This has been characterized for Cx43, in which interactions between the cytoplasmic loop (CL) and the C-terminal domain (CT) mediate channel gating by pH and other factors. The CL-CT mediated regulation of Cx43 is crucial for its physiological function, and is a therapeutic target for cardiovascular pathologies. However, the role of CL-CT interactions in modulation of other connexin channels, just as likely to be biomedically important, and their mechanism of action have not been explored. We have found that Cx26 channels are modulated by CL-CT interactions. Cx26 and Cx43 are representative members of the two largest families of connexins. Though structurally analogous, the effects of CL-CT interaction on Cx26 channel function seem to be fundamentally different from those in Cx43, suggesting that CL-CT interaction is a common modulatory mechanism in connexins, yet operate in connexin-specific ways. We propose to elucidate the molecular mechanisms of CL-CT control of channel function and properties using Cx26 and its closely related isoform Cx32. Cx26 is the only connexin channel for which there is a high- resolution structure, making it the basis for structure-based studies of all connexin channels. The proposed studies explore the basis and mechanisms of CL-CT interactions and channel properties they modulate, using strategies successfully applied to other channels, including use of competing peptides, engineered disulfide linkages, macroscopic and single channel recordings, and mutational analysis. The experiments utilize intact channels, complemented by peptide NMR. We propose to (a) determine the involvement of CL-CT interactions in channel gating, (b) identify the sites of CL-CT interactions, and (c) determine how CL-CT interaction and its effects are altered by mutations that cause human disease. Cx26 and Cx32 are widely distributed in the body. Mutations of Cx26 are responsible for over half the inherited sensorineural deafness worldwide, and also cause serious disfiguring skin disorders. Mutations of Cx32 cause a peripheral demyelination. In both connexins, many disease-causing mutations are positioned to affect CL-CT interaction. Both connexins are implicated tumor progression and a wide variety of pathological and physiological processes. The combination of a high resolution structure, a large number of disease causing mutations and the extensive experience of both PIs in studying gating, permeability and regulation of Cx26 and Cx32 channels provide a basis for productive, comprehensive investigation of CL-CT interactions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Administrative Core provides overall coordination, logistical support, and financial accounting for all cores and research projects. Among the tasks the Administrative Core will be responsible for are: bookkeeping and accounting; provision of grants management services; procurement of services, supplies, and equipment; coordination of and assistance in preparing progress and fiscal reports; maintenance of IRB approvals; and preservation of copies of Conte Center records and data sets. Collaborative education for the next generation of young scientists and physicians will be implemented to train students to become well versed in the art and science of interdisciplinary investigations that study genomic variations and environmental signals that promote mental health, and onset and progression of mental disorders. This is the major training goal of the center.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (adapted from the applicant's description): The overall hypothesis of this proposal is that alterations in myocyte calcium handling are centrally involved in the abnormal contractility of failing myocytes and contribute to the progressive decline of cardiac function in CHF. Two specific hypotheses will be tested. The first hypothesis is that the \"signature\" defects in calcium handling in CHF are due to an increase in the relative transport rate and capacity of the Na/Ca exchanger (NCX) with respect to the sarcoplasmic reticulum (SR) rather than an increase or decrease in the absolute rate or capacity of either transport mechanism. This will be tested by (1) measuring the calcium transport capacities of the SR and NCX in single myocytes from normal, hypertrophied and failing feline hearts and from non-failing, failing and left ventricular assisted device-supported (LVAD) human hearts, and (2) manipulating the calcium transport capacity of the SR and/or NCX in feline and human myocytes transfected with SERCA2a, native or mutant phospholamban (PLB), and/or NCX. The second hypothesis is that calcium influx via reverse-mode NCX makes a greater than normal contribution to excitation-contraction (EC) coupling and SR calcium loading because NCX density increases and the density of the L-type calcium channels and t-tubules decreases in CHF. This will be tested by (1) determining if calcium influx via reverse mode NCX modulates calcium-induced calcium release (CICR), (2) determining if calcium influx via reverse mode NCX replaces calcium entry via L-type calcium channels as the primary source of trigger calcium in CICR, and (3) measuring the amount of L-type calcium current, NCX current and SR calcium loading that is lost when T-tubules are disrupted by osmotic shock from the surface membranes of non-failing and failing feline and human myocytes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objectives of the research supported by this grant are to determine the novel chemical mechanisms by which vitamin B6 (PLP), Vitamin B12 (adenosylcobalamin), S-adenosylmethionine (SAM), and iron-sulfur clusters function in enzymatic reaction. The conventional role of PLP biology is to stabilize carbanionic intermediates in enzymatic reactions. A major focus of the research supported by this grant is the elucidation of the role of PLP in catalyzing reactions that involve free radical-paramagnetic intermediates. PLP-facilitated radical reactions appear to take place in reactions of aminomutases such as lysine 2,3-aminomutase and lysine 5,6-aminomutase. Radical intermediates of substrates and vitamin coenzymes will be characterized spectroscopically, and the structures of the enzymes that catalyze their reactions will be elucidated. The roles of SAM and [4Fe-4S] clusters in the initiation of radical formation will be unmasked and characterized chemically, spectroscopically, and kinetically. The relationship between the actions of adenosylcobalamin in Vitamin B 12-dependent and SAM/[4Fe-4S]-dependent aminomutases will be explored. Novel chemistry in the actions of vitamins B6 and B12, SAM, and [4Fe-4S] will be discovered. New beta-aminoacids will be produced by engineering of the genes producing aminomutases. Aminomutases are important in amino acid metabolism and the biosynthesis of antibiotics such as Streptothricin F, Myomycin, Blasticidin S, and Taxol. The contributions of beta- amino acids produced by lysine 2,3-aminomutase and arginine 2,3-aminomutase to the functions of antibiotics are not known. The beta-aminoacids may become significant in drug development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We will study the importance of two cellular receptors for herpes simplex 1 (HSV-1), herpes viral entry mediator (HVEM) and nectin-1, in the infection of the cornea and the establishment of latency by using mice knocked out (KO) for these receptors. Studies in cell culture and in vivo murine models of HSV infection indicate that HVEM and nectin-1 are the most efficient at mediating entry and most important in HSV pathogenesis. Reactivation of HSV-1 can lead to recurrent disease in the form of oral ulcers or more serious disease including encephalitis and herpes stromal keratitis (HSK). The precise mechanism of HSK pathogenesis is not fully understood, but factors known to contribute to disease include viral replication and the resulting immune response. We plan to study the outcome of corneal inoculation as well as examine which receptors function as HSV-1 entry mediators in susceptible tissues (corneal epithelium, corneal stroma, TG neurons, non-neuronal cells of the TG, immune cells, etc.). To fully understand disease pathogenesis and develop therapeutics, it is essential to understand the target tissues of HSV-1 in the eye. Our proposed studies will lay the foundation for further studies to determine whether infection of specific cell types, such as cells of the immune system, are important in HSV pathogenesis in the eye. By determining the precise cells infected by HSV in an ocular infection and the receptors utilized by the virus, novel treatment regimens can be developed that target those receptors. Such targeted therapies might attenuate primary infection and/or reduce inflammatory immune cells thereby preventing devastating sequelae including HSK, blindness, and encephalitis. In Aim 1, we will characterize the role of HVEM and nectin-1 in primary HSV-1 infection of the cornea by determining whether HVEM and/or nectin-1 are required for primary HSV-1 infection of the cornea. We will also evaluate the importance of input viral load on the development of zosteriform disease and determine which cells in the eye and TG are infected during primary HSV-1 infection and whether or not those cells are expressing HVEM and/or nectin-1. In Aim 2, we will determine whether HVEM and/or nectin-1 are necessary for HSV latent infection of the trigeminal ganglion. PUBLIC HEALTH RELEVANCE: Our specific aims are to identify the importance of the cellular receptors, HVEM and nectin-1, for HSV-1 infection of the eye. But more importantly how the use of these receptors contribute to herpes stromal keratitis (HSK). An understanding of the role for these receptors in HSK and how they contribute to disease will provide insight for the development of novel therapeutics for the treatment HSK which is the most frequent cause of corneal blindness in the US.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Head and Neck Squamous Cell Carcinoma (SCCHN) is one of the most common cancers worldwide and a significant cause of cancer morbidity and deaths. Although SCCHN is unique among solid tumors in that the majority of patients present with local disease, metastasis remains a critical problem for SCCHN treatment: patients without metastases can be cured; patients with metastases cannot. Even for local disease, despite improvements in outcome with intensive, multimodality therapies, such treatments are highly toxic, poorly tolerated and only partially efficacious. Long-term survival is only ~ 50%, with most patients dying from their cancers. There is an urgent need to uncover the critical genetic alterations in SCCHN and to determine how these impact patient outcome and metastatic risk. To search for genes that are critical for head and neck squamous carcinoma (SCCHN) biology, prognosis and therapy, I have initially used high resolution single nucleotide polymorphism (SNP) arrays to detect copy number alterations in 684 human cancer cell line of diverse tissue origin, including SCCHN. This analysis has identified recurrent homozygous deletions involving >200 genes. Among these, PARD3, encoding a critical regulator of cell polarity, is disrupted in squamous carcinoma cell lines and primary tumors, including from the head and neck region. Since abnormal cell polarity has been linked to increased migration and metastasis, these findings may implicate loss of function of PARD3 or its downstream effectors in the risks of recurrence and metastasis of a subset of these cancers. The overall goals of this proposal are: (1) to further characterize the role of PARD3 in SCCHN tumorigenesis, migration and metastasis; and (2) to capture and genotype circulating tumor cells (CTCs) from the peripheral blood of patients with SCCHN, to determine if CTCs can identify patients at risk for metastases before metastatic disease is clinically apparent. The experiments will utilize primary tumors and peripheral blood from patients treated for SCCHN, as well as PARD3 loss-of-function and gain-of-function squamous cell lines for in vitro and in vivo studies to identify the cancer-related phenotype(s) controlled by PARD3 and the domains and activities required for tumor suppression. By characterizing PARD3 loss in the tumor type in which inactivating mutations have arisen as a primary event during tumor formation, this work will directly implicate alterations in cell polarity in the development and invasive and metastatic potential of these cancers. By capturing and molecularly characterizing CTCs in actual SCCHN patients undergoing treatment, this work will permit the real time in vivo analysis of tumor cells to quantify clinical risk and guide treatment. As a result, these studies may lead to improved biomarkers and suggest potential new targets for therapies, thus providing benefit to public health by improving the care of patients with these cancers. PUBLIC HEALTH RELEVANCE: Although cancers of the head and neck pose a significant health problem worldwide, how they develop is incompletely understood, existing treatments are only partially effective and safely measuring whether treatments actually target the tumor cells in the patients being treated is very difficult. We have discovered frequent loss of a single gene, PARD3, in human head and neck cancers that could permit cells to loosen from each other, move through the blood to different sites and thereby evade detection and resist treatment. By determining whether patients possessing cancers which have lost PARD3 do worse with treatment, how PARD3 contributes to cancer formation and spread, and whether the tumor cells present in patients' blood can be captured and used to measure the benefit of treatment, the experiments outlined in this proposal may lead to better and less toxic treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this research is to examine the antecedents, process and consequences of divorce using samples of black and white males and females from divorcing and intact families located in metropolitan Cleveland. The study uses a prospective design in which the two samples, each of approximately 200, are being followed for a 3 1/2 year period. The divorcing are interviewed 3 times: at time of filing; six months after settlement; six months to a year after the second interview. The intact family sample is interviewed twice: simultaneously with the first and third divorce interviews. Among the study objectives are: (1) to indicate which periods during the divorce process produce the highest rates of disturbance for individuals of varied characteristics; (2) to indicate what factors aid or hinder socialization to the roles associated with the divorced status; (3) to explore the influence of human support systems such as kin, friends, voluntary associations and service agencies in easing or exacerbating the process of adjustment; and (4) to compare the divorced to members of the intact family sample in order to determine the relative influence of divorce as a crisis event on health status on role functioning. The analysis involves within and between-groups comparisons and employs such techniques as analysis of variance and multiple regression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Substantial evidence suggests that activity plays a crucial instructive role in shaping the structure and connectivity of the central nervous system, but the molecular mechanisms that translate developmental activity patterns into lasting structural changes in the brain remain open mysteries. Neurotrophins have long been implicated as candidate signaling molecules that potentially mediate activity- dependent development, but their in vivo roles have been difficult to predict based on conventional genetic approaches or culture studies. By using a novel genetic mosaic approach to generate small groups of labeled homozygous mutant cells in intact animals, this proposal will dissect the role of activity and neurotrophin signaling in brain development in vivo, with single-cell resolution. Specifically, the proposal will combine inducible genetic strategies and genetic mosaic analysis to test the function of neurotrophin receptors p75, TrkB, and TrkC in development and refinement of central somatosensory circuitry. The somatosensory system relies on activity for proper structural maturation of both axons and dendrites, and thus represents an excellent opportunity to test key hypotheses in neurotrophin signaling, activity-dependent development, and neural circuit formation. As p75 and Trk receptor dysfunction contributes to neurological disorders including Alzheimer's disease and schizophrenia, potential results will also have broad implications for our understanding of neural circuit maintenance and repair, as well as therapeutic approaches to neurodevelopmental and neurodegenerative diseases. PUBLIC HEALTH RELEVANCE: This proposal will study the molecular mechanisms by which activity influences nerve cell structure during brain development. Proper development of cell shape and connectivity in the central nervous system is critical for perception and cognition. Accordingly, studies in nerve cell morphogenesis will considerably aid our understanding of how the brain wires itself during development, how our early experiences can generate long-lasting changes in our brains, what happens when nerve cells are injured by neurodegenerative disease or trauma, and how we can repair nervous system damage.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Eukaryotic cells dedicate significant resources to the deployment of proteins to the membrane compartments that comprise the secretory pathway. Biogenesis of all membrane proteins starts with the folding and assembly of newly synthesized proteins within the endoplasmic reticulum (ER), often with the aid of cellular chaperones. Cells must strike a precise balance between ensuring that only fully folded proteins are allowed to leave the ER and avoiding the accumulation of misfolded proteins within the ER lumen. Uptake of newly synthesized cargo molecules into ER-derived transport vesicles only occurs once proteins are fully folded. These transport vesicles, known as COPII vesicles for the cytoplasmic coat proteins that drive membrane curvature and select cargo, thus play a critical role in regulating forward transport of new proteins. We study the close relationship between protein folding and packaging into ER- derived COPII vesicles in the model organism, Saccharomyces cerevisiae. Using a combination of genetics and biochemistry, we aim to define the cellular machinery that acts at the interface between protein folding and ER export. A model for examining this process is the yeast ABC transporter, Yor1, a plasma membrane protein that acts as a drug pump to clear toxic substances from the yeast cytoplasm. Yor1 is a homolog of the human cystic fibrosis transmembrane conductance regulator (CFTR), defects in which cause cystic fibrosis. Deletion of a Phe residue in Yor1, equivalent to the major disease-related mutation in human cystic fibrosis, causes Yor1, like mutant CFTR, to be ER-retained and degraded by the cytoplasmic ubiquitin/proteasome pathway. Thus Yor1 is a useful model that allows the direct comparison of the intracellular itineraries of native and aberrant forms of a single protein. This research proposal consists of four specific aims. (1) To define the molecular mechanisms that drive uptake of Yor1 into COPII vesicles and assess how protein folding influences this event. (2) To determine how cellular chaperones contribute to Yor1 biogenesis and assess how the kinetics of chaperone/client interactions may influence COPII binding and thereby regulate ER export. (3) To identify and characterize novel factors that may facilitate Yor1 biogenesis, including specific membrane chaperones and more general folding factors. (4) To determine the mechanisms by which the unfolded protein response improves the folding and/or transport of misfolded proteins. Ultimately, a better understanding of cellular machinery that acts to regulate protein folding and forward transport may lead to novel therapeutic approaches to treat the many diseases associated with aberrant protein folding within the secretory pathway. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. N-acetyl-L-cysteine (NAC), a natural antioxidant, improves the efficiency of chelating agents used against heavy metal poisoning and facilitates the excretion of Cr(III), Cd(II) and Pb(II), but not of Hg(II). Recent bio-tests on NAC complexes with transition biometals (Zn2+, Co2+, Co3+, Fe2+) showed significant anti-inflammatory activity. The goal of our proposed research is to study the structure of NAC complexes with heavy metals, and transition biometals, in solid state and in aqueous solution at physiological pH and pH = 8.5, using different [NAC]/[Mn+] ratios and [Mn+] = 10 and 100 mM. In this study, we will combine the results from different spectroscopic techniques, such as extended x-ray-absorption fine structure (EXAFS), multi-nuclear NMR, Raman and IR, to identify the local structure around the central metal ion in these complexes. We will use the method that we have developed for EXAFS data analysis of mixtures for similar Cd(II), Hg(II) and Pb(II) complexes with cysteine and penicillamine (3,3'-dimethyl cysteine), including Principal Component Analysis (PCA) of the EXAFS spectra. The aim of the current study is to provide a basis for better understanding of the detoxification treatments and the anti-inflammatory activities with the use of cysteine derivatives as complex-forming agents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "As a participating institution in the Primary Breast Cancer Therapy Group (NSABP), we are contributing data from patients entered into Protocol No. 4 of the group, which is a protocol to evaluate the relative efficacy of radical mastectomy, total mastectomy with radiation and total mastectomy alone in patients with clinically negative axillary nodes. A comparison between radical mastectomy and total mastectomy with radiation is made in those with positive axillary nodes. To date, over 900 patients have been entered into the protocol by 35 institutions of which ours is one. The present application seeks funding to permit continued patient accrual into the present study so as to bring it to completion and to permit participation in the development and implementation of new protocols which are to follow. It is premature to make statements relative to the various treatment modalities. Interesting data, however, has been obtained relative to ancillary material being collected. It has been demosntrated that patients randomized into various treatment categories are similar according to a variety of criteria. Moreover, information has been accumulated relative to the pathology of breast cancer. The submittal of data has been satisfactory. If patient accrual continues at the present rate, it should be adequate to complete the study (accrual) within the next year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ixodes scapularis ticks transmit Borrelia burgdorferi as well as several other pathogens of public health importance. Feeding ticks cut into host tissues and create a feeding pool from which they continuously feed for days. Host immune responses interfere with tick feeding by clogging the feeding site and producing molecules that are noxious to the tick. The host complement system is likely to pose special problems for feeding ticks because complement can directly damage non-self tissue as well as recruit arms of the immune response. The focus of this proposal is anti-complement proteins produced by I. scapularis. We plan to test the hypothesis that anti-complement proteins facilitate tick feeding and pathogen transmission. Under aim 1 of the proposal we plan to functionally characterize two novel proteins designated Salp9 and Salp20 that are highly homologous to a known tick anti-complement protein. Under aim 2 we propose to screen tick cDNA libraries prepared from salivary glands and guts for novel anti-complement proteins. Proteins produced by ticks are attractive vaccine targets because such vaccines have the potential to block tick feeding and pathogen transmission. Under aim 3 we plan to test anti-complement proteins as vaccines that block feeding and/or pathogen transmission. Our industry partner in these studies is L2 diagnostics, which will assist with further development of tick anti-complement proteins as drugs and vaccines.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract The development of human enteroids as a model to study the human intestine offers tremendous opportunities to study the pathogenesis of infectious enteric disease. We propose to combine the substantial expertise from three institutions with long and impressive histories in the investigation of enteric diseases to exploit this powerful model to study the pathogenesis of four major etiologic agents of diarrheal disease: three pathotypes of diarrheagenic E.coli (enteroaggregative, enterohemorrhagic and enterotoxigenic E. coli) and Shigella. Investigators from the University of Maryland, Johns Hopkins University and the University of Virginia will collaborate in this Program Project Grant (PPG). The leadership team of this PPG comprises three internationally renowned experts in enteric diseases, with complementary training in microbiology, gastroenterology, molecular physiology, and pediatric infectious diseases. They will direct a multidisciplinary team of co-investigators with expertise in cell biology, molecular pathogenesis, and mucosal immunology. The overall goal is to increase understanding of the pathophysiology and potential treatments of these four important pathogens. The proposed studies will use normal human mini-intestines, called enteroids or colonoids, grown on monolayers to develop models that mimic human disease. Examination of pathophysiologic aspects common to the diseases studied will serve to integrate the projects. These aspects include the role of mucins, bacterial proteases called SPATES, enterotoxins and secreted cytokines and are in addition to pathophysiologic aspects specific to each infection. In addition, the contribution of cells involved in innate immunity will be examined by co-culture of the enteroids/colonoids with human macrophages, neutrophils and dendritic cells. These studies using human mini-intestines offer the possibility of revealing insights in disease pathophysiology that are specific to normal human intestine rather than the animal models and cancer cell line models used until now. Each of the four projects focuses on a specific pathogen: enteroaggregative E. coli, Shigella, enterohemorrhagic E. coli, entertoxigenic E. coli. Besides an Administrative Core, there is an Enteroid Core that provides human enteroids/colonoids and growth media, instructs all projects on how to produce enteroid/colonoid monolayers and an Immunology Core that measures cytokines and chemokines, and isolates human macrophages, neutrophils and dendritic cells. The Enteroid and Immunology Cores work together to develop co-culture systems of innate immune cells and enteroids/colonoids that will be used by the projects. The investigators will regularly interact by monthly joint laboratory meetings. The proposed project will yield many significant new insights into enteric disease caused by these important pathogens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Western immunoblot for the detection of 14-3-3 protein in the cerebrospinal fluid of human patients with sporadic or familial spongiform encephalopathy and chimpanzees with experimentally induced TSEs has provided for a highly sensitive and specific test in support of a clinical diagnosis of these diseases. However, it is not known how early in the course of these infections the protein becomes detectable in the CSF. We have demonstrated in spider monkeys that neuropathological lesions precede clinical signs and have hypothesized that 14-3-3 proteins are detectable due to neuronal loss at a rate sufficient to outpace the normal turnover and resorption of CSF. We have detected these proteins in the CSF of chimpanzees inoculated with CJD several weeks before clinical signs become recognizable. In humans with very early behavorial changes the test has been negative but repeat collections as the disease progresses the test becomes positive. To more definitively determine the earliest possible detection of 14-3-3 proteins in CSF we have inoculated ten chimpanzees with sporadic, iatrogenic and familial strains of CJD and are colelcting CSF and blood at intervals of every six weeks. Concurrently we have determined the stablility of 14-3-3 protein and have shown that the protein in CSF is stable at room temperature and 4C for over 35 days and indefinitely at 20C.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this project is to understand at the molecular level the mechanism by which steroid hormones affect the expression of the genome of their target cells. We have chosen to investigate the cytolytic response of murine lymphoid cell lines to glucocorticoids because this system presents unique and desirable features. Since variants resistant to the steroid can conveniently be isolated, this system offers a genetic approach to analyze the steps involved in the response. A large collection of lymphoid cell variants resistant as the result of a defect in the glucocorticoid receptor is already available. We propose to use a variety of procedures to search for new types of resistant variants. The existing collection of resistant variants, and all new variants that will be isolated, will be characterized genetically and biochemically. The genetic characterization includes complementation analysis in cell hybrids and linkage studies. The biochemical tests involve assays for analysis in cell hybrids and linkage studies. The biochemical tests involve assays for glucocorticoid receptor and for a putative nuclear acceptor site for the receptor. Enucleated cells will be prepared to examine the role of the nucleus in the lymphocytolytic response, and ouabain resistant variants will be isolated to examine a possible relationship to glucocorticoid sensitivity. We will also pursue the biochemistry of the glucocorticoid receptor. This receptor will be purified and its molecular weight, isoelectric point and subunit structure examined. We will prepare a specific anti-receptor serum. This serum will be used, in particular, to detect the production of a protein crossreacting with receptor in \"receptorless\" variants, to purify the receptor by immunoadsorption, and to examine its conformational change and structure. The anti-receptor serum will also serve as a tool to compare the receptor of our sensitive lymphoid cell lines with that of lymphoid cells resistant to the steroid as the result of normal differentiation, and of other types of mouse cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The broad objective of the proposed research is to evaluate the potential usefulness of the ferret as an experimental animal for work on the relation between changes in brain morphology and cognitive processes. It is hoped that the capability to vary the degree of chemically induced micrencephaly in an otherwise normal animal and then to observe the ensuing degree of impairment in higher mental process will provide a means of study of a form of mental deficiency. Specifically, the proposed work is aimed at determining whether the performance of ferrets made micrencephalic by prenatal treatment with methylazoxymethanol is impaired on problems that appear to require symbolic (abstractive) processes (viz., acquisition of oddity and matching-from-sample learning sets).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Biofilms found on mammalian tissue surfaces contain complex mixtures of bacterial species growing within a glycocalyx matrix. Species in biofilms have shown remarkably greater resistance to antibiotics than when grown in a planktonic state. Surprisingly little is known about the effect of systemically administered antibiotics on the microbial composition of naturally occurring, complex biofilms. Thus, the long-term objective of this proposal is to determine the effect of antibiotics on the microbial composition of biofilms as they exist in vivo and to determine if the proportion and nature of antibiotic resistant taxa is affected. Subgingival biofilms that occur on teeth will be employed as a model system because of their complexity, ready accessibility and known microbial composition. Specific Aim 1 will examine the effects of 3 systemically administered antibiotics on the microbial composition of subgingival biofilms in adult humans with periodontitis. All subjects will receive scaling and root planing and will be randomly assigned to one of 4 groups of 36 subjects each receiving one of doxycycline, amoxicillin, metronidazole or control. 28 subgingival biofilm samples will be taken in each subject at baseline, at selected time points while the agent is being taken, the same time points after cessation of the agent as well as at 3, 6 and 12 months. These samples will be evaluated individually for their content of 40 subgingival species using checkerboard DNA-DNA hybridization. Data will be evaluated longitudinally and compared with clinical parameters. Specific Aim 2 will examine the proportion and nature of subgingival species that are resistant to 4 mug/ml of the test antibiotic at the same time points. Subgingival biofilm samples will be plated on media with and without the test antibiotic and resistant isolates identified using DNA probes. Data from this investigation will indicate the kinetics of suppression of species in subgingival biofilms during antibiotic administration, the kinetics of repopulation after antibiotic withdrawal as well as the nature and proportion of subgingival species that are resistant to the antibiotics at different time points. The data should indicate the effects of antibiotics on the microbial composition of complex ecosystems such as those found in biofilms and be useful in guiding and interpreting in vitro studies of mechanisms of antibiotic resistance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We continue our long-term study of the regulation of ribosomal RNA transcription in the yeast, Saccharomyces cerevisiae. Ribosomal RNA is transcribed by RNA polymerase I (po1I) and comprises over half of the total RNA of the cell. Both the cis-acting DNA elements, as well as the trans-acting protein factors which control po1I transcription have largely been identified in past work. The challenge now is to understand how these DNA and protein elements function to produce ribosomal RNA in strict co-ordination with the growth rate of the cell. Because ribosomal RNA production is so important to the economy of the cell, it is subject to regulation at many different levels. Thus, experiments are proposed to study regulation at the level of 1) the polymerase itself, 2) the pre-initiation complex that forms at the promoter, 3) chromatin which must be cleared for the pre-initiation complex to form. Other experiments will survey all of the po1I transcription machinery during a major metabolic shift (glycerol/ethanol to glucose) to determine which of these potential regulatory levels are actually utilized. Ribosomal RNA is a major and essential component of all living cells. Alterations in po1I transcription are not known to be associated with any disease state, but understanding how ribosomal RNA production is co- ordinated with cell growth is one of the major unsolved problems of cell biology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "LIPOPOLYSACCHARIDE INDUCED NEUROINFLAMMATION SELECTIVE INVOLVEMENT OF ARACHIDONIC ACID IN NEUROINFLAMMATION In a rat model of neuroinflammation, produced by a 6-day intracerebral ventricular infusion of bacterial lipopolysaccharide (LPS), we reported marked disturbances in brain arachidonic acid (AA, 20:4n-6) metabolism. In the present study, we demonstrated that parameters of brain docosahexaenoic acid (DHA, 22:6n-3) metabolism were unaffected in this model. Selective targeting of brain AA metabolism with non-steroidal antiinflammatory or other drugs should be considered for treating human brain diseases associated with neuroinflammation (Rosenberger et al., 2010). ANTI-INFLAMMATORY EFFECT OF ASPIRIN ON BRAIN ARACHIDONIC ACID METABOLITES Pro-inflammatory and anti-inflammatory mediators derived from arachidonic acid (AA) modulate peripheral inflammation and its resolution. Aspirin (ASA) is a non-steroidal anti-inflammatory drug (NSAID) that switches AA metabolism from prostaglandin E (PGE) and thromboxane B (TXB) to lipoxin A (LXA) and 15-epi-LXA. It is unknown whether chronic therapeutic doses of ASA are anti-inflammatory in the brain. We hypothesized that ASA would dampen increases in brain concentrations of AA metabolites in a rat model of neuroinflammation, produced by a 6-day intracerebroventricular infusion of bacterial lipopolysaccharide (LPS). In rats infused with LPS (0.5 ng/h) and given ASA-free water to drink, concentrations in high-energy microwaved brain of PGE, TXB and leukotriene B (LTB) were elevated. In rats infused with artificial cerebrospinal fluid, 6 weeks of treatment with a low (10 mg/kg/day) or high (100 mg/kg/day) ASA dose in drinking water decreased brain PGE, but increased LTB, LXA and 15-epi-LXA concentrations. Both doses attenuated the LPS effects on PGE, and TXB. The increments in LXA and 15-epi-LXA caused by high-dose ASA were significantly greater in LPS-infused rats. The ability of ASA to increase anti-inflammatory LXA and 15-epi-LXA and reduce pro-inflammatory PGE and TXB suggests considering aspirin further for treating clinical neuroinflammation (Basselin et al., 2011a). LITHIUM MODIFIES BRAIN ARACHIDONIC AND DOCOSAHEXAENOIC METABOLISM IN RAT LIPOPOLYSACCHARIDE MODEL OF NEUROINFLAMMATION. Neuroinflammation, caused by 6 days of intracerebroventricular infusion of a low dose of lipopolysaccharide (LPS;0.5 ng/h), stimulates brain arachidonic acid (AA) metabolism in rats, but 6 weeks of lithium pretreatment reduces this effect. To further understand this action of lithium, we measured concentrations metabolites generated from AA and docosahexaenoic acid (DHA), in high-energy microwaved rat brain using LC/MS/MS and two doses of LPS. In rats fed a lithium-free diet, low (0.5 ng/h)- or high (250 ng/h)-dose LPS compared with artificial cerebrospinal fluid increased brain unesterified AA and prostaglandin E2 concentrations and activities of AA-selective Ca2+-dependent cytosolic phospholipase A2 (cPLA2)-IV and secretory sPLA2. LiCl feeding prevented these increments. Lithium increased brain concentrations of lipoxygenase-derived AA metabolites, 5- hydroxyeicosatetraenoic acid (HETE), 5-oxo-eicosatetranoic acid, and 17-hydroxy-DHA by 1.8-, 4.3- and 1.9-fold compared with control diet. Lithium also increased 15-HETE in high-dose LPS-infused rats. This study demonstrated, for the first time, that lithium can increase brain 17-hydroxy-DHA formation, a precursor of antiinflammatory resolvins, indicating a new antiinflammatory therapeutic action of lithium (Basselin et al., 2010). PARKINSON DISEASE MODEL UPREGULATED BRAIN ARACHIDONIC ACID ENZYMES IN RAT MODEL OF UNILATERAL PARKINSON DISEASE We had reported that arachidonic acid (AA) signaling is upregulated in the caudate-putamen and frontal cortex of unilaterally 6-hydroxydopamine (6-OHDA) lesioned rats, a model for asymmetrical Parkinson disease. In the present study, we demonstrated that this upregulation was associated with increased expression of two enzymes involved in AA metabolism, cytosolic phospholipase A2 (cPLA2) and cyclooxygenase (COX)-2, ipsilateral to the lesion in the caudate putamen and frontal cortex. This confirms that the tonically increased ipsilateral AA signal in the lesioned rat corresponds to upregulated cPLA2 and COX-2 expression within the AA metabolic cascade;such changes, if present in Parkinson disease, may contribute to symptoms and pathology of this disorder (Lee et al., 2010). HIV-1 DEMENTIA MODEL IMAGING NEUROINFLAMMATION WITH ARACHIDONIC ACID IN HIV-1 TRANSGENIC RAT Human immunodeficiency virus (HIV)-1 associated infection involves entry of virus-bearing monocytes into brain, followed by microglial activation, neuroinflammation, and upregulated arachidonic acid (AA) metabolic enzymes. The HIV-1 transgenic (Tg) rat, a noninfectious HIV-1 model, shows neurologic and behavioral abnormalities after 5 months of age. We used our in vivo imaging method with quantitative autoradiography to demonstrate that brain AA metabolism was elevated in 6-7 month old unanesthetized HIV-1 Tg rats. Brain activities of cytosolic phospholipase A2 (cPLA2-IV), secretory sPLA2, and calcium independent iPLA2-VI, which release AA and docosahexaenoic acid from membrane phospholipids, and concentrations of proinflammatory prostaglandin E2 and leukotriene B4, also were elevated, consistent with neuroinflammation and increased AA metabolism (Basselin et al., 2011b). We now plan to use our clinical method of positron emission tomography with 1-11CAA, to test whether brain AA metabolism is upregulated in HIV-1-infected patients as a marker of neuroinflammation (AG000148). INCREASED INFLAMMATORY AND ARACHIDONIC ACID CASCADE MARKERS, AND REDUCED SYNAPTIC PROTEINS, IN BRAIN OF HIV-1 TRANSGENIC RAT Cognitive impairment has been reported in human immune deficiency virus-1 (HIV-1-) infected patients and in the HIV-1 transgenic (Tg) rat. We hypothesized that this impairment in the rat could be linked to neuroinflammation, disturbed brain arachidonic acid (AA) metabolism, and synapto-dendritic injury. To test this, we measured protein and mRNA levels of markers of neuroinflammation and the AA cascade, as well as pro-apoptotic factors and synaptic proteins, in brain from 7- to 9-month-old HIV-1 Tg and control rats. Compared with control brain, HIV-1 Tg rat brain showed immunoreactivity to glycoprotein 120 and tat HIV-1 viral proteins, and significantly higher protein and mRNA levels of (1) the inflammatory cytokines interleukin-1&#61538;and tumor necrosis factor alpha, (2) the activated microglial/macrophage marker CD11b, (3) AA cascade enzymes: AA-selective Ca2+-dependent cytosolic phospholipase A2 (cPLA2)-IVA, secretory sPLA2-IIA, cyclooxygenase (COX)-2, membrane prostaglandin E2 synthase, 5-lipoxygenase (LOX) and 15-LOX, cytochrome p450 epoxygenase, and (4) transcription factor NF-&#61547;Bp50 DNA binding activity. HIV-1 Tg rat brain also exhibited decreased levels of brain-derived neurotrophic factor and drebrin, a marker of post-synaptic excitatory dendritic spines. In summary, HIV-1 Tg rats show elevated brain markers of neuroinflammation and AA metabolism, with a deficit in several synaptic proteins. These changes are associated with viral proteins and may contribute to cognitive impairment. The HIV-1 Tg rat may be a useful model for understanding progression and treatment of cognitive impairment in HIV-1 patients (Rao et al., In press).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "From a clinical perspective, understanding and manipulating the dynamics of T cell development may offer key insight in to the progression of various autoimmune diseases as well as present potential treatment options with regards to cancer immunotherapy and vaccination. Although existing single cell approaches are now elucidating the significance of heterogeneity, the way in which T cells evolve to achieve particular phenotypic states remains unclear. We are proposing to use novel microfluidic platforms for dynamically monitoring the response of individual CD8+ T cells over several generations to investigate differentiation, plasticity and metabolism. To accomplish this, we will utilize hydrodynamic traps to dynamically interrogate several generations of a single T cell's progeny by implementing standard immunofluorescence techniques on-chip. Since fluid surrounding the cells can be rapidly and frequently exchanged without perturbing growth, we will be able to monitor expression of cell surface markers and deliver drugs that alter cellular metabolism at precise time points. As a compliment to biochemical markers of lineage we will also measure multigenerational growth rates of single activated T cells in order to provide a physical measurement of the cell's metabolic state.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Investigating the key variables that affect exercise adoption and exercise maintenance for cancer survivors is important to begin to address the plethora of negative consequences of an inactive lifestyle on cancer survivorship. Identifying these variables and how they differ in specific disparate cancer survivor populations is an integral step in developing culturally competent behavioral interventions for adopting and maintaining a physically active lifestyle. The objective of this particular application is to examine differences in the determinants of exercise behaviors between Hispanic and non-Hispanic cancer survivors; and to conduct an efficacy culturally tailored intervention pilot specific for Mexican-American Latina breast cancer survivors. PUBLIC HEALTH RELEVANCE: There is a critical need to understand how to get people to adopt a more active lifestyle to help offset illness and increased risk for disease from a physically inactive lifestyle. This study will investigate factors that affect beginning and maintaining a more physically active lifestyle. The study will look to see if there are differences in these factors between Hispanics and non-Hispanics and based on those differences conduct a trial to increase physical activity specific for Hispanic breast cancer survivors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The continuation of work in this area has revolved about the analysis of lymphocyte responsiveness in vitro to a putative suppressive immunogenic virus (rubeola) and the meaning of this putative suppression in relation to observations within the clinical disease in man. In addition, work has been performed utilizing laser cytometry as a method of quickly and accurately evaluating cell change in vitro as a measure of cell responsiveness to specific antigen, mitogen or allogeneic cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To recognize and interact with objects in an object-filled environment, an accurate perceptual representation of the surfaces that define the objects is essential. The surface representation process integrates local feature information from the early cortical processes to form surfaces. Complicating the process of representing surfaces, however, is the fact that most objects overlap one another, resulting in the occluded parts of the objects' surfaces not being encoded by the early cortical processes. Thus, to accurately represent surfaces, the surface representation process uses an arsenal of perceptual rules derived from environmental constants even as it relies on the early features of the non-occluded parts. But how these perceptual rules are implemented, at which stage of the surface representation process, and how top-down attention modulates the representation of both the occluding and occluded surfaces are not well understood. In light of these, our proposal addresses three fundamental issues in representing surfaces. The issues are: A. The boundary contour and surface property information in amodal surface interpolation B. The operational constraints in surface formation during binocular viewing C. Inhibitory mechanism and plasticity in binocular surface perception These issues will be investigated using psychophysical methods on human observers with normal vision, and those with significant sensory eye dominance. Observers in the latter group tend to suppress the image viewed by the non-dominant eye when the two eyes see different images; a situation almost resembling the extreme condition afflicting strabismic amblyopia patients. In this proposal, we will test and evaluate a new perceptual learning paradigm aimed at reducing sensory eye dominance and improving perception. Therefore, the outcomes of our investigations will not only further the knowledge of perceptual and attention mechanisms in perceiving surfaces, but also has a clinical significance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In order to understand such important processes as enzyme-substrate binding and antibody-antigen recognition, numerous investigators have synthesized and studied artificial hosts like water soluble cyclophanes. Some of these hosts have bound guests with high affinity constants but many improvements are needed. We intend to synthesize and study a particularly rigid \"molecular tweezer\" which will \"sandwich\" aromatic guests between two DNA intercalators. The complexation is expected to be very efficient and with a predictable geometry. We also believe that the approximate binding constant can be predicted using a simple cooperative model. Molecular tweezers with functional groups converging on the binding site will be investigated. These will be the first hosts which include their guests using both hydrogen bonding and hydrophobic forces resulting in improved guest discrimination. The specificity in binding mono-nucleotides will be investigated. In addition to host-guest chemistry these molecular tweezers are expected to be novel probes of DNA structure. A tremendous array of medicinals, including the clinically useful anticancer agent adriamycin, contain flat aromatic chromophores which bind to DNA by intercalation. Many aspects of the intercalation processes are not well understood including the important neighbor exclusion principle. Our molecular tweezers are the first bis-intercalators to force chromophores into adjacent sites. We will investigate chromophores of different \"widths\" to determine the origin of the neighbor exclusion principle. Those molecular tweezers which obey the principle will be unable to act as mono-intercalators and therefore unable to bind the internal sites of the DNA helix. We will demonstrate that these molecules are able to selectively bind to the ends of DNA helices which are not constrained by neighbor exclusion. This type of selective binding has not been seen before and should have a number of important applications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project concerns mechanisms of demyelination in human disease and continues to focus on the human polyomavirus JC (JCV), the etiologic agent of progressive multifocal leukoencephalopathy (PML). PML is a fatal demyelinating disease which complicates about 5% of AIDS cases, and for which there has been no treatment known. Work this year has emphasized the detection of the virus by polymerase chain reaction (PCR) in PML tissues, in normal human brain tissues, in brain tumors, in human kidneys, in urine in infected hamster tissues, and in transgenic mice. (Parts of this work are reported under related projects in this Section.) We found that JCV DNA could be detected in 33 of 46 (72%) of sections from PML blocks. We have also found that JCV sequences could be detected in a paraffin section which had been scraped from slides following immunocytochemical staining. Detection of JCV in that biopsy tissue allowed confirmation of suspected PML which has been successfully treated with high dose AZT. This allows reexamination of large numbers of archival PML materials, even those which have already been utilized for conventional pathological studies. In a related project, two genotypes of JCV have been demonstrated following sequencing of a 610-bp fragment comprising the VP1 gene. This information has allowed design of type-specific primers which can also be applied to archival CNS tissues in paraffin. As these two types of JCV do not differ serologically, this approach is the only means to differentiate these JCV types, the clinical significance of which is currently unknown. This work has demonstrated that PCR is a powerful technique to detect JCV DNA sequences in sections from CNS paraffin blocks-allowing confirmation of diagnosis with great sensitivity, allowing distinction from SV40 DNA sequences with virus-specific primers, and allowing typing of JCV DNA as JCV-1 or JCV-2. In collaboration with a laboratory at Penn State university, JCV DNA sequences have also been demonstrated in frozen tissues from non-PML brains without evidence of demyelination or other pathology. This finding suggests that some cases of PM L may represent the reactivation of JCV which was already latent in the brain prior to the acquisition of HIV infection or another immunosuppressive condition.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "1. The role of foxa2 in the survival of dopamine neurons. [unreadable] [unreadable] Animals that have only one copy of the foxa2 gene show spontaneous loss of dopamine neurons with age. This mouse will likely be very widely used because although many mutations have been identified that contribute to Parkinsons disease and related disorders, it has been difficult to reproduce the specific loss of dopamine neurons in an animal model. This animal has the potential to teach us a number of important lessons: why some dopamine neurons are more at risk than others, why the disease is progressive and why dopamine neurons are sensitive to mutations in genes that are widely expressed? [unreadable] [unreadable] Much of what is known about the relationship between the various symptoms in Parkinsons patients and the underlying defects in the nigrostriatal system comes from lesion studies in the rat. The foxa2 heterozygous mouse allows us to study for the first time, in an animal model, motor and cognitive behaviors in the context of a progressive neurodegenerative process. We are currently focused on the cellular role of foxa2 in dopamine neurons. In brain slices, we have shown that the cellular location of the foxa2 gene is controlled by inputs that signal health or stress. This result provides a way measuring the signaling logic that controls the survival of dopamine neurons. [unreadable] [unreadable] 2. Supporting dopamine neuron survival in vivo.[unreadable] [unreadable] We have shown that a single intra-ventricular injection of Notch ligands alone or in combination with other angiogenic factors promotes widespread activation of the stem cell niche in the adult brain and rescues dopaminergic neurons in a model of PD. These data suggest vascular cytokines promote regenerative responses to brain injury. A major goal of our group is to set up in vitro assays that predict the activation of the stem cell compartment in vivo. In this project, we will use our growing understanding of survival signaling in the stem cell niche to rescue injured dopamine neurons in vivo. This approach may allow a more rapid transition to clinical application than cell replacement therapy. In the past year, we have shown that the major regenerative features found in the adult rat brain are also present in adult monkeys. This result encourages our continued belief in the clinical potential of this approach.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall commercial aim of this project is to discover and develop novel therapeutic drug leads against diverse therapeutic targets from symbiotic marine dinoflagellate algae using multidisciplinary approaches. Using newly developed cell-based screening technologies, novel biochemical structures of antiviral and anticancer drug leads will be identified in extracts of cultured symbiotic strains of dinoflagellates isolated from diverse marine invertebrates. Culture protocols will employ various media and stress conditions to induce metabolite production. Based on knowledge of the known bioactive metabolites from free-living dinoflagellates, many or most of the bioactive leads from symbiotic strains are expected to be polyketides. Degenerate PKS primers will be used to amplify PKS gene sequences. Specific marine dinoflagellate polyketide synthase (PKS) probes will be designed and utilized to identify PKs sequences in both cultured (Phase I) and uncultured (Phase II) symbiotic dinoflagellates separated from invertebrate hosts. The PKS sequences will be cloned, sequenced, and ultimately expressed (Phase II) in a new recombinant system. This innovative research should not only yield novel drug leads, but it should also validate our methods for developing PKS libraries from the large number of uncultured (including uncultuurble) marine dinoflagellates and other microorganisms in the oceans. PROPOSED COMMERCIAL APPLICATIONS: Discovery and heterologous expression of polyketide synthase genes from symbiotic marine dinoflagellates for commercial-scale fermentation of new polyketide-based therapeutic agents for the treatment and cure of cancer and viral diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this Phase 1 SBIR is to design and test a prototype of the Channel Laser Stimulator capable of stimulating single cells or part of the cell. This tool is needed to study dose dependency of heat/capsaicin-sensitive cells/neurons and \"calcium\" channels. Understanding how these \"calcium\" channels work and principles of their kinetics is important in neuroscience in general and especially in therapeutics and drug development. Further, the development of this instrument will enable the manipulation of cell activity in culture or in tissues with light, a method that could lead to important new applications for regulating cellular or physiological processes in vivo. This device may also lead to novel methods of drug screening and development as it could provide a rapid, well calibrated, and reproducible method of delivering a thermal pulse to cells or purified ion channel protein in single or multi-well format. We will design and test a Channel Laser Stimulator that (A) will deliver a consistent light intensity directly to the target-membrane and channels of the cell; (B) will enable the operator to precisely control the heat stimulus and thus activate some (predictable) amount of channel activity; and (C) will enable the operator to calibrate heat stimulus by power and pulse duration of the laser pulses. We will design this prototype of the Channel Laser Stimulator with parameters optimized for single cell level research, based on (1) the design of an infrared laser diode pain stimulator, developed by LASMED, and (2) the results of preliminary testing. We will consult with a leading scientist in the field of molecular investigations of thermo sensitive ion channels in the mammalian nervous system. All necessary tests to verify performance of the Channel Laser Stimulator will be carried out in the laboratory of this scientist by his personnel.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We are carrying out studies of the histone modifications over the insulin gene locus and its neighborhood in human islet cells, in an attempt to identify long range regulatory influences that may affect insulin gene expression. We have identified an extended domain of open chromatin structure. In addition, we are carrying out measurements of long range physical contacts within the nucleus between the insulin promoter and other genomic sites. We are particularly interested in the possible influence of downstream imprinted loci on insulin expression. We have also collaborated with the laboratory of Dr. Marvin C. Gershengorn in studies of the histone modifications over the insulin locus in human islet-derived precursor cells (hIPCs). We have shown that these cells, although they have lost expression of insulin, retain to a large extend the same modifications found in the active beta cells of islets, strongly supporting the idea that they are poised for expression of insulin. In contrast, other cell types show no such modifications.[unreadable] [unreadable] We are now exploring the long-range interactions between the human insulin locus and other sites in the genome, in order to determine the role of such interactions in insulin gene expression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Family planning programs have become critical elements of most developing country population control policies. Despite the generally widespread acceptance of the key role of family planning programs in reducing fertility, there is little consensus regarding the programs importance relative to the roles of general economic development and improving women;s status. To address this dialogue, we propose to investigate the following questions: To what extent can be recent fertility decline in Indonesia be explained by the following factors: improved development, and improvement in women's education and economic status? Which elements of the Indonesian family planning program have the greatest impact on fertility: the distribution information, education and communication activities? Do the effects of family planning program activities vary for different groups? For example, does fertility decline more quickly when public family planning program infrastructure is expanded in communities that are in earlier stages of the fertility transition that when infrastructure is expanded in areas at later stages of the transition? How have the expansion of the public family planning program and private family planning services, economic development, and improvements in women's status influenced the major proximate causes of fertility: contraceptive use, marriage and postpartum amenorrhea? Are there time lags in the impact of family planning program components on fertility and its proximate causes? To what extent do expansion of public program components, better access to private family planning services, contraceptive methods to more effective methods? In other words, to what extent do family planning programs affect contraceptive mix versus prevalence? Our methods unify he demographic proximate determinants and economic reduced-form models of fertility. Applications of the proximate determinants model for program evaluation have often yielded different conclusions about family planning programs than have applications of reduced-form \"demand for children\" models. We demonstrate the compatibility of the two approaches and develop methods to directly compare their results. We will also address the statistical problem raised by the fact that policy maker allocate program inputs for optimal impact, rather than randomly. To control for this endogeneity of program placement, we will use a community fixed effects estimator. The results are interpreted as the effects of changes in the program inputs on changes in fertility within communities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "a. Search for XMRV in prostate tumors Several recent papers reported the detection of a new MLV, xenotropic murine leukemia virus-related virus (XMRV), in human prostate tumors (e.g., Urisman et al., PLoS Pathog. 2:e25, 2006; Schlaberg et al., PNAS 106:16351-16356, 2009). In collaboration with Drs. Karen Sfanos and Angelo De Marzo (Johns Hopkins School of Medicine), we surveyed nearly 800 prostate tumors for XMRV, using a combination of duplex real-time PCR and immunohistochemistry (IHC). Our studies used 22Rv1 cells, which are known to be infected with XMRV (Knouf et al., J. Virol. 83: 7353-7356, 2009), as positive controls, and uninfected human cells, such as 293T or HeLa cells, as negatives. Every PCR well was simultaneously analyzed for viral sequences and for CCR5, a single-copy human gene, as a test of the quality and quantity of sample DNA. Dilution of 22Rv1 DNA into HeLa DNA showed that we could detect the XMRV sequences in 1.5 22Rv1 cells, even in the presence of a 10,000-fold excess of HeLa cells. Our IHC assays used a rabbit antiserum raised against HPLC-purified Moloney MLV p30CA and another against HPLC-purified Moloney MLV gp70SU. Controls included not only 22Rv1, but also 293T cells transfected with a molecular clone of XMRV or mock-transfected. In all cases, the 22Rv1 cells and the XMRV-transfected cells stained with both antisera whereas the negative cells did not. Neither of these well-controlled assays gave any indication of XMRV infection in any of the prostate tumor samples. We concluded (Aloia et al., Cancer Res. 70:10028-10033, 2010) that the positive reports from other laboratories likely represented false positives. For example, positive PCR results can result from contamination with minuscule amounts of mouse DNA, and positive IHC results can arise if the antisera react with host-cell, as well as viral, antigens. Subsequent developments have revealed that XMRV was originally formed during xenotransplantation of the 22Rv1 tumor line, and that humans are almost certainly not infected by this virus (Paprotka et al., Science 333:97-101, 2011; Sfanos et al., Nat. Rev. Urol. 9:111-118, 2012). b. Development of a new MLV infectivity assay and further applications of the iGLuc infectivity assay strategy As part of our continuing studies on XMRV, we adapted a retrotransposition assay (Curcio and Garfinkel, PNAS 88:936-940, 1991; Heidmann et al., PNAS 85:2219-2223, 1988) for detection of replication-competent gammaretroviruses. In this assay (called iGLuc), an MLV-based vector is placed into 293 cells. The vector carries a Gaussia luciferase gene in reverse orientation, interrupted by an intron in forward orientation. Only when this vector has been rescued by a competent helper virus and has progressed through the entire retroviral life cycle is the Gaussia luciferase expressed. This work was performed in collaboration with Drs. Gisela Heidecker, Vineet KewalRamani, and the late David Derse (HIV Drug Resistance Program, National Cancer Institute; see Mazurov et al., PLoS Pathog. 6:e1000788, 2010). Although other reporters can be used in this assay, the Gaussia luciferase offers unique advantages: it is extremely bright, rendering the assay very sensitive, and it is secreted, so that the assay can be performed directly on culture fluids. It is easily performed in a 96-well format. Together with Drs. Sfanos and De Marzo, we screened 70 widely used cell lines for the presence of MLVs, using both PCR and IHC assays (Sfanos et al., PLoS One 6:e20874, 2011). Three lines were found to be producing replication-competent MLVs. The viruses were characterized in considerable detail: two were found to be derived from the endogenous MLV Bxv-1, whereas the third was related to another xenotropic MLV. Their replication-competence was confirmed by the iGLuc assay. In some cases, the MLVs infected the human cells during the xenotransplantation passages used to establish the cell lines; in others, the infections probably represent cross-contamination that occurred in other laboratories. We believe that the iGLuc strategy for detection of retroviral replication can have wide application. We have collaborated with Dr. Kenneth Cornetta (Indiana University) to demonstrate the utility of the iGLuc assay for testing gene-therapy vector preparations for replication-competent gammaretroviruses (Aloia et al., Gene Therapy, in press, 2012), and we intend to do the same for HIV-1-based gene-therapy vectors. In collaboration with Dr. Heidecker, we are developing the reagents for assaying mouse mammary tumor virus; a rapid, sensitive infectivity assay should greatly facilitate future in vitro studies on the biology of this virus. c. Characterization of an HIV-1 entry inhibitor Several years ago, we and colleagues at the NCI-Frederick described a small-molecule entry inhibitor of HIV-1 entry, named Stibavirin or NSC13778 (Yang et al., J. Virol. 79:6122-6133, 2005). This compound is already FDA-approved for other uses. To gain further insight into its mechanism of action, Dr. John Mellors (University of Pittsburgh) has now selected HIV-1 mutants partially resistant to the drug. We have shown that the changes he observed in and near the V3 loop of gp120 are responsible for this resistance. Several kinds of data show that Stibavirin blocks entry by binding to CD4, preventing gp120 from binding. It seems possible that this type of inhibition could be a useful addition to antiviral therapy. The mechanism of the resistance is being analyzed by molecular modeling in collaboration with Drs. Rick Gussio and Tam Nguyen (Developmental Therapeutics Program, National Cancer Institute). Patent Linked to Project: U.S. Patent #7,572,828: Identification of Anti-HIV Compounds Inhibiting Virus Assembly and Binding of Nucleocapsid Protein to Nucleic Acid; issued August 11, 2009; Robert Shoemaker, Michael Currens, Alan Rein, Ya Xiong Feng, Robert Fisher, Andrew Stephen, Shizuko Sei, Bruce Crise, Louis Henderson, and Karen Worthy. This patent describes a class of compounds with anti-HIV-1 activity, which are under investigation for use in antiretroviral therapy. [Corresponds to Rein Project 4 in the October 2011 site visit report of the HIV Drug Resistance Program]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The investigators and staff of the Immunodeficiency-Cancer Registry seek funds to host a conference on the role of immunodeficiency and cancer. The proposed evening conference will be held in conjunction with the annual spring meetings of the Society for Pediatric Research and the Society of Clinical Investigation in Washington D.C., May 1986. The purpose of the conference will be (1) to provide up-to-date information on several major topics pertaining to the association between immunodeficiency and cancer and (2) to explore the relevance of studies in patients with naturally-occurring immunodeficiencies who develop tumors, to cancer in patients with iatrogeneic or acquired immunosuppression as well as \"seemingly\" nonimmunodeficient persons. Case material from the Immunodeficiency-Cancer Registry will provide an introduction to discussion of the pathophysiological, cytogenetic, and virologic correlates of tumors in patients with naturally-occurring as well as therapy-induced immunodeficiencies. Experts in several areas (e.g., epidemiology, genetics, virology, and/or molecular biology), who have also had experience with immunodeficient patients or their tumors, will be invited to make formal presentations on the state-of-the art and future directions. Several clinicians who have had experience with cancer in immunodeficient hosts will be invited as discussants (rapporteurs). It is hoped that attendees, primarily clinical researchers, will gain new enthusiasm for studying prospective patient material, that new collaborations between basic scientists, pathologists, and clinicians will follow, and that the Immunodeficiency-Cancer Registry, as well as the nonprofit lay organization, Immunodeficiency Foundation, will facilitate these developments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A new proteomics technology utilizing multidimensional (2D) ion mobility spectrometry separation steps combined with mass spectrometry will be developed to characterize the human plasma proteome. The new instrumentation provides sufficient peak capacity such that a high-throughput analysis of the plasma proteome can be performed without the need for a LC separation step. Thus the measurement can be performed on a timescale that is 30 times faster. The instrumentation will be used to generate Addressable Digital Array Maps (ADAMs) for human plasma analyses. These searchable databases will be utilized in comparative proteomics studies (Phase II) to create a cardiovascular disease ADAM (map). The rapid, high-throughput measurement provided by the 2D IMS instrumentation is groundbreaking in that it is now possible to perform plasma profiling studies on a timescale necessary for population proteomics studies. Such studies are required to identify and score multiple biomarkers to populate the cardiovascular disease ADAM. In the future the map will be used to provide an accurate assessment of health for customers in our personalized medicine business model. The research described in this proposal involves the development of instrumentation required to perform population proteomics studies in a timely fashion. These studies will allow the development of disease biomarker databases that can be used in assessments of disease predisposition for our personalized medicine business model. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The present proposal is to record intracellularly from cells in the posteroventral and dorsal cochlear nuclei, parts of the first integrative stage in the mammalian auditory pathway, in brain slice preparations. Brain slice preparations allow stable intracellular recordings to be made while controlled manipulations can be made of: (1) intracellularly injected current, (2) the precise temporal patterning of auditory nerve input, (3) the ionic and pharmacological environment and (4) the physical presence of tissue which might provide synaptic inputs. The voltage changes of cells in response to injected current reveal the intrinsic electrical characteristics of cells. Synaptic responses to electrical stimulation of the auditory nerve show what the properties are of synapses between auditory nerve fibers and postsynaptic cells. If there exists neuronal circuitry in the tissue of the brain slice, the synaptic responses to electrical stimulation of the auditory nerve will consist of short latency responses mediated directly through primary afferent innervation followed by longer latency responses mediated through interneurons. By cutting slices which include different parts of the cochlear nuclear complex, the position of interneurons can be used to separate synaptic inputs. Pharmacological blockers can be used to separate inputs and to obtain clues about the identity of neurotransmitters. Once separated, the characteristics of synaptic inputs can be studied unambiguously. Trains of electrical shocks will be used to give a rough indication of how the various neuronal pathways function in vivo. The physiological characteristics of cells will probably differ. After learning to differentiate cells physiologically, intracellular dye injections will be used to correlate their physiological with their morphological characteristics. To ensure that the tissue is not degenerating, both physiological and anatomical characteristics will be used to assess the tissue. In viable slices, the input resistance of cells should be high and the electrical characteristics and synaptic responses should be stable over hours. Cells injected intracellularly with horseradish peroxidase should look like Golgi-stained tissue fixed in situ.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sjogren's syndrome in man is characterized by dry mouth and eyes often leading to stomatitis, increased incidence of caries and periodontal disease, and severe ocular problems. The salivary and lacrimal glands are infiltrated by T and B lymphocytes and antibodies to organ specific antigens can be found in the serum of patients with the disease. Experimental autoallergic sialadenitis (EAS) in the Lewis (LEW) rat shares many histopathologic features with Sjogren's syndrome and will serve as our model for the study of pathogenetic factors in the disease. It is hypothesized that EAS is due to the exposure of cell associated molecules in salivary tissues to an altered or poorly regulated immune system. This exposure, in turn, initiates a T cell mediated, autoimmune response that leads to the destruction of the salivary parenchyma. We have confirmed the reproducibility of the LFW rat model of EAS and now propose to use it to answer the following questions: 1) What is the nature of the target antigen(s) in EAS? 2) Where is the target antigen(s) in EAS located? 3) What are the abnormalities in the cellular and/or the humoral immune system that are important in the pathogenesis of EAS? 4) Can procedures that modify and/or alter the immune system affect the induction or induce reversal of EAS? We will improve the induction and immunization protocol and determine if there are clinical symptoms that correlate with the lymphocytic infiltrates seen in the glands. We will identify, isolate and localize the target antigen(s) immunocytochemically and by using established cell fractionation and protein purification techniques. Immunological analysis will include: identification and characterization of the autoantibodies and mononuclear cells present in the glands during EAS; attempts to induce the disease by adoptive transfer of immune cells and/or serum, and; attempts to inhibit the disease by modulation of the immune system and/or the target antigen(s) with monoclonal antibodies or immunomodulating agents. These results should provide a comprehensive analysis of EAS in LEW rats and may provide important insights into the pathogenesis of Sjogren's syndrome in man.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "I will determine whether different RNA polymerases characterize the different morphological and functional stages of development in insects to account for the differential transcription that appears to occur. The multiple RNA polymerases will be characterized, isolated and their physicochemical properties studied so as to not only determine the nature of changes that occur during development and differentiation, but also to correlate the in vivo patterns of RNA synthesis to the variations in their levels, relative proportions and distribution. Whether both the hormones ecdysone and juvenile hormone bring about their profound effects via RNA polymerases and if so, how, will be studied in order to understand the mechanism of action of these hormones at the molecular level. The biochemical manifestations at the molecular level during the transition of cells and tissues from growth to cessation of growth and virtual non-growth to growth as occurs during induction and break of diapause in insects will be studied with particular emphasis on RNA polymerases and RNA synthesis. The existence of modifying factors which alter the transcriptive specificity, cause inhibition or activation in activity of core RNA polymerases of even cause interconversion between different molecular forms of a specific polymerase during growth and differentiation will be looked for in order to explain selective transcription by a limited number of RNA polymerases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary objective of this project is to secure tissue from CaP patients (during the course of their diagnosis and treatment) for the purpose of biological studies designed to characterize cancers with respect to their biological potentials. Frozen samples as well as paraffin-embedded samples of prostate tissue will be provided for study at the University of California-Davis laboratories for DNA quantitation. It is expected that the results obtained will be helpful in the prognostication aspects of the CaP population. Ethnic differences may be defined by the studies also. Approximately 100 cancer patients are expected to enter the study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term goal of the proposed project is to develop a transgenic rodent wherein activation of apoptosis can be imaged non-invasively. Strict coordination of proliferation and apoptosis is essential for normal physiology. An imbalance in these two opposing processes results in various diseases including AIDS, neurodegenerative disorders (Alzheimer's disease), myelodysplastic syndromes (Aplastic anemia, thalassemia), ischemia/reperfusion injury, cancer and autoimmune disease among others. Objective imaging of apoptosis will be a major advancement not only in the screening and validation of novel therapeutic molecules for the above diseases but also in the evaluation of therapeutic success or failure of current and future therapeutic treatment paradigms. In phase I of this proposal we will construct and test in a tissue culture a recombinant DNA molecule that codes for an inactive luciferase protein which during apoptosis becomes a functional luciferase enzyme thus enabling imaging of apoptosis. The ability to image apoptosis non-invasively and dynamically over time will be an invaluable resource to pharmaceutical companies for in vitro high throughput screening of compounds with pro- and anti-apoptotic activity, but also for target validation in vivo. PROPOSED COMMERCIAL APPLICATION: Apoptosis has been implicated in a wide variety of human diseases. The development of drugs that specifically target enzymes involved in the apoptotic cascade is a vibrant and dynamic research area. The ability to non-invasively image the effectiveness of these drugs in vivo would be a breakthrough that would have significant market applications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the adrenal medulla and sympathetic neuron intracellular vesicles called chromaffin granules are involved with the biosynthesis, storage, and secretion of catecholamines. The membranes surrounding these chromaffin granules uniquely contain a single b-type cytochrome of which neither the electron donor nor acceptor is known. The membranes also contain an NADH oxidoreductase activity which is inhibited by sulfhydryl-reactive reagents but which has not been further characterized. The cytochrome and NADH oxidoreductase will be isolated and purified from the chromaffin granule membrane using detergent extraction, ion exchange, hydrophobic, gel filtration, and affinity chromatography. These enzymes will be characterized in terms of molecular weight, isoelectric point, amino acid composition and amino terminus using standard standard techniques. The presence of discrete catalytic and membrane binding domains will be investigated using limited proteolytic digestion. The catalytic site orientation of the enzymes relative to the interior or exterior of the granule membranes will be determined using permeant and impearmeant labeled amino, carboxyl, and sulfhydryl reagents. The binding of these enzymes to artificial lipid membranes will be characterized in terms of protection from proteolytic cleavage. Electron transfer between the purified NADH oxidoreductase and cytochrome will be investigated with the enzymes reconstituted in phospholipid vesicular membranes containing lecithin with varying proportions of lysolecithin, cholesterol, and ethanolamine plasmalogen. A search for the function of the electron transfer system in the chromaffin granule membrane will begin with attempts to demonstrate electron transfer from the purified cytochrome to purified dopamine-beta-hydroxylase which is normally bound to the interior surface of the granule membrane. As a preliminary to this latter work the binding of dopamine-beta-hydroxylase to artificial phospholipid membranes will also be attempted.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronic fatigue syndrome has received considerable attention in lay and medical media. This syndrome reportedly leads to significant physical and psychological debility in a large segment of the population. Specific diagnostic and treatment regimens have been proposed and are being administered to patients. Yet, there remain few data which can validate the existence of the syndrome as distinct from a wide variety of other, largely psychiatric, disorders, and little understanding of its pathogenesis. The pilot feasibility study proposed in this application seeks to clarify and extend an understanding of the role of hypothalamic-pituitary-adrenal (HPA) axis dysregulation in the pathogenesis of this disorder. The overall objective of this protocol is to test the hyposthesis that patients with chronic fatigue syndrome manifest a characteristic neuroendocrine defect, namely, a reduction in adrenal glucocorticoid secretion mediated by a failure in the central release of corticotropin-releasing hormone (CRH). Specifically, we will confirm and extend our preliminary findings of impaired activation of the HPA axis in patients with chronic fatigue syndrome by examining the patterns of pulsatile secretion of ACTH during periods of maximal and minimal activity of the axis. The results of this proposal will be used to demonstrate the feasability of the pulsatility methodology and provide further pilot data which will enhance the likelihood for success of our federal grant application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This renewal application for the Pediatric AIDS Clinical Trials Unit at the University of Miami School of Medicine proposes the conduct of clinical trials in pregnant women, children and adolescents with HIV infection. We propose to enroll 100 patients into various protocols placing special emphasis on the areas of perinatal transmission of HIV infection, antiretroviral therapy in pediatric populations, and the prophylaxis and treatment of opportunistic infection in children. This program, consisting of a pediatric unit, an obstetrical unit and a virology core laboratory has a distinguished record over the past five years. The program faculty have provided scientific input into the conduct of clinical trials both nationally and locally and have pioneered the treatment of children and pregnant women with HIV infection through both Phase I and Phase II/III trials. Over 90% of the women and children recruited into treatment protocols have been members of minority groups. The extensive infrastructure of social and medical services available at the medical center has facilitated the conduct of trials. In this proposal,the addition of a new subunit at the Children's Diagnostic Center in Fort Lauderdale is proposed. This subunit is located 25 miles to the north of Miami and would allow access to clinical trials for a large number of infected children. The virology laboratory component has participated in research protocols in pediatric patients since 1983 and is fully certified by the ACTG. In addition, there is an ACTG certified immunology laboratory on campus which is proposed as an immunology core. Three developmental grants in each of the stipulated areas and applications for core virology and immunology laboratories accompany the core application. Long term goals of this project include the interruption of perinatal transmission of HIV, the preservation of the immune system, delayed progression of disease and prolonged survival. The strengths of this program lie in its already existing collaborations among the program faculty, their scientific expertise, research experience and publication record, the prior experience in clinical trials, the extensive supportive infrastructure for patient care, the extensive experience in caring for women and children with HIV infection, the integration with existing community programs, and the large number of patients available for recruitment into trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Rheumatoid arthritis (RA) is an inflammatory autoimmune disease that causes chronic pain and joint destruction in approximately 1.3 million people in the United States. While drugs to treat RA exist, better treatments strategies are needed to prevent and treat this debilitating disease. As with a variety of autoimmune disorders, there is evidence of an elevated level of tryptophan catabolism in RA patients. This is indicative of activation of the enzyme indoleamine-2,3-dioxygenase (IDO), which initiates the breakdown of tryptophan. IDO is an immune regulatory enzyme that has been implicated in the inflammatory process associated with tumor formation. Because IDO is thought to be immunosuppressive, it has been generally assumed that inhibiting IDO would exacerbate classic autoimmune disorders such as RA. However, data from patients has suggested the opposite, that IDO activity may actually be associated with the development of disease symptoms. Consistent with this, inhibitors of IDO reduce autoantibody titers and alleviate arthritis symptoms in a B cell-mediated mouse model of RA. These data support a role for IDO in exacerbating the disease and further implicate IDO in driving autoreactive B cell responses. This proposal will explore the hypothesis that IDO does not simply promote the general suppression of immune responses, as currently thought, but rather plays a more complex role in modulating inflammation by directing the immune profile of B cell responses. Drug-like inhibitors of IDO and mice that are genetically deficient in IDO, together with the K/BxN mouse model of RA, will be used as tools to determine the mechanism by which IDO inhibition alleviates arthritis development. In this proposal, we will 1) define how the inflammatory features of arthritis induced in mice are inhibited by targeting IDO activity;2) determine whether IDO-driven B cell activation is unique to the K/BxN model or is a universal phenomenon;and 3) determine whether inhibition of IDO can be used as a co-therapeutic with current RA therapies, to increase their efficacy and reduce their side-effects in the alleviation of arthritis. The long-term goal of this project is to define the mechanism by which IDO modulates the immune response leading to autoimmunity and provide new insights into strategies that can be used to manipulate this pathway to reduce or prevent debilitating disease. If successful, the results from this proposal will suggest that targeting IDO has the exciting potential to lead to a new approach for the prevention and treatment of RA and other autoantibody mediated diseases. PUBLIC HEALTH RELEVANCE: Understanding the factors that contribute to the initiation and propagation of autoimmunity is important in the design of therapeutic strategies for the prevention and treatment of autoimmune diseases such as rheumatoid arthritis. The studies in this proposal will provide insights into the mechanism by which the immune system becomes activated to induce inflammation in the joint and identify key targets to prevent the resulting debilitating disease. In the future, this strategy could lead to a new approach for the prevention and treatment of rheumatoid arthritis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "An obstacle to the use of adenovirus as a vaccine carrier is the induction of neutralizing antibodies to hexon, the major protein component of the icosahedral capsid. The Program Project is designed to exploit the fact that human subjects have no preexisting neutralizing antibodies to chimpanzee adenovirus 68 (AdC68) that would limit its efficacy. Nevertheless, subsequent use of AdC68 for booster immunization would be limited by antibodies generated in response to the primary dose. The solution lies in using structural knowledge to design and create a series of novel AdC68 derivatives with modified neutralization epitopes. Three-dimensional crystal structures are known for human adenovirus types 2 and 5 hexons, and the overall virion architecture is known from cryo-electron microscopy. Comparison of all known hexon structures and sequences shows that there is a common molecular fold that provides an accurate and representative model for designing new adenovirus vectors. The hypervariable regions of AdC68 hexon are likely to contain the epitopes recognized by neutralizing antibodies. In this project, molecular modeling will suggest sites where modifications can be made without structural disruption. The AdC68 hexon crystal structure will be determined to confirm and refine the structural predictions. Simultaneously, the AdC68 hexon epitopes responsible for antibody neutralization will be identified experimentally by generating a panel of anti-hexon neutralizing monoclonal antibodies (mAbs). The panel will be used to find neutralization escape mutants by screening AdC68 virions with randomly-introduced mutations in the epitope regions. In addition, random pentapeptide insertions will be made in the hexon hypervariable regions to identify sites that tolerate extensive structural modification. The crystal structures for escape mutant hexons will be solved to refine structural information for each epitope. The combined information will be used to synthesize variants with disrupted epitopes. Finally, \"artificial serotypes\" with multiply-disrupted epitopes will be tested against the antibody panel and )olyclonal neutralizing sera to demonstrate their efficacy as booster vaccine carriers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The flow-through centrifuge eliminates complication arising from rotating seals. Preliminary studies on plasmapheresis demonstrated negligible platelet injury and no evidence of hemolysis during 12 hours of operation. The system may provide a broad application to cell washing and elutriation, zonal centrifugation and countercurrent chromatography.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objective of this project is the evaluation of the potential of liposomes as carriers of anti-tumor agents. Several model systems of antibody directed liposomes are analyzed to obtain information on the generation of functionally active targeted liposomes. The properties of the cell-liposome interactions are investigated in the presence of serum proteins. The in vitro and in vivo efficiency of drugs entrapped in targeted liposomes is evaluated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Two major research projects will be undertaken: (1) a clinical study to elucidate the role of alterations of the various calcium and bone regulating hormones in the pathogenesis of diabetic osteopenia, and 2) a project to develop a radioimmunoassay for the isoenzyme of alkaline phosphatase derived from bone. The RIA will be used to assay serum samples from patients with the various forms of hypophosphatasia - a crippling inborn error of metabolism - to gain insight into the clinical and genetic variability of this disorder. Juvenile and adult diabetics will be studied longitudinally on the Clinical Research Center, Barnes Hospital so that changes in circulating parathyroid hormone, calcitonin, vitamin D metabolites, estrogens, etc. can be contrasted with studies of bone density and gastrointestinal calcium absorption. Differences in the degree of osteopenia, known to differ in various diabetic populations, should be reflected in dietary calcium absorption and circulating serum 1,25(OH)2D levels. Identification of hormonal differences between diabetics with and without osteopenia should provide clues to the pathogenesis of this frequent complication and suggest possibilities for corrective therapy. Normal fetal and neonatal bone will be extracted and purified for development of an RIA for normal bone AP isoenzyme. Routine methods will be used to develop the RIA once purified antigen is available. The RIA will be used to study the variability in clinical expression of this disorder, the variability in its genetic transmission, to identify \"carriers\", and in prenatal diagnosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Leishmaniasis is a major public health problem in large parts of the world, due in part to the lack of a vaccine and inadequate chemotherapy. Studies of the immune responses in humans and mice following Leishmania infection have provided an understanding of many of the cells and cytokines that contribute to the control of this disease. Nevertheless, a vaccine for human leishmaniasis does not exist. Understanding how memory T cells develop will be crucial in the development of such vaccines. Two types of memory T cells have been described: T effector memory cells, which produce effector cytokines and migrate through the tissues, and central memory T cells that do not produce effector cytokines and migrate through lymph nodes. C57BL/6 mice infected with Leishmania major are immune to rechallenge after they resolve their infections, but this immunity was thought to be dependent upon residual parasites. Now an attenuated L. major parasite that is eliminated after eight weeks has been shown to stimulate the development of long-lived protective memory T cells. These cells have the characteristics of central memory T cells, and the studies in this proposal will characterize these cells and assess whether we can generate memory T cells with an effector phenotype. The studies will include a comparison of the ability of a Listeria expressing a leishmanial antigen and L. major to stimulate effector memory T cells, assessment of the role that parasite persistence plays in blocking the development of effector memory cells, and how dendritic cells influence memory T cell generation. The experiments utilize state-of-the art tools that will allow qualitative and quantitative assessment of memory T cell development. Overall, the experiments described in this proposal will determine what type of immunologic memory can be induced in leishmaniasis, which will provide direction for the field in establishing what are reasonable goals for a leishmaniasis vaccine. PUBLIC HEALTH RELEVANCE The development of a vaccine for human leishmaniasis is a major priority, but our understanding of how to stimulate the memory T cells required for vaccine-immunity is limited. This proposal will determine how memory T cells develop, are maintained and are regulated using an experimental model of the disease. The results from these experiments will provide a foundation for the development of a successful leishmaniasis vaccine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The physiological function and the mode of regulation of transglutaminases are being studied as to their role in the formation of temporary tissue matrix (fibrin-connective tissue matrix) during tissue or bone fracture repair and in the modulation of specific cellular processes. The physiological significance and biochemical mechanism of factor XIIIa (plasma transglutaminase) and red blood cell transglutaminase catalyzed cross-linking of plasmin inhibitor (a2PI) to fibrin and other matrix proteins are under investigation both in vivo and in vitro. The cross-linking of a2PI to fibrin plays a major role in stabilization of a temporary matrix which is vital for the initial phase of cell migration and proliferation and wound sites. In cells activated by mitogens and in transformed cells, several different molecular forms of transglutaminases have been identified. These enzymes are enriched in the plasma membranes and nuclear fractions in proliferating cells and appear to modify specific proteins in the membrane fractions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aim of this RO3 proposal is to examine the ability of cyclin D1 and other cell cycle regulatory proteins to promote hepatocyte proliferation in culture and in vivo. Mature hepatocytes retain the ability to function as stem cells and can repopulate the live after hepatic injury. Previous studies have suggested that up-regulation of cyclin D1 plays an important role in governing progression of hepatocytes through G1 phase of the cell cycle. To test whether cyclin D1 is capable of promoting mitogen-independent hepatocyte proliferation, we have constructed a replication-defective adenovirus encoding this protein (ADV-D1). Our preliminary studies indicate that ADV-D1 but not a control adenovirus, promotes cell cycle progression in primary hepatocytes in the absence of growth factor, and induces hepatocyte proliferation on extracellular matrices that are normally growth-inhibitory. Pilot in vivo studies indicate that ADV-D1 effectively transfects hepatocytes in the intact animal. These results suggest that transfection of hepatocytes with ADV-D1 can promote hepatocyte proliferation independent of extracellular signals, and warrants testing as an agent to promote liver regeneration. The three Specific Aims of this study will examine the mechanisms by which cyclin D1 promotes hepatocyte growth, and will examine the ability of other G1 phase cyclins and E2F transcription factors to promote hepatocyte cell cycle progression. These studies are intended to provide unique mechanistic insight in the regulation of hepatocyte growth, and to identify candidate proteins for gene targeting strategies to therapeutically enhance liver regeneration.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Short REM latency is heritable for narcolepsy and depression. Individuals with narcolepsy almost all express HLA DR2/DQw1. If short REM latency in depression is associated with HLA expression, it would indicate that REM sleep itself may be regulated by genes in or around the MHC and/or that MHC-related genes may be involved in depression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is a rapidly increasing awareness of the importance of titin in causing neuromuscular disorders (titinopathies), two of which we focus on in this work, centronuclear myopathy (CNM) and hereditary myopathy with early respiratory failure (HMERF). Titinopathies frequently present early-onset muscle weakness and respiratory difficulty but understanding their underlying mechanisms is held back by our still limited understanding of the functional roles of titin in skeletal muscle. Titin comprises the third myofilament of muscle and spans along the sarcomere, from Z-disk to M-band. Titin?s I-band region functions as a molecular spring that generates passive stiffness with recent studies indicating that passive stiffness affects active force at sub- maximal activation levels. However, it is not known how important titin?s stiffness is to overall skeletal muscle health and if altered stiffness can cause a myopathy. That the importance might be high is suggested by our pilot studies that reveal deranged titin stiffness and reduced active tensions in titinopathy patients with CNM. Aims 1 and 2 address how altering titin-based stiffness affects both passive and active muscle properties and if it can be disease-causing. We will perform mechanical studies on biopsies from titinopathy patients (focus on CNM) as well as study two contrasting mouse models in which the stiffness of titin?s spring region is either increased (PEVK truncation) or reduced (inactivation of Rbm20). The working hypothesis is that altering titin?s spring region alters both passive muscle stiffness and active tension and that this causes myopathy. Aim 2 also studies a novel mouse model that mimics CNM with splice site mutations in titin?s spring region and we will examine the functional consequences at the RNA, protein, structural and functional levels. We also study the A-band segment of titin, specifically the C-zone. This zone has not been studied, it is clinically important as this is where a large number of disease-causing mutations are found. We investigate in Aim 3 a mutation in the C-zone exon 343 which causes HMERF, a myopathy with respiratory muscle involvement that can be fatal. Using a novel HMERF mouse that we made the mechanistic basis of the disease will be studied. Through excision of the mutated titin exon 343, the therapeutic potential of exon skipping for treating HMERF will be tested. With its basic science and translational goals and its in-depth and integrative approach, this application seeks to continue our track record of cutting-edge titin research. Powerful techniques and novel mouse models are in place, pilot data support the guiding hypotheses, and our research team is highly experienced. The proposal will greatly enhance insights in titin biology, titin?s role in muscle disease, and titin?s potential as a therapeutic target.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "\"Prepostaglandin endoperoxide\" is the proposed pivotal intermediate in prostaglandin biosynthesis. The ultimate goal of this research is preparation in the laboratory of this intermediate, and study of its bioconversion to prostaglandins. It is expected that this research will provide a better understanding of the biosynthesis of prostaglandins. The initial objective is the synthesis of 2,3-dioxabicyclo (2.2.1) heptane and simple derivatives. A study of the chemistry of these endoperoxides will follow. Then biogenetic type syntheses of this novel structure type will be sought. The recent isolation of minute quantities of a preprostaglandin endoperoxide lead to the discovery of physiological activity which significantly exceeds that of the related prostaglandin derivatives. Thus, it is quite possible that the endoperoxides may constitute a class of ultra- potent medically useful substances, e.g. for the treatment of heart disease. The total synthesis proposed herein would provide material in sufficient quantities for research on the medicinal utility of endoperoxide derivatives and ultimately for medicinal application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Heart failure (HF) is characterized by an elevation in sympathetic tone. The mechanisms responsible for the sympatho-excitation of HF are not completely understood. Recent studies from this laboratory have shown that the cardiac \"sympathetic afferent\" reflex is enhanced in dogs with pacing-induced HF. The mechanisms by which this enhancement occurs are unclear. There is an enhancement in afferent fiber sensitivity to bradykinin and capsaicin. Preliminary evidence from this laboratory suggests that an enhanced central gain of this reflex is, in addition, responsible for the augmentation of this reflex. Furthermore, we have shown that central angiotensin II (Ang II) is at least one mediator for this enhancement. A second mechanism which may explain the increased gain of he cardiac sympathetic afferent reflex in HF is a decrease in nitric oxide (NO) production in several central sites which regulate sympathetic outflow. We hypothesize that both an increase in central Ang II and a decrease in central NO contributes to the increase in the sensitivity of the cardiac sympathetic afferent reflex and to the tonic sympatho-excitatory state in dogs with HF. Therefore, the specific aims of this project are to: 1) determine if the central gain of the cardiac sympathetic afferent reflex in dogs with HF is related to increased levels of central Ang II or to changes in Ang II type1 receptor density or both, 2) determine if acute and chronic central administration of the Ang II receptor antagonist, losartan and L-158,809 and/or NO donors prevent or reduce the enhancement of the cardiac sympathetic afferent reflex in dogs with HF, 3) determine if bradykinin prostaglandins and NO are mediators of the enhanced sensitivity of cardiac sympathetic sensory endings in dogs with HF, and 4) determine if chronic thoracic sympathetic deafferentation alters the time course and/or magnitude of the sympatho-excitatory response during the development of pacing-induced HF. These studies integrate into the overall scope of this Program Project in that the regulation of sympathetic outflow in HF is likely to be mediated by a variety of peripheral inputs with important modulation from central substances. The cardiac sympathetic understanding of neuro-humoral regulation in this disease state should include this potentially potent reflex.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Median survival of high-grade gliomas remains less than a year despite multi-modality treatment. Cure is considered anecdotal. These tumors usually have extended beyond the limits of a complete surgical resection and a dose of conventional external beam radiotherapy has been limited by surrounding brain tolerance. We believe that we can achieve a higher radiation dose to the tumor by placing radioactive seeds of Iodine 125 directly into the tumor bed, with a sharp fall-off of radiation to the surrounding normal brain. Hopefully this will achieve a much better therapeutic ratio, especially when delivered at low dose rates.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term goal of the Applicant's research program is to explore the molecular mechanisms underlying the etiology of age-related macular degeneration (AMD) and develop novel target-based therapeutic strategies. This K99/R00 grant will facilitate the transition for the Applicant to become an independent investigator at the Vanderbilt Eye Institute, which has a rich environment of scientific collaboration and nurturing career development of junior scientists. AMD is the leading cause of blindness in elderly Americans. The majority of AMD patients has atrophic (dry) form of the disease and has only limited treatment options at the present time. Atrophic AMD is likely resulted from gene/environmental interaction causing progressive degeneration of the retinal pigment epithelium (RPE). Aging is the most well defined environmental risk factor of AMD. We hypothesize that the mammalian target of rapamycin (mTOR)-mediated signaling pathway plays key roles in controlling the aging process of the RPE. The hypothesis is supported by recent literature data suggesting that mTOR plays key roles in integrating various environmental signals and linking them to altered tissue function and organism's life span. In our preliminary studies, we found that rapamycin reversed the senescent phenotype of primary human RPE cells in vitro. To further test our hypothesis, we have proposed three specific aims in this application. Aim 1 is to determine how the aging process regulates mTOR pathways in the RPE. Aim 2 is to determine how modulating the mTOR signaling affects RPE aging in vitro by a potential mechanism of regulating autophagy. Aim 3 is to determine whether mTOR regulates aging of the RPE in vivo using SOD1 knockout mice which develop AMD-like phenotype in the retina. During the mentored phase, the Applicant will conduct the proposed experiments under the supervision of a mentoring committee, which is consisted of mentors with expertise in AMD etiology and pathogenesis (Dr. Paul Sternberg), animal models of neurodegeneration in the retina (Dr. David Calkins) and mTOR/autophagy (Dr. Lu Bo). Members of the committee will meet regularly, monitor the research progress and assist the Applicant to advance her career towards independence. Research at the R00 phase will test the potential therapeutic effects of mTOR inhibitors in an animal model relevant to AMD. Results from these studies will provide novel mechanistic information on aging and age-related degeneration of the RPE and retina. PUBLIC HEALTH RELEVANCE: This project focuses on characterizing a novel signal transduction pathway which may control the aging process of the retina and contribute to age-related macular degeneration (AMD). Results from the studies can potentially be translated into new therapeutic approaches for treating the atrophic form of AMD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": ": The overarching goals of the Lincoln University- Fox Chase Cancer Center Partnership in Cancer Research and Training are to create a cadre of well trained and experienced cancer researchers at Lincoln University oriented toward reducing cancer health disparities, to foster such research at the Cancer Center, and to create a pipeline of students with laboratory experience who choose to further their education in cancer biology, population science, and medicine. This cooperative effort will accomplish the following objectives: 1) to expose Lincoln University faculty and students to research conducted at Fox Chase in the areas of cancer biology and population science; 2) to enhance Fox Chase and Lincoln's collaborative research efforts and networking opportunities through increased participation in scientific conferences; 3) to introduce basic cancer principles through workshops, seminars, lectures, community/university outreach, and academic instruction; 4) to publish joint research publications; 5) to incorporate team-based projects that will generate a steady pipeline of students to continue advanced studies in the nation's top graduate institutions; and 6) to create competitive minority scientists that are successful in receiving funding for addressing cancer health disparities. [unreadable] [unreadable] The Partnership will conduct its activities over four years in three stages. The first year will be a planning stage, where an Administrative Leadership Group, Research Training and Research Project committees will be established, and Internal and External Advisory Boards constituted. The second year will be the priority-setting stage during which time a competition for joint pilot research projects will be held, and the training program established. The third and fourth years will constitute the implementation stage, wherein two pilot research projects and a research training program will be conducted. Summative activities of the Partnership will be the development of competitive research and training grants directed by a team of Lincoln faculty who have achieved leading roles in this endeavor. [unreadable] [unreadable] The ultimate ambition for this partnership is to establish a strategic academic collaboration with Lincoln University to implement research and education that highlights populations with health disparities. The instruction of minority and majority scientists at Lincoln and Fox Chase about cancer health imbalances will increase the sensitivity toward this issue and augment the number of capable researchers available to address this present dilemma. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Enterococci are commensal organisms that inhabit the gastrointestinal tract and rarely cause infections in the healthy host. However, in immunocompromised individuals or in individuals on antibiotic therapy, these organisms can cause serious disease. Hospital-acquired infections by Enterococcus faecalis, especially those associated with intravascular catheters, biliary stents and other implants have increased dramatically in recent years. This trend has likely resulted from high level antibiotic-resistance within this genus coupled with the acquisition of virulence determinants and the ability to form biofilms. It is recognized that biofilm associated bacteria are phenotypically different from planktonic cells and tend to be more resistant to antibiotics and to host immune clearance mechanisms. Innate immunity plays a crucial role in the host response to microbial infections but little is known about the immune response to enterococci and enterococcal biofilms. Our laboratory has identified and characterized two important biofilm enhancing loci in E. faecalis;one specifying the enterococcal surface protein Esp and the other the Bee pilus. Both these factors significantly enhance biofilm formation and adherence of E. faecalis to abiotic surfaces in vitro, but much remains to be learned about their functions in vivo. Our underlying hypothesis in this proposal is that the host innate immune response to biofilm and planktonic E. faecalis is quite different and we will examine this through two specific aims. In aim 1, we will compare in vitro the interaction of professional phagocytes and enterococcal biofilms versus planktonic cells from two different genetic backgrounds, to document bacterial survival, host cell response and influence on biofilm architecture. In aim 2, we propose to examine host-pathogen interactions in vivo using two murine models of biofilm-associated infections. The novelty of these studies will be the utilization of a positron emission tomography /computed tomography (PET/CT) approach to image and follow the time course of infections in real time. These exploratory studies are designed to lead to a better understanding of the host response to biofilm-related enterococcal infections and could ultimately lead to new therapeutic strategies to prevent serious enterococcal infections. PUBLIC HEALTH RELEVANCE: Hospital-acquired infections by antibiotic-resistant Enterococcus faecalis have risen dramatically in recent years. Greater understanding of the host defense mechanisms that protect against invasive enterococcal disease could provide new ways of controlling enterococcal infections.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The objective of this study is to determine the role of caveolae in airway remodeling in asthma. This chronic inflammatory disease of the airways affects an estimated 15 million people in the United States and 100-150 million people worldwide. Among this population at risk, a subset of subjects will develop irreversible remodeling and airway obstruction. Airway remodeling is characterized by an abnormally large number of myofibroblasts in the subepithelial basement membrane associated with significantly increased deposition of connective tissue proteins leading to a thickening of the lamina reticularis. Interleukin 4 (IL-4), a Th2 cytokine important in the control of allergic response, has also been proposed as an effector influencing airway remodeling. We have recently demonstrated that IL-4 regulates the expression of caveolin-1 (cav-1). Cav-1, main structural and functional protein of caveolae, notably regulates transforming growth factor (TGF)-beta activity by dampening its transduction signal process initiated by the TGF-beta receptor complex. TGF-beta1 is a pivotal pro-fibrotic cytokine in asthma and fibrosis. Our goal, therefore, is to demonstrate that Th2 cytokines, and more specifically IL-4, mediate caveolae deficiency by regulating cav-1 gene transcription in fibroblasts resulting in enhanced TGF-beta responses in airway remodeling. We will: 1) Evaluate the hypothesis that the reduced expression of cav1 is associated with the expression of inflammatory markers, more specifically IL-4 and TGF-beta, and the development of airway remodeling in allergen challenged mice. 2) Investigate the hypothesis that among the Th2 cytokines expressed in asthma, IL-4 regulates caveolin-1 expression and caveolae formation;3) To test the hypothesis that the reduced expression of caveolin-1 protein enhances TGF-beta effects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our goal is to identify the biological parameters that control myelin metabolism. We are using an in vitro culture system of suspended, spherical cellular aggregates prepared from fetal mammalian central nervous systems. With these cultures we are following the development of myelin synthesis by measuring several myelin membrane specific molecules such as sulfatides, myelin basic protein and myelin associated enzymes. Preliminary studies indicate myelin synthesis does occur in these cultures and follow the same temporal pattern normally occurring in vito. Cultures from animals exhibiting genetically determined demyelinating disease (e.g., quaking and jumpy mice) will be analyzed for the same parameters and compared to normal cultures. Experiments will be performed to test the effects of hormones on myelination in these cultures. Measles virus, because of its purported relationship to multiple sclerosis, will be carefully investigated for its affects on this myelinating system. In addition, we will determine if any protein kinase activity is associated with the measles virus as well as characterize the protein substrates of the phosphotransferase enzyme(s). These brain aggregate culture studies should increase our understanding of the biological signals and mechanisms controlling myelination and may lead to the development of novel preventative, diagnostic and therapeutic methods treating demyelinating disease, such as multiple sclerosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Current methodologies for diagnosing infection with Neisseria gonorrhoea require specimens that are obtained by invasive means: urethral swabs from men and cervical swabs from women. These specimens are usually cultured onto Thayer-Martin media; alternatively a variety of other methods can be employed to diagnose gonorrhea, but none is as sensitive as culture. The objective of this proposal is to create an inexpensive, rapid, simple and non-invasive immunologic test for gonorrhea. It must be as sensitive as culture and applicable for use in developing countries. The project will be accomplished through 3 specific aims: (l) to optimize an already existing prototype assay called the HygEIA GC Test to use with non- invasive specimens, and to convert this ELISA format to an immunochromatographic (ICT) format with a membrane read-out. Initially, the assay will be developed using two existing antibodies, one directed against the H8 antigen (a current component of the HygEIA GC Test) and a gonococcal anti-lipooligosaccliaride antibody that reacts broadly with N. gonorrhoea; (2) to create polyclonal antibody libraries using a molecular cloning system that produces antibodies that recognize multiple epitopes on gonococcal surfaces exclusively; simultaneously these libraries will be immortalized and can be utilized time and again to produce, antibodies with reproducible specificities and; (3) to use patients at risk for gonorrhea to test prototype formats. Through overlapping strategies, beginning with a prototype that already functions, it is anticipated that a final product will emerge.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cdk5 regulates pain signaling: Our present studies are primarily focused on delineating the roles of Cdk5 in the central and peripheral nervous system with a special emphasis on nociception and pain. We have earlier identified expression of Cdk5 and p35 in nociceptive neurons, and discovered that this expression is modulated during peripheral inflammation. We also discovered that induced inflammation results in increased calpain activity in sensory neurons, which activates cleavage of p35 to p25 and subsequently increases Cdk5 kinase activity. The p35-/- mice, which exhibit significantly decreased Cdk5 activity, showed delayed responses towards painful thermal stimulation compared to their wild-type controls. In contrast, mice overexpressing p35 with elevated levels of Cdk5 activity were more sensitive to painful thermal stimuli than controls. We analyzed TRPV1 for potential phosphorylation by Cdk5 and found that Cdk5 can directly phosphorylate TRPV1 at threonine 407, and this in turn modulates agonist-induced calcium influx. We also found that inhibiting Cdk5 activity resulted in attenuation of capsaicin-induced calcium influx in cultured DRG neurons, and this attenuation was reversible. These observations suggest that Cdk5-mediated phosphorylation of TRPV1 is important for capsaicin-mediated calcium influx through this receptor. Since Cdk5-/- mice die perinatally, we generated primary nociceptor-specific Cdk5 conditional knockout (COKO) mice to identify the precise role of Cdk5 in primary afferent pain signaling. In the basal state, the conditional knockout mice had significant hypoalgesia, confirming the direct role of normal Cdk5 activity in primary afferents. Collectively, our findings show a novel molecular mechanism for the functional regulation of TRPV1 by Cdk5 and suggest that Cdk5/p35 may be a target for development of analgesic drugs.[unreadable] [unreadable] The aim of our current study in FY08 was to identify the proinflammatory molecules that regulate Cdk5/p35 activity in response to inflammation. We constructed a vector that contains the mouse p35 promoter driving luciferase expression. We transiently transfected this vector in PC12 cells to test the effect of several cytokines on p35 transcriptional activity and Cdk5 activity. Our results indicate that tumor necrosis factor-alpha (TNF-alpha) activates p35 promoter activity in a dose- and time-dependent manner and concomitantly upregulates Cdk5 activity. Because TNF-alpha is known to activate ERK1/2, p38 MAPK, JNK, and NF-kB signaling pathways, we examined their involvement in the activation of p35 promoter activity. MEK inhibitor, which inhibits ERK activation, decreased p35 promoter activity, while the inhibitors of p38 MAPK, JNK, and NF-kB increased p35 promoter activity, indicating that these pathways regulate p35 expression differently. The mRNA and protein levels of early gene response 1 (Egr-1), a transcription factor, were increased by TNF-alpha treatment, and this increase was dependent on ERK signaling. In a mouse model of inflammation-induced pain in which carrageenan injection into the hind paw causes hypersensitivity to heat stimuli, TNF-alpha mRNA was increased at the site of injection. These findings suggest that TNF-alpha-mediated regulation of Cdk5 activity plays an important role in inflammation-induced pain signaling.[unreadable] [unreadable] Phosphoproteomic analysis of Cdk5 targets: The human genome encodes over 500 different protein kinases, the key regulatory enzymes that catalyze the phosphorylation of proteins at about 100,000 different sites to reversibly control their functional activities. Defects in specific protein kinases have been linked to over 400 diseases, and about 25% of all pharmaceutical industry research and development is now focused on the discovery and evaluation of protein kinase inhibitors for therapeutic applications. Cdk5 has become a target of high interest to the drug industry because of its key role in neuronal homeostasis. So far more than 40 different Cdk5 substrates have been identified, and abnormal Cdk5 activity has been implicated in several disease processes, including neurodegenerative disorders, cancer, and diabetes. However, a global profiling of protein phosphorylation mediated by Cdk5 is still not available. Our current knowledge about such profiling comes from experiments performed in different laboratories mainly based on 2-dimensional gel electrophoresis or yeast 2-hybrid screening. Because of the limitations of these techniques at the point of validation of targeted proteins, we took a different approach to resolve this issue. We compared phosphorylation status and total protein levels of 258 different proteins by simple Western blotting analyses of Cdk5-/- and WT wild-type brains. The antibodies used in this analysis are already proven to be highly specific for their targeted sites in different biochemical pathways. We based our selection of these proteins on some known and predicted functions of Cdk5 and further categorized them into 6 different groups. The first group contained 25 different proteins involved in apoptosis, the second consisted of 77 different other kinases, the third consisted of 39 different substrates for these kinases, the fourth consisted of 43 different proteins involved in the cell cycle, the fifth contained 37 different proteins involved in numerous neurobiological functions, and the sixth consisted of 37 different proteins controlling different kinase pathways. This phosphoproteomic screening gave us a broad overview of Cdk5 targets involved in these biochemical pathways and cellular processes.[unreadable] [unreadable] We will develop strategies to help us identify the role of Cdk5 in tooth pain and develop potential analgesics. Work on other potentially important questions involving Cdk5 have been shifted to ongoing major collaborations with a number of leading laboratories in neurobiology. This shift will benefit us in our increasingly predominant program on Cdk5 and pain signaling.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this trial is to determine the toxicities of SPI-77 when given concurrent with radiotheraphy in patients with advanced head and neck cancer; to determine the maximum tolerated does (MTD) of SPI-77 when given once every 2 weeks for 2 and 3 doses concurrent with radiotheraphy; to determine the pharmacokinetics of SPI-77; and to gather preliminary response information that may result from the treatment of SPI-77 when given concurrent with radiotheraphy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objectives of this research are to develop monoclonal antibodies to distinct determinants on human prostate carcinoma-associated antigens (PCAA) in order to: (1) develop double determinant immunoassay (DDIA) to study the shedding of PCAA by prostate cancer cells in vitro and in vivo; (2) study the cellular heterogeneity of human prostate tumor cell lines and tissues; and (3) analyze the molecular heterogeneity of PCAA. In the next year, efforts will be focused on the following activities: (1) production of new antibodies to distinct determinants of PCAA will be continued using new immunization procedure, e.g., by injecting mice with purified PCAA covered with one antibody to the antigen; (2) the DDIA will be developed with the antibodies that have already been obtained in the past year that show preferential activity for prostate carcinoma cells to study the shedding of PCAA by cultured prostate tumor cells; (3) the same antibodies will be applied in immunohistochemical study of human prostate tissue using peroxidase staining methods; and (4) the antibodies will be used in immunochemical analysis of the PCAA by two-dimensional gel analysis, i.e., isoelectrofocusing and SDS polyacrylamide gel electrophoresis. (AG)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this research program is to understand the roles of key coagulation and fibrinolytic factors in inflammatory host defense and bacterial virulence. Since a rigorous understanding of these complex processes can only be achieved in an in vivo experimental setting, our proposed studies focus on innate immune surveillance and bacterial pathogenesis in gene-targeted mice with specific alterations in core hemostatic factors. The aims of this project center on the following specific hypotheses: i) host factors mediating fibrin deposition and dissolution are important determinants of the inflammatory response in vivo, ii) bacterial factors known to bind/activate host prothrombin, fibrinogen and plasminogen serve to subvert host defense, iii) leukocyte engagement of fibrin within challenged tissues is an important cue in \"target recognition\" leading to the implementation of effective antimicrobial functions, and iv) hemostatic factors control leukocyte activation events as well as macrophage egress from inflammatory sites into lymphatics. These hypotheses will be tested through detailed studies of S. aureus virulence/host defense in mice lacking fibrin(ogen) or expressing mutant forms of fibrinogen that either lack the leukocyte integrin receptor alphaMbeta2 binding motif, lack platelet integrin receptor alpha(IIb)beta3 binding motif, or cannot be converted to a fibrin matrix (Aim I). Further, the role of procoagulant and fibrinolytic factors in establishing S. aureus virulence/host defense will be determined through investigator-imposed genetic changes in prothrombin, plasminogen and their respective microbial-derived activators: coagulase and staphylokinase (Aim 2 and 3). Finally, the mechanistic role of neutrophils and macrophages in the fibrinogen/prothrombin-dependent bacterial clearance will be established in the context of S. aureus peritonitis (Aim 3). The proposed studies will provide a detailed understanding of the cross-talk between the hemostatic and inflammatory systems in the implementation of effective antimicrobial response in vivo. Further, these studies may illuminate therapeutic strategies centering on specific hemostatic system components that are effective in the treatment of life-threatening microbial infection/sepsis. Lay Description: The bacterial species that are the most common causes of human suffering and death have uniformly evolved the means to engage coagulation factors, apparently as a means of subverting host defense. The goal of this research is to develop a detailed mechanistic understanding of the role of host hemostatic factors in bacterial virulence and the inflammatory response as a step toward novel therapeutic interventions for life-threatening infectious disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will examine how specific neuronal connections are established between vestibular sensory organs and their central nervous system targets. In adult vertebrates, primary vestibular afferent axons serving the different vestibular end organs form distinct though overlapping regions of axonal arborization in the vestibular nuclei. While the presumed role of this specific connectivity in sensory processing is obvious, little is known about the developmental mechanisms that are responsible for its establishment. The proposed experiments will be a first step towards analyzing the development of organotopic specificity in the primary vestibular afferent projection to the vestibular nuclei. Vital-dye fiber-tracing techniques will be used to label populations or single axons in the zebrafish, Brachydanio rerio. In the initial experiments, the pattern and extent of axonal growth from each vestibular sensory organ in young zebra fish will be examined in fixed specimens both in whole-mounts using confocal microscopy and in cryostat sections. In addition, the dynamic rearrangements of axons in each projection will be studied in living animals using confocal and low light level video microscopy. Such direct observations will establish the normal time-course of axonal patterning, and permit simple perturbation experiments that will test the relative roles of axon-axon and axon-target interactions in forming specific connections. These experiments will be the first to follow the development of the neuronal connections that subserve vestibular senses. At present, little is known about the relative roles of inherent positional labels, environmental influences and patterns of neuronal activity in the formation of the specific neural connections. It is not clear to what extent the lessons learned from studies of the visual system can be applied to the vestibular system. Thus, without such knowledge of vestibular development, it is difficult if not impossible to completely understand its improper development observed in birth defects or its abnormal function after trauma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We will study a relatively genetically homogeneous population to test the hypotheses that genetic factors are linked to schizophrenia. The study will focus on two populations. The first is from the Azores, a nine island archipelago in the Atlantic ocean that is a Portuguese state. The Azores have a centralized health system. All ten psychiatrists on the islands are collaborating with us on this project. The second population is from continental Portugal. The majority of the Azorean population is derived from this population base. Families with multiple affected members with schizophrenia, will be studied employing both parametric and non-parametric analytic strategies. We projected approximately 100 families segregating for schizophrenia, including over 300 affected family members. A complementary strategy will be used to study candidate loci. We will study a sample of 225 subjects suffering from schizophrenia and their parents (Total n=675) employing the haplotype relative risk and the transmission/disequilibrium test strategies. This strategy insures that we control for all ancestry for each subject, using the uninherited haplotype derived from the two parents. The third sample will include all other Azorean patients with schizophrenia, who agreed to participate in hopefully, achieving close to complete ascertainment of patients with schizophrenia in the Azores. This will be a valuable sample for linkage disequilibrium approaches given the nature of the Azorean population and provide us with a unique epidemiologic frame. These complementary strategies will allow us to cross validate any positive results. The careful diagnostic definition of phenotype will be based on detailed structured clinical data employing the Diagnostic Interview for Genetic Studies (DIGS), which we have translated into Portuguese. Our project is designed to capture a very complete history of the patients illness, as well as to be able to follow most subjects prospectively for a long period of time. This will be extremely valuable for achieving diagnostic certainty and minimizing false positives. An important new addition to this proposal is the Whitehead/MIT Center for Genome Research, that will perform a genome-wide scan and collaborate on all data analysis for the project.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is porposed to study cell mediated immunity in transplantation. The interaction between sensitive lymphoid cells and antigen will be investigated under defined in vitro conditions in man and in animals. Factors which may influence the expression of cellular immunity, such as uremia, will be investigated. In these studies, the migration inhibition assay will be used as an in vitro correlate for cell mediated immunity. It is also planned to study the effect of soluble mediators of cellular immunity on the behavior of skin homografts in rodents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Epilepsy is a neurological disorder in which normal brain function is disrupted as a consequence of intensive burst activity from groups of neurons. Synchronized population spikes are key concomitants to seizure, but the phenomenon has remained a paradox because it cannot be explained by any known neuronal synaptic mechanism. Several lines of evidence suggest a key role of glutamate in the pathogenesis of depolarization events, which in turn trigger synchronized firing. The observation that astrocytes release glutamate via a regulated Ca2+ dependent mechanism prompted us to hypothesize that glutamate released by astrocytes plays a causal role in epileptogenesis. Our recent study snowed that chemoconvulsive agents including 4-AP and bicuculline triggered TTX-insensitive paroxysmal depolarization shifts in hippocampal slices which were closely correlated with astrocytic Ca2+ oscillations. Photolysis of caged Ca 2+ in astrocytes, but not in neurons, was sufficient to trigger local depolarization events. Furthermore, agents that blocked astrocytic glutamate release reduced epilepiform activity, with no effect on baseline EEC in adult rats. The next critical step is to expand the analysis to reactive astrocytes in epileptic animals. We propose here to analyze astrocytic Ca2+ signaling in epileptic mice with cranial window using 2-photon imaging concomitant with EEC recordings. We will correlate astrocytic signaling in reactive astrocytes with neuronal firing in epileptic mice. Reactive astrocytes are easily identified in live exposed cortex based upon the intensity of GFP emission in transgenic mice expressing GFP under the GFAP promoter. The hypothesis that the efficacy of antiepileptic drugs is better correlated with the potency by which they reduce astrocytic Ca 2+ signaling and glutamate release, than with their direct effects on synaptic transmission, will be tested. These experiments offer a new conceptual and operational approach to understanding the cellular basis of seizure disorders. If a dysregulation in Ca2+ signaling in reactive astrocytes indeed proves causal in epileptogenesis - as our preliminary data strongly suggest - then the implications of this new perspective to pharmacotherapy could be profound. By more specifically targeting the glial cause of neuronal excitability, we might be able to achieve more specific, less variable and less toxic treatment options for patients with epilepsy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will capitalize on, extend, and enhance the Chicago Community Adult Health Study(CCAHS), (P50 HD 38986) which is designed to become a major prospective multi-level study of the impact of individual and social environmental factors on health, their role in understanding socioeconomic and racial- ethnic disparities in health, and the biological and behavioral pathways that are involved. This project has completed a major survey of a probability sample of 3105 adults age 18+ in the city of Chicago, with a response rate of 72% and including physical measurements of height, weight, waist, hip length, and blood pressure. In addition, saliva and/or a blood sample have been collected for 661 people (60% of respondents in a subsample of 80 of the 343 neighborhood clusters (NCs) covering the entire city of Chicago, developed and characterized by the Project on Human Development in Chicago Neighborhoods (PHDCN) (http://phdcn.harvard.edu/), from which the CCAHS sample is drawn. In addition to utilizing existing and collecting new archival data on these areas, the CCAHS carried out Systematic Social Observations of 1664 blocks containing sampled households for the study. Thus, the current proposal seeks to utilize all these data to extend and enhance the CCAHS as follows: 1) to estimate via state of the art multi-level analysis the role in health and health disparities of the immediate social contexts or \"neighborhoods\" in which people live; 2) to estimate how a wide range of aspects of immediate residential contexts and broader social contexts (assessed by multiple methods and measures and considered both singly and in interaction with each other and individual and household factors) affect health and health disparities and help to understand or flesh out the role of social context in health; 3) to estimate via innovative new methods the role in health and health disparities of the broader social contexts which surround more immediate residential contexts; and 4) to begin to convert the existing cross-sectional study into a longitudinal and prospective study via respondent tracking and mortality follow-up, which will allow initial prospective analyses of the predictive association with mortality of individual, household, and contextual psychosocial risk factors for health assessed in the CCAHS, as well as laying the foundation for a second wave of data collection in 2008-9, which will be funded by a subsequent competing renewal proposal, and allow for prospective analyses with respect to the full range of health outcomes in CCAHS. This study will importantly advance the understanding of socioeconomic and racial/ethnic disparities in health - a major priority of the Public Health Service and National Institutes of Health for the new millenium.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Each year in the United States, 5-20% of the population display symptoms of infection with the influenza virus. More than 110,000 of these persons are hospitalized and about 36,000 die from complications. While the spread of infection is generally avoidable through immunization, vaccine may be in short supply, logistics may limit distribution, compliance and lack of concern limit the immunized population and the vaccines are not as efficacious in the very old and very young. This year, a highly virulent avian influenza strain, designated H5N1, has spread rapidly throughout Asia and into Europe. Reports of human infections with H5N1 have increased the concern that this strain, or a recombinant with a virus that commonly infects humans, may expand its host range to infect humans and result in a global pandemic. A moderate pandemic could kill more than 30 million people around the world. A vaccine could be available within six months of recognition of a pandemic. However, influenza vaccine production is limited - only 300 million doses can be manufactured worldwide due to the difficulty in cultivating virus and capacity will take years to expand. Further, in the event of any widespread disease, rapid distribution to the population, insufficient healthcare personnel to administer the vaccine, lack of compliance due to needlestick phobia, and other issues may result in a poorly immunized population. The research proposed by Rocky Mountain Biosystems, Inc. in this application will demonstrate an innovative solution to achieve an immunized, protected population, addressing issues of limited supply, lack of access and compliance. The Self Administered Vaccination Electromechanical (SAVE) device gently and quickly reduces the stratum corneum, administers efficacious and consistent amounts of vaccine in minutes, and avoids creation of \"sharps\" that constitute biohazardous waste. The device is designed to increase the efficacy of drugs and vaccines, while reducing dosage and extending vaccine supplies. The device is small, inexpensive, self-administrable and can be stockpiled for rapid deployment in the case of a pandemic. The device is expected to 1) extend the vaccine supply as much as 20-fold by adaptation to the more efficient transcutaneous route of vaccine administration, 2) greatly improve compliance through self-administration without needles, and 3) be capable of rapid deployment to the populace. Furthermore, SAVE's transcutaneous delivery is expected to enhance cellular immunity versus intramuscular (IM) inoculation, potentially improving immunity in the very young and old. The SAVE technology may also extend the benefits of transdermal drug delivery to new drugs, such as the growing number of macromolecule drugs and fragile protein or peptide bond biopharmaceuticals. In this proposal, the investigators propose to build pre-clinical prototypes of the patch and test it in vitro and in vivo in a swine model, as well as testing impedance in humans. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Positron emission tomography is a sensitive and specific noninvasive approach for delineating abnormalities of myocardial perfusion and for characterizing the efficacy of interventions designed to enhance nutritive flow. Nonetheless, the use of PET has been limited by the availability of cyclotron-produced flow tracers. Accordingly, development of generator- produced, positron-emitting flow tracers is highly desirable. In the initial two years of this award, we developed and implemented approaches to quantify regional myocardial perfusion with 82/RbCl and 62/Cu- pyruvaldehyde bis(N/4-methylthiosemicarbazone) (PTSM). We demonstrated however that 62/Cu-PTSM binds to human albumin limiting the ability to quantify flow after pharmacologically-induced hyperemia. In the proposed continuation we plan to evaluate the myocardial kinetics of two alternative 62/Cu flow tracers: n-propylglyoxal bis(thiosemicarbazone) (Cu-n-PrTS) and its ethylglyoxal congener (Cu-ETS) (tracers which we have demonstrated in preliminary studies permit quantification of regional myocardial perfusion over a wide range of flows and which do not exhibit avid binding to human albumin) is isolated perfused rabbit hearts in which flow can be altered precisely. We will then evaluate these tracers in intact dogs with PET during selected physiological and pathophysiological conditions including permanent and transient ischemia and coronary hyperemia and compare estimates of regional flow to those obtained with microspheres. Estimates of absolute regional flow with 82/Rbcl and PET, validated during the current grant interval in experimental studies, will be extended to evaluations in humans compared with estimates made with H/2/15/O. Since recent studies have suggested that the backward efflux of 82//RbCl initially trapped by the myocardium (k/2) may delineate viable myocardium (heart muscle that will improve function after revascularization procedures) from nonviable myocardium, an additional goal of the proposed continuation is to evaluate the ability of k/2 (estimated with a compartmental model) to predict viable myocardium in dogs with transient myocardial ischemia referenced to sequential echocardiographic measurements of regional function. The most promising approaches will be evaluated in normal human volunteers and in patients with coronary artery disease. Mathematical models will be evaluated with computer simulations and error analysis and modified as required. The results of this research will enable PET centers to use generator- produced, positron-emitting flow tracers to quantitatively estimate regional myocardial perfusion and, potentially, to delineate myocardial viability.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The initiation mechanism of DNA replication in Escherichia coli is unstable and inactivated after having used only once. Thus, the initiation of a new round of replication does not occur in the absence of protein or RNA synthesis. This inactivaton of DNA initiation activity appears to be a built-in property of the hypothetical DNA replication complex since an altered replication apparatus can be formed after a period of thymine starvation, leading to continued DNA replication in the absence of protein or RNA synthesis (Stable DNA Replication). The long-range goal of the proposed research is to determine the molecular mechanism of this inactivation reaction. The objective is to isolate mutants which are capable of stable DNA replication without the inductive treatments such as thymine starvation and to use these mutants as tools for genetic and biochemical analysis of the regulatory mechanism. In fact, we have already isolated such mutants (Sdr-c mutants) and partially characterized some of them. In this proposed research, we will determine whether the regulatory mechanism is indispensable for normal growth of the cell. To this end, we will attempt to isolate non-leaky Sdr-c mutants after M5 phage mutagenesis. Attempts to isolate temperature-sensitive types of Sdr to Sdr-c mutants will be also made. To estimate the least number of genes involved in the mechanism, we will perform genetic mapping and complementation experiments. The biological and biochemical properties of the DNA synthesized by the Sdr-c mutants in the absence of protein and/or RNA synthesis will be examined using transformation and DNA-DNA hybridization techniques. Finally, to determine the replicon specificity of the sdr mutations, we will examine the replication of one of the stringent type of plasmids, F plasmid, in Sdr-c mutants in the absence of protein and/or RNA synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dry eye is a common problem with a severe impact on the quality of life and potential vision-threatening complications. It often results from lacrimal insufficiency caused by immune-related processes, as in Sjogren's syndrome, or by hormone changes associated with aging and various physiological states. One critical hormonal influence on the lacrimal glands appears to be prolactin. Studies with human subjects, hypophysectomized rats, transgenic mice, and acinar cells in primary culture indicate that prolactin can impair lacrimal function, even at serum concentrations within the range of normal values. Moreover, the source of the prolactin that impairs lacrimal gland function may be the lacrimal glands themselves. The lacrimal glands express prolactin mRNA and protein, which may act as an autocrine or intracrine factor that in some circumstances may interfere with secretion. This project will use lacrimal acinar cells in primary culture to answer the following questions: 1. Does locally expressed prolactin act as an autocrine/intracrine factor that supports secretory functions at normal concentrations and impairs them at excessive concentrations? Neutralizing antibodies and antisense reagents will be used to minimize actions of locally expressed prolactin. Expression constructs will be used to overexpress prolactin. Acinar cell morphology, carbachol-induced protein secretion, expression of polymeric immunoglobulin receptors, and expression of ion transport proteins will be evaluated for changes related to altered prolactin expression. 2. Do altered forms of prolactin (16 kDa and phosphorylated 24 kDa) that have inhibitory effects in other cells inhibit lacrimal secretory function? The effects of overexpressed and added forms will be tested as in Specific Aim 1. 3. Does lacrimal prolactin act as a paracrine factor contributing to autoimmune activation? Antisense reagents and expression constructs will be used to suppress or enhance acinar cell prolactin expression, and the modified cells will be tested for their ability to promote proliferation of autologous lymphocytes in mixed cell reactions. This work will advance our understanding of significant mechanisms in lacrimal physiology, and it will have immediate implications for the direction of other studies now in progress. It also has the possibility of stimulating much work well beyond its present scope, including epidemiological studies aimed at identifying sub-populations of lacrimal deficiency patients with different etiologies, followed by design of appropriate, highly specific therapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will test the ability of anti-inflammatory agents to block decrease in blood flow directly measured in the skin, deep tissue and bone marrow of subjects with sickle cell anemia. We hypothesize that therapeutic interventions that interrupt adhesive interactions between sickle erythrocytes, leukocytes, platelets and vascular endothelium will lessen the vaso occlusion, hemoglobin desaturations, and release of mediators of vascular damage induced by intermittent hypoxia. While sickle cell anemia (SCA) is thought of as acute episodes of marked red blood cell (RBC) sickling, sickle-related vaso occlusion occurs continually even dudng non-crisis periods. The cumulative effect of these processes is end-organ damage. Recent data from animal models underscore the importance of leukocyte adhesion to the vascular endothelium in the genesis of sickle vasoocclusion. It is therefore likely that therapies that interfere with the formation of the adhesive interactions between leukocytes, sickle red cells and vascular endothelium will decrease end-organ damage. RBC sickling is triggered by hypoxia. Sleep studies in SCD children demonstrate that 10 to 40 episodes of desaturation occur each night with oxygen saturation dropping to 75 to 85 percent. We directly measured decreases in blood flow in response to nitrogen-induced hypoxia in subjects with sickle cell disease. These decreases are six-times greater than normal controls (p<.001). We will develop this model of vaso occlusion in humans and extend the studies to include measurement of hypoxia-induced changes in inflammatory mediators, markers of vascular damage, and measures of deep tissue blood flow. These parameters will be measured in response to nitrogen-induced hypoxia as well as hypoxic episodes naturally occurring during sleep. We will then use anti-inflammatory agents to block the changes in blood flow and increase in markers of vascular damage after detailed characterization of the N2 challenge and sleep hypoxia models. If these agents are successful, we will have direct evidence in humans that inflammation plays a role in vaso occlusion. Furthermore, this model may serve to test candidate treatments for sickle cell disease. While not within the scope of the present proposal, we anticipate that therapeutic interventions that abrogate hypoxia-induced blood flow decreases and changes in markers of vascular damage detected in these human models will decrease frequency of crisis and lessen the degree of end-organ damage if tested in larger clinical trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this project, mechanisms of normal cellular immunity and tumor immunology are studied: immunological surveillance of cancer patients and the effects of chemotherapy and immunotherapy on the immune response are explored. We are characterizing acute lymphocytic leukemia according to the immunological subtypes of lymphoblasts and the capacity of lymphoblasts to express immunoregulatory helper and/or suppressor functions in vitro. The role of B cell alloantigens in lymphoblastic leukemia is also under investigation. Particular emphasis is placed on immunologic study of the monocyte-macrophage system (including development of new clinical assays of human monocyte function) in malignancies and immunodeficiency diseases, in the newborn, and in malignant histiocytosis. We are also investigating serum suppressors of cellular immunity in cancer patients, and the role of immune adjuvants and their ability to stimulate granulocyte recovery following marrow-suppressive chemotherapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The family of receptors involved with cellular adhesion has been termed integrins because of the predicted function of these proteins in integrating the extracellular matrix with the cytoskeletal framework. The prototypic integrin, the fibronectin receptor, mediates the interaction of cells with fibronectin. The purpose of this study is to elucidate the cytoplasmic interactions of the fibronectin receptor and ultimately to characterize the molecular basis for the fibronectin receptor-cytoskeleton association. The work proposed herein will concentrate on identifying and characterizing intracellular proteins that interact with cytoplasmic domains of each subunit of the fibronectin receptor. Each of the two cytoplasmic domains of the receptor possesses the potential to interact with intracellular proteins. Synthetic polypeptides representing the individual cytoplasmic domains will be coupled to Sepharose and used as affinity matrices for isolation of binding proteins from detergent extracts of cells or tissue. Preliminary chromatography experiments using an affinity matrix of a synthetic peptide representing the cytoplasmic domain of the fibronectin receptor beta subunit coupled to Sepharose has been used to identify a novel protein. This protein, which we have named fibulin, may mediate cytoplasmic connections of the fibronectin receptor as well as the other adhesion receptors that share the beta subunit. Such connections potentially include an interaction with the cytoskeleton. In addition, this molecule may be involved with receptor signal transduction and modulation of receptor function. A significant portion of the work proposed in this study will concentrate on characterization of the structure and biological function of fibulin. A complete amino acid sequence of fibulin will be determined from a combination of protein and cDNA sequencing. Particular emphasis will be placed on characterizing the interaction of fibulin with the fibronectin receptor, with itself, and with other cytoplasmic proteins. Using cDNA and antibody probes the expression of fibulin will be examined in both normal and tumor cells. Additional affinity chromatography experiments are proposed to identify and protein(s) that might interact with the fibronectin receptor alpha subunit cytoplasmic domain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research is directed at understanding the molecular mechanisms by which the papovaviruses, polyoma and SV40, cause major changes in host gene expression. Research will focus primarily on understanding the viral mediated regulation of one particular viral induced enzyme, dihydrofolate reductase (DHFR). For these studies we have isolated methotrexate resistant cells in which DHFR is a major gene product, accounting for over ten percent of the soluble protein. This 200-fold increase in the level of DHFR in these cells is accounted for by an increase in the level of DHFR mRNA and the number of DHFR genes. Proposed experiments are designed to understand 1) polyoma mediated induction of DHFR synthesis, 2) the role of T-antigen in the regulation of DHFR gene expression during abortive infection by SV40, and 3) the relationship between viral mediated control of DHFR synthesis and the control by cell cycle, serum growth factors, and cyclic nucleotides. An important feature of the proposed research is the use of methotrexate resistant cell lines in which one of the viral induced enzymes, DHFR, is a major cellular protein. This feature will allow the use of currently available techniques to study changes in the regulation of DHFR gene expression that occur as a result of infection by polyoma or SV40. As a detailed investigation of the effect of papovaviruse infection on the regulation of host gene expression these studies should lead to a better understanding of how these and other DNA tumor viruses transform cultured cells to a malignant phenotype and cause tumors in susceptible animals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposal is to study the local immune response in chronic periodontal disease. The research plan contains three stages: 1) Explant cultures of gingival tissue were to be used to examine de novo biosynthesis of immunoglobulins, complement components and other proteins. The immunoprecipitation of biosynthetically 14C labeled proteins. 2) Determine specificities of gingival antibodies RIA, ELISA, modified Jerne plaque or passive hemaglutination. 3) Isolate T and B cells for gingiva and characterize their cell surface receptors and blastogenic responsiveness.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (adapted from abstract): Cystic fibrosis (CF) is the most prevalent autosomal recessive lethal genetic disease in the U.S. with 5% of the population carrying a mutant CFTR gene. The long term goals of this proposal are to understand how mutant CFTRs fail to function correctly in CF patients and how these failures might be repaired. The deltaF508 mutant of CFTR is retained in the endoplasmic reticulum (ER) and thereby fails to traffic properly to the apical membrane of epithelial cells. While the mechanism of this trafficking failure may involve recognition of improperly folded CFTR by protein chaperones, preliminary data indicate that phospholipid interactions with the first nucleotide binding fold domain (NBF-1) might also play a role in CFTR trafficking. Dr. Pollard and his colleagues have therefore hypothesized that aberrant interactions between the mutant NBF-1 domain and specific phospholipids might contribute to the ER retention and degradation, and thus cause the disease. To test this hypothesis, the investigators propose: 1) to determine the specificity of phospholipid interaction with NBF-1 and the alteration of this specificity by the deltaF508 mutation by measuring lipid interaction with wild type or mutant NBF-1 using biophysical assays; and 2) to show that NBF-1 induces permeability changes in membranes, and that deltaF508 mutation changes the lipid specificity. This property will be used to develop a screening assay for identifying drugs that might affect mutant CFTR trafficking in CF patients; 3) to show that changes in the phospholipid specificity of NBF-1 have direct consequences for CFTR trafficking in vivo. This aim will be approached by analyzing wild type and deltaF508-CFTR trafficking in a cell line which is temperature sensitive for lipid biosynthesis. The significance of this proposal is that it suggests a novel approach to the mechanism of CF, involving a trafficking defect affected by phospholipid interactions with the mutant protein. Identifying this defect can provide targets for repair. If successful, this approach will set the stage for development of therapeutic means for correcting the aberrant interactions between mutant CFTR and specific phospholipids.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract This renewal proposal supports the maintenance of Cornell?s long standing and successful Molecular Biophysics Training Grant Program. This program provides vigorous interdisciplinary training merging the fields of physical and biological sciences. The program, now in its twenty-seventh year, continues to advance the training of our most motivated and well-qualified students by focusing their graduate academic work around a core curriculum and creating an engaging, cohesive community. The trainees may have undergraduate degrees in either the physical or biological sciences and must have been admitted to the Graduate School at Cornell for training leading to the Ph.D. in a Graduate Field of one the thirty-one participating faculty members who span nine departments, seventeen fields, and four Colleges. All participating faculty are associated with Cornell?s well established and continuously growing Biophysics Program, and have well-funded quality research programs either in physics with strong biological applications or in biology with strong physical connections. The overall research interests of the faculty are broadly distributed and include: structure and function of proteins and other macromolecules using theoretical approaches, synchrotron X-ray crystallography, electron spin resonance spectroscopy, and multidimensional NMR; single molecule studies of dynamics of molecular motors; structures and molecular mechanisms of cell membranes, receptors, and neurotransmitters and associated cellular functions; materials and technology developments including nonlinear laser microscopy, steady-state and time resolved spectroscopy and imaging, single channel recording, nanophotonic optical tweezers, and nanofabrication. Through collaborations and University center facilities, Cornell offers bountiful opportunities for innovation and creative research. The program supports 11 trainees each year, with each individual supported for a maximum of three years. Trainees undertake interdisciplinary studies with advanced courses in mathematics, quantum mechanics, statistical thermodynamics, biochemistry, molecular and cell biology, computation and instrumentation, and other special topics. In addition, students participate in multiple program functions including a weekly Biophysics Colloquium, Summer Student Seminar Series, a yearly campus-wide Biophysics Symposium, and a yearly campus-wide Responsible Conduct of Research Symposium. These activities serve to provide a strong, unified program identity within the broad interdisciplinary structure of the program. Thesis research and collaborations in the laboratories of the participating faculty complete preparation for a career of teaching and research in molecular biophysics. Through these experiences, trainees gain exposure to, and experience in, interdisciplinary biomedical research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We recently discovered a tumorigenic factor interactome connected through the tumor suppressor microRNA-198 in human pancreatic cancer (PC) patient samples and confirmed the tumor suppressive roles of miR-198 in PC animal models. We found that miR-198 is downregulated in PC and is involved in an intricate reciprocal regulatory loop with mesothelin (MSLN), which represses miR-198 through NF-kB-mediated homeobox transcription factor POU2F2 (OCT-2) induction. Furthermore, miR-198 repression leads to overexpression of pre-B-cell leukemia homeobox factor 1 (PBX-1) and valosin-containing protein (VCP). The dysregulated PBX-1/VCP axis leads to an increase in tumorigenicity. Reconstitution of miR-198 in PC cells results in reduced tumor growth, decreased metastasis, and increased survival through direct targeting of MSLN, PBX-1, and VCP. Our preliminary data strongly suggest the significant role of miR-198 and this interactome in PC pathogenesis. In addition, we found that miR-198 can sensitize PC cells for gemcitabine killing because miR-198 effectively downregulates VCP expression and inhibits autophagy maturation in PC cells. In this proposal, we hypothesize that miR-198 and this interactome could serve as a potential prognostic marker and the miR-198 replacement therapy could attack this tumorigenic network through a central vantage point and improve therapeutic efficacy in pre-clinical animal models. Three specific aims are proposed: 1). Demonstrate the significant role of miR-198 and the tumorigenic factor interactome in human PC clinical prognosis in a large cohort of PC patient samples; 2). Design and characterize MSLN-specific targeted miR-198 nanoparticles for specifically delivering miR-198 to PC cells; and 3). Demonstrate the therapeutic efficacy of MSLN-targeted miR-198 replacement in PC patient-derived xenograft (PDX) mouse models. The project will substantially contribute to PC research and will have an enormous impact on clinical practice for patients with PC.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a competitive renewal of an Independent Scientist Award (K02 MH01153) for Paul Worley M.D. Dr. Worley is currently an Associate Professor in Departments of Neuroscience and Neurology at The Johns Hopkins School of Medicine and he devotes full time effort to basic research. The focus of research is the molecular basis of protein synthesis-dependent neuronal plasticity. During the tenure of the previous award period, the laboratory has cloned and characterized several novel brain immediate early genes (IEGs) including Cox-2, Homer, Rheb, Narp and Arc. Analysis of the function of these genes has lead to important new insights into how neurons response to activity and this information is broadly relevant to cortex development, drug addiction and memory. The K02 Award has substantially contributed to Dr. Worley's career by assuring stable salary support and allowing him to commit full effort to research and career development. The research plan has been successful based on record of publications and current support by grants (RO-1) from NIHM, NIDA, NINDS and NIA. The Institution is supportive of continued full time research commitment by the P.I. as evidenced by expansion and renovation of laboratory and office space. Plans for future career development will extend the focus of analysis of IEGs to include studies of cellular physiology and system level function. The Research Plan describes our proposed studies of Homer that will examine its role in regulating cellular calcium dynamics and activity-dependent behaviors, including cocaine sensitization. Planned activities make full use of the outstanding opportunities for collaboration and career development at Johns Hopkins School of Medicine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The marginal cell of the stria vascularis is associated with the production of the endocochlear potential and potassium-rich endolymph. These facilitate the mechanoelectrical transduction in the sensory hair cells. With the whole-cell recording method we previously had found that an inward current of the marginal cell is blocked by micromolar concentrations of amiloride; the dose is consistent with amiloride- sensitive sodium channels. Immuno-cytochemical study showed that immunoreactivity to antibodies raised against sodium channels from the bovine kidney, is distributed in marginal cells as well as vestibular dark cells. These observations are consistent with the importance of amiloride-sensitive channels in absorbing sodium ions from the endolymph to maintain its unique ionic composition. Preliminary studies of ion channel regulations in these cells were carried out by examining immunocytochemical localization of various GTP-binding proteins.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although the Warburg effect has long been known as the hallmark characteristic of cancer cells, its exploitation for cancer therapy has never been successful because of failure in identifying an appropriate target protein. Recent studies suggested PFKFB3, which causes a massive increase in F-2,6-P2, the most potent activator of mammalian glycolysis, as the most causative protein of the Warburg effect. We hypothesized that a drug-like PFKFB3 inhibitor will prevent onset of the cancer-specific glycolysis by preventing the F-2,6-P2 surge and, eventually, induce apoptic death of cancer cells. To test this hypothesis, we propose to develop a drug-like PFKFB3 inhibitor. To obtain the molecular basis of the target, which is necessary for design of the desired inhibitor, we determined the crystal structure of human PFKFB3 and suggested a molecular model of its catalytic mechanism. We also performed an initial drug discovery study and obtained several promising PFKFB3 inhibitor molecules. Taking advantage of the progress we have made, we now initiate a full-scale study to develop a drug-like PFKFB3 inhibitor and test if it can cause inhibition of cancer glycolysis and, ultimately, apoptic death of cancer cells. To obtain the molecular templates for PFKFB3 inhibitors, virtual screening and subsequent functional screening of database compounds will be carried out. Hit compounds will be selected on the bases of binding affinity, selectivity, amenability to chemical modification, and structural novelty. The selected hits will be optimized to be drug-like via chemical modification. During this optimization step, new molecules will be synthesized from the hit templates and tested regarding their potency, cell- permeability, stability, and target-selectivity on cultured cancer cell lines. The compounds selected from this test would be considered as the drug-like PFKFB3 inhibitors. Finally, the biochemical and pharmacological properties of these final lead molecules will be investigated to test our hypothesis. This project is aimed to develop a novel drug-like compound with potential of anti-cancer drug. This compound will be designed to inhibit cancer glycolysis, which has never successfully exploited for cancer therapy. Thus, successful completion of this project will have an immediate impact on development of new class cancer drug and new strategies in cancer therapy. PUBLIC HEALTH RELEVANCE: This project is aimed to develop a novel drug-like compound with potential of anti-cancer drug. This compound will be designed to inhibit cancer glycolysis, which has never successfully exploited for cancer therapy. Thus, successful completion of this project will have an immediate impact on development of new class cancer drug and new strategies in cancer therapy. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We aimed to identify the reproductive targets of pollutants by studying high exposures. We entered into a collaborative agreement with the Ukrainian Institute of Pediatrics, Obstetrics and Gynecology and the University of Bristol in 1993 to conduct the research described here. The Ukraine Placental Study had the following goals: 1. To document the levels of common pollutants experienced by pregnant women in two urban areas, using environmental measurements and biological markers 2. To describe the reproductive outcomes of these women 3. To determine what relationship, if any, exists between the estimates of exposure and the outcome of the pregnancies. PAHs. Concentrations of PAHs were measured in placentas from 200 women from two cities in Ukraine, Kyiv and Dniprodzerzhinsk. The participants had no special exposures and were chosen from among subjects in an ongoing study of reproductive health. All seven of the polycyclic aromatic hydrocarbons were found in all placentas, with the sole exception of benzo(a)pyrene in one placenta. Chrysene was present at the highest concentrations, with median 1.38 ng/g dry weight. Dibenz (a,h)anthracene and benzo(g,h,i)perylene had the lowest concentrations; each had median 0.73 ng/g dry weight. Concentrations in Kyiv were slightly higher than those in Dniprodzerzhinsk, but the difference was significant only for dibenz(a,h)anthracene. Dibenz(a,h)anthracene and benzo(g,h,i)perylene increased significantly with maternal body mass index, but other PAHs showed no such pattern. Hospital admissions. We investigated the reasons for hospital admissions in a sample of deliveries from Ukraine, a country of the former eastern bloc. We enumerated all hospitalizations among 3,099 women who delivered live singletons of at least 20 weeks gestation in two urban areas of Ukraine and abstracted data from their medical records. . More than a third of the women were admitted to hospital during their pregnancy, and 91 % of the admissions were for a pregnancy complication, primarily threatened abortion or early labor. Median length of stay for all admissions was 12 days. The ratio of admissions to deliveries was 52 per 100.The Ukrainian ratio of hospitalizations to deliveries is notably higher than any that have been published in studies from the United States and Australia, reflecting patterns of care that stress hospital?based treatment. This high ratio does not necessarily mean that Ukrainian women are sicker, although that may be the case. Preterm birth. The economic, social and health problems faced by former eastern bloc countries after the demise of the Soviet Union are unique in the recent history of Europe. We conducted a study in two urban areas of Ukraine, asking if the traditional predictors of preterm delivery continue to be associated with risk under these conditions. Subjects were pregnant women with LMP between 12/25/92 and 7/23/94. Self-completed questionnaires and the medical record provided data. We compared 137 spontaneous preterin deliveries with 2886 full term births, using all established risk factors for which we had data. Maternal age was the variable most strongly related to preterm birth. Being 18 or less had an OR of 3.7; being 30+ had an OR of 2.5 relative to the reference group of age 25-29. Placental complications and pre-existing hypertension had ORs of 2.7 and 2.3 respectively but the confidence interval included 1.0. Low net pregnancy weight gain (less than 10 kg) was significantly associated with preterm birth, but the rate of net weight gain was not. Marital status and educational category were only weakly related. Comparison with England. We compared the rates of spontaneous preterm birth in Avon County, England and in two urban areas of Ukraine, using the same study methodology in both sites and accounting for differing in maternal risk factors. We used data from the European Longitudinal Study of Pregnancy and Childhood; eligibility required residence in geographically defined areas and last menstrual period in a specific time window. Data were collected by questionnaire and medical record abstraction between 1992 and 1995, using comparable instruments and methods in both countries. Preterm births were classified into those that were spontaneous, and those that involved medical intervention. Rates of spontaneous preterm birth were compared in the two sites, taking account of maternal characteristics. Placenta and birth weight. We investigated whether the difficult conditions in the former eastern bloc in the last decade might have altered the weight of the placenta or its relationship to other measures of fetal size from what has historically been reported. Placentas were obtained from 1621 singleton births of at least 28 weeks gestation in a Ukrainian city during 1993-94, using a systematic protocol. Information on maternal characteristics was obtained from two questionnaires completed by the mothers. Data on pregnancy complications were abstracted from the medical record, as were absolute measures of birth size (infant weight, infant length, crown-rump length, and head circumference). We examined the relationship of placental weight and ratio to these variables. The range of placental weight was 100 to 1000 grams, with a mean of 470 g. Mean placental ratio was 13.9%. Placental weights increased and placental ratios decreased with gestational age. Larger ratios were related to larger maternal BMI. Absolute measures of infant size and placental weight were mutually positively correlated. Placental ratio, infant length, and ponderal index were nearly uncorrelated. Ukrainian placentas were of similar absolute and relative weight as those historically reported, as was their relationship to other indicators of infant size. Alcohol and breastfeeding. We aimed to replicate a previous study of one-year olds that reported deficit in motor development associated with moderate alcohol use during lactation, using different but comparable population. The mental development of 915 18-month-old toddlers from a random sample of a longitudinal population-based study in England was measured using the Griffiths Developmental Scales. Frequent self-administered questionnaires during and after pregnancy provided maternal data. The dose of alcohol available to the lactating infant was obtained by multiplying the alcohol intake of the mother by the proportion of breast milk in the infant's diet. We compared this dose with the Griffiths scales, taking into account potential confounding variables in the sample. ACE inhibitors. We explored the association between enalapril exposure and adverse outcomes in pregnancy, taking into account other possible risk factors. We analyzed a series of all usable cases reported to the FDA between 1986 and 2000 in which enalapril was a suspect drug for the observed adverse outcomes (N=110). Parameters of exposure and reported outcomes as well as information on potentially confounding variables were systematically abstracted from this series by a single physician. Because exposure to ACE inhibitors after the first trimester of pregnancy had been associated with adverse outcomes in the existing literature, we divided the cases into those exposed in the first trimester only (considered as the baseline group) and cases exposed beyond or after this time. Frequency of reported adverse outcomes in the second group was compared with those in the baseline group; odds ratios were computed, taking account of potentially confounding variables by logistic regression where appropriate. Dr. Little has retired; Beth Gladen (DIR/BB) will assume role of PI on this Project.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Work will be carried out on two aspects of the biology of Trichinella spiralis: 1) Immunobiology of T. spiralis infection in rats and mice. The antigens responsible for induction of protective immunity are contained in secretory granules, which are localized to the stichocyte cells. These antigens will be fractionated and characterized by a variety of chemical and physical techniques. The detection system to be used for these antigens will include Ouchterlony analysis and in vitro bioassay for adult worm fecundity. The stage of the parasite most susceptible to the effects of protective immunity will also be investigated. 2) The Nurse-cell parasite relationship. The nuclear and cytoplasmic events leading to the formation of the Nurse cell (i.e. modulated infected muscle cell) will be investigated utilizing biochemical and cytological techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this project is to develop a number of organoironcarbonyl reagents to be used in the synthesis of a number of natural products which are important in the health sciences. The type of organoironcarbonyl complex on which our work has focused thus far are tetracarbonyliron complexes of alkenes. In one phase of this work we have already prepared a number of alkenes with geminal carbonyl substituent, including methylenemalonates and alpha-acylacrylates, which are capable of forming stable tetracarbonyliron complexes. These alkenes and the corresponding complexes are being developed as reagents for the construction of polycyclic fused ring natural products. A specific objective of this part of our work is the total synthesis of estrone, a centrally important steroidal hormone. In another phase of this work, a new approach to macrolide synthesis is being developed through the use of tetracarbonyliron complexes of acrylate esters which incorporate a potential internal nucleophile. Formation of the macrolide ring involves generation of the internal nucleophile with base followed by intramolecular nucleophilic addition of nucleophile to the acrylate ester. The scope of this reaction type is currently being established after which the method will be applied to the synthesis of specific naturally occurring macrolides.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development of new female-initiated HIV prevention methods, including oral and topical microbicides, is urgently needed to stem the growing epidemic among adolescent women. Despite the disproportionate impact of HIV/AIDS on adolescent women, they have been markedly under-represented in HIV prevention trials. For example, only one phase III microbicide trial has attempted to recruit adolescents under the age of 18, even though sexual debut and high risk sexual behavior may occur years earlier. There are important scientific, ethical and programmatic reasons to consider recruitment of adolescent women. Nevertheless, challenges to recruiting and retaining adolescent women in HIV prevention trial research will be numerous. This proposed study will conduct formative community-level research and a pilot prospective clinic-based study in India and Tanzania to systematically examine the challenges and opportunities related to recruitment and retention of adolescent women aged 15-21 in clinical microbicide trials and make specific recommendations about whether and how such trials should be adapted to accommodate adolescent participation. The specific aims of this study are: Aim 1: To identify the HIV risk characteristics of adolescent women and determine whether these risks differ by younger (15-17) versus older age groups (18-21) and by country context. Risk characteristics of interest include: types and patterns of sexual partnerships;types of sexual risk-taking and risk-reduction behaviors;perceptions of risk for HIV, sexually transmitted infections (STIs), and other negative health outcomes;and psychosocial and environmental factors contributing to risks; Aim 2: To evaluate the legal, socio-cultural and service delivery factors that hinder adolescent women's participation in microbicide trials and make specific recommendations to enhance their participation; Aim 3: To examine and compare adolescent women's (aged 15-17 and 18-21) understanding of and ability to adhere to the requirements of trial participation in the context of a prospective clinic-based study;and Aim 4: To determine adolescent women's acceptability and use of a proxy gel, including circumstances in which participants apply the gel;reasons for non-use of gel;negotiations about gel use with partners;and the influences of gel use on sexual satisfaction for themselves and their partners.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Obesity and cognitive dementia may seem to have little in common other than both can have severe health consequences and both have so far been highly resistant to therapeutic interventions. Traditionally, investigators have viewed the problems of obesity and metabolic syndrome, on one hand and cognitive dementia on the other, as involving distinct etiologies, that target different underlying behavioral and biological functions, and that rely on largely separate brain structures and circuits. However, recent epidemiological findings point to a link between obesity in mid-life and the emergence of Alzheimer's disease and other serious cognitive impairments in the aged. These findings have encouraged the hypothesis that obesity may be causally-linked to cognitive impairment. Using a rodent model, this present project will explore (the perhaps counter-intuitive) idea that excessive energy intake leading to obesity and related metabolic disorders (e.g., diabetes, hypertension) is a consequence of interference with learning and memory processes. Based on the results of prior support for our work, we have developed a model that describes the learning and memory mechanisms that underlie the ability of animals, including humans, to use internal cues corresponding to \"satiety\" to signal when food and cues related to food will be followed by appetitive or desirable postingestive ( i.e., post oral) stimulation. Our research indicates that this type of signaling can be degraded by (1) the consumption of sweet no- or reduced-calorie foods/fluids that may weaken t he normally strong predictive relationship between the orosensory properties of food and the caloric or nutritive consequences of eating;(2) consuming diets that impair the functioning of a brain structure (i.e., the hippocampus) that underlies the ability of rats to use their satiety cues to predict when eating food will or will not produce reinforcing postingestive outcomes. A result of degrading the operation of these learning mechanisms is excess intake and body weight. The present research will examine whether learned control mechanisms develop without explicit laboratory training, whether consumption of high-fat diets interferes with these mechanisms, and whether this interference has potential to produce significant weight gain. These studies promise to yield important new information about the role of dietary factors in both obesity and cognitive decline. PUBLIC HEALTH RELEVANCE: Obesity and cognitive decline are both serious threats to the public health and to the of quality life. This project will explore the possibility that both of these disorders have common environmental origins and brain substrates. The idea that disruption in a specific learning process gives rise to obesity will also be examined.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neurons communicate with each other in the brain through specialized junctions, called synapses. During brain development, numerous new synapses are established and new synapses continue to form throughout life. The long-term goal of the research proposed in this application is to determine the molecular basis of synapse formation in the vertebrate brain. The first proteins have now been identified that organize synapse formation and development. One such protein is SynCAM 1, a synaptic cell adhesion molecule that connects pre- and postsynaptic sides. Importantly, SynCAM 1 induces the formation of new, fully functional excitatory synapses between neurons. It is highly expressed in the developing brain during intense synaptogenesis, indicating a broad function for this molecule in synapse formation. Such synaptogenic functions have been validated in cultured neurons and in vivo. The objective of this application is to define the signaling pathways through which SynCAM 1 organizes synapses and determine how other trans-synaptic proteins act in concert with it. The central hypothesis of this application is that SynCAM signaling organizes developing synapses and regulates synaptic function at later stages. To attain the objective of this application, three specific aims will be pursued. The first aim of this application is to determine the intracellular signaling pathways through which SynCAM 1- mediated synaptic adhesion instructs synapse development, focusing on changes in the synaptic cytoskeleton. Second, it will be analyzed how trans-synaptic interactions act in concert to assemble synapses and shape their structure. Third, it will be determined to which extent SynCAM 1 functions in vivo together with other synaptic adhesion molecules to organize synapses. These experiments involve the biochemical characterization of SynCAM binding partners and their activities. Functional analyses of SynCAM interactions will be performed by quantitative immunocytochemistry, imaging of synapses in cultured hippocampal neurons, and electrophysiological recordings. In addition, the in vivo relevance of these interactions will be tested using structural and functional studies of synapses, including ultrastructural analyses, electrophysiological recordings, and behavioral analyses. Achieving these goals is important for human health, as altered synapse organization affects the wiring of neuronal circuits and synaptic plasticity. These changes are associated with alterations in human behavior, the ability to learn and remember, and addiction to drugs of abuse. Furthermore, deficits of synapse formation likely underlie neurodevelopmental disorders such as autism. In summary, this application aims to identify the molecular interactions involved in synapse formation. The progress under this application will allow testing to which extent these synapse-organizing processes are affected in disorders of the human brain and whether they represent novel points of therapeutic intervention. PUBLIC HEALTH RELEVANCE: Nerve cells communicate with each other in the brain through specialized junctions, called synapses. These junctions form in the human brain soon before birth, and changes in this process impair the wiring of the brain and can cause mental retardation. This research program is relevant to public health because it will analyze how nerve cells connect to each other in the healthy brain, allowing us to understand what steps go wrong in developmental disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Influenza A viruses exhibit extreme diversity as exemplified by the multiple serotypes of the hemagglutinin (HA, H1-H18) and neuraminidase (NA, N1-N11) surface antigens. To date, only three of the possible 198 combinations found in avian and other animal reservoirs have been associated with human pandemics (H1N1, H2N2, H3N2). However, ever increasing anxiety about the potential for infections by infections of zoonotic origin have heightened concern about emergence of a human transmissible virus that could devastate the human population. The emergence of H5N1, H6N1, H7N7, H7N9, H9N2, and H10N8 viruses in the human population are constant reminders of this possibility. Influenza B viruses with its two distinct lineages further increase the health and economic burden of seasonal influenza worldwide. This proposal seeks to elucidate, at the structural level, key sites of vulnerability on influenza virus that can be utilized to develop therapeutics as well as improved vaccines. Antibody-mediated neutralization of influenza virus is a complex combinatorial problem for the human immune system as it is presented with diverse, highly variable and constantly evolving viruses. While neutralizing antibodies against human flu are traditionally regarded as being strain specific, recent advances have shown that much broader responses can be mounted that give valuable insights into conserved sites of vulnerability. We are therefore amassing compelling evidence that a sustained, cross- serotype response can be mounted against influenza and this vital information can now be harnessed for design of small molecules, peptides, and proteins to target these key sites of vulnerability, thereby blocking influenza infection. No effective drugs are currently available for preventing the entry of influenza virus. Thus, we will elucidate common features for recognition of sites of vulnerability of pandemic and emerging influenza viruses from crystal structures of diverse HAs with broadly neutralizing antibodies and sialosides that are mimics of the natural receptor. A combined biophysical, biochemical, and chemical approach employing state- of-the-art structural biology, glycan arrays, and chemical biology will be used to provide key insights into influenza virus neutralization that enable design of novel therapeutics to control and combat future influenza pandemics and seasonal epidemics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The three areas of OCD research are: Suicide Risk in the Medical Setting Suicide is an international public health threat. In the U.S., it is the 2nd leading cause of death for youth and the 10th leading cause of death for adults. As suicide risk screening becomes a national priority for medical settings, non-mental health clinicians require valid, psychometrically-sound screening instruments for assessing suicide risk. However, there are very few such mental health assessment instruments that have been validated empirically with medically ill patients in hospital settings. Recognizing this gap early and capitalizing on clinical and research expertise within our group, we began developing a risk of suicide screen for pediatric patients in the emergency room in 2008 which resulted in a validated tool, the ASQ (Ask Suicide Screening Questions). The ASQ has been translated into Spanish, French, Dutch and Chinese and is currently being implemented in emergency departments in the U.S. and abroad. The tool is available on the NIMH website at http://www.nimh.nih.gov/news/science-news/ask-suicide-screening-questions-asq.shtml for public use. Since the development of the ASQ, the focus of our ongoing suicide screening studies has been to develop and validate brief screening instruments that can be administered by non-mental health clinicians in other medical settings and with specific patient populations. We currently have 5 IRB-approved protocols addressing suicide screening in different medical settings (NCT00623493, NCT01517126, NCT02140177, NCT02050867, NCT02830334). We are addressing areas of need, including pediatric and adult emergency department patients, and pediatric and adult medical inpatients. Our research is current with the February 2016 Joint Commission (JC) Sentinel Event Alert calling for suicide risk screening of all medical patients in a hospital setting and is consistent with NIMH's commitment to join with the National Action Alliance for Suicide Prevention to reduce the suicide rate by 20% over the next 5 years. In addition, our research team is working closely with NIH CC leadership in order to meet the JC recommendations; we are utilizing our research to inform implementation of suicide risk screening at the NIH CC and hospitals nationwide. Mental Health Aspects of Coping with Chronic Medical Illness Born out of our clinical work on the PCLS and neurocognitive assessment service for the past decade, a natural research focus has emerged from our collaborative work with other Institutes at NIH. For example, working with the Pediatric Oncology Branch we began to study distress and its correlates in medically ill children, particularly those with life-limiting illnesses, through 3 protocols over the past 5 years (NCT00824278, NCT00969579, NCT02423031). The collaboration has been highly productive leading to the development of the first advance care planning guide for adolescents and young adults with life-limiting illnesses, Voicing My CHOiCES http://www.agingwithdignity.org/forms/voicingmychoices.pdf. This planning guide is available at www.agingwithdignity.org Another study along these lines (NCT01778478) focuses on how to screen and diagnose psychiatric disorders in young adults (ages 18-25) with and without cancer. The study examines the feasibility of using the structured psychiatric diagnostic interview, Kiddie-Schedule for Affective Disorders and Schizophrenia Present and Lifetime version (K-SADS-PL), in this age group and evaluates whether the parent interview affects diagnostic findings. In addition, the study assesses psychosocial developmental milestone acquisition among young adult cancer patients compared to their healthy peers. Neuropsychiatric Aspects of Immune and Infectious Disorders (including HIV/AIDS), Genetic, and other Disorders The OCD Neuropsychology Unit is involved in a broad range of research protocols that are studying cognitive and emotional functioning in various diagnostic groups. These research protocols include genetic disorders (such as Mobius syndrome, methylmalonic academia, Cerebral Autosomal Dominant Arteriopathy with Sub-cortical Infarcts and Leukoencephalopaghy (CADASIL)), infectious and immune diseases (HIV, progressive multifocal leukoencephalopathy, Ebola, anti-NMDA receptor encephalitis), brain tumor (re-irradiation effects), and chronic fatigue syndrome. A large focus of these neuropsychiatric studies is HIV/AIDS. HIV-related research activities in the OCD fall into two categories: 1) NIH Intramural NeuroHIV Program, and 2) Collaborations with multi-site research networks, including an NIH DoD HIV/AIDS associated neurocognitive disorder protocol. In 2008 the NIH, led by NIAID, initiated a partnership with the District of Columbia (DC) government with the goal of reducing the impact of HIV on DC, which had almost a 3% prevalence rate of HIV infection, the highest of any city in the US. The program was also created to develop research infrastructure in the city to generate knowledge and change public policy as well as to develop research across NIH. In alignment with the NIMH's commitment to reduce the burden of mental illness in the HIV/AIDS population, NIMH, NINDS and NIAID have developed 2 intramural protocols (NCT01875588; NCT01692236) investigating HIV-related neurocognitive disorders, which are a clinical challenge and threat to the long-term health of people living with HIV/AIDS. In addition, we have worked to assess the mental health needs of HIV patients and build an interdisciplinary approach to HIV and mental health in DC, leading to funded projects being initiated at other DC institutions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Data on daily climatic conditions as reported from approximately 300 weather stations have been obtained from the National Climatic Center, and linked to daily mortality data for the period 1972 through 1977. An age-specific analysis of average daily mortality at various levels of climatic variables will be performed in order to ascertain relationships between age and climatic conditions. Climatic variables will include temperature, relative humidity, rapid changes in temperature, departures from normal temperatures, and sustained heat waves and cold fronts. Levels at which hypothermia and hyperthermia are more likely will be investigated in greater detail. Concomitant information on socioeconomic status and geographic location of the cases will also be considered.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In immunologic lung disorders such as sarcoid activated alveolar macrophages are key to the pathophysiology of the alveolitis. These disorders can be successfully treated with systemic corticoids. Since therapy is often compromised by toxic side effects, localized inhalation therapy would be optimal. However, due to the poor aqueous solubility of corticoids, no inhalable dosage form currently exists which is optimally suitable for small particle nebulization to alveolar lung regions, where the inflammation occurs. We propose a novel therapeutic approach to such immunologic lung disorders: Alveolar delivery of liposome-incorporated corticoids by aerosol in order to promote corticoid uptake by alveolar macrophages, and thus modulate the inflammatory alveolitis at the macrophage level. The overall objective of this project is to initiate research that will lead to improved inhalation therapy of immunologic lung disorders, using sarcoid as an immunologically well-defined and corticoid- responsive example. The rationale is that corticoid inhalation could be more effective by targeting liposomal drug directly to alveolar lung regions and to macrophages. While optimal therapeutically effective levels could be established, systemic drug levels, thus side effects would be significantly reduced. Prolonged retention of liposome-corticoid in the lung may result in prolonged therapeutic efficacy at the cellular level. We propose to (1) design lyophilized liposome-corticoid dosage forms suitable for small particle aerosolization; (2) compare in vitro human macrophage uptake and receptor binding of liposome-corticoid and corticoid in solution; (3) determine the efficacy of liposome-corticoids on regulation of human macrophage immune functions n vitro; and (4) determine the in vivo distribution of liposome corticoids in rat lung, plasma and extrapulmonary tissues.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is part of a larger endeavor whose aim is to systematically investigate executive control functions, studying their development, neural bases, and genetic & neurochemical modulation from infancy through old age. The focus here is on inhibition of attention, inhibition of action, and cognitive flexibility during early development. It is hypothesized that: (1) Errors made by children of 3-10 years reflect problems with flexibly switching. Even 3-year-olds will succeed at steady-state attentional inhibition (selective attention) and steady-state action inhibition (inhibition of old stimulus-response mappings). Lags of up to 3-6 years will be found, however, between when children can first show inhibition in steady-state and when they can switch back and forth. (2) This progression does not depend on improved memory; even the younger children will be able to state the correct rule on each trial. It does depend, though, on improvements in the ability to execute the mental computations necessary to translate abstract rules into practice. The core problems in the development of executive control functions are the abilities to flexibly switch mental settings and to flexibly manipulate information in one's mind, not inhibition or memory per se. (3) Many of the errors made by infants and preschoolers reflect difficulty in grasping that two things are conceptually connected if they are not physically connected (e.g., not part of the same entity), and the flip side, difficulty grasping that attributes of a single entity can be separated and the entity conceptually redescribed from different perspectives. Thus, while switching attentional focus and response mappings is difficult, when the stimulus dimensions are separated, children will succeed at such switching at a very young age. The proposed research will test these and other hypotheses, charting the development of inhibitory control and cognitive flexibility in the early years of life. This will provide valuable insights into why children have difficulties, conditions for optimizing their performance, and benchmarks by which to assess children's development or deficits. A more refined understanding of executive control functions will assist efforts to understand how these are differentially affected in disorders such as Attention Deficit Hyperactivity Disorder, Obsessive Compulsive Disorder, schizophrenia, addictions, and autism. Insights into the conditions under which children can succeed may yield procedures that might prove helpful to those afflicted with disorders affecting these critical functions. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The training proposed in this NRSA contains both a practical research and didactic component. The broad aim of the research component is to determine the extent to which the presence and severity of metabolic syndrome (MetS) contributes to aging-related functional decline, and to examine whether this relationship is mediated by excess inflammation. Because maintenance of physical function is a critical factor in the ability of older adults to remain independent, the research component of this project is highly relevant to the mission of the NIA. The project will be accomplished via secondary analyses of data collected in the Health ABC Study, a prospective cohort of 3,075 well-functioning white and black elders (70-79 years), well characterized for MetS, with several indices of physical function and inflammation. Specifically, this project will (1) determine whether the presence and severity (number of components) of MetS is associated with lower physical function at baseline and with a decline in physical function over time in older adults; (2) determine whether the presence and severity of MetS is associated with inflammation at baseline and with changes in inflammation over time in older adults; and (3) determine whether, and to what degree, inflammation reduces the effect of MetS on physical function. Statistical methods to be used include logistic regression and mixed effects modeling, as well as mediation analysis. This project will provide a practical opportunity for the candidate to utilize the statistical and epidemiological skills she will learn as part of the proposed coursework (described below). The research will also provide the candidate with direct, practical experience in data management and longitudinal data analysis. For the didactic component, Dr. Beavers will complete a series of courses offered through the Master's program in Clinical and Population Translational Sciences at the Division of Public Health Sciences and Translational Science Institute at Wake Forest University in statistics, epidemiology, and clinical trials, such as Foundations of Clinical and Translational Science, Epidemiology, Clinical Trial Methods, and Applied Linear Models. Completion of these courses will provide a more comprehensive understanding of statistical and epidemiological concepts and clinical trial methodology, allowing Dr. Beavers to be able to conceptualize, plan, and execute clinical research in her future career. The research findings will provide better characterization of the relationship between MetS, inflammation, and physical function, which may assist in the development of preventive and therapeutic efforts for older men and women with functional disability, while the coursework in Clinical and Population Translational Sciences offers a unique opportunity to maximize the candidate's post-doctoral training at Wake Forest University and develop competencies in conduction and translation of clinical and population research. PUBLIC HEALTH RELEVANCE: Maintenance of physical function is a critical factor in the ability of older adults to remain independent. Better characterization of the relationship between common preventable conditions, such as metabolic syndrome, and physical dysfunction is critical to efforts aimed at preventing disability. Given the growing demographic of persons aged 65 years and older, coupled with the economic and emotional burden related to age-associated loss of physical function, prompt identification of modifiable functional decline risk factors is of considerable public health concern.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is for continuation of collaborative research in the natural history, epidemiology, biology, and treatment of childhood cancer at the Children's Hospital & Medical Center (CHMC) in Seattle, Washington as a full member institution of the Childrens Cancer Group (CCG). The specific aims of this proposal are: 1) to develop better treatment of childhood cancer; 2) to develop a better understanding of the biology of childhood cancers; and 3) to develop a better understanding of the natural history, epidemiology, and late effects of treatment of childhood cancer. CHMC will pursue these aims by contributing patients to phase III randomized intergroup and CCG trials, by participating in CCG pilot therapeutic trials that will form the basis for future randomized trials, by participating in new agent trials, by contributing specimens and data for biology, epidemiology and late effects studies, and by providing leadership in the scientific and administrative efforts of CCG. During the first three years of the last grant period, 1989,1990 and 1991, two hundred-forty-eight patients were entered onto therapeutic studies, 181 onto epidemiology and late effects studies, and 36 onto biology studies. At the end of 1991, there were 838 patients in active follow-up after completing a CCG therapeutic study. In the first 8 months of 1992, following Dr. Miser's arrival at CHMC, 97 patients were entered onto CCG therapeutic protocols (12 patients per month) and 83 patients onto non- therapeutic protocols (10 patients per month). CHMC is an important contributor to the leadership of CCG: Dr. Chard, former Associate Chairman for leukemia studies and former vice chairman of CCG, is the principal investigator, a member of the nominating committee, and an important group leader. Dr. Miser is a member of the CCG Executive Committee; Associate Chairman for CCG solid tumor studies; Chairman of the Bone Tumor Strategy Group; Chairman of CCG-7881, Chairman of CCG-8605, Chairman of the CCG pilot osteosarcoma program, and a member of a number of other therapeutic and strategy committees. Dr. Geyer is a member of the Brain Tumor Strategy Group, Chairman of CCG-9921, Chairman of CCG-0896, Chairman of CCG-B921, and a member of several other brain tumor committees. Dr. Bernstein is a member of the biology committee and principal investigator of the ANLL reference laboratory. Dr. Pendergrass is Chairman of CCG-8602 and a member of a number of epidemiology committees. Drs. Milstein and Berger are members of brain tumor committees. Dr. Schaller is a member of the surgical and neuroblastoma committees. Dr. White is a member of the imaging committee. Dr. Breiger is a member of the psychology committee.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Structural studies we have conducted to date on highly purified porcine relaxin indicate that it has a molecular weight of approximately 6,700 and consists of two non-identical chains (alpha and beta) of similar size which are linked by disulfide bond(s). We propose to determine the amino acid sequence of the porcine relaxin chains and the location of the disulfide bond(s). We then propose to determine that portion of the relaxin molecule responsible for its biological properties--the so-called biologically active site. We propose to locate the tissues responsive to relaxin in the rat. We will employ a sensitive and precise radioimmunoassay for relaxin to determine factors which influence the secretion of porcine relaxin immediately preceding parturition. BIBLIOGRAPHIC REFERENCES: O.D. Sherwood, K.R. Rosentreter, and M.L. Birkhimer, \"Development of a Radioimmunoassay for Porcine Relaxin Using 125I-Labeled Polytyrosyl-Relaxin,\" Endocrinology 96:1106, 1975.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the proposed effort, GW Medical Technologies will asses the benefits, risks, and effectiveness of a peripheral blood lymphocyte (PBL) simulation index diagnostic test for Alzheimer's disease (AD). The company's test detects AD by evaluating a patient's stimulation index against a predetermined cutoff, where the stimulation index is defined as the ratio CD69 presentation on mitogenicly stimulated peripheral blood lymphocytes to the CD69 presentation of un-stimulated lymphocytes. In preliminary studies the company found a statistically significant difference (p<0.05) in the mean stimulation index of AD patients and cognitively intact controls and was able to identify samples from AD \"probable\" patients with high sensitivity and selectivity (83% and 96% respectively). However, these early results are suspect due to the limitations of the original test methodology and protocol. Early testing relied on manual processes for data collection, transfer and analysis, which could introduce significant errors. Early testing also used a small sample size (n=72) and did not include samples from patients with other forms of dementia. In the proposed effort, the company addresses all three of these limitations by developing automated routines for data collection, handling and analysis and by conducting a randomized, blind clinical study (n=300) that includes control samples from both cognitively intact individuals and patients with other forms of dementia. Should this study prove successful, a this test could provide a preclinical diagnosis of AD, improve patents' quality of life, and reduce the cost of care.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term goal of this project is to identify biological roles for mammalian glycans in development and cancer. Glycosyltransferase gene mutants of mice, embryonic stem (ES) cells and CHO cells will be used to investigate the basis of cell-type specific blocks to differentiation and transformation caused by alterations in specific glycans. Our recent studies show that 1) oocyte-specific knockout of complex and hybrid 7V-glycans compromises ovulation and oocyte developmental competence, but fertilization, blastogenesis and implantation proceed in the absence of these glycans; 2) the bisecting GlcNAc on complex W-glycans of non-hepatocyte glycoprotein(s) promotes hepatocarcinogenesis and liver regeneration after partial hepatectomy; and 3) deletion of mouse protein O-fucosyltransferase 1 generates a canonical Notch signaling phenotype identifying the Pofutl gene as an essential component of the Notch signaling pathway. We now propose in Specific Aim 1 to investigate the molecular basis of the requirement for complex and hybrid A/-glycans in oogenesis, and to identify roles of mucin O-glycans and Ofucose glycans in early development after oocyte-specific deletion of the core 1 p3GalT-l (Clgaltl) or Pofutl genes. In Specific Aim 2 the hypothesis that a triantennary, GlcNAc-terminating complex N- glycan is required for spermatogenesis will be investigated by deletion of a novel GlcNAc-transferase gene that is expressed almost exclusively in testis and by eliminating complex and hybrid vV-glycans from spermatocytes, spermatogonia or Sertoli cells. The Pofutl gene will also be deleted in spermatocytes and spermatogonia or Sertoli cells to identify roles for Notch signaling in spermatogenesis. In Specific Aim 3 the hypothesis that the Mgat3 gene may function as a tumor suppressor in mammary gland will be tested in the MMTV-Polyoma middle T (PyMT) mammary tumor model which metastasizes to lung and in cell signaling assays. In addition, roles for N- and 0-fucose glycans and Notch signaling will be investigated in mammary gland development and transformation by selective deletion of the Mgatl and Pofutl genes in mammary epithelia before puberty using MMTVCre, or during pregnancy using whey acidic protein (WAP) Cre recombinase.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this project is better understanding of the contribution of prostaglandins, thromboxanes and other products of arachidonic acid in human airway function. This goal will be approached by in vitro studies of excised human lung tissue and supplemented by analogous studies of guinea pig lung. Studies will focus upon: (a) regulation of the tension of pulmonary smooth muscle by arachidonate metabolites; and (b) the role of prostaglandins and thromboxanes in the bronchospastic response accompanying anaphylaxis in sensitized lung tissue. Two experimental models will be employed simultaneously to pursue these questions. These are: (a) organ bath studies of the smooth muscle response of both lung parenchyma and large airways, both separately and together, under various experimental conditions; and (b) chopped lung model of anaphylaxis using passively sensitized human lung chiefly, but also actively sensitized guinea pig lung fragments. Radioimmunoassay for PGE, PGF2a, TxB2, PGD2 and the prinicpal metabolite of prostacycline (6-oxo-PGF1a) will be used to develop a profile of arachidonate products during antigenic challenge of sensitized human and guinea pig lung. Specific experimental protocols will evaluate the effect of basic physiologic parameters on PG production, the pharmacology of smooth muscle response to exogenous prostaglandins and thromboxanes, the effect of anti-inflammatory drugs (including anti-inflammatory steroids) on large and small airway sensitivity to smooth muscle agonists, and confirmation of preliminary evidence for the existence of a prostaglandin releasing factor derived from lung mast cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A number of studies have identified patterns of parenting that are associated with early childhood problems of offspring, including maladaptive temperament traits, aggression, and attachment problems. These early traits are harbingers of more frank mental disorder in later childhood. Recent investigations of the parenting-offspring temperament connection have suggested that these relationships come about from multiple influences, including reciprocal causation or feedback, and probably common genetic and cultural influences as well. The proposed study takes advantage of a large longitudinal data base to add information uniquely useful for making these distinctions in a three generational design. Data gathered since childhood on a large general population sample of children are to be used in models estimating the influences on parenting and child rearing practices used with their offspring, and the origins of the relationships between offspring temperament and parenting variations. The longitudinal follow-up of the parents and offspring over the transition year from infancy to early childhood (9 to 24 months) will allow the development of models in which the relative magnitude of various influences on these relationships can be estimated and tested. Comparisons of such effects in first-born nd second-born children from the same families, assessed at fixed ages of the offspring, will permit tests of hypotheses regarding the origins of within-family differences in parenting and offspring temperament. Biological markers will be examined for consistency with the theoretically proposed dimensions of temperament.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gammaherpesviruses (GHVs) establish lifelong chronic infections that place the host at risk for numerous cancers. During chronic infection, GHVs express viral gene products that stimulate host-cell proliferation and differentiation, processes thought to facilitate long-term latent persistence and contribute to tumorigenesis. However, GHVs are not acutely transforming, and cancer is rare given the high incidence of infection among adult humans, estimated at more than 90% for Epstein-Barr virus. This suggests that host cells are equipped with an intrinsic resistance to GHV-driven proliferation and cellular immortalization, but few host molecules capable of mediating this effect are known and whether these molecules functionally restrict GHV persistence and disease in vivo has not been established. Experiments described in this proposal seek to identify and confirm the in vivo relevance of host molecules with the capacity to limit GHV chronic infection and disease in three Specific Aims. Based on preliminary studies that employed murine gammaherpesvirus-68 (MHV68) infection of mice, experiments described in Specific Aim 1 test the function of host tumor suppressor protein p53, a critical molecule for preventing neoplastic disease, as an innate barrier to MHV68 latent persistence and cellular transformation. Experiments presented in Specific Aim 2 seek to define mechanisms whereby p53 limits MHV68 latent infection. Experiments in Specific Aim 3 will provide the first in vivo test of the long- standing hypothesis that latency-associated nuclear antigen, a conserved GHV disease determinant capable of inhibiting p53 functions in tissue culture, is necessary to overcome a p53-mediated restriction to enable long- term latent infection. These experiments harness the powerful mouse and MHV68 genetic systems to address a question that is fundamental to our understanding of the GHV-host dynamic. Further, we anticipate that results of this work will inform new therapeutic approaches aimed at enhancing p53 functions to limit or prevent GHV-related cancers, especially in high-risk patients such as HIV-infected individuals or solid-organ transplant recipients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Emerging evidence suggests that racial barriers in the patient-physician relationship may be an important source of disparities in the quality of health care. These barriers include cultural dissimilarity between doctors and patients; social discomfort or distrust when interacting with people from other racial or ethnic groups (\"interracial anxiety\"); and negative racial bias or stereotyping of minority patients by physicians. Training in \"cultural competence\" has become the primary vehicle for reducing the impact of racial barriers betweens patients and physicians. To date, however, there has been limited empirical evidence to inform the content of cultural competence training. Specifically, there has been scant research on the nature and influence of specific racial barriers in the patient-physician relationship, and on the knowledge, attitudes, and skills needed to reduce them. The objective of this developmental grant application is to develop and test tools necessary to measure racial barriers and cultural competence in the patient-physician relationship, as a first step towards assessing their role in causing and mitigating, respectively, racial disparities in health care quality and outcomes. Our specific aims are to: 1) develop a psychometrically sound instrument measuring cultural competence among physicians; 2) test the reliability and validity of scales measuring interracial anxiety among physicians; and 3) test the validity of a computerized, interactive tool measuring racial bias among physicians. We will use a multi-step approach to achieve these aims, including expert scientific review of draft survey items, cognitive interviews with practicing physicians, a national physician survey, and psychometric analysis. Developing and testing measures of cultural competence and racial barriers in the patient-physician relationship will provide essential tools necessary to advance the study of racial disparities in health care. These measures will serve at least three crucial functions. First, they will allow us to empirically assess the role of physician bias, and other racial barriers in the patient-physician relationship, in explaining disparities in the quality of health care. Second, they will allow us to determine which (if any) facets of the broad concept of cultural competence reduce racial barriers and in turn reduce health care disparities. Both of these steps will help inform the content of cultural competence training and other interventions to reduce disparities. Finally, rigorously developed measures of racial barriers and cultural competence will provide needed evaluation tools for judging the effectiveness of cultural competence programs, many of which are currently being developed under NIH sponsorship. The aims of this project are to develop and test tools needed to explore the role of racial barriers in the patient- physician relationship as a source of disparities in the quality of health care, and the role of cultural competence among physicians as a means to mitigate those barriers. These tools are needed to inform and evaluate future interventions to reduce pervasive racial disparities in health care quality and outcomes. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This P41 Biotechnology Resource Grant application seeks renewed support for our successful efforts to develop and apply innovative neuroimaging technologies within the highly integrated multimodal framework of the Center for Functional Neuroimaging Technologies (CFNT). The overall goal of this established NIBIB Biomedical Technology Resource Center (BTRC) is to provide advanced technology resources to more closely examine, and thus better understand, the human brain in health and disease. To this end, we seek to develop new techniques and advance existing technologies to acquire and analyze functional images of the working brain, with unprecedented physiological precision and spatiotemporal resolution, and to deploy these innovative tools to promote investigation of complex neuroscientific questions. Through coordinated research and development, collaborative research, service use, training and dissemination activities, our BTRC has built a standard of excellence in developing, sharing, and supporting the use of multimodal imaging tools that have consistently advanced capabilities for research that spans many basic science and clinical domains. Central to this effort are our four Technology Research and Development (TRD) projects to improve and extend techniques for non-invasive magnetic resonance image analysis (Project 1) and acquisition (Project 2), electromagnetic source imaging (Project 3), and optical neuroimaging (Project 4). Directly motivating the Aims of these TRD projects is a strong network of Collaborative Projects, which both challenge the TRDs to continue to innovate the next-generation neuroimaging tools, and reciprocally, employ the new tools we develop to drive their own research forward in new directions. Another essential element in this framework is the extensive application of our resources by a wide and diverse Service Users community. Finally, the TRDs, Collaborative and Service Projects contribute to the BTRC's Training and Dissemination mission, which additionally includes multiple dedicated Fellowships and Workshops, strong web presense, and important industrial partnerships, providing multiple channels to share the knowledge needed to apply the tools we develop with the scientific community locally, nationally, and internationally.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have used recombinant DNA techniques to construct a series of vectors which function in both Escherichia coli and mammalian cells. These constructs fall into two categories: shuttle vectors and mammalian expression vectors. Shuttle vectors are designed to permit efficient bidirectional transfer of genes between Escherichia coli and mammalian tissue culture cells. We have applied bacteriophage lambda site-specific recombination mechanisms towards developing a new class of such vectors. Mammalian expression vectors are designed to facilitate study of eukaryotic promoters, activators, and other transcriptional regulatory elements. We have constructed expression vectors which permit the Escherichia coli enzyme chloramphenicol acetyltransferase to be used as a rapid, sensitive and specific assay for function of such elements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Objective To characterize the prevalence and causes of endometriosis in the aged female monkey, and to assess immune alterations associated with endometriosis. In the course of a study on the long-term effects of dioxin exposure, Drs. R. Bowman and S. Rier discovered that female monkeys exposed to either 5 or 25 ppt TCDD for 4 years were at greater risk for developing endometriosis as they aged (Rier et al., 1993, Fundamental and Applied Toxicology). 71% of females in the high dose group and 43% in the low dose developed endometriosis, in contrast to 33% of the control subjects. These females are now approximately 24 years of age, and provide a unique opportunity for assessing endocrine and immune sequelae associated with this chronic disease. We have been evaluating lymphocyte cytolytic responses and in vitro cytokine production in these subjects. In addition, we have been investigating the natural prevalence of the disorder throughout the colony by means of an in-depth analysis of necropsy records over the last 15 years. Recently, we have identified certain family lineages of monkeys at greater risk for this gynecologic condition. Future studies will focus on predictors of disease in offspring in these high- and low-risk families. Key words endometriosis, aging, inflammation, cytotoxic responses, female toxicology", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mycobacterium tuberculosis has a unique repertoire of virulence mechanisms and generates a diverse spectrum of clinical disease. M. tuberculosis has two large families of proteins, the PE and the PPE proteins, which are likely to be critical to virulence. The PE and PPE proteins are rare in nonpathogenic mycobacteria but account for four percent of the M. tuberculosis genome. Many are secreted or surface localized and thus perfectly positioned to play critical roles in host-pathogen interactions. It has been difficult to use genetics to define the function of these proteins because of the sheer number of potentially redundant gene products. However, the fact that PE and PPE proteins are secreted in the absence of a signal sequence for the general secretory pathway suggests that there is an alternative secretion system for these proteins. In Aim 1, we will identify and target this secretion pathway in M. tuberculosis. We expect that by disrupting the appropriate processing of some or all PE and PPE proteins, we will define the contribution of these proteins to the growth and virulence of M. tuberculosis. In the Aim 2, we will study the relationship between PE and PPE protein secretion, post-translational modification and the immunogenicity of these proteins. This work will refine our model of how PE and PPE proteins generate antigenic diversity in M. tuberculosis. Together, these data will dramatically increase our understanding of PE and PPE proteins and provide a foundation for future studies of their function and role in pathogenesis [unreadable] [unreadable] Despite the fact that tuberculosis kills millions of people annually, little is known about how it causes disease. Here we will develop tools to define the function of a unique family of mycobacterial proteins, the PE and PPE proteins that are likely to be critical to virulence. This work will provide a foundation for understanding the role of these proteins in the interaction between M. tuberculosis and the infected host. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary purpose ofthe Biostatistics Core (Core D) is to continue to provide biostatistical expertise for the design and analysis of all the protocols contained in the Program Project Grant. The Specific Aims of Core D are: Aim I. Assist in the design of experimental protocols and insure the proper sample size is selected to provide reasonable power Aim II. Facilitate data transfer and data quality assurance for all projects Aim III. Analyze and interpret the results from data gathered from the experiments Aim IV. Assist in the preparation of statistical components of presentations, reports and manuscripts Aim V. Develop and disseminate new biostatistical approaches for the analysis of the data; and Aim VI. Participate in discussions and present statistical seminars As we have done in the past, every new experimental protocol will be written in conjunction with a biostatistician from Core D who will provide design expertise and sample size justification for the animal care protocol. Members of the Core will attend the weekly Department of Hypertension and Vascular Research staff meetings. This will facilitate introductions to new PPG staff and also enable the scheduling of biostatistical seminars to cover topics of interest. We anticipate that the efforts of the Biostatistics Core staff (50%) will be divided equally with all four projects contained in this application. We reiterate that the presence of a Core provides availability of a set of biostatisticians with dedicated time and multiple years of experience with the Program Project. This availability is essential to the continuing success of the grant. Although located in a separate building the Biostatistics Core staff are a short walk from the Hypertension and Vascular Research Division, always available by e-mail, and always available for face-to-face meetings to discuss project design and contribute to the preparation of both manuscripts and grants. RELEVANCE (See instructions): The presence of biostatistical expertise is synergistic with the scientific expertise of the investigators in the Program Project. Their presence improves the scientific output from the PPG.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed interdisciplinary training program at the University of Minnesota combines graduate training in the computational, chemical, physical, and engineering sciences with graduate training in neuroscience. Neuroscience is a highly interdisciplinary field that uses a variety of experimental approaches to understand the development, structure, and function of the nervous system. As neuroscience matures, the need grows for quantitative modeling, physical and chemical insights, advanced technologies, and state-of-the-art hardware and software that the computational, chemical, physical and engineering sciences can provide. Therefore, interdisciplinary graduate training is needed to take the maximum advantage of these opportunities. The proposal incorporates the strengths, resources, and administrative structures of several existing graduate programs and the University of Minnesota Supercomputing Institute with an interdisciplinary faculty with diverse research interests, to provide a new paradigm in graduate education. We propose to offer 5 three-year Fellowships each year to attract outstanding pre-doctoral students. Enrolling in existing degree programs in Biomedical Engineering, Chemistry, Computer Science, Mathematics, Neuroscience, Physics and Scientific Computation. The Fellows will be trained across disciplines using a variety of tools including special interdisciplinary coursework, research rotations, dual thesis advisors, special seminars and symposia and unique training opportunities. Each trainee's thesis work will cross the disciplines of neuroscience and the physical/computational sciences. An advisory system will help guide students through the program. Also, several mechanisms are proposed to evaluate the effectiveness of the training program. The trainees will receive instructions in the responsible conduct of research. The proposal documents the efforts that the training program and University will make to ensure that traditionally underrepresented students are recruited and included in the program. On completion the trainees will be prepared for research careers in academia, industry and government. The overall goal is to train the next generation of scientists who can bridge the gap between biology and the physical/computational sciences. Broader impacts include advancing our understanding of the brain, cross-fertilization of the disciplines, and establishing a new model for interdisciplinary graduate training.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During the past three years the applicant and his students have developed highly specific methods for the isolation of native plasminogen from human, bovine and ovine plasma. Their methods, which are now used worldwide, demonstrated for the first-time that affinity chromatography can be used to purify a proenzyme. The applicant proposes during this grand period to (1) develop affinity chromatography methods for the isolation of human and bovine plasmin, (2) to attempt to obtain crystals of human and bovine plasminogen and plasmin and (3) study further the mechanism of the activation of native plasminogen from human, bovine, and other species as time permits.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is an application for renewal of a longstanding program for predoctoral training in Pharmacological Sciences. Eight training slots are requested to support students during their first two years. The program seeks to meet the need for well-trained scientists who can maintain rapid progress in applying advances in biology to medicine. Rigorous training in molecular biology, genetics, biochemistry, structural biology, and cell biology, as well as pharmacology, forms the foundation of the program. These many disciplines and others such as systems biology and physiology are reflected in the research activities and classes offered to the students. The training program is a specialized area of interest within the Biological and Biomedical Sciences (BBS) graduate program at Harvard Medical School. The training program draws its faculty members from various basic science and clinical departments. The central department for this program is Biological Chemistry and Molecular Pharmacology, although faculty members from other departments play important roles in the training program. The various departments and the training grant faculty are highly interactive. The research activities of the training grant faculty span a broad spectrum of pharmacological sciences with multiple areas of research strength. Students in the program are closely advised and monitored, both before and after starting dissertation research. In their first year, they take core courses covering multiple disciplines in basic biomedical sciences, and a course that stresses reading original research papers and critical thinking. They are required to take a core pharmacology course. They go on to take advanced courses in pharmacology and in areas relevant to pharmacological sciences including human biology, which also stress critical and quantitative thinking. Full time dissertation research follows course work, laboratory rotations, and qualifying examinations. Students receive training in teaching. They participate in multiple other important activities of the training program including a seminar series, a yearly Symposium, and a journal club. This training plan should ensure the strengthening of a program that aims to train students to go on to match or even exceed the accomplishments of previous trainees who now fill leadership positions in pharmacological sciences.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal is to investigate the regulatory role which extracellular matrix (ECM) exerts on type II pneumocytes. The extracellular components of the lung have long been known to influence the physical/mechanical properties of the tissue and thereby to play an important role in the function of normal and diseased tissue. alterations in the content and/or organization of connective tissue elements are involved in pulmonary fibrosis and emphysema, both of which are of extensive clinical significance. Recent reports of the effects of ECM on cellular proliferation and differentiation make it clear that ECM constituents cannot be regarded as an inert scaffold, but that they exert significant effects at the metabolic level. The overall goal of the proposed research is thus to investigate the hypothesis that the ECM of the lung exerts a regulatory effect on differentiated metabolism and function of type II pulmonary epithelial cells. The approach involves definition of the effects of biologically significant ECMs on type II cell morphology, as well as on retention of a typical metabolic profile. Specific metabolic steps at which these effects are exerted will be identified and these changes will be attributed to specific ECM components and/or characteristics. ECMs to be investigated range from well-defined materials, including collagens, laminin and fibronectin, to biological matrices of complex composition, including \"biomatrix\" isolated from whole lung and \"matrix gel\" elaborated by an EHS-tumor. Parameters to be monitored, which appear to provide an index of differentiated type II cell function, include cell viability, morphology, size and shape; synthesis and secretion of phospholipids and proteins associated with surfactant production; the presence of typical cytoskeletal and/or membrane characteristics, including expression of specific keratins and activit;y of Beta-adrenergic receptor-ligand interations; and the quantity, activity ratio and isozyme pattern of the cyclicAMP-dependent protein kinase system. Further investigations will extend these baseline observations to examine the effects of paraquat-induced oxidant damage on ECM function and on the interaction of type II cells with their substratum. Together, these experiments will define the biochemical mechanisms by which ECM components influence maintenance expression and physiological control of differentiated metabolic function of type II cells, as well as how these influences may be affected by oxidant damage.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our previous research established the pattern of acquisition of cell surface receptors during normal human myeloid differentiation. These membrane markers, now being used to examine leukemic myeloid differentiation are detected on cells from only a minority of patients. The objectives of this proposed research are: (1) to induce differentiation in freshly isolated human acute nonlymphoid leukemic cells (ANLL) by incubation with known inducers of differentiation and examine cells for differentiation, viability and recovery; (2) to assess differentiation by cell surface markers for IgG and complement, morphology, and cytochemistry; (3) to investigate whether 5'-nucleotidase and microviscosity can be used as new markers of myeloid membrane maturation; (4) to induce membrane differentiation using HL-60 cells (a human acute promyelocytic leukemia cell line) and clone these cells to examine the mechanism of induced membrane marker expression; (5) to investigate whether chemotherapeutic drugs also will induce differentiation in both HL60 and freshly isolated leukemic cells; (6) to compare in vitro induction of differentiation to in vivo effects using a diffusion chamber technique in mice; and (7) to compare in vitro chemotherapeutic results with in vivo clinical response in patients receiving the same agents. The overall objective of this research is to provide a useful in vitro model to aid in the diagnosis, treatment, and prognosis of ANLL.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "SUMMARY Metastases of tumors are associated with more than 90% of cancer deaths. Despite years of therapeutic development, mortality has improved only incrementally by few months at best. The preclinical development phase for modern metastatics currently misses critical quantitative information on metastatic progression. Multi-point in vivo observation of early metastasis and quantitative assessment of its development would enable unprecedented precision of longitudinal control. PhotoSound Technologies, Inc. proposes a novel platform for in vivo molecular imaging that addresses the critical barrier in quantitative preclinical imaging of metastasis via contrasted dual-modality 3D imaging approach. The proposed project promises to enable in vivo quantification of numbers and volumes of early metastatic tumors, which is critical for preclinical development of future anti-metastatics. The platform integrates a 3D photoacoustic tomography into a multi-modality imaging platform with a coregistered 3D fluorescence unit (PAFT) and a switchable optical nanoprobe targeted to the studied metastatic cells. The nanoprobe has capability to activate optical and fluorescence contrasts upon external illumination with safe levels of pulsed laser radiation, and it was designed to maximize benefits of dual-modality PAFT imaging. The fluorescence imaging component of PAFT is used to boost detection sensitivity by providing low-resolution spatial constraint for the distribution of activated nanoprobes, which are then precisely mapped in 3D by photoacoustic imaging component. The ultimate objective is to maintain the molecular sensitivity of state-of-the-art fluorescence techniques, while boosting spatial resolution of the detected metastasis 10-fold. Current trends on $1.5B market of in vivo small animal imaging favor commercial introduction of the proposed multi-modal imaging platform, which is designed for table-top application and has a 3D anatomical reference component implemented through a photoacoustic unit. PAFT could be also used as a universal instrument for 3D functional imaging of volumetric blood content and oxygenation without a need for any contrast agent, imaging of various NIR absorbing probes and bioluminescent cells. Such versatility would be attractive for Animal Research Facilities engaged in a broad spectrum of fundamental and preclinical imaging studies. Phase I project will demonstrate feasibility for the PAFT-nanoprobe imaging platform to detect and quantify early metastasis in a preclinical murine model. The focus of Phase II will be the development of a commercial imaging platform that is optimized to monitor the in vivo effects of anti-metastatic therapies. When our system is benchmarked against the current standards of optical imaging, we expect to find a 10x increase in spatial resolution enabling detection of metastases separated by only 0.5 mm, as well as quantitative assessment of individual tumor volumes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gene therapy using adenoviruses is limited by the inability to selectively deliver DNA to specific targets. Retargeting can be accomplished by introducing fibroblast growth factor (FGF) onto viral capsids through its conjugation to neutralizing anti-viral antibodies. As a result, vectors bind to cells not through their native receptors, but rather through FGF receptors overexpressed on tumors and proliferating endothelium. In addition, an increased transduction efficiency is accomplished, which we hypothesize results from the high affinity of FGF receptors (Kd= approximately 2 pM) and their ability to traffic to the nucleus. In this application we propose to examine the mechanisms of FGF retargeting in vitro. Specific aim one will define the roles of FGF and adenoviral receptors in cell surface binding and transduction. Specific aim two will confirm the intracellular pathways of virus internalization and gene delivery. Phase II studies would use this information to rationally develop in vivo models for examining the influence of FGF-retargeting on viral toxicity and immunogenicity, and for the development other ligand-viral conjugates. The data generated will validate the use of growth factor retargeting for gene therapy, and establish the groundwork required for its use in clinical trials. PROPOSED COMMERCIAL APPLICATION: The proposed studies are designed to develop therapeutic products for gene therapy. Initial drug candidates will target adenoviruses to growth factor receptors overexpressed on tumor cells and associated vasculature, while at the same time eliminating normal viral tropism for uninvolved organs. Targeted delivery will allow transduction of tumors with cytotoxic proteins and other anti-proliferative agents, and accomplish antitumor activity by destruction of tumor cells and/or proliferating endothelium. This approach will allow application of the growing number of adenoviral vectors to cancer gene therapy by eliminating their major limitation, namely non-targeted delivery to normal tissues.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The main objective of our application to NIH is to establish the Chicago Tri-lnstitutional Center for Chemical Methods and Library Development (CTCMLD), which is designed to address the fundamental challenges of high-throughput organic synthesis and to provide major advances that will significantly facilitate the development of small-molecule libraries for broad biological screening. Our CTCMLD application is strongly supported by the three participating institutions and the Chicago Biomedical Consortium, which provided a $2 Million Lever Award to match the NIH funding. The CTCMLD will have several major goals listed below. Center Objective 1: Conduct four highly integrated projects that are designed to significantly facilitate the generation of chemical diversity and assembly of high-quality small-molecule libraries. The four proposed projects are: (a) practical synthesis of new nitrogen-based heterocyclic libraries (leaders: Scheldt and Gevorgyan); (b) multi-component assembly of polyketide libraries (leader: Yamamoto); (c) assembly of scaffold-unbiased small-molecule libraries (leader: Kozmin); and (d) development of tailored surfaces for new reaction development, miniaturized library synthesis and small-molecule immobilization (leader: Mrksich). The scientific projects will be carried out at the highest level, resulting in the development of an arsenal of new synthetic methods, which in turn will enable the generation of approximately 20,000 new compounds of 20 mg scale in highly efficient, parallel manner. Center Objective 2: Training the next generation of scientists, who will develop and arsenal of new methods and strategies for high-throughput organic synthesis. Center Objective 3: Establish core research facility for library synthesis, characterization and cheminformatics. The Core facility will have three highly integrated components: High-Throughput Synthesis Component located at the University of Chicago, Hit-to-Lead Development Resource located on Northwestern campus and Cheminformatics Core at the University of Illinois at Chicago. Objective 4: Provide broad outreach to the biology community enabling rapid and efficient screening of new compound libraries. In addition to providing new compounds for the NIH Roadmap Molecular Libraries Initiative, we will collaborate with three major high-throughput screening centers in Illinois, and many individual biology laboratories. The increased connectivity and synergy resulting from the work of the CTCMLD will represent a major advance in the Chicago area's capabilities and a paradigm shift in how the Chicago academic community will approach and solve problems at the interface of chemistry, biology and medicine. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long time goal of our studies is to delineate the molecular mechanism that regulates the immune defense against invading pathogens. The study proposed is focused on the role of the family transcription factors of interferon regulatory factors (IRF) in the innate and acquired responses to viral infection. These factors were shown to participate not only in the induction of Type I IFN genes but also in the induction of other cytokines, chemokines and genes directly involved in the antiviral and anti-inflammatory responses. Over the past years we have sequentially isolated IRF-3, IRF-7 and IRF-5 and have shown that these factors serve as direct transporters of virus induced signaling. The function of these factors is not redundant since these factors are expressed in different cell types and stimulate a profile of distinct genes. Among these three factors IRF-7 plays a limiting role in the induction of IFNa that was shown to be important both for the innate and adaptive immune responses. It is our hypothesis that these three IRFs, especially then IRF-7, are regulating both of these immune responses. The key objective of this application is to validate this hypothesis and to determine the molecular mechanism by which IRFs activates the targeted genes in response to pathogens. [unreadable] The study has three aims. [unreadable] In Aim#1 we shall examine the molecular mechanism involved in the IRF-7 mediated activation of antiviral genes. [unreadable] In Aim#2 we shall determine whether Toll 3 and Toll 9 responses to dsRNA and CpG DNA respectively results in the activation of IRF-3, IRF-5 and IRF-7. [unreadable] In Aim #3 we shall determine whether the stimulation of macrophages and dendritic cells by virus, dsRNA and CpG DNA targets similar or distinct set of antiviral genes. [unreadable] We believe that the basic understanding of the role of IRF factors in pathogen induced cellular responses will provide a new therapeutic platform for the treatment of the immune and inflammatory disease. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Physician Postdoctoral Research Training in Perioperative Medicine (PPRTPM): More than 41,000 anesthesiologists practice in the US, but only a small number are physician-scientist researchers in this field of anesthesiology/perioperative medicine. In FY13 fewer than 120 MD or MD PhD anesthesiologists were PI on an NIH grant. Nevertheless, anesthesiology is a requisite component of every medical center, hospital and outpatient surgical facility because of the 40 million surgical procedures performed annually. Advances in pharmacology, imaging, and technologies used in interventional/procedural medical care make the field of anesthesiology/perioperative medicine rich with research career development opportunities. The goal of this program is to address the as yet unmet need to train more committed physician-scientist anesthesiologists. PPRTPM program direction, aims and objectives: Program leadership will be provided by Dr. David Eckmann, serving as contact PI, with further leadership provided by additional Executive Committee members Dr. Gordon Barr, Dr. Roderic Eckenhoff and Dr. Andrew Ochroch. All are members of the University of Pennsylvania Department of Anesthesiology and Critical Care. Their responsibilities will be focused on directing four theme-based research training tracks, three of which are devoted to laboratory research trainees and one of which is devoted to clinical research trainees gaining research training in perioperative medicine. The aims of the PPRTPM are to: Identify, recruit and foster research trainees, both anesthesiology residents and clinical sub-specialty fellows willing to commit to training and career development in perioperative medicine research Match up trainees strengths and interests with mentoring teams Provide guidance for structured learning opportunities Maximize the opportunities for mentored research and career mentoring The objectives of the PPRTPM are to: Train a cadre of committed physician-scientist anesthesiology researchers to advance the field of perioperative medicine research. Provide these individuals with the skill sets and foundation for career advancement Encourage leadership and innovation PPRTPM goals-to pursue the aims and objectives through a training program consisting of: Didactic opportunities, including core requirements and courses designed to provide research skills Seminars, workshops and a journal club focusing on research and progress in the field Mentoring with a team approach, mentor training and scholarship oversight Programmatic interactions with mainstream research through local/national professional interactions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The rapid accumulation of genome sequences and protein structures during the last decade has been paralleled by major advances in sequence database search methods. The powerful Position-Specific Iterating BLAST (PSI-BLAST) method developed at the NCBI formed the basis of our work on protein motif analysis. In addition, Hidden Markov Models (HMM) and protein structure comparison methods were applied. During the last year, we made further progress in detailed analysis of the classification, evolution, and functions of several classes of proteins. Specifically, we studied in detail the protein domains that are involved in eukaryotic RNA interference mechanisms and showed that the protein machinery of eukaryotic RNAi was pieced together from ancestral archaeal, bacterial and phage proteins that are involved in DNA repair and RNA processing. We also used computational methods to identify a novel prokaryotic toxin-antitoxin systems that are predicted to function via RNA binding or cleavage and other, diverse mechanisms. We explored the evolution of the eukaryotic phagocytosis system and demonstrated the presence of actin-related domains in a distinct set of crenarchaeota and korarchaeota, suggesting that the archaeal ancestor of eukaryotes might have already possessed a primitive form of branched cytoskeleton that facilitated engulfment of other prokaryotes. We further investigated the evolution and genomic context of prokaryotic homologs of the eukaryotic argonaute protein (the key component of the RNAi system) and showed that the genes encoding these proteins tend to be located within \"genomic islands\" that also contain many other genes for various defense systems. These findings suggest that the prokaryotic homologs of argonaute are components of a novel defense system that also includes a variety of other putative nucleases that we identifies using sensitive methods of domain analysis. We additionally contributed to experimental and structural analysis of a variety of domains previously identified in our computational studies, in particular, the key components of the prokaryotic CRSIPR-associated acqauired immunity system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Congestive heart failure is a major public health problem affecting millions of patients in the United States. Because heart transplantation can only be offered to a minority of these patients, pharmacological therapy remains the mainstay. Survival has improved with the use of current pharmacological therapy, but nevertheless mortality is high in patients with severe congestive heart failure. If beta-blockers reduce mortality by 25% or more, this finding would have major implications in the care of the patients with this common condition. This project seeks to evaluate the efficacy of beta-blockers to improve the survival of a common clinical condition with a high mortality rate, Class III-IV congestive heart failure. Bucindolol is an investigational non-selective beta-blocker with mild vasodilator properties. It has already been used in several hundred patients with heart failure and has been extremely well tolerated. Patients are randomized between Bucindolol and plcebo, titrated upward to the maximal tolerated dose of Bucindolol and followed for 4.5 years.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study is designed to examine the relative value of different self-reward strategies in the long-term maintenance of breast self-examination (BSE) practice among women trained to do BSE. These self-reward maintenance procedures will be implemented by breast screening educators coordinated through the Community BSP Office in a Florida community beginning Year 2 of the Program Project. The three self-reward maintenance conditions will be: (a) self-reward instructions and materials delivered at the end of the BSE training session with no monthly prompts to encourage their use; (b) self-reward suggestions delivered by the BSE Maintenance Coordinator through the mail each month contingent upon evidence of BSE performance; and (c) external, monetary rewards and self-reward suggestions delivered through the mail on an intermittent schedule contingent upon BSE practice. Subjects will be 1800 women who attend a community-based breast screening training session in the study site where community-based BSE training efforts are underway. Training groups will be randomly assigned to conditions urging a randomized block design for six months until 1800 women have been enrolled in the study. Subjects will receive their respective self-reward manipulation for one year, followed by a second year in which all external rewards and mailed monthly self-reward prompts will be withdrawn. BSE frequency and quality, and self-reward use will be assessed through a telephone survey of all women in the three conditions at one year following BSE training and again at two years following initial training. Additionally, for a sub-set of the women in each condition, proficiency will be assessed in personal interviews using silicone breast models at each of these two measurement times.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Biomedical and rehabilitation research depends increasingly on state-of-the-art imaging modalities to visualize cells, tissues, and organs. At VAPAHCS, our research program that depends on cutting edge imaging includes the study of such important diseases and disorders for the Veteran population as Alzheimer's disease, traumatic brain injury, cancer, autoimmune disorders, degenerative disorders, vascular disease, spinal cord injury, and traumatic injuries to muscle, bone, and tendon. For so many lines of research, disease modeling and experimental therapeutics have been revolutionized by the development of imaging systems that allow for the tracking of cells in vivo in living animals over time. This approach has been used extensively at VAPAHCS for the past 10 years for the analysis of such processes as non-invasive disease monitoring, stem cell therapies, cell transplantation, tumor growth, in vivo gene expression analysis, immune cell tracking, and neurogenesis, and remains a critical element of our research program. The Perkin/Elmer IVIS Spectrum In Vivo Imaging System is a top-level research instrument that combines cutting edge technology with quality and exceptional ease of operation. This instrument will replace an outdated version that is almost ten years old and failing regularly despite routine maintenance. Furthermore, the new system features includes a camera of enhanced sensitivity providing increased spatial resolution and software that provides more powerful image processing and analysis for the key functionalities of Bioluminescence Imaging (BLI), Fluorescent Light Imaging (FLI), Cherenkov light, 3D tomography, and Spectral Unmixing. The current, outdated equipment is used up to 10 hours a day, including weekends, by investigators at VAPAHCS. The ability to replace our outdated and failing system with a new and updated system would greatly facilitate and accelerate the research program at VAPAHCS. We have outlined an administrative process to oversee the use and maintenance of the new equipment, and we outline a plan for institutional support to provide the resources for equipment service over the next ten years. Overall, the purchase of a new Perkin/Elmer IVIS Spectrum In Vivo Imaging System is critically important for the ongoing biomedical and rehabilitation research at VAPAHCS in order to allow investigators to continue to be at the forefront of research in their respective fields with access to state-of-the-art equipment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During development, sensory experiences produce synaptic modifications that specify the capabilities and limitations of brain function in adults. In adults, very similar modifications appear to be the substrates of learning and memory. Therefore, a question of great significance is how synapses in the brain are modified by sensory experience. Mechanisms for long-term synaptic depression (LTD) and potentiation (LTP) have been identified, as has a mechanism for regulating the conditions for LTP induction. However, it remains to be determined the extent to which these mechanisms contribute to naturally occurring modifications in the brain. The mouse visual cortex provides an excellent model system for addressing this question. Visual cortex is well-known to be modified by simple manipulations of experience, such as depriving one eye of vision, and the mouse can be genetically modified to test specific hypotheses about molecular mechanisms. Moreover, while this type of plasticity changes over the course of postnatal life, it persists in adult mouse visual cortex and therefore can provide insight into how age alters the qualities of synaptic plasticity in the cerebral cortex. The aims of this proposal are to determine (1) the qualities of visual cortical plasticity across the life-span, (2) the contributions of LTD mechanisms to deprivation-induced response depression in vivo, (3) the contribution of LTP mechanisms to experience-dependent response potentiation in vivo, and (4) if changes in NMDA receptor subunit composition are permissive for experience-dependent response potentiation in vivo. These aims will be accomplished by taking advantage of the special expertise of this Center in mouse genetics, chronic recording, and the molecular bases for synaptic plasticity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lymphedema (LE) following treatment for breast cancer is the most common form of secondary LE in the industrialized world. It occurs in 20% to 87% of patients following treatment for breast cancer and results in significant disability. At the present time, the definitive phenotypic, genotypic and epigenotypic predictors that place patients at highest risk for the development of LE are not known. Therefore, the specific aims of this study, in a sample of patients following treatment for breast cancer, are to: determine genetic predictors of LE using a candidate gene approach and evaluate for epigenetic changes, as measured by DNA methylation and subsequent gene expression, in candidate genes associated with the diagnosis of LE. The secondary aims of this study are to: evaluate for latent classes of women with distinct phenotypic predictors of LE; and evaluate for differences in symptoms, functional status, and QOL outcomes between women with and without LE and among the latent classes with LE. The results of this study will provide new information on the underlying mechanisms for LE and allow for the development and testing of novel approaches to prevent or reduce the negative effects of LE.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The central hypothesis of the project is that deubiquitinating enzyme UCH-L1 is an essential regulator of vascular lesion formation. The proposal will uncover a novel mechanism that UCH-L1 serves as a key molecule in assembling inflammatory signaling complex thereby fine tuning vascular inflammatory responses and lesion formation. It is our view that the outcome will provide novel insight into the understanding of the complex sequelae of inflammation in vascular lesion formation. Our hypothesis will be tested by uitilzing UCH-L1 gain- and loss-of-function approaches in vitro and in vivo to address three specific aims as follows; Aim 1. Define an essential role of UCH-L1 in regulating VSMC inflammation in vitro Aim 2. Define molecular mechanism of UCH-L1-mediated inhibition of VSMC inflammation in virto Aim 3. Determine an essential role of UCHL1 in regulating VSMC inflammation and vascular lesion formation in vivo", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this research is to determine the neural basis of aggressive behavior, first in cats, ultimately in monkeys, with a view to clarifying the neural basis of aggressive behavior in man. We propose to observe naturally occurring aggressive behavior in cats with a view to determining the forms its takes, and to secure experimental control over its occurrence. We use aggressive behavior elicited by electrical stimulation of the brain to help us determine the mechanisms, and anatomical and physiological techniques to identify them. This knowledge will serve as the basis for the investigation of the aggressive behavior that occurs naturally. BIBLIOGRAPHIC REFERENCES: Huang, Y. H. and Flynn, J. P. Unit activities in the hypothalamus and midbrain during stimulation of hypothalamic attack sites. Brain Research 93: 419-440, 1975. Flynn, J. P. and Brandler, R. J. Patterned reflexes during centrally, elicited attack behavior. In: The Neural Bases of Violence and Aggression, University of Texas Press, 1975.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this application is to request NCRR matching funds for the construction of a state-of-the-art research and training facility for the Psychology Department of Tufts University. The Psychology Department has 13 researchers engaged in a variety of collaborative bio-behavioral research endeavors focused on basic and clinical research questions in biological, cognitive and social psychology. These researchers have vibrant, well funded and growing research programs addressing important mental health issues including learning disabilities, dug addiction,, nutrition affects on behavior, anxiety disorders, infant motor and social development, mental retardation and social communication. However, their efforts are severely hampered by an aged/inadequate research and administrative facility, no capacity to expand, and by being housed in three by buildings The proposed plan addresses these problems by consolidating the department into two neighboring 21st century research facilities. The new 28,800 square foot Psychology Research Building (for which funds are being requested) will contain laboratories for researchers engaging in human research, for conducting neurochemical and neuroanatomical studies as well as offices for all faculty members, graduate students, post-docs, research staff and administrative staff. The second building, Bacon Hall (for which funds are not being requested), will continued to serve as the primary laboratory facility for faculty members who use experimental animals in their research programs. The consolidation of faculty, students, research and administrative staff into a new building, and the proximity of the two Psychology Department facilities will allow for much greater efficiency as well as future expansion for current and anticipated new faculty.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to develop a unique skin coating formulation for use as a combination occlusive dressing and drug delivery vehicle as a topical treatment for psoriasis, a disease which affects 1% to 3% of the world s population. Currently used occlusive dressings are associated with local side effects and have limited patient acceptability because of poor cosmetic properties. The specific aims of the project are * to optimize skin coating formulations for use as a sustained-release steroid treatment for psoriasis * to evaluate skin coating formulations with encapsulated steroids in vivo in an animal model * to evaluate skin coating formulations with encapsulated steroids in vivo in a human clinical trial on psoriasis patients. PROPOSED COMMERCIAL APPLICATION: The cost of psoriasis care in the United States is estimated to be between $1.6 and $3.2 billion annually. By combining occlusion and drug delivery in a convenient, cost effective one-step process, we seek to improve outcomes and decrease the cost of treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sensory hypersensitivity is commonly seen in FXS patients and the FXS mouse model - the Fmr1 knockout (KO). Recent data suggests that this abnormality stems from hyperexcitability in sensory circuits. We have established that cortical microcircuits are hyperexcitable in the Fmr1 KO mouse model, and that sensory responses are enhanced in Fmr1 KO mice and FXS patients. Thus, investigation of sensory sensitivities is clinically relevant, but perhaps more important is the promise of sensory system studies to advance understanding of the mechanisms and consequences of hyperexcitability in neocortical circuitry that could represent a primary pathophysiological factor impacting the development of a wide range of perceptual, cognitive, and language skills in FXS. Further, we have identified biochemical signaling mechanisms that may underlie hyperexcitability involving processes that we and others have uncovered that can be examined in detail in KO mouse models and tested in FXS patients to develop a foundation for novel therapeutic development. The striking consistency of findings across levels of investigation and species offers an unprecedented opportunity to investigate mechanisms of brain dysfunction in a mouse disease model and translate it directly to patients - a multidisciplinary mission that is ideal for a Center environment. Our Center is organized to pursue precisely this aim with a tightly integrated and highly novel scientific program of translational research. Project 1 (Huber/Gibson; UTSW; co-investigators) will determine the cellular, molecular and synaptic mechanisms of auditory neocortical dysfunction using in vitro brain slices in FXS mouse models. Project 2 (Razak/Etheii/Binder; UCR; co-investigators) will study auditory sensory processing deficits in vivo in FXS mouse models, test mechanisms, and examine developmental and structural correlates of these deficits. Project 3 (Sweeney/Byerly, UTSW, co-investigators) will investigate auditory cortical processing deficits using novel neurophysiological strategies in individuals with FXS. All Projects will examine candidate mechanisms of sensory hyperexcitability with an acute pharmacological probe strategy to test mechanisms of interest in parallel studies of mice and patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the proposed program we plan to study novel human glycosyltransferases. Much emphasis will be made on the identification and characterization of enzymes present in ovarian epithelial linings and fluids. Synthesis of various oligosaccharides which can be effectively employed as acceptor-substrates for certain glycosyltransferases have already been accomplished in our laboratory. A number of these synthetic substrates have proven to be very useful and valuable acceptors for glycosyltransferases from sources other than human. Our main strategy for the characterization of the product resulting from the reaction of a glycosyltransferase and its acceptor involves the synthesis of the expected saccharide which can be used as a reference compound. The structure of the saccharide isolated from enzymatic product will be established by n.m.r., mass spectroscopy, and methylation studies. We plan to employ glycosidases for the structural studies of the new product. Adequate use of newly acquired HPLC instrumentation will be made to facilitate these investigations. The proposed studies are directed toward the characterization of the spectrum of glycosyltransfeerases which are responsible for elongation of carbohydrate units O-glycosidically linked to Serine or Threonine. We are concentrating on the study of glycosyltransferases which have not been identified but whose existence is inferred from structural studies and the \"one enzyme-one linkage\" concept. Our studies will further elucidate the biosynthetic pathway of glycoconjugates. It is anticipated that glycosyltransferases of defined specificity, as proposed in this program in conjunction with glycosidases, may be of general value in further understanding the roles of specific oligosaccharide structures in the complex mileu of cell-surface glycoconjugates.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aim of this application is to develop a rapid, easy-to-use, and inexpensive multi-sample diagnostic system to identify sexually transmitted infection (STI) pathogens, such as Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) in non-traditional healthcare settings. According to the CDC, chlamydia and gonorrhea, caused by C. trachomatis and N. gonorrhoeae, respectively, are the first and second most frequently reported STIs in the US and likely the rest of the world. These STIs are often under-diagnosed, leading to delayed treatment, continued spread and higher public healthcare costs (~$16 billion annually). As a result of their prevalence and long term health consequences, there is a need for rapid, sensitive methods of detecting STIs in order for patients to get results and treatment immediately. It is important that patients receive results and initial consultation or treatment during the first patient visit, as they rarely return for a second visit. Point-of- care (POC) tests are an important strategy to address the STI epidemic, and because many affected by STIs (young and/or poor) often have limited access to traditional healthcare, POC tests that can be performed in non-traditional settings (university clinics, community public health care clinics, jails, detention centers) could greatly reduce STIs. The aim of this application is to develop a rapid, easy-to-use, inexpensive diagnostic system for identifying STI pathogens such as CT and NG. The system will combine AI Biosciences, Inc.'s novel nucleic acid extraction cartridge and a rapid real-time thermal cycler to form a multi-sample system for use in many non-traditional healthcare settings. In Phase I, a high performance, low-cost self-contained nucleic acid extraction and purification cartridge will be developed to perform a 10 min. extraction. The extracted NA samples will be placed in an thin-walled PCR card and amplified using an innovative water based cycler to perform rapid (20 min for 40 cycles) multiplexed real-time polymerase chain reaction (PCR) for CT and NG targets. We will demonstrate that the entire assay can be completed in less than 30 min. The proposed work will be carried out in collaboration with STIs and fluidic/thermal characterization experts. Our sample preparation cartridge and the PCR technologies are highly suitable for parallel, automated sample processing. This is a substantial advantage over other integrated systems being developed as all of them can only process one sample at a time, making several units necessary to meet the minimum throughput demand. The successful development and commercialization of our technology will impact how sophisticated molecular diagnostic assays can be implemented in non-traditional healthcare settings.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gene delivery to the liver for inherited metabolic disorders provides the opportunity for specific treatment of two forms of glycogen storage disease (GSD). Glycogen accumulation due to single enzyme defects represents the prototype of metabolic storage disease. A detailed biochemical understanding and availability of mouse models for GSD Ia, and GSD II make these conditions excellent examples for correction of the metabolic defect in the liver by gene replacement therapy. In order to demonstrate the feasibility of this approach, Dr. Byrne and his colleagues propose to investigate the AAV-mediated delivery of the gene for glucose-6-phosphatase (G6P) and, acid alpha-glucosidase (GAA) in animal models of GSD I and II. The proposed study is designed to examine the ability of AAV to direct the sustained hepatic expression of G6P and GAA in murine models GSD. The vector has been shown to yield high-level, long-term expression of a number of therapeutic proteins without eliciting a clinically significant immune reaction. The investigators have recently demonstrated high-efficacy gene transfer of GAA into embryonic tissues, cultured adult and neonatal rat cardiomyocytes, as well as, adult rat heart, and murine skeletal muscle in vivo using this approach. Substantial preliminary evidence in his laboratory and others demonstrates the utility of over-expression of therapeutic proteins in hepatic tissues. In principle, this system has the capability to deliver the therapeutic protein to all tissues via secretion from the hepatic platform. Additionally, correction of G6P deficiency examines the important question of direct correction of a microsomal enzyme defect by hepatocyte transduction. They now propose to evaluate this potential by assessing the effectiveness and biological impact following AAV-mediated reconstitution of G6P and GAA in animal model of GSD. The effectiveness of several hepatic specific promoters will be tested in vitro using immortalize cell lines from the representative mouse models. One of the important considerations of systemic delivery of corrective vectors will be evaluated by new technologies using MR imaging and MR spectroscopy. The efficiency of processing and targeting of lysosomal enzymes will be examined by strategies which allow for the augmentation of the phosphotransferase enzyme involved in lysosomal enzyme secondary processing or by anti-sense of critical proteins which control lysosomal targeting and thereby result in a preference for the secretory pathway. These studies will yield important new information in establishing a clinically relevant treatment for these fatal diseases and add new understanding to the basic pathophysiology of GSD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Use of Engineered Nervous Tissue Constructs to Repair Extensive Nerve Injury Each year approximately 360,000 people in the United States suffer a peripheral nerve injury, which is a leading source of lifelong disability. While a primary strategy to repair major peripheral nerve injury is to bridge the damage with autologous nerve grafts, producing nerves of sufficient length and number has posed a significant challenge. The so called gold standard in peripheral nerve graft repair (i.e. the autologous nerve graft) is limited by the time consuming harvesting of donor nerves and complications arising from the harvesting surgery. In addition, most alternative methods currently used for nerve graft repair (e.g., synthetic tubes) are limited in the length that they can span to promote repair and are typically used for gaps of less than 2 or 3 cm. Here, we propose to utilize a novel tissue engineering technique to create transplantable nervous tissue constructs for major peripheral nerve repair. The key of this procedure is to use a specially designed microstepper motor system to produce continuous mechanical tension on axons spanning two populations of neurons in culture. As our central hypothesis, we propose that engineered living nervous tissue constructs will promote recovery after major peripheral nerve injury by providing a living labeled pathway to guide host axons from the proximal nerve stump across large nerve lesions to reinnervate the target tissue. In addition to providing a pathway through the injured gap, we also propose that axons from the construct will grow into both the proximal and distal nerve stumps. In the first part of this proposal, we will repair acute extensive acute nerve injury in an animal model using our novel living engineered nervous tissue constructs. In the second part of this study, we will attempt major reconstruction of brachial plexus injury in rodents, spanning the forelimb from the vertebrae to the paw using our living nervous tissue constructs. We will directly compare the outcome of animals receiving the nervous tissue construct containing the elongated dorsal root ganglion cell cultures with groups receiving conventional autologous nerve reconstruction (reverse autologous graft), repair with a synthetic tube alone (material substrate control), no repair (no treatment control), immunosuppression control (receiving daily cyclosporine A injection) and physical rehabilitation control. In the third part of this study, we will assess the immunogenicity of our nervous tissue construct. We will investigate the hypothesis that the lack of MHC I expression in our elongated DRG neurons is the main reason that immunorejection does not occur in transplanted hosts. If successful, our novel tissue engineered nerve construct could revolutionize methods for major nerve reconstruction by providing laboratory grown 'off-the-shelf' living nerves ready for transplant in patients with extensive peripheral nerve injuries. PUBLIC HEALTH RELEVANCE: In this study we propose to use a fundamentally novel tissue engineering technique to create nerve constructs for repair of extensive nerve injury in an animal model. If successful, our findings could benefit patients with devastating nerve injuries by providing laboratory grown 'off-the-shelf' living nerves ready for transplant.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project focuses on a protein phosphatase that interacts with the phosphorylated form of RLK5, a receptor-like protein kinase from Arabidopsis thaliana. RLK5 has features characteristic of the growth factor receptor kinases, including an extracellular domain of tandem leucine-rich repeats, a transmembrane domain, and a C-terminal serine/threonine protein kinase catalytic domain. Numerous serine/threonine receptor kinases have been identified in higher eukaryotes including the TGF-?/activin receptor superfamily in animals and the receptor-like kinases from higher plants. However, the mechanisms by which these receptors transduce a signal is unknown. Interaction cloning was used to identify an Arabidopsis protein which associates with the catalytic domain of RLK5. This protein, which has been termed KAPP for kinase associated protein phosphatase, consists of three functional domains: an N-terminal type one signal anchor, a central kinase interaction (KI) domain responsible for interaction with RLK5, and a C-terminal region with homology with type 2C ser/thr protein phosphatases. KI domain association requires phosphorylation of RLK5 as shown by in vitro binding assays. Interaction of a protein with the phosphorylated form of a receptor protein kinase is reminiscent of the interaction of SH2 domain-containing proteins with tyrosine kinases. SH2 domains bind with high affinity to specific phosphorylated tyrosine residues of active tyrosine kinases and are key components of many intracellular signal transduction cascades. Interaction of the KI domain of KAPP with phosphorylated RLK5 suggests that this may be a general mechanism for receptor protein kinase-mediated cellular signaling common to both receptor tyrosine and receptor serine/threonine protein kinases. The autophosphoryl-ation sites in the catalytic domain of RLK5 will be mapped with MALDI-MS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Candida albicans is the most common fungal pathogen of humans. Its clinical importance has increased in recent years as the number of immunocompromised patients (AIDS, transplant recipients, cancer, corticosteroid treated, etc.) has expanded and the incidence of opportunistic infections has escalated. The long-term objectives of this grant are to understand the fundamental mechanisms by which pathogenic fungi communicate with each other and with their environment within human hosts, and how host hormones may regulate critical fungal processing during infection. We have described the presence of steroid binding proteins (SBPs) in C. albicans and other pathogenic fungi. These SBPs exhibit high affinity for selected mammalian steroid hormones, particularly estrogens and corticosteroids. We have also demonstrated functional responses in the fungi subsequent to hormone treatment by steroids that bind to the SBPs. These findings have led us to raise 4 hypotheses: (1) the SBPs represent hormone receptors in the fungi; (2) the fungi have endogenous hormones to match each SBP; (3) since the host hormones bind to the SBPs, the fungus responds to the hormonal milieu of the host, thus mediating physiological changes in the invading pathogen; and (4) the SBPs are evolutionarily related to the steroid hormone receptors of higher organisms. We postulate that the host-pathogen interaction contributes to differences in susceptibility to infection depending on the hormonal status of the host (e.g. gender, menstrual stage, pregnancy, treatment with birth control pills or corticosteroids, etc.). The grant will focus on the corticosteroid binding protein (CBP) and the estrogen binding protein (EBP) found in c. albicans. The aims of the grant will be: (1) to clone the genes encoding the SBPs and raise antibodies to the proteins; (2) using these molecular probes, to define the location and function of the SBPs in C. albicans; (3) among other actions, to evaluate the role of SBPs in regulating cyclic AMP, dimorphism and cellular adhesion, actions that would likely affect fungal pathogenicity; and (4) to explore the evolutionary relationship of the Candida SBPs to homologs in other pathogens, both fungal and protozoan, as well as to other species up the evolutionary tree including human. Our long-range goal is to understand better the biology of pathogenic fungi so that advances can be made in the elucidation of the molecular basis of the epidemiology of human infection and improved methods can be developed for the diagnosis and treatment of fungal disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cognitive memory and habit formation are two qualitatively different learning processes based on separate neural systems, a cortico-limbic and a cortico-nonlimbic system, respectively. To see how emotional and social behavior develop in animals whose infantile global amnesia might persist from infancy through adulthood, we have prepared monkeys with neonatal limbic lesions and followed their behavioral development. Animals with neonatal removal of cortical area TE, a higher-order visual station linked to both learning systems, serve as controls. The results indicate that neonatal TE lesions leads to a transient impairment of habit formation three months later (compared to permanent impairment seen with the same lesion in adults), whereas both neonatal and adult limbic lesions leave habit formation intact. Interestingly, data on both normal and operated infants suggest that development of the nonlimbic habit system is sexually dimorphic, and that this is due to the high testosterone levels present in male infants before and shortly after birth. At ten months of age, the infants with limbic lesions show impairment in memory formation, whereas the operated controls show significant functional sparing (compared to those that received the same lesions as adults). These findings point to greater compensatory potential after neonatal cortical than after neonatal limbic removals, indicating that association areas of the cortex are immature at birth, and may thus possess greater plasticity than limbic structures. Direct evidence of neocortical immaturity in the macaque has been provided by our neurobiological studies on opiatergic and cholinergic receptor distribution and on metabolic activity. Finally, early damage to the limbic memory system produces later socio-emotional abnormalities that are similar in many respects to the behavioral syndrome seen in autistic children.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is an open-label multiple dose study in which patients with advanced solid tumors will receive a single dose of losoxantrone as 14C-labeled compound during their first course of treatment. Blood, urine, and feces will be collected for analysis. Following enrollment, patients will receive up to 8 courses of losoxantrone, once every 3 weeks, depending on response to treatment and tolerability.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This R13 application proposes to support a 2-day annual meeting to enhance the collaborative network, and maximize the long-term national impact of the National Consortium for Multicultural Education for Health Professionals (Consortium). Funded in 2004 by NHLBI as a 5-year competitive training award program (K07) for U.S. medical schools, the Consortium uses the approach of addressing disparities by integrating cross- cultural education into health professional training. We propose to build on our success by implementing a meeting focused on the role of reflective practice to support health professional education in cultural competency and health literacy. To reduce costs, increase participation, and maximize collaboration, the meeting will be scheduled to coincide with the American Public Health Association annual meeting to be held November 2014 in New Orleans, Lousiana. The goal of this conference is to provide an opportunity for experts in cultural competency and health literacy curriculum development to network with leaders in public health education, and researchers on the impact of unconscious bias on health outcomes to further build upon the exchange of ideas which began recently among research and practice professionals in cultural competency and health literacy. Specific aims include: 1) create a network of experts in cultural competency and health literacy curriculum development and research, 2) identify common competencies, approaches and assessment methods that can be applied to education and research in cultural competency for multiple health professions; and 3) advance research and publication potential of the combined fields of cultural competency and health literacy. The meeting comprises a: 1) scientific panel session (n=30-100) on use of reflective pedagogy to teach about and assess unconscious bias that impacts patient health outcomes, (2) Learning Institute (skills- building workshop) on preparing submissions to MedEdPortal with the Association of American Medical Colleges (n=20), and 3) working session (n=30) to bring together experts from different health professions to design curriculum addressing unconscious bias for an interprofessional setting.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract Phasing, defined as the estimation of haplotypes from diploid genotype data, is a fundamental problem in medical and population genetics. Phasing is a key preprocessing step for genotype imputation algorithms employed in genome-wide association studies of diseases and complex traits, and is also important for mapping molecular QTL using allele-specific reads, detecting clonal mosaicism, inferring population structure, and detecting natural selection. Considerable resources have been invested into developing accurate phasing algorithms, but currently, unsolved challenges include: (i) incorporating large reference panels, such as the Haplotype Reference Consortium, to improve phasing accuracy (reference-based phasing), and (ii) phasing extremely large cohorts using within-cohort data (cohort-based phasing). Here, we propose an exploratory two-year research program, in which we will develop methods and software for both reference-based phasing, and cohort-based phasing, using a new data structure based on the Positional Burrows-Wheeler Transform (PBWT). We aim to make fast and accurate phasing methods and software freely available to all researchers via public phasing servers. We will also explore the early and conceptual stages of developing PBWT-based methods for reference-based imputation as well. Our team has multiple strengths: our statistical and computational expertise; our track record of producing practical, publicly-available software packages for a broad range of applications in statistical genetics that are widely used by the community, and our data-driven approach to methods research. We will guide our methods development using data from 500,000 samples from the UK Biobank, and will work closely with the Haplotype Reference Consortium (see letters of support).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our aim is to understand chemotactic migration in cells from a biochemical standpoint. To this end we have investigated this process in both phagocytic cells and connective tissue-type cells. Studies on the regulation of phagocyte chemotaxis, using formylated peptides attractants, have indicated that a tumor-derived peptide inhibits migration by reducing receptor availability, inhibiting hydrolysis of the attractant, and decreasing membrane lipid turnover. Receptor mediated internalization of attractant may be required for chemotaxis since inhibitors of translutaminase affect both processes in a well-correlated manner. Glucocorticoids, anti-inflammatory agents, appear to exert their effect by inducing the synthesis of a phospholipase A2 inhibitor. Actual locomotion involving microtubules may depend upon attractant-stimulated addition of C-terminal tyrosine to tubulin. Studies on chemotaxis in connective tissue-type cells have shown that fibroblasts respond to the cell binding domain of intact fibronectin and to platelet-derived growth factor; that angiogenesis factors induce endothelial cell migration, and that transformed endothelial cells respond to angiogenesis factors, platelet-derived growth factor, and fibronectin.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this research is to study the chemical and structural properties of the ACTH receptor system in adrenal membranes. ACTH analogs containing fluorescent probes will be used for spectroscopic measurements of the binding interactions. Other analogs will be synthesized for use as ACTH inhibitors. The structural relationships between the hormone receptor and adenylate cyclase will be studied in attempts to solubilization and fractionation of membrane components. Ectopic receptors for hormones in a rat adrenal carcinoma will be characterized by measurement of their binding affinities for peptide hormones and catecholamines.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY The secretory pathway employs vesicle transport to provide a linear pathway for export of cellular products and distribution of membrane and organelle components throughout the cytoplasm. Many diseases, including neurodegeneration, involve disruption of the biosynthetic secretory pathway and unresolved secretory stress--making it essential to understand how secretion is up- and down-regulated under different physiological conditions. While the basic engines of vesicle budding, docking and fusion have been identified, little is known of how they are tuned to respond to physiological conditions and stresses. In mammals, ER-to-Golgi transport, which represents the rate-limiting step in the secretory pathway and the step most relevant to transport-related diseases, has been extensively characterized in vivo and reconstituted in vitro. In broad terms, ER-to-Golgi transport has been shown to comprise: 1) cargo sorting and vesicle budding mediated by the COPII vesicle coat; 2) homotypic COPII vesicle tethering and fusion mediated by tethers and SNAREs to form pre-Golgi organelles called vesicular tubular clusters (VTCs); and 3) VTC-mediated cargo sorting and transport along microtubules leading to fusion with the Golgi. Little is known about how these processes are adjusted dynamically to match secretory output rates with the needs to enforce secretory quality control, avoid ER stress, and keep pace with secretory protein biogenesis and cell growth. One key aspect to regulation of ER-to-Golgi transport that has become apparent in recent years is the role of ER luminal calcium. Calcium, when released from the ER by channel proteins appears to interact with penta-EF hand proteins (PEFs) in the cytoplasm that bind to the COPII coat and modulate its assembly and the rate of cargo egress from the ER. However, the mechanisms of these proteins to produce different secretory outcomes and how they are integrated with ER calcium homeostasis and calcium channels are not understood. This project will employ kinetic assays of ER-to-Golgi transport in intact mammalian cell lines, live-cell calcium measurements, and in vitro studies of PEF protein interactions with vesicle machinery to elucidate how how calcium channels, PEF proteins, and COPII components are integrated to dynamically regulate secretion rate. These studies will have wide significance because proper regulation of secretion is fundamental to key cell survival and stress pathways.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neurons communicate by establishing connections, or synapses, with specific partners. Mechanisms that control the fidelity of these choices are poorly defined but are likely to depend on specific gene regulation. In the nematode, C. elegans, we have shown that the UNC-4 homeoprotein and transcriptional co-repressor, UNC-37/Groucho, function in VA motor neurons to block inputs normally restricted to sister VB motor neurons. We developed cell specific microarray profiling methods to identify UNC-4 regulated genes in this pathway. One of these targets, CEH-12, is homologous to the conserved homeodomain transcription factor, HB9, a known determinant of motor neuron fate in mammals. We used a powerful new strategy for visualizing motor neuron inputs to confirm that CEH-12/HB9 is in fact a VB gene normally repressed by UNC-4 to prevent the adoption VB-type inputs. These experiments also showed that this CEH-12 function is restricted to VA motor neurons in the posterior nerve cord and therefore suggest that UNC-4 must regulate other downstream genes to control synaptic inputs to anterior VAs. RNAi and genetic tests of additional candidate UNC-4 target genes from our microarray data sets have revealed that UNC-4 also negatively regulates the Frizzled protein and Wnt receptor, MOM-5. We hypothesize that a posterior source of Wnt signal acts through MOM-5 to drive ectopic expression of CEH-12 and VA miswiring in unc-4 mutants. A major goal of this project is to test this model by defining the molecular components of this Wnt signaling cascade and its mechanism of action. The existance of A/P gradients of Wnt signaling in the vertebrate spinal cord could be indicative of a similar Wnt dependent mechanism for specifying A/P differences in motor circuit connectivity. To identify unc-4 target genes that function in anterior VA motor neurons, we used high throughput genetic screens to isolate new mutations that \"suppress\" the Unc-4 movement defect. VA inputs in these mutants will be assessed with GFP labeled gap junction and synaptic proteins to confirm unc-4 pathway function. The DNA sequences of these UNC- 4 targets could be used in the future to identify similar genes in mammals where their roles in synaptic specificity can be explored. Thus, our work with a simple model organism is likely to provide important clues to fundamental processes governing the development of complex neural circuits in the the human spinal cord. PUBLIC HEALTH RELEVANCE Nerve cells (neurons) project elongated processes (axons) from the brain into the spinal cord to make connections or synapses with motor neurons that drive body movements. To facilitate the identification of genes that control the specificity of these connections, we are using the nematode, C. elegans, a model organism with a simple, well-defined nervous system. The results of this work should reveal similar human genes with crucial roles in the creation spinal cord circuits and therefore potentially contribute to the development of treatments for spinal cord injury and dysfunction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hepatitis C virus (HCV) is a major cause of community-acquired viral hepatitis. Prototype strains of the various genotypes of HCV, including some of those discovered in this laboratory, are being biologically amplified in chimpanzees, packaged and distributed for use as challenge inocula in studies of passive and active immunoprophylaxis, etc. Full-length cDNA clones of HCV (genotypes 1a, 1b and 2a) have been constructed and transcribed RNA used to transmit hepatitis C to chimpanzees by in vivo hepatic transfection. Chimpanzees, transfected with infectious cDNA clones of HCV, are being followed to determine the natural history of infection. Infectivity pools have been prepared from chimpanzees infected with monoclonal HCV (derived by in vivo transfection with RNA transcripts of infectious cDNA); these have been titered for infectivity in other chimpanzees. In addition, the availability of infectious cDNA clones of HCV has permitted for the first time a mutational analysis of genomic regions. For example, individual portions of the 3' NCR have been deleted from the full-length clone and the resultant deletion mutant clones inoculated into chimpanzees by intrahapatic transfection. Certain regions of the NCR have been identified as critical for in vivo replication of HCV. We have constructed an infectious cDNA clone of GB virus-B (GBV-B), a monkey virus that is the closest relative to HCV. In addition, we have prepared challenge pools of GBV-B and have determined the infectivity titer of these in tamarins. We plan to use the GBV-B tamarin system to study characteristics of the virus that it shares with HCV, which must be studied in chimpanzees. In other studies, we have constructed chimeric genomes from infectious cDNA clones of HCV and bovine viral diarrhea virus. These genomes can replicate in transfected cells but the resultant viral products cannot assemble into infectious virus in the absence of helper virus.We have determined the genetic heterogeneity of HCV isolates that were recovered from patients who were infected following transfusion. The sequence of the hypervariable region and adjacent portions of envelope proteins 1 and 2 were determined for multiple clones obtained from patients who had fulminant hepatitis, from patients who convalesced following acute hepatitis and from patients who progressed to chronic hepatitis C. Distinctive patterns of dynamic change in sequence of clones during the first several weeks of infection were observed. Patients with fulminant or resolving hepatitis had few changes in the sequences of clones, whereas there were many changes in the sequences of clones from patients who progressed to chronic hepatitis. Thus, the outcome of an HCV infection could be predicted in the first few weeks of the infection", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, isocitrate dehydrogenase and malic enzyme share a common pyridine nucleotide (NADP) as coenzyme. Activities of these enzymes are coordinated to generate adequate pools of NADPH for lipid biosyntheses. Biochemical comparisons of the parameters of pyridine-linked dehydrogenases suggest a common evolutionary origin of these enzymes. In Drosophila melangaster there appears to be two forms of NADP-isocitrate dehyrogenase and there may be two genes controlling the activity of this enzyme. A molecular characterization of this gene(s) and the transcript(s) made from it is proposed as the initial step in understanding regulation of this gene-enzyme system. Expression libraries of Drosophila cDNA will be screened for sequences specific for IDH. The identity of these cloned sequences will be verified by in situ hybridization to salivary gland polythene chromosomes and immunoprecipitation of in vitro translated hybrid selected mRNA. Clones which are identified by these techniques will be used to isolate corresponding sequences of genomic DNA. Restriction analyses will compare cDNA and genomic sequences. The developmental expression of these sequences will be determined by Southern analysis. Hybridization of strand specific transcripts of cDNA to \"Northern\" blots of appropriately staged pupal mRNA will allow determiniation of the orientation of transcription. Future experiments would include S1 nuclease mapping and primer extension analysis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Most of the human population world-wide has been infected by herpes simplex viruses. Following the initial lytic infection, HSVs establish permanent latent infections within sensory neurons. Reactivation of latent virus not only results in viral disease (new infections, blindness, and encephalitis) but also contributes to HIV infection, diabetes, cardiovascular and neurodegenerative diseases. No effective vaccine is available and no therapy eliminates latency or prevents reactivation. The long-term goal of this project is to find interventions for recurrent HSV episodes by defining mechanisms that control establishment and reactivation of HSV-1 latency. The gene expression cascade during HSV-1 lytic infection begins with activation of immediate-early (IE) gene transcription by the virion protein VP16 with host factors Oct-1 and HCF-1. In contrast, the initial events in the reactivation from latency are still poorly defined. The central hypothesis of this proposal is that regulation o both VP16 expression and activity underlie the establishment of latency and reactivation from latency. These two levels of control involve multiple positive and negative inputs to allow or inhibit viral replication in the sensory neuron in vivo. Aim 1. This project will determine the mechanism of de novo VP16 gene activation and silencing in sensory neurons in vivo. The working hypothesis is that the VP16 gene in the HSV-1 genome can be regulated by action of neuron-specific and stress-responsive promoter elements and corresponding transcription factors either to allow or inhibit lytic replication upon initial infection or exit from latency. Uing recombinant viruses in a mouse model of infection and latency, we found that expression of VP16 is both necessary and sufficient to trigger the exit from latency and we identified a neuron-specific promoter for the VP16 gene. We will test whether this promoter controls the entry into lytic phase infection during acute infection and during reactivation. We will determine whether the predicted transcription factors bind to the various elements of this promoter to positively or negatively regulate VP16 gene expression, whether singly, in combination, or in competition. Aim 2. This project will define the VP16 coactivator interactions essential for VP16-dependent exit from latency and identify mechanisms regulating these interactions in vivo. Our data strongly suggest that the exit from latency by HSV is regulated by CK2 mediated phosphorylation and that this phosphorylation may be also competitively regulated by O-GlcNAcylation (a PTM that regulates signaling in response to nutrients and stress). Our goal is to elucidate the functions of each of these PTMs in viral latency and to define the roles of their crosstalk in regulating immediate early gene expression of viral proteins through VP16 transactivation. The outcomes of this work will identify transcription factors or protein modifying enzymes that could be targets for future development of therapeutic interventions for HSV reactivation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Since its inception in 1984, the goal of the CSHL FACS Shared Resource has been to give Cancer Center members access to state-of-the-art flow cytometry equipment and support. The Resource has evolved with emerging technologies and changing research needs. In 1984, the primary use was separation of cells by DNA content or by expressed cell-surface antigens. The advent of fluorescent markers (such as the green fluorescent protein GFP) ushered in a new phase in FACS usage, as Cancer Center members used GFP to track transfected cells, and to isolate particular populations of cells from complex mixtures. Increases in the speed and efficiency of sorting by FACS have enabled completion of experiments that once would have been impossible or impractical[unreadable]such as the isolation of large quantities of tumor or stem cells for genomic and genetic analyses. The recent introduction of the new generation of proteins that fluoresce in various colors and are less toxic than their predecessors, changed the requirements for flow cytometers and increased the demand for the Resource. In response to these demands, the FACS Shared Resource has undergone an extensive series of changes[unreadable]in location, instrumentation, and personnel[unreadable]designed to expand FACS capabilities at CSHL, and improve the efficiency and reliability of flow cytometry for Cancer Center members.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Oral pre-exposure prophylaxis (PrEP) against HIV infection has demonstrated efficacy in men who have sex with men, serodiscordant couples, and young heterosexuals; however, efficacy was not shown in two studies of women. Lack of adherence is the leading hypothesis for these discrepant findings. Adherence outside of rigorous clinical trials is unknown and is essential to know in order to determine whether PrEP will be an effective public health strategy. The proposed study will determine if HIV-uninfected people in serodiscordant couples outside a clinical trial will adhere adequately enough to PrEP to be protected against HIV transmission. Approach: This study builds on our experience conducting the Adherence Sub-study within the Partners PrEP Study (a phase III clinical trial of tenofovir and emtricitabine/tenofovir PrEP) and will be integrated into a recently funded study of a public health delivery model of antiretroviral therapy-based HIV prevention strategies involving 500 serodiscordant couples in Uganda and Kenya (R01MH095507, PI: Baeten). Our Specific Aims are a follows: 1) Assess the level, trajectory, and correlates of PrEP adherence in research-naive discordant couples receiving PrEP in a public health delivery model with adherence counseling practical for scale-up; 2) Determine the relationship between patterns of adherence and risk exposure outside of a clinical trial; and 3) Determine if the level and patterns of real world adherence and risk exposure determined in Aims 1 and 2 substantially alter current estimates of PrEP effectiveness and cost-effectiveness. Innovation: We will: 1) characterize PrEP adherence in serodiscordant couples recruited into a demonstration project of PrEP delivery involving limited adherence counseling that is practical for scale-up settings; 2) use a novel conceptual framework to understand PrEP adherence behavior; 3) use text message reports of sexual behavior combined with electronic adherence monitoring to characterize temporal patterns of sexual exposure and PrEP adherence; and 4) develop models of the tenofovir drug concentration-time relationship, using prospectively collected samples, which can be used to determine which patterns of adherence are likely adequate for protection against HIV infection. Investigators: Dr. Jessica Haberer is a new investigator with significant experience studying adherence to antiretroviral therapy and PrEP in sub-Saharan Africa. Her team includes expertise in clinical epidemiology (Drs. Jared Baeten and Connie Celum), adherence measurement (Dr. David Bangsberg), qualitative analysis (Dr. Norma Ware), tenofovir pharmacokinetic modeling (Drs. Craig Hendrix and Ayyappa Chaturvedula), statistical analysis (Dr. Deborah Donnell), cost-effective modeling (Dr. Tim Hallett), and sub-Saharan African research experience (Drs. Nelly Mugo and Elly Katabira).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (based on applicant's abstract): The research contained in this application will focus on host factors that modulate HIV infection. Specifically, the application will concern the role of the human fetal thymus and placenta in in utero HIV infection and the unexpected consequence of early spontaneous abortions suggested by evidence gathered on Baylor grant R01 AI32466-04, \"Role of the Placenta in Transmission of HIV.\" The long-term objective of this research application is directed at answering critical questions of the importance of host factors in the early HIV immunopathogenesis in vertically-acquired HIV infection acquired in utero. The specific aims are to: 1) explore the role of the thymus in early rejection of the HIV-infected fetus in first and second trimester spontaneous abortions in HIV-infected women, 2) determine the role that cytokines in the placenta play in fetal loss in HIV infection, and 3) compare the phenotypic/genotypic characteristics of HIV in the early trimester aborted HIV-infected fetus to those in the mother. The applicant proposes to explore the immune mechanisms of rejection of the fetus, possibly mediated by factors (inflammatory cytokines, interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha) derived from the placenta or by the fetus as a consequence of HIV infection. The project will utilize an existing model of HIV infection, a fetal thymus organ culture (FTOC) system. This model system of HIV infection will allow the careful interpretation of the importance of timing of HIV infection and viral burden/viral strain in early fetal death. The methods to be used include: 1) recruitment of HIV-infected pregnant women in a Pediatric/Obstetrical Research Center; 2) pathological examination of fetal and maternal products of conception, including in situ hybridization studies; 3) denaturing gradient gel electrophoresis, single strand conformation polymorphism, gene sequencing, and T cell and macrophage/monocyte tropism studies of HIV strains recovered from fetus and mother; 4) measurement of mRNA and protein product of interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha; and 5) HIV infection (and pathological consequences) in an FTOC system in vitro.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although there is an important genetic contribution to the normal range of variation seen in human behaviors as well as human behavioral disorders, very little is known about the genes that influence behavior in any organism. In particular, there are currently few model vertebrate systems in which the study of behavioral variation in a natural, outbred population can be genetically dissected. The recent development of genetic and genomic tools for the threespine stickleback (Gasterosteus aculeatus) presents an opportunity to study the genetic basis of behavioral variation in a well-defined ecological and evolutionary context. The mating behaviors of the threespine stickleback are perhaps the best-studied of any animal system. Previous work has shown that divergence in male and female mating behaviors leads to reproductive isolation between stickleback populations that have adapted to living in two different habitats ('benthic'and 'limnetic') in a single lake. There are multiple lakes in British Columbia in which these benthic and limnetic 'species pairs'have evolved, and similar changes in mating behavior have occurred independently in each lake. The aim of this study is to investigate the genetic underpinnings of behaviors involved in reproductive isolation between benthic and limnetic sticklebacks from two lakes (Priest and Paxton) using quantitative trait locus (QTL) mapping. These experimental populations will be raised and behaviorally evaluated under fully natural conditions in outdoor ponds. By investigating the genetic basis of behavioral phenotypes involved in reproductive isolation (e.g., male nesting behavior and female mate preferences), this work will test whether parallel species pairs of sticklebacks in isolated lakes share similar genetic architectures for convergent behaviors involved in reproductive isolation. This project will also test whether QTL influencing behaviors involved in reproductive isolation map disproportionately to genomic regions affecting traits known to be important for mate selection, such as body size and shape. Importantly, the QTL mapping experiments proposed here will serve as a first step in the identification of genes that underlie behavioral differences in natural stickleback populations. Results in sticklebacks are likely to have important implications for understanding the genetic basis of behavioral variation and behavioral disorders in humans because many of the genetic and neural pathways that underlie behaviors are likely to be conserved between fish and humans. In addition, humans and sticklebacks have migrated and adapted to new environments over a similar evolutionary timeframe, suggesting that studying the genetic architecture of behavioral variation in sticklebacks has direct relevance to similar studies in humans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The main purpose of this project is to test the efficacy of epsilon-aminocaproic acid administered during the 14 day period immediately after the bleeding episode. The activities of the last year have been mostly those of data collection; the randomized portion was suspended because EACA was shown to be superior at the time. Supporting data are included.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cerebral malaria (CM) is a severe and potentially fatal neurological manifestation of disease caused by infection with Plasmodium falciparum. Despite effective anti-malarial therapy, approximately 25% of CM survivors develop long-term neurological deficits, such as memory loss and motor coordination impediments. The mechanisms that mediate this neurocognitive impairment are not well understood. Neuronal injury has been associated with the neurological deficits in several neurodegenerative diseases and may contribute to the impairment seen in CM. In this regard, damage to neuronal axons has been observed in both human and murine experimental CM (ECM). Furthermore, improper regulation of tau protein, an axonal protein important for microtubule stability and cytoskeletal organization, has been demonstrated in mouse and human disease. The neuronal injury and neurocognitive impairment observed in CM may result, in part, from abnormalities in tau. We hypothesize that pathological tau is one of the main mediators of neuronal damage and subsequent neurocognitive decline in CM. To address this hypothesis, we propose the following specific aims: AIM1: To characterize the role of tau in ECM. Aim1a: To examine the localization and secretion of hyperphosphorylated tau in the brains of mice with ECM and compare our findings to mice with malaria-associated severe anemia and uninfected mice. Aim1b: To characterize the contribution of tau to the neuronal damage and the subsequent neurocognitive deficits which occur during ECM using both a monoclonal antibody against phosphorylated tau and tau-knock-out (KO) mouse experiments. AIM2: To determine the role of tau in an in vitro human CM model. We will challenge cultured immortalized human neurons with supernatants obtained from blood-brain barrier co-cultures of astrocytes and endothelial cells (with and without P. falciparum exposure) and determine the effects on neuronal morphology and function as well as tau protein. We will also quantify secreted and intracellular tau levels and examine whether targeting the protein immunotherapeutically with PHF-1 antibody reverses neuronal damage. Our goal for this project is to further establish tau as a significant contributor to the pathogenesis of CM. Targeting this protein therapeutically may prevent both damage to neurons as well as the subsequent neurocognitive impairment which occurs during disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the proposed program is to address several technical feasibility questions (in Phase I), and then demonstrate (in a subsequent Phase II) incorporation of Sonetics'groundbreaking CMUT-in-CMOS ultrasound transducer technology into a novel, high-performance 2D ultrasound array suitable for commercial applications. Phase I specific aims will investigate bandwidth improvement for 2D array elements, scaling and enhancement of on-chip readout circuits, and reduction of dielectric charging effects related to device membrane sealing layers. This phase will culminate with the fabrication and characterization of a 64-element 2D array prototype, which will be used to evaluate the imaging potential of a larger scale array. Full implementation into a prototype scanhead (including a 128x128 element CMUT array) suitable for real-time 3D imaging would be demonstrated in Phase II, with the long-term goal of commercializing the technology (as well as other CMUT-in-CMOS-based products) for the ultrasound equipment market, forecast to be $3.75B globally by 2010. The academic segment of this market would benefit from the practical realization of fully-populated 2D ultrasound arrays, which will improve the quality and speed of 3D imaging for disease diagnosis. As well, the potential of CMUT-based scanheads to reduce the largest cost element of many state-of-the-art ultrasound systems, while simultaneously exhibiting improved imaging capability, will provide increased opportunity for researchers and students to undertake real-time, in vivo studies of living systems-something 3D ultrasound is uniquely positioned to do from a cost, availability, and convenience perspective (vs. MRI, PET, or CT, for example, all of which require expensive, dedicated facilities and technicians). The clinical market stands to gain similarly, as 3D ultrasound provides safe, convenient, real-time imaging technology-enabling improved triage and better patient outcomes, lower-cost imaging, and reduced staffing needs, all of which will serve the U.S. healthcare industry well as it seeks to control skyrocketing costs. PUBLIC HEALTH RELEVANCE: Potential benefits to public health from the successful development of Sonetics'novel ultrasound transducer technology include: improved availability and affordability of high-quality 3D medical imaging for disease diagnosis;improved medical training as low-cost 3D ultrasound enters the classroom;and lowered health-care costs for society as a whole, as better ultrasound improves clinical triage and patient outcomes. Furthermore, ultrasound is the only technology with the potential to become a convenient, safe, real-time imaging tool for use in limited-budget facilities such as lowincome health clinics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Anxiety and affective disorders constitute a group of heterogeneous illnesses that are common and show significant heritability. Considerable work has focused on identifying the genes involved in anxiety and affective disorders providing interesting leads, but no definitive answers. One of the most prominent childhood risk factors for the development of these illnesses is behavioral inhibition (BI), a temperamental disposition characterized by extreme shyness and inhibition in response to novel situations or strangers. Using young rhesus monkeys, we have developed a model that is analogous to childhood BI and have demonstrated that individual differences in monkey BI are significantly heritable. In addition, we have used functional imaging to identify the brain regions associated with BI. In this project we will measure BI, associated physiological parameters, and functional brain activity in a large multi-generation pedigree of rhesus monkeys. These phenotypes will be used in whole genome linkage analyses to investigate the genetic basis of brain mechanisms underlying anxiety and depression. By combining proven approaches to the study of the genetics of complex disease with simultaneous analysis of intermediate brain reactivity phenotypes, our novel strategy using young rhesus monkeys will: 1) identify novel genes that influence BI, 2) quantitate the influence of genetic variation on individual differences in reactivity of the neurocircuitry underlying emotion, and 3) determine specific genes that are involved in mediating both individual differences in BI and increased reactivity of emotion-related brain circuits. These studies are not feasible in rodent species or humans. The proposed experiments provide an invaluable opportunity to identify novel genetic factors that play a major role in the development of human anxiety and affective disorders, results that will be immediately relevant to children at risk for the development of psychopathology. PUBLIC HEALTH RELEVANCE Anxiety and affective disorders constitute a group of mental illnesses that can be inherited. Using young rhesus monkeys as subjects, this study will: 1) identify genes that influence behavioral inhibition, 2) quantitate the influence of genetic variation on individual differences in reactivity of emotion-related brain circuits, and 3) determine specific genes that are involved in mediating both individual differences in behavioral inhibition and increased reactivity of emotion-related brain circuits. The proposed experiments provide an invaluable opportunity to identify genetic factors that play a major role in the development of human anxiety and affective disorders, results that will be immediately relevant to children at risk for the development of psychopathology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposed project intends to address challenges identified by and realize the full potential of two earlier SEPA-funded projects: a project completed in 2012, Education for Community Genomic Awareness (Grant Code: R25RR022703); and our currently project (2014- 2019), A New Genomic Framework for Schools and Communities (Grant Code: 8 R25 GM129186-05). These projects focused on addressing the critical need for modern genomics curriculum for middle and high school students by connecting classroom instruction with the community to give both students and community members opportunities to understand, explain and apply ideas about health-related phenomena to their lives tied to gene-environment interactions, natural selection, and evolution. The implementation of these projects has produced evidence of success in student learning and interest, it has also produced three challenges to the realization of their full potential: 1) Aligned with the Next Generation Science Standards, these units are vastly different than those teachers are familiar with and making community connections are also difficult to orchestrate; 2) The potential for maintaining and enhancing student engagement was limited by short periods during the school year and lack of continuity from year to year; 3) The extent of support provided by the project to schools suggests the need to develop a model for sustained support from internal sources once the grant period is over. This research educational program plan outlines a novel design for science education that combines three components: 1) high quality curriculum materials, 2) innovative professional learning, and 3) a community health/science education consortium. These components work together to support and sustain genomic and environmental health learning and inspire interest in careers for underrepresented middle and high school students both in and out of school. The result will be a new, innovative, sustainable educational program with the following characteristics: 1) ?Health in Our Hands?, a coherent set of curricula for secondary grades that maximizes deepening of student understanding; 2) A ?Community of Practice? linking teachers with informal educators, learning new teaching practices, making seamless connections between the classroom and community, and enabling students to apply what they have learned to real-world health issues; 3) ?Greater Flint Health in Our Hands? consortium of health-related organizations, providing the mentors and the community-based activities, and support for future growth of the learning program after the expiration of this grant.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent genetic evidence indicates that the integrin class of fibronectin-binding adhesion receptors (5E1 and others) can regulate both the form and function of the heart. Integrin ligation drives recruitment of a number of structural and signaling molecules to the ventral plasma membrane collectively termed a \"focal adhesion\" which serves to link the force-generating actin cytoskeleton inside the cell to the extracellular matrix (ECM), and to coordinate activation of downstream signaling pathways. The non-receptor tyrosine kinase, Focal Adhesion Kinase (FAK) is strongly activated by both integrins and growth factors, and is a likely candidate to integrate downstream signals from these diverse pathways during growth and development. Indeed, germline deletion of FAK results in mesodermal defects and embryonic lethality between E7.5-10 similar to the phenotype observed in both fibronectin-, and D5-null mice. Although a direct role for FAK in cardiac development has yet to be examined, hearts from FAK-null embryos revealed a lack of separate mesocardial and endocardial layers, indicative of a defect in cardiomyocyte maturation. Interestingly, recent work by our group and others clearly indicate that FAK is activated in cultured cardiomyocytes by a variety of hypertrophic stimuli including, phenylephrine (PE), endothelin I (ET-1), angiotensin II (AII), and hypo-osmotic stress, and that increased cardiac FAK activity is observed in vivo in hypertrophic hearts. The idea that FAK activation plays a direct role in the development of cardiomyocyte hypertrophy is evident from our seminal findings that the activation of FAK is required for PE-stimulated hypertrophy of cultured cells and similar findings from others that FAK is required for maximal ET-1 and stretch-induced hypertrophy in vitro. The experimental goals of this proposal are to test the hypothesis that FAK regulates cardiac development and pathological hypertrophy in vivo and to identify the FAK-dependent signaling pathways involved in these processes. We will generate genetically modified mice in which FAK will be deleted in a temporal and cardiac-restricted fashion using Cre/LoxP technology to examine a functional role for FAK in cardiac growth. We will also establish a cardiac cell culture model to identify FAK-dependent signals and target genes that are differentially regulated by hypertrophic stimuli. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goals of this research as stated in the grant title are the optimization of hygroscopic aerosol delivery to the human bronchial tree. We are achieving these goals by a combination of experimental studies on physical models of the bronchial airways, theoretical calculations of aerosol drop size and size distributions effected by evaporation, development of aerosol evaporators, and ultimately testing of the delivery system on human lungs and on patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Excessive alcohol consumption remains a leading cause of preventable death in the United States. Alcoholic liver disease (ALD) is a major cause of morbidity and mortality from heavy alcohol drinking and is the most prevalent cause of liver disease-related death. Despite increased understanding of the many cellular and molecular events occurring in ALD, the causal mechanisms remain elusive, especially with regards to the factors responsible for disease progression and severity. To advance mechanistic understanding of this serious liver disease, the current proposal will investigate the importance of circadian clock disruption and `time-of-day' as critical risk factors for ALD. These views stem from the growing recognition that circadian clock disruption is linked to numerous pathologies and diseases. One essential function of the molecular circadian clock is to provide a selective advantage of anticipation, allowing for rapid and temporally appropriate adaptation to metabolic stress, immune challenges, and environmental insults. When these processes fail, disease ensues. We propose that perturbation of circadian clocks underlie many of the metabolic and inflammatory events contributing to alcohol-induced liver injury. As such, we will determine the role of the hepatocyte clock and the monocyte clock in alcohol-induced liver injury. In support of this concept, we have found that the circadian clock is significantly altered in livers of chronic alcohol-fed mice, genetic disruption of the liver clock increases levels of hepatic inflammatory mediators, and alcohol-induced liver injury and steatosis are elevated in mice with a disrupted liver clock. Furthermore, studies show that disruptions in normal time-of-day patterns in alcohol drinking increases risk for ALD and binge alcohol drinking is now recognized to be harmful to health. Collectively, these observations have led us to hypothesize that circadian clock disruption exacerbates alcohol-induced liver injury and inflammation. Similarly, we propose that the magnitude of alcohol toxicity is dependent on the time of day of alcohol ingestion. We will test these hypotheses through two specific aims. In Aim 1, we will mechanistically show the critical role of cell autonomous clocks in chronic alcohol- induced liver injury by using two genetic mouse models, the hepatocyte-specific BMAL1 knockout mouse and the monocyte (myeloid cell)-specific BMAL1 knockout mouse. In Aim 2, we will use the `chronic + binge' alcohol model and determine the `window' or time of day that the liver is most sensitive to alcohol binge toxicity. Successful completion of this project will reveal the importance of cell-specific circadian clocks in alcohol-mediated tissue injury and inflammation and show that `time-of-day' is a significant risk factor for liver injury from binge alcohol drinking. Our long-term goal is that the scientific knowledge gained from these pre- clinical animal studies will lead to future translational investigations using various chronobiology-based therapeutic approaches for treatment of ALD and other related liver diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Huntington's disease (HD) is a dominantly inherited, incurable neurodegenerative disease affecting primarily the striatum and corticostriatal pathway. Sex differences in mice and humans with HD suggest that gonadal steroid hormones may play a role in the onset and progression of HD. Neuroprotective effects of 17--3-estradiol (E2) suggest that it may contribute to these sex differences. The goal of the proposed research is to characterize the effects of E2 on neural pathophysiology and the behavioral phenotype of HD in female mice. Because HD results in damage and altered neural activity of the corticostriatal pathway, it is likely that cortical dysfunction contributes to excessive neuronal activity and a deficit of the antioxidant vitamin, ascorbate (vitamin C; AA) in the striatum. E2 effects on glutamate (GLU) transmission may normalize cortical neuronal activity in female HD mice preventing striatal AA loss and resulting in later onset and/or slower progression of HD in females. Electrophysiology and voltammetry will be used to record neural activity in the prefrontal cortex (PFC) and monitor AA release in the striatum of behaving HD mice and wild-type controls (WT) receiving either ovariectomy (OVX) + .placebo, OVX + E2, or sham surgery + placebo. During cortical recordings, mice will be engaged in a reversal learning task as this is a cortico- striatal dependent task involving PFC. One mechanism of E2 neuroprotection may be via upregulation of glial GLU transporters such as GLT1. This may prevent the GLU excitotoxicity believed to occur in HD and increase GLU clearance in female HD mice. Immunohistochemistry and behavioral assessments will be used to determine whether E2 upregulates GLT1 in the cortex and striatum, improving the behavioral phenotype of HD in female mice. Results of this study will indicate whether E2 may protect female HD mice from striatal AA loss, normalize cortical neuronal activity and improve the behavioral phenotype associated with HD. Furthermore, results will suggest whether E2 effects on GLU transmission contribute to sex differences observed in HD mice. HD affects -30,000 people in the United States and -150,000 more are at risk for inheriting the gene. If E2 is neuroprotective in HD models, it may be a suitable therapy, particularly for women with HD. Furthermore, evaluation of the role of E2 in regulating motor circuits in the brain will provide a better understanding of how this hormone regulates normal and pathological motor control. This is particularly important because more than 40 million U.S. citizens are affected by movement disorders. E2 likely plays a critical role in motor function and neuroprotection and may represent a therapy for movement disorders and neurodegenerative conditions that is currently not well investigated. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Comparison of potency tests of recombinant FVIII manufactured by Miles and by Baxter in the one and two stage assays and the chromogenic assay indicates an extraordinary degree of both interassay and intro-assay variability. Differences between individual runs of same assays can deviate up to 100% from the labelled potency. Predilution of both products in either albumin or factor VIII deficient plasma also affects the potency, with those samples prediluted in deficient plasma having overall higher values. It is planned to do in vivo half life and recovery studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Control of the primary tumor is a major goal of radiotherapy for cancer. Escalating the dose delivered to the tumor provides a method to improve local control. For lung cancer patients in particular, respiratory-induced organ motion has impeded safe dose escalation. Methods to compensate for this motion or to immobilize the tumor have been developed in recent years. However, these new technologies have not been applied in concert with reduced margins that would enable dose escalation due to the lack of data characterizing uncertainty in respiratory-induced organ motion. It is critical to characterize the uncertainties associated with tumor immobilization to enable the use of appropriate margins. It is our hypothesis that the combination of image guidance techniques and integrated active breath hold radiotherapy will enable characterization and reduction of the geometric uncertainties due to respiratory-induced organ motion. Active breathing control (ABC) has been shown to be a safe and effective means of tumor immobilization for breast cancer patients which allows for the reduction of the dose to normal tissue structures such as the heart and lungs. For lung cancer patients, however, the ABC technique must be adapted to increase compliance for patients with pulmonary compliance issues. Furthermore, the integration of image guidance techniques with ABC radiotherapy enables the characterization and reduction of daily setup variation and immobilization uncertainty. The specific aims of this project are to (1) Measure the random and systematic uncertainties of tumor immobilization for integrated active breath hold radiotherapy. (2) Evaluate a model image-guidance strategy with respect to the presence of these uncertainties and to design a treatment margin to compensate for these uncertainties. (3) Quantify residual setup error in a small population of patients. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT This proposal is for a phase I/II fast track project for the STTR program with the main goal to develop a test for high sensitive detection of tau oligomers in biological fluids, which could be used for the biochemical diagnosis of Alzheimer's disease (AD) and related tauopathies. AD is the most common dementia in the elderly population and one of the leading causes of death in the developed world. One of the main problems in AD is the lack of an early, sensitive and objective laboratory diagnosis to identify individuals that will develop the disease before substantial brain damage. Compelling evidences point that the hallmark event in AD is the misfolding, aggregation and brain accumulation of amyloid-beta (A?) and tau proteins. Recent evidences suggest that A? pathology is the primary driving force of the disease initiation, but this is accomplished by induction of tau hyperphosphorylation, misfolding and aggregation, leading to the neurodegenerative cascade. Tau aggregation follows a seeding-nucleation mechanism and involves several intermediates, including soluble oligomers and protofibrils. Recent evidence has shown that tau oligomers are circulating in biological fluids and these structures appear to be key for inducing brain degeneration in AD. Our working hypothesis is that detection of misfolded tau oligomers circulating in blood may be the basis for an early biochemical diagnosis for AD. Our approach is to use the functional property of misfolded oligomers to seed the aggregation of the monomeric protein as a way to detect them. For this purpose, we have developed the protein misfolding cyclic amplification (PMCA), which represent a platform technology to detect very small quantities of seeding-competent misfolded oligomeric proteins associated with various protein misfolding diseases. Currently, PMCA has been adapted to detect misfolded prion protein implicated in prion diseases in various biological fluids, including blood and urine and more recently soluble oligomers composed of A? and ?- synuclein in cerebrospinal fluid (CSF) of patients affected by AD and Parkinson's disease, respectively. The major goal of this project is to adapt the PMCA technology for specific and highly sensitive detection of misfolded tau oligomers in human CSF and blood plasma, perform studies of specificity and sensitivity using large number of samples coming from patients affected by AD and other tauopathies as well as to evaluate the utility of tau-PMCA for monitoring disease progression. The results generated in this project may lead to the first biochemical test for diagnosis of AD. The studies included in this project will constitute the basis for regulatory approval of the test that Amprion will commercialize.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The main intention of this grant is to increase our understanding of the effect of ginseng as an anti-bacterial agent against Pseudomonas aeruginosa pathogenesis in chronic lung infection the most common cause of morbidity and mortality among patients with cystic fibrosis (CF). Although antibiotic therapy, mostly with a mixture of aminoglycoside and Beta-lactam antibiotics, is used to combat the infection, treatment often fails, owing to the emergence of alginate-producing P. aeruginosa isolates. The genetic and molecular mechanism of alginate overproduction has been elucidated in great detail. Yet, a possible therapeutic intervention has been elusive. Recent studies using animal models (described in detail later) have shown that ginseng might play a role in enhancing immune response and bacterial clearance. This research proposes to use genetic and molecular biological techniques to systematically analyze the effect of ginseng against Pseudomonas aeruginosa. Specifically this research will investigate the following: I. Analysis of the ginseng extracts. II. Determine whether ginseng modulates (down- or up-regulates) the alginate genes in vitro. III. Identify novel genes that are modulated by ginseng. Understanding the mechanism of ginseng action would be a key factor if we want to promote this alternative or supplementary therapy. This is vital to devising efficient antipseudomonal therapy and eradicating complications resulting from P. aeruginosa infections among patients with CF.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal requests support for a Keystone Symposia meeting entitled Neurodegenerative Diseases: The Molecular and Cellular Basis for Neurodegeneration, organized by Ted M. Dawson, Virginia M.-Y. Lee and Stephen M. Strittmatter, which will be held in Taos, New Mexico from February 21 - 26, 2011. Neurodegenerative diseases are chronic age-dependent progressive disorders that are substantial and growing health problems, which exert a tremendous toll on the patient, family, health system and society as a whole. Accordingly, there is an urgent need to identify therapies that slow and/or reverse the progression of these disorders. Research in neurodegenerative disorders is providing tremendous advances in the molecular understanding of these disorders. New insights in cell biology, biochemistry, genomics and proteomics into these illnesses are leading to mechanism-based therapies and new tools and biomarkers to study disease progression and therapeutic efficacy. In this meeting an emphasis will be placed on understanding new molecular and common mechanisms of disease in Alzheimer's Disease (AD), Parkinson's Disease (PD), triple repeat diseases, frontotemporal lobar dementia and others. This meeting will provide a forum to bring together basic and translational scientists from both academia and industry to share data, identify key areas to focus future research, and to create new collaborations in the translational neuroscience of neurodegenerative disease. PUBLIC HEALTH RELEVANCE: According to the World Health Organization, neurologic disorders are the leading cause of death, disability and loss of quality of life worldwide. In the US, neurological disorders affect approximately 50 million Americans and cost an estimated $400 billion annually in medical and related expenses. Neurodegenerative diseases are projected to triple over the next several decades as the percentage of elderly increases dramatically.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This overall program project in trauma is designed to quantitatively evaluate various biochemical and physiological parameters in the critically ill patient. The pathophysiology of trauma has been expressed by a number of overt clinical changes manifested by the patient which have served to emphasize the severity and seriousness of trauma, \"the forgotten disease\". This program has been conceived to explore in depth the clinical changes with the hope of providing a better rationale for the treatment and care of such patients. The core program includes a Metabolic Facility which provides the basis for the constant clinical care of the patient and for research by the basic scientist. The program encompasses study areas to evaluate in critically ill patients the following: (1) biochemical indices related to nutritional status, (2) trace element requirements, (3) immunological function related to trauma and nutrition, (4) metabolism of carbohydrate using 14C-glucose, (5) gluconeogenesis using 14C-L-alanine and 14C-glycerol, and (6) whole body protein metabolism using 15N-L-alanine and 14C-L-leucine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Patient falls and pressure ulcers are designated as \"nursing-sensitive quality indicators\" by national groups such as the American Nurses Association and the National Quality Forum. Yet research linking the occurrence of falls and pressure ulcers to differences in nurse staffing has yielded equivocal findings. Unreliable measurement of adverse events and inadequate measures of nurse staffing could account for the inconclusive findings. Prior studies have analyzed these effects at hospital and nursing unit levels, but never simultaneously. Differences in the nursing practice environment may moderate the link between staffing and adverse events, although this has not been tested. The proposed study builds on evidence developed by this research team that nurse staffing and the practice environment influence patient outcomes. This study advances our understanding of the relationship between staffing and patient adverse events by 1) calculating adverse events using secondary data from observational prevalence studies and incident reports, 2) using a comprehensive set of nurse staffing measures that account for qualitative as well as quantitative differences among RNs, 3) examining these relationships at both the nursing unit and hospital levels, and 4) exploring the influence of a formally-recognized favorable nursing practice environment on these relationships. Multi-level models will be used to predict the effects of staffing on rate of falls at the nursing unit level and pressure ulcers at both the patient and the nursing unit levels using data from 4,300 nursing units in 650 community hospitals in the ANA-sponsored National Database of Nursing Quality Indicators. American Nurses Credentialing Center Magnet Designation status will be used to differentiate the nursing practice environment across hospitals and explored as a hospital-level factor moderating the associations between staffing and outcomes. This study will strengthen the evidence base on how nurse staffing patterns and practice environments support the quality and safety of patient care.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The CAPRISA Clinical Trials Unit for AIDS/Tuberculosis Prevention and Treatment (CAPRISA CTU), strategically located in KwaZulu-Natal, South Africa, at the epicenter of one of the world's most severe HIV and tuberculosis (TB) epidemics, has recently made significant scientific contributions on tenofovir gel and pre-exposure prophylaxis, HIV-TB treatment, prevention of breastfeeding transmission, and immunity with broadly neutralizing antibodies. The scientific team includes investigators who have served at the highest levels in Network leadership. During the current funding cycle, 2,854 participants have been enrolled in 25 protocols to date, with retention rates usually exceeding 90% and data quality scores exceeding 95%. The CAPRISA CTU comprises 4 components; firstly, experienced leadership, senior investigators and CTU coordinator; secondly, an administrative and governance component; thirdly, 10 research support cores; and fourthly, 4 Clinical Research Sites (CRSs). The eThekwini CRS for vaccine, microbicide and adult HIV treatment studies, is located in central Durban adjoining a local clinic that annually treats 40,000 patients with sexually transmitted infections (HIV prevalence of 59.3% (CI: 56.5-62.0) and HIV incidence rate of 6.4 per 100 person-years (CI: 2.6-13.2)) as well as 3,500 new TB cases, two thirds of whom are co-infected with HIV. At the rural Vulindlela CRS for integrated prevention, vaccine and microbicide studies, the HIV prevalence among young women is 35.7% (CI: 32.7-38.8) and the HIV incidence rate is 6.5 per 100 personyears (CI: 4.4-9.2). In this community, HIV incidence is 10.2 per 100 person-years (CI: 4.1-20.9) in women under 20 years. The Springfield CRS for adult HIV/TB treatment studies is located at a regional referral hospital that treated 2,359 MDR-TB and 206 XDR-TB patients (71% co-infected with HIV) in 2010. The Umlazi CRS for maternal/pediatric HIV studies is at a 1,200 bed hospital that provides pre-natal services to about 12,000 women annually (HIV prevalence: 39% (CI: 36.7-41.5)). The vertical transmission rate is 2.4% (CI 1.1-4.5) at birth but increases to 6.6% (CI 5.3-8.2) at 6-8 weeks post-partum due to breastfeeding. The CAPRISA CTU headquarters is located at the Nelson R Mandela School of Medicine and houses the Administration as well as the laboratory, pharmacy, data management and IT, community engagement, evaluation and quality assurance, financial management, bioethics, communication, regulatory compliance, and training Cores. The CTU's organizational structures (Leadership Group, Executive Committee and Community Advisory Board) and communication tools (regular meetings, video conferences, monthly newsletters and website) enable effective communication, management and governance in the unit. Overall, the CAPRISA CTU has diverse well-characterized high-risk populations, well established clinical facilities, accredited laboratories, pharmacies, and data management systems, strong community linkages, and extensive experience in conducting clinical trials, together with a track record of scientific innovation available to support the 5 Networks in developing new approaches to HIV and TB prevention and treatment. RELEVANCE: The overall goal of the Centre for the AIDS Program of Research in South Africa (CAPRISA) Clinical Trials Unit for AIDS/Tuberculosis Prevention and Treatment (CAPRISA CTU) is to advance the scientific agendas and research contributions of each of the five NIAID Clinical Research Networks (CRNs) focused on 1) adult HIV therapeutic strategies, 2) strategies to address HIV in pediatric and maternal populations, 3) integrated HIV prevention strategies, 4) microbicides. and 5) vaccines", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Osteoporosis has become a major health problem as the life expectancy of the general population has risen rapidly in recent years. Post-menopausal women are at greater risk because of accelerated bone loss induced by estrogen deficiency superimposed on age related bone loss. One of the important etiological factors in postmenopausal osteoporosis is the interaction between estrogen and PTH. PTH is known to be an important initiator of bone remodeling and persistent elevation of PTH, such as in hyperparathyroidism, presents as a risk factor for the development of osteoporosis. However, intermittent administration of PTH has been shown to be a promising regimen for improve bone mass in both the animals and humans. The dichotomy of this issue is not fully understood, especially in the estrogen deficient population. This project attempts to understand further the dualistic role of PTH as a catabolic and anabolic hormone in animal models of estrogen deficiency. The proposed study will have the following specific aims. Specific Aim 1. To differentiate the anabolic action of continuous elevation of PTH from its catabolic action. Specific Aim 2. To demonstrate the interactions of dietary calcium intakes with continuous and intermittent PTH administration under estrogen deficiency state, and Specific Aim 3. To study the interactions of PTH, cytokines and anti-resorptive agents at both tissue and subcellular levels in a mouse model of estrogen deficiency.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Tumor specific therapeutic antibodies direct the immune system to eliminate or prevent growth of tumor cells. Passive and active humoral immune responses are mediated via interactions between the IgG Fc domain and specific Fc receptors (FcR) present on the surfaces of effector cells. Proof of principal experiments show that it is the interaction between tumor specific antibodies and the FcR activating receptors on the surface of immune system effector cells that mediate tumor cell depletion. Modulation of the binding activity of the Fc domain to satisfy specific Fc receptor binding parameters will enable genetically engineered antibodies to act as efficient antitumor drugs. Novel Fc mutations that enhance the affinity of the Fc domain to FcgammaR3A and decrease the affinity to FcgammaR2B may play an important role in development of new therapeutics. These novel scaffolds could be ligated to Fabs that target tumor specific antigens and ultimately used as therapeutics to mount a successful cytotoxic response against tumor cells. Recent advances in proteomics and cDNA microarray technologies have aided in the identification of a number of tumor specific antigens. These peptides have been used for the development of antibodies that target specific cancer cells. To enhance the immune response against tumor cells we have developed a novel genetic screen to identify Fc mutants that preferentially bind to the activation receptor FcR3A. Further research and development of these Fc mutants should lead to therapeutic antibodies with increased capacity to trigger antibody dependent cytotoxic reactions against tumor cells. This proposal describes an experimental paradigm to identify Fc mutations, characterize the mutations in functional assays, and ultimately test therapeutic antibodies harboring the best Fc mutations in animal model studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Abstract Traumatic brain injury (TBI) represents a signature injury of the Iraq and Afghanistan conflicts. TBI causes severe persistent disability including cognitive impairment and mental health problems, resulting in loss of productivity and quality of life for the young Veterans returning from the wars. The long-term health care costs of combat-related TBI have created a huge financial burden and generated serious public and personal crises in the United States. To date, evidence-based treatment for TBI recovery is not yet available. Considering the fact that TBI Veterans are currently in the chronic phase of the disease, there is a critical need to develop neurorestorative strategies for brain repair in the chronic phase of TBI. Our recent studies have revealed that a stroke-damaged brain is repairable in the chronic phase by the combination of two essential hematopoietic growth factors, stem cell factor (SCF) and granulocyte-colony stimulating factor (G-CSF) (SCF+G-CSF). Using the same approach, our preliminary studies have demonstrated the neurorestorative efficacy of SCF+G-CSF in brain repair in the chronic phase of TBI. However, it remains unclear how SCF+G-CSF repairs the brain in the chronic phase of TBI and whether SCF+G-CSF treatment could repair a TBI-damaged brain in a delayed chronic phase. The objective of this research proposal is to address these unanswered questions. Based on our recent publications and preliminary data, we hypothesize that SCF+G-CSF repairs a brain in the chronic phase of TBI through neural network rewiring, which is accomplished by its direct regulation of neurons and its indirect effects via bone marrow-derived monocytes/macrophages (BMDM). Using the approaches of live brain imaging, transgenic mice and targeted knockout mice, neurobehavioral assessments, and cell signaling, the central hypothesis will be tested and the objective of this application wil be achieved by accomplishing the following 3 specific aims: Aim 1 will determine the contribution of SCF+G-CSF on brain repair in the delayed chronic phase of TBI, Aim 2 will define the role of neural network rewiring in SCF+G-CSF-induced brain repair in chronic TBI, and Aim 3 will identify the involvement of BMDM in SCF+G-CSF-induced brain repair in chronic TBI. This project is innovative in the unique approach of using hematopoietic growth factors to repair the brain in chronic TBI, which is originally created by our group. This study will significantly advance current knowledge in rehabilitation research for TBI. This research is highly compatible with the objective and scope of the RR&D priority areas for supporting the development of therapeutic strategies on neural plasticity in chronic TBI repair. The contribution of this research is in keeping with the VA mission to ensure that Veterans achieve maximal recovery from combat-related neurotrauma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Flow Cytometric immunophenotyping is a sensitive technique for analysis of benign and malignant tumors. We are studying the refinement of this technique and its application to diagnosis and measurement of prognostic markers in different systems. We are studying the flow cytometric immunophenotype of CLL and correlating the expression of specific antigens with morphology, cytogenetics and clinical course. Data from this study may provide prognostic markers for this disease. We have refined methods for flow cytometric monitoring of patient specimens for apoptosis induced by chemotherapy and have been able to detect apoptosis in specific cell populations. This has allowed us to demonstrate different sensitivities of specific cell lineages to chemotherapeutic agents. We have initiated a study of clonal cytotoxic T-cell populations that arise in patients with B-cell neoplasia to determine prognostic importance as well as resulting difficulties in minimal residual disease detection. As a result, we have found that clonal cytotoxic T-cell proliferations occur with surprising frequency in hairy cell leukemia. The laboratory has an ongoing interest in detection of minimal and residual lymphoma. We are using multiparametric approaches to improve the sensitivity of detection of monoclonal B-cell populations. By targeting abnormal patterns of antigen expression (eg CD10, CD5, CD23, FMC7, or abnormal intensity of antigen expression) by neoplastic B-cells in light chain detection, we are attempting to detect very small numbers of neoplastic B-cells among admixed polyclonal B-cells. The laboratory is studying specific Flow Cytometric markers of various lymphoma sub-groups to improve diagnostic accuracy. The Flow Cytometry Laboratory is developing methods for Flow Cytometric analysis to detect EBV gene expression in the research and clinical setting.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Insulin resistance is the common factor among metabolic diseases that include obesity, type 2 diabetes and steatohepatitis. Hyperinsulinemia is an early indicator of insulin resistance. Although in many instances hyperinsulinemia reflects impaired insulin response, considerable data support the alternative view that hyperinsulinemia causes insulin resistance. Studies supported by this grant have pioneered the biochemical, physiological, and genetic identification of a CEACAM1-dependent signaling pathway that regulates hepatic insulin clearance. We have shown that CEACAM1 regulates peripheral insulin action by promoting hepatic insulin clearance. Using mice with liver-specific inactivation or global null mutation of Ceacam1, we have shown that impairment of insulin extraction causes chronic hyperinsulinemia and leads to insulin resistance by down-regulating insulin receptors and promoting de novo lipogenesis in liver. Based on preliminary data that underscore the central role of CEACAM1 in pathways regulating insulin action, lipid synthesis, and inflammation, we now expand our work to investigate the cause-effect relationship between hyperinsulinemia and insulin resistance in the context of diet-induced obesity. We show that loss of hepatic CEACAM1 is associated with human and rodent obesity, and is an early correlate of diet-induced obesity in mice. Thus, feeding mice a high-fat diet reduces hepatic CEACAM1 levels by > 50% to cause hyperinsulinemia and insulin resistance. Conversely, Ceacam1 overexpression in liver protects mice against insulin resistance and visceral obesity. This suggests that hyperinsulinemia plays a detrimental role in the pathogenesis of diet-induced obesity and insulin resistance. We now propose to test the hypothesis that reduction in hepatic CEACAM1 underlies the causative role of hyperinsulinemia in diet-induced insulin resistance. To this end, we will in Aim 1, investigate whether preventing hyperinsulinemia curbs diet-induced insulin resistance in a model of hepatic Ceacam1 gain-of-function, and we will systematically dissect which aspect of the metabolic syndrome (insulin resistance, dyslipidemia, hepatosteatosis, and inflammation) is prevented by the sustained expression of CEACAM1. In Aim 2, we will investigate the mechanism by which free fatty acids acting through PPARa cause transcriptional repression of Ceacam1. In Aim 3, we will investigate whether abolishing Ceacam1 regulation by PPARa preserves CEACAM1 expression, and protects against diet-induced insulin resistance. We will generate knock-in mice in which the PPARa response element in the Ceacam1 promoter has been mutated, and test whether this genetic manipulation confers protection against insulin resistance induced by high-fat diet. This should provide a critical test of the causative effect of hyperinsulinemia on insulin resistance, and identify CEACAM1 as a tractable drug target for the development of medications to combat altered metabolic conditions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This new institutional grant requests funds for four postdoctoral positions for training and research in neuroimaging. Two general classes of neuroimaging will be emphasized: radiotracer and nuclear magnetic resonance. The specific imaging methods are described by the following acronyms: PET (positron emission tomography, MRI (magnetic resonance imaging), fMRI (functional magnetic resonance imaging), and MRS (magnetic resonance spectroscopy). These imaging methodologies offer real promise for the expanded clinical utility of neuroimaging in the diagnosis, treatment, and understanding of the pathophysiology of major neuropsychiatric disorders. However, these new methodologies typically derive from complex, multidisciplinary sciences, which thereby create a challenging information barrier to their further development and application in medical practice. The purposes of the proposed program are the following: - Provide formal course training to postdoctoral fellows, with an emphasis on imaging and clinical research methodology, and instruction in their underlying multidisciplinary sciences. - Provide mentored training for fellows to apply these new methods to the interdisciplinary advance of our understanding of the function of the brain in health, substance abuse, and other psychiatric diseases. - Provide an integrated research experience for the fellows in both imaging applications and methodology. All fellows will be expected to complete at least one research project with an applications and one with a methodology mentor during their training period. The integration of training in methodology and imaging applications will be enhanced by the organization of this program as two interactive tracks of approximately equal size: Methodology and Application. We expect that the majority of fellows in the Methodology track will have Ph.D. degrees, and those in the Application track, primarily M.D. degrees. Each fellow will have a primary and secondary faculty mentor to represent the two tracks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] The PIs of this application have developed techniques for producing and studying atherosclerosis using funds from the first AMDCC program. Specifically, we have found that streptozotocin-treated mice develop increased atherosclerosis in the presence of a transgene for human aldose reductase (hAR). We have also noted that hearts from these mice have areas of cardiac apoptosis. In addition, we have developed novel methods to study atherosclerosis regression that can be applied to studies of lesions in control and diabetic mice. [unreadable] Aim 1 To create new mouse models of diabetic cardiovascular disease: We propose to create two new genetically altered mice. Aim 1a is to use the tet on system to allow expression of hAR in a time dependent manner. This system will allow us to test whether hAR expression in established lesions alters plaque morphology. These animals can also be used to produce tissue specific expression of hAR. Aim 1b is to produce mice with expression of hAR in cardiomyocytes. These mice, we hypothesize, will develop cardiomyopathy with diabetes. [unreadable] Aim 2 To study the development of vascular lesions in diabetic mice: Mild diabetes due to deficiency of Pdx1 or high fat diets did not alter atherosclerosis in Ldlr-/- mice. In addition, Pdx1 did not affect regression after transplant of arteries containing atherosclerosis. We will use two additional methods to generate hyperglycemia, Akita and high fat diets on the FVB background, in Ldlr-/- mice [unreadable] hAR. Increased vascular disease in STZ-treated hAR mice could result from greater monocyte/macrophage accumulation in lesions, or could be secondary to a defect in lesion regression. Both processes will be studied in vivo and mechanistic information obtained by studying gene and protein expression. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposed research is to investigate family factors and other social contexts in relationship to the perception and use of drugs among African-American and white adolescents. Such information will be useful for the design and implementation of preventive intervention programs for black adolescents. The task of designing preventive interventions for drug problems for black youth is hampered by the fact that more research is needed. While several studies have provided evidence concerning drug use and related problems among black youth, questions remain about some of the factors hypothesized to be critical to the phenomenon of drug use. The proposed research requests two years of funding to investigate differences in the use of drugs and alcohol by black and white adolescents. The proposed study has three broad aims. The first is to investigate the role of parenting behavior and practices for black and white adolescents as a factor in the use of drugs. The second is to investigate how parenting behavior and practices in the context of different family structures may be a factor in the use of drugs for black and white adolescents. Because of possible links with family socioeconomic status (SES), we will also investigate family SES as a factor in adolescent drug use. The third aim is to investigate the possible mediating role of peer use, religiosity and neighborhood cohesion between parenting behavior and practices and drug use for black and white adolescents. This proposed research is structured as a secondary data analysis of existing survey data. The data we propose to use to examine these issues is from the National Longitudinal Study of Adolescent Health (AddHealth), a nationally representative study of adolescents in grades 7 through 12 in the United States in 1995. The study was designed to investigate adolescent health, health behavior and relationships, and other issues. The data are longitudinal in design, and two waves of study data are available. Approximately 20,000 adolescents were surveyed, and additional data is available from the parents of the adolescents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "CLINICAL RESEARCH MANAGEMENT OFFICE The Clinical Research Management Office (CRMO) is the shared resource ofthe Kimmel Cancer Center (KCC) that supports physicians, scientists and other staff in the development, implementation, and conduct of clinical trials. The CRMO provides a broad range of services tailored to the specific needs of investigators, the KCC, and consortium partners. These services include supervision of activities related to trial registration and conduct, quality assurance, and monitoring of KCC institutional, pharmaceutical and cooperative group clinical trials. The CRMO also serves as a resource for the KCC community and patients by providing relevant information regarding clinical trial availability. Specific aims of the CRMO include: Orientation of new faculty to KCC, CRMO and (TJU) policies and procedures related to clinical research cilitate the development of KCC investigator initiated clinical trials Coordinate the preparation and submission of studies for review by the Multidisciplinary Groups (MDG), Clinical Cancer Research Review Committee (CCRRC) and the Thomas Jefferson University Institutional Review Board (IRB) Collect, abstract, maintain and update data specific to patients entered on clinical trials Coordination of study visits and procedures related to clinical trials Assure timely and accurate preparation and submission of all regulatory reviews (e.g. new submissions, amendments, annual reports, safety information, etc. to the IRB and FDA) Serve as a liaison between the TJU IRB, FDA or other regulatory bodies, and KCC investigators Assure accurate entry of patients participating in KCC clinical trials in a centralized database Assist in the maintenance of an accurate shared drive repository for complete protocols and informed consents. To improve easy access to viewing clinical trials Participate in the integration and activation of OnCore clinical trials management system Assure appropriate integration of consortium partners into the conduct of clinical research", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The mechanism underlying the high incidence of hypertension (HTN) in obesity and in noninsulin dependent diabetes mellitus (NIDDM) has not been clarified. Our hypothesis is that hyperinsulinemia and insulin resistance contribute to the pathogenesis of HTN of these states. We suggest that insulin increases blood pressure by enhancing sympathetic nervous system (SNS) outflow by altering sodium balances via absorptive processes including activation of Na+/K+ ATPase, by modifying the renin-angiotensin system, and by altering vascular responsivity through changes in Mg++ and through mitogenic effects on vessels. In addition, HTN per se causes insulin resistance and hyperinsulinemia, thus establishing a vicious cycle of events. Nine projects to explore these relationships are planned: 1) In a clinical module, a group of patients will be studied over time who are lean or obese, NIDDM or normal, and hypertensive or normotensive. Glucose and insulin kinetics, SNS, Na+ handling, renin, and various eicosanoids will be measured and correlated. 2) The effect of insulin and the hypertensive state on SNS will be studied both centrally and peripherally in Wistar-Kyoto and SHR rats. 3) The effect of insulin on vascular and cardiac cell growth in culture and on Na+/H+ antiporter and Na/K ATPase activity and gene expression will be quantitated both in vivo and in vitro. 4) The effect of insulin on vascular reactivity and angiotensin II (AII) and norepinephrine action will be studied. 5) We will determine by the role of Magnesium deficiency on vascular reactivity, platelet aggregation, and response to AII. 6) Systemic and renal production of prostacyclin and lipoxygenase (LO) products will be measured in the various clinical groups, during insulin clamp studies, and in a continuum of NIDDM patients with micro and macroalbuminuria. Using renin secreting cells and tissues as a specialized vascular model, the effect of the diabetic state on eicosanoid metabolism, AII action, and role of various growth factors on renin secretion will be explored. 7) Prorenin processing in NIDDM will be determined at the cellular and molecular level. 8) The effects of HTN and insulin on key cardiac protein gene expression and cell mitogenesis will be quantitated. 9) The role of the microcirculation or insulin sensitivity and receptor biology will be researched. The interaction of the various investigators, the presence of the clinical modules and animal core will allow us to extend knowledge and treat this attractive hypothesis in a manner that a series of independent projects could not accomplish.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The main objective of this investigation is to determine the optimal number and the optimal size of radioactive microspheres that should be injected into small animals to measure cardiac output and regional distribution of blood flow. The optimal number and optimal size will be based on: (1) The number of spheres which can be injected to give reproducible determinations of cardiac output and regional distribution of blood flow to organs or portions of organs without subsequent damage or impairment of flow to the terminal vasculature. (2) The most appropriate size of spheres for individual organs of small animals with minimal impairment of flow in that organ and with minimal escape from the systemic to the pulmonary circulation. Other objectives will be to determine 1) What percent of microspheres become lodged in vessels in the first circulation and how and where the remaining spheres eventually stop, 2) If the radioactivity varies significantly in any tissue over a time span from the first minute after injection to several hours after injection, 3) If microspheres significantly alter circulation of blood in the terminal vasculature.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this grant proposal is to map the frontal eye fields (FEF) in monkeys by microstimulation and to determine the neuronal connections of this cortical area by light microscopy. The FEF will be defined as the area which yields low threshold (20 - 50uamp) eye movement responses to stimulation. Injections of tritiated amino acids, (3H) in an FEF microstimulation site will label axons and terminals autoradiographically by anterograde axonal transport of labeled proteins. Nerve cell bodies projecting axons to FEF will be labeled by retrograde axonal transport of horseradish peroxidase (HRP), from FEF microstimulation sites. Anteorograde transport of HRP will be used as well to label FEF and other relevant axonal projections. Combined use of anterograde and retrograde labeling techniques in the same experiments will demonstrate interconnections between FEF and other functionally related areas, particularly the inferior parietal lobule (IPL) and superior colliculus (SC). FEF neurons sending axon collaterals ot two or more terminal sites will be identified by double retrograde labeling experiments using HRP and tritiated, inactive HRP (3HRP).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Description: (Taken from the Investigator's Abstract) The Pilot Projects Program is the responsibility of the Deputy Director. Once a year, an announcement will be made regarding the availability of funding; in addition, special requests for applications addressing targeted research areas may be issued. Center members and others outside the Center who wish to develop collaborations with Center members will be eligible to apply. Proposals will be reviewed by scientists not competing for pilot project funding. These reviews will be discussed by the Center's Internal Advisory Committee, which will make final decisions. Recipients will be required to write annual progress reports, and will be contacted beyond the end of the funding to document publications, grants, and collaborations that resulted from the pilot project. Pilot Project proposals will initially be reviewed by the Scientific Coordinator, Center Director, and the Deputy Director to determine whether they are appropriate to undergo the full review. Three reviewers will score each proposal and write a brief critique. Reviewers from affiliates of the Center, Center members not submitting a proposal, and scientists outside the Center will be sought for this purpose. External Advisory Committee members may be approached to review or suggest possible reviewers for the proposals once they have been received. The reviews will be completed within one month of receipt. All appropriate proposals will be scored for 1) relevance to the Center?s mission, 2) likelihood to result in a successfully funded application, 3) innovation, 4) appropriateness of laboratory and statistical methods and 5) collaborations within or among the Research Cores (for Center members) or with the Research Cores (for non-members). The scores will be tabulated by the Scientific Coordinator and the scores and critiques will be provided to the Internal Advisory Committee, which will discuss the projects and make the final funding decision. During its deliberations, the Internal Advisory Committee may take other considerations into account, besides the formal review criteria. These may include: impact on the junior investigator?s career development, utilization of the Facility Cores, potential impact on the Center as a whole, or other concerns.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development and spread of drug resistance in malaria parasites has become a major obstacle in the treatment and control of a disease that causes approximately 300 million infections and up to 3 million deaths per year. Artemisinin, and its derivatives, offer new hope in the effective treatment of malaria. This class of drugs rapidly clears clinical symptoms and parasites, including those that are multi-drug resistant. Unfortunately, when these drugs are used alone, > 40% of cases will produce recrudescent infections. Unlike recrudescence following treatment by other anti-malarial drugs, parasites appearing after artemisinin treatment remain susceptible to artemisinin. Our preliminary data suggests that P. falciparum parasites have a unique mechanism to survive artemisinin treatment: The drugs induce a dormant ring stage parasite in which growth is arrested for several days before the parasites recover and grow normally. This project aims to investigate the rate at which dormant parasites develop and recover following treatment with various artemisinin derivatives in vitro. In addition the duration of dormancy will be estimated. The role of dormancy in vivo will be investigated in an animal model. Physiological, cellular and molecular characterization of the dormant parasites will be performed to identify determinants/markers for dormancy and establish the mechanism(s) by which dormancy occurs. Artemisinin combination therapy (ACT) has been strongly recommended by WHO as a strategy to reduce recrudescence and to combat widespread resistance to all other cheap, available antimalarial drugs. We will determine if coadministration of drugs is effective in killing dormant parasites, or if the elimination half-life of the combination drug is the key factor in the success of combination therapy. We will supplement the experimental plan by using mathematical models of the in-host dynamics of P. falciparum infections to explore factors that may influence the formation of dormancy and potential ways to reduce parasite recrudescence following treatment with artemisinin. Although conventional resistance to artemisinin drugs has not yet been observed in the field, experiences with all other antimalarial drugs indicate the significant risk of parasites developing resistance to artemisinin drugs. We have developed artemisinin resistance in P. falciparum in our laboratory and will characterize the cellular and molecular mechanism(s) associated with resistance. These include transcriptional, translational/post-translational changes and mutations in potential drug transporters. This component of the project will elucidate possible mechanisms by which parasites develop resistance to this class of drug and the role of artemisinin-induced dormancy in the process. The results of this project will provide valuable information regarding the mechanisms of treatment failure for artemisinin drugs. The results will aid the formulation of optimal ACT regimens, improved treatment outcomes for malaria patients and defined strategies of preventing the development of resistance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Adapted from applicant?s abstract): It is hypothesized that the white blood cell genomic response can be used to deduce the presence of neuronal injury due to acute neurological diseases, and that the blood genomic response patterns can be used to differentiate between the diseases causing the neuronal injury. Our preliminary data using microarray technology show unique patterns of gene expression by lymphocytes of adult rats subjected to ischemic strokes, hemorrhagic strokes, status epilepticus, hypoxia, hypoglycemia and sham-surgeries as compared to untouched controls. The first Aim of this proposal will determine whether short durations of global cerebral ischemia, focal cerebral ischemia (transient ischemic attack), hypoglycemia and seizures produce different white blood cell genomic responses in rats that can be used to differentiate between these conditions hours to days later. The second Aim will determine whether long durations of global ischemia, hypoglycemia and status epilepticus regulate specific genes in white blood cells in response to the diffuse neuronal injury caused by all of these conditions, and whether these genes can serve as indicators of the diffuse neuronal injury. The genomic expression of neutrophils, lymphocytes and whole blood will be examined at various times after cerebral ischemia, insulin-induced hypoglycemia, seizures and status epilepticus. Genes regulated in the different white blood cells by these conditions will be correlated with the presence of diffuse neuronal cell death in brain using TUNEL staining. The third set of Aims will determine whether the same genes regulated in white blood cells of rodents following single seizures and status epilepticus are also regulated in the white blood cells of men and women patients following seizures and status epilepticus. These studies will also determine whether blood genomic responses can be used to distinguish whether patients have had seizures, pseudoseizures or syncope, and whether some of the neuronal injury-related genes regulated in the blood of rodents with status epilepticus are regulated in patients with status epilepticus. Genes regulated more than two fold on microarrays will be confirmed by quantitative RT-PCR for all of the aims. The goal is to objectively differentiate seizures, syncope, global cerebral ischemia, hypoglycemia, and transient ischemic attacks hours to days after they occur; and to begin to identify blood genomic markers of neuronal death associated with acute neurological diseases that might also be useful in chronic neurological diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To support the NIH National Center for Biotechnology Information to maintain a national database of genomic and other data sources on resistant bacteria as outlined in the National Action Plan to Combat Antibiotic Resistant Bacteria. This effort addresses the growing problem of antimicrobial resistance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The high affinity receptor for IgE on mast cells and basophils plays a central role in immediate hypersensitivity reactions. Crosslinking of bound IgE by polyvalent antigen leads to aggregation of the receptors and cellular secretion of both preformed and newly synthesized mediators of inflammation. The molecular mechanisms by which aggregation of the receptors generates cellular responses are still largely undefined. During the past year we have continued to employ mutated receptors in order to analyze which portions of the receptor participate in its principal functions. Our new results show: 1) None of the cytoplasmic domains are required for receptors to exhibit detergent-insolubility after aggregation. Indeed, lipid-anchored ectodomains of the alpha subunit are sufficient to demonstrate this effect. 2) A receptor-less mast cell-like line has been successfully transfected with various mutant receptors and studied with respect to the early biochemical events associated with aggregation of the wildtype receptors for IgE. Truncation of the cytoplasmic domain of the alpha subunit or at the amino-terminus of the beta subunit failed to inhibit signalling. Truncation of the gamma chains also led to loss of functions. In this case the receptors for IgG were also inactivated possibly because the functionally active receptors are of the type that contain gamma subunits. Surprisingly the inactive IgE receptors were found to contain normal amounts of the wild type (endogenous) gamma chains. This suggests that the transfected mutant gamma chains had a negative dominant effect.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our application addresses broad Challenge Area (04): Clinical Research and specific Challenge Topic, 04- HD-102: Development of Pediatric Medical Devices. The severity of hemiplegia due to damage in the brain in both the pediatric and adult populations stems not only from the loss of neurons killed by the vascular insult but also from the loss of function in viable but dormant neurons in the affected hemisphere. The latter problem may be avoidable and, to a certain extent, correctable. The down-regulated excitability in these neurons results from compensatory overuse of cortical motor neurons in the non-stroke hemisphere, which 1) suppresses motor neurons in the stroke hemisphere through inhibitory transcallosal projections, and 2) competes with and suppresses the functional maturation of corticospinal projections from the stroke hemisphere through its ipsilateral projections to the paretic hand. The strategy in rehabilitating pediatric hemiplegia is to overcome this \"developmental disuse\". Therefore, in pediatrics, where there may be greater potential for neuroplasticity, the challenge is to restore excitability and voluntary control of these down- regulated neurons. Disruption of the excitability in the non-stroke hemisphere, with resultant increased excitability (disinhibition) in the stroke hemisphere, can be achieved with low-frequency repetitive transcranial magnetic stimulation (rTMS), a noninvasive method of stimulating targeted areas of the brain. Furthermore, the disruptive effects of low-frequency rTMS can be increased and prolonged by preceding it with 6-Hz priming stimulation. We have recently demonstrated the safety of this priming/low-frequency rTMS in adults with stroke and we are currently engaged in an NIH-funded study exploring the effectiveness, mechanism and safety of serial treatments of rTMS combined with motor learning training. Very little research, however, has been attempted on interventions to promote brain reorganization and recovery in hemiplegia in children. Thus, this study will determine the effectiveness, mechanism, and safety of 5 treatments of 6-Hz primed low- frequency rTMS applied to the non-stroke hemisphere and combined with constraint induced therapy (CIT) to promote recovery of the paretic hand. Fifteen children with pediatric hemiplegia, age 8 -16 years, at each of two pediatric medical facilities (total N = 30), will be randomly assigned to one of two treatment groups that will receive treatment for 2 weeks. The rTMSreal/CIT group will receive alternating days of the real rTMS and CIT for 5 treatments of each. The rTMSsham/CIT group will receive alternating days of sham rTMS with real CIT for 5 treatments of each. Subjects will be tested at pretest and posttest. The hypotheses are: 1) the rTMS treatment will show no serious side effects, 2) both groups will show improvement in hand function but the rTMSsham/CIT group will show significantly greater improvement, 3) the rTMSreal/CIT group will also show significantly greater improvements in brain reorganization measured by paired-pulse TMS testing, cortical silent period testing, and fMRI. The proposed research is important because very little research on rehabilitation-induced brain reorganization has been done in pediatric hemiplegia. It is innovative because it applies a technique never used before, i.e. 6-Hz primed low-frequency rTMS combined with CIT. The potential impact of this research is a radical change to pediatric rehabilitation that accomplishes a higher functional recovery. This project has the potential to discover a new method of promoting brain reorganization and recovery of hand function in children with hemiplegia. Very little research has been done on children with hemiplegia that addresses brain reorganization. Noninvasive brain stimulation using repetitive transcranial magnetic stimulation, when combined with Constraint Induced Therapy, may show a potent synergism that produces unprecedented rehabilitation gains. Pursuing this knowledge will help to promote higher quality of life in thousands of children each year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Polyphenols are common constituents in botanical extracts available in over-the-counter nutraceutical preparations as well as being found in different fruits and vegetables, olive oil, and beverages like red wine and tea. Based on a growing body of evidence it is thought that the chemoprotective effect of polyphenols involves the inhibition of reactive oxygen species (ROS) generation rather than simply scavenging the radicals. Recent studies indicate that NADPH oxidase is the major source of ROS in the three principal cell types of the blood vessel wall (e.g., endothelial cells (EC), vascular smooth muscle cells (VSMC), and adventitial fibroblasts). There has been growing interest in the vascular NADPH oxidases, largely because it is recognized that oxidative stress plays a critical role in the pathogenesis of vascular diseases, such as atherosclerosis. Preliminary studies performed in our group and others have demonstrated that ortho- methoxyphenols, such as apocynin, are activated by peroxidases in vivo and that their oxidation products appear to act as inhibitors of NADPH oxidase assembly and activation, although the exact identity of the oxidation products and the mechanism of inhibition have not been fully elucidated. Such information is critical for the ultimate development of safe and effective inhibitors of vascular NADPH oxidases. In the present application, we propose to obtain structural information on the active metabolites of apocynin-like compounds and the biochemical reactions by which they bind to and inhibit the assembly and activation of the NADPH oxidase complex. Specific goals of this R21 application are to: (1) Elucidate the chemical structures of the active metabolites of apocynin that bind to endothelial p47phox, p67phox, and p22phox peptide sequences and confirm that these compounds also bind to the same domains on the p47phox and p67phox protein subunits of NADPH oxidase; (2) Determine whether apocynin oxidation products disrupt specific protein-protein interactions that are involved in the assembly of mature NADPH oxidase. The outcome of these studies will be the foundation of a more detailed understanding of the chemical structure and molecular process by which apocynin-like compounds inhibit the assembly and activation of NADPH oxidase. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Base-selective heavy atom labels for electron microscopy-based DNA sequencing Project Summary/Abstract The development of inexpensive and rapid DNA sequencing technology remains a major challenge of broad scientific interest. Preliminary work at Halcyon Molecular has shown that transmission electron microscopy (TEM) can be used to obtain ultra-fast ultra-low-cost DNA sequences. Since efficient electron scattering to a detector is highly dependent on atomic number (Z), it is possible to label single stranded DNA (ssDNA) with heavy atoms. To test the limits of this trend, we propose a multipronged approach to selectively prepared metal-DNA base pair complexes. Our effort will be synergistic, taking advantage of the experience of the Toste group in organometallic and heavy atom cluster synthesis, and the capabilities of Halcyon Molecular in manipulating DNA and performing TEM. For this proposal, we are focusing on the selective labeling of DNA bases and the development of an appropriate assay to evaluate our success. Two general synthetic methods will be investigated in order to develop distinct labeling protocols. First, triosmium (ZOs = 76), tetrairidium (ZIr = 77) and trigold (ZAu = 79) clusters tethered to a group that selectively react with (alkylating reagents) or bind (platinum diamine complexes) purine bases will be explored. Incorporation of gold (ZAu = 79) and mercury (ZHg = 80) atoms through direct metal-metal bonds to the osmium atoms will also be explored. In this case, the labels would appear as intense spots in the TEM spectra. For the complimentary pyrimidine label, osmium tetraoxide bipyridine will be the selective binding agent thymine and cytosine. Using the bipyridine ligand as a scaffold for functionalization, additional osmium, platinum (ZPt = 78) or uranium (ZU = 92) atoms may be incorporated. A linear arrangement of metal atoms would allow a positional vector to be drawn towards the corresponding base. Proof-of-concept experiments will be performed using nuclear magnetic resonance (NMR) spectroscopy using individual DNA bases. If successful, testing will be performed on single DNA strands and sequenced using TEM. The success of these methods will enable the base-selective labeling of DNA with metal atoms and help develop ultra-fast ultra-low-cost DNA sequencing technology. The assembly of a whole human genome with our pilot-scale instrument can demonstrate TEM sequencing's potential for high consensus accuracy, extremely long (>150kb) reads, and lack of sequence specific bias in molecule deposition and readout. The subsequent, commercial availability of whole human genome sequencing using this technology (with expected >99.9999% consensus accuracy and completeness in <10 minutes/genome, at a cost of <$100) will enable new opportunities in translational medicine and foster many new discoveries by NIH investigators. PUBLIC HEALTH RELEVANCE: The proposed program aims to develop heavy atom organometallic compounds for ssDNA base-selective labeling for use in ultra-low-cost DNA sequencing technology based on single- atom sensitivity transmission electron microscopy (TEM).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract For patients with severe pancreatitis refractory to medical and endoscopic therapy, total pancreatectomy (TP) with islet autotransplantation (IAT) may be considered. While 90% of TPIAT recipients have some function of the transplanted islet graft, only about 1/3rd come completely off insulin. The long-term goal of the proposed research is to develop new therapies that will increase the number of patients who are non-diabetic following islet autotransplant. Such therapies may also benefit recipients of islet allotransplant for type 1 diabetes. Following islet transplantation, the islets must acutely survive the stress of the procedure, and then they must engraft in the liver and establish a vascular supply. The greater the functional islet mass engrafted, the lower the risk of post-operative diabetes. It has been estimated that more than half of the islet mass may be lost in the early post-transplant period in islet transplant recipients. Beta cell apoptosis is common during the first month post-transplant and is upregulated in the presence of inflammatory cytokines such as TNF?. Thus, a major contributor to islet loss is the inflammatory damage sustained by the transplanted islets in the early post-transplant period; we propose to directly target this destructive process. Two promising anti-inflammatory therapies are available to address this problem: (1) the TNF? inhibitor etanercept and (2) the serine protease inhibitor alpha-1 antitrypsin. Both agents are commercially available for clinical trials. Proof-of-principle for etanercept has been demonstrated in type 1 diabetic allotransplant recipients, in whom a 10 day course of etanercept early post-transplant significantly improved long-term insulin independence, due to better survival of the transplanted beta cell mass in the engraftment period. Alpha-1 antitrypsin (A1AT) reduces inflammatory cytokines, protects against cytokine-induced beta cell apoptosis, and prolongs islet graft survival in mice and intraportal IAT non-human primates. This initial 3-arm drug-treatment clinical trial will investigate the use of Etanercept and A1AT to improve IAT function at 90 days and 1 and 2 years post-TPIAT compared to standard care. Forty-five patients undergoing TPIAT will be randomized 1:1:1 to receive either: 1) etanercept (50 mg on day 0; 25 mg on days 3, 7, 10, 14, and 21), 2) alpha-1 antitrypsin (90 mg/kg IV days -1, +3, 7, 14, 21, 28) or 3) standard care. Patients will have mechanistic assessments drawn in the early post-operative period including inflammatory cytokines and chemokines and measures of beta cell loss. Metabolic testing will occur at 90, 365, and 730 days post-TPIAT, including mixed meal tolerance testing, IV glucose tolerance testing, and glucose-potentiated arginine-induced insulin secretion (GPAIS). The latter measures the maximally stimulated acute insulin response (AIRmax) as the best estimate of islet mass and the primary endpoint (at day 90) for this study. Results will be used to select the most promising agent for future study in a randomized, blinded multi-center clinical trial.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aryl hydrocarbon receptor (AhR) is an orphan nuclear receptor and a central mediator of the effects of an entire class of environmental toxicants. The AhR has an undefined role in normal and disrupted embryogenesis as well. Reproductive senescence is accelerated by 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD, dioxin), the specific and potent AhR ligand, at environmentally relevant exposures and one consequence is a decline in oocyte and subsequent embryo quality. As women increasingly delay attempts at pregnancy into their 30s and beyond, any acceleration of reproductive aging has dire consequences for the possibility of successful pregnancy and the risk of birth defects. Furthermore, early reproductive senescence in women is associated with decreased lifespan and quality of life. Although the effects of AhR ligands on oocyte numbers have been studied extensively, little is known about the AhR, oocyte quality and subsequent organization of the early embryo. In our preliminary work, we have identified early chromosomal and cytoskeletal defects that may be pathognomonic for exposure of oocytes and embryos to AhR ligands such as dioxins; this can explain the decreased fertility resulting from chronic exposure with age. The objective of this project is to identify checkpoints of oocyte development and early embryogenesis that are regulated by AhR ligands. Our central hypothesis is that environmental exposure to AhR ligands reinforces the decline in oocyte and embryo quality with age. This work, once completed, promises new insights to prevent the loss of fertility in polluted environments and slow the loss of fertility with age. The research team will achieve these experimental goals through the following specific aims: Specific Aim 1: Determine the effect of defined AhR activation on critical periods of oocyte maturation and early embryogenesis in vitro. This aim encompasses the most detailed investigation into the impact of AhR activation or antagonism on the loss of oocyte and embryo quality to date. Endpoints include embryonic morphology, cytoskeleton and chromatin and its epigenetic modulation in oocytes and embryo. Specific Aim 2: Determine the impact of chronic, environmentally relevant AhR activation and antagonism on oocyte and embryo quality during normal and accelerated reproductive senescence. Oocytes and embryos from young and middle aged rats will be assessed for morphological and cytoskeletal conformation and chromatin remodeling and epigenetic modification and imprinting from oogenesis to early embryogenesis. This will provide insight into decreased oocyte and embryo quality and fertility with age and the importance of the AhR pathway in the aging ovary and oocyte. This work is novel through use of realistic exposures to AhR agonists and a focus on subtle measures oocyte and embryo quality and epigenetics. These studies explore a novel diagnostic indicator of compromised oocyte and embryo quality due to exposure to toxic AhR ligands. This project will provide crucial knowledge for the identification and prevention of infertility and birth defects due to the AhR pathway in older mothers. Hundreds of manmade compounds including dioxins, polychlorinated biphenyls and carcinogens in cigarette smoke act on cells through the aryl hydrocarbon receptor (AhR) pathway. We have recently reported detrimental effects of AhR activation on aging of the ovary and a number of defects in eggs and embryos from females exposed to AhR ligands. This project will provide crucial knowledge for the identification and prevention of infertility and birth defects due to the AhR pathway in older mothers. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Substance use disorders among criminal offenders present a unique challenge to criminal justice and public health systems. These systems must balance punishment and rehabilitation while simultaneously dealing with addiction - a chronic, cyclical, and multidimensional disease that is pervasive among criminal offenders and requires long-term monitoring and extended care protocols (McLellan et al., 2000;Scott et al., 2003). Women offenders, approximately 70% of whom have substance dependence or abuse issues (BJS, 2002), are of particular concern from economic, clinical, and criminal justice perspectives as they represent the fastest growing subgroup in local jails (BJS, 2003). Women offenders are also more likely to have co-occurring mental health problems that further complicate their rehabilitation and recovery process upon reentering society (CASA, 2010). Moreover, the social and economic implications of addiction and criminality extend to the children of women offenders. The need to address substance use within the women offender population has generated interest in Recovery Management Checkups (RMCs), an evidence-based approach to promoting long-term recovery that provides clients with periodic monitoring, feedback, and linkage with treatment as needed. The Recovery Management Checkups for Women Offenders (RMC-WO) intervention is currently testing the long-term effectiveness of the RMC model with a group of women offenders coming out of jail. These interventions are designed to reduce substance use, HIV-risk behaviors, and criminal recidivism. Very little economic research has been conducted on community re-entry and long term monitoring models for women offenders that have the potential to improve long-term outcomes and lower total costs to society. In response to the pressing need for economic evaluations of RMCs, the proposed research project will estimate the costs, cost effectiveness, and net benefits of RMC-WO relative to a control condition. The specific aims of this research are as follows: 1. Estimate the economic cost (total and per client) of the RMC-WO intervention. 2. Estimate the cost effectiveness of RMC-WO relative to the control group over the three-year follow-up period for core outcomes (substance use, HIV risk behaviors, illegal activity) and ancillary outcomes (treatment engagement and treatment retention). 3. Obtain or derive appropriate monetary conversion factors and estimate the economic benefits of RMC- WO relative to the control group over the three-year follow-up period to form benefit-cost ratios and net benefit estimates. Successfully fulfilling these aims will not only advance scientific knowledge of the overall impact of addiction interventions for women offenders but also improve clinical practice by increasing our understanding of the financial considerations of implementing RMC-WO on a broader scale. PUBLIC HEALTH RELEVANCE: Despite the progress and increasing use of economic evaluation methods in addiction research, rigorous economic evaluation of post-release interventions for women offenders remains rare. From a public health perspective, women offenders are of special concern because they are a rapidly growing subpopulation among all offenders and because they are vulnerable to numerous forces that carry a high personal and social cost. The proposed study will conduct the first comprehensive economic analysis of a randomized control trial of Recovery Management Checkups for Women Offenders (RMC-WO) to examine the cost effectiveness and net social benefits of this intervention and to provide treatment providers, criminal justice agencies and policy makers with empirical evidence of the economic impact of RMC-WO.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Epidemiological data demonstrate that aerobic exercise training can dramatically reduce cardiac mortality even in patients with pre-existing cardiac disease. The mechanisms responsible for this cardio protection remain largely to be determined. It is probable that exercise-induced changes in cardiac autonomic regulation play a major role in the improved cardiac mortality. Cardiac autonomic balance is altered by cardiac disease and the patients with the greatest changes (i.e., decreased parasympathetic and/or increased sympathetic activity) are also at the greatest risk for sudden death presumably due to ventricular fibrillation (VF). Exercise training can increase parasympathetic and decrease sympathetic activity and could thereby reduce mortality. Therefore, the proposed studies will test the central hypothesis that exercise training augments parasympathetic and/or reduces cardiac sympathetic activity and thereby protects against VF. Specific Aim #1 will test the hypothesis that exercise training alters cardiac autonomic balance in animals susceptible and resistant to VF. Specifically, cardiac autonomic balance will be evaluated in animals either resistant or susceptible to VF before, during and after the completion of an exercise conditioning program (8-10 weeks of daily treadmill running). The autonomic response to two different physiological stressors, submaximal exercise and acute myocardial ischemia, will be evaluated. Cardiac autonomic balance will be evaluated by pharmacological tests (agonist dose response, effects of selective antagonists), baroreflex sensitivity, and time series analysis of R-R interval variability. Specific Aim #2 will test the hypothesis that the cardiac autonomic changes induced by exercise training are responsible for the protection noted for this intervention. The effects of parasympathetic activity will be evaluated with a cholinergic antagonist. Thus, if parasympathetic enhancement is responsible for the protection, then atropine should reinstate VF in the susceptible animals. Specific Aim #3 will test the hypothesis that exercise training can reverse the increased Beta-adrenoceptor responsiveness that we have shown to occur in dogs that become susceptible to VF following myocardial infarction. Ventricular contractile responses to Beta1- and Beta2-adrenoceptor stimulation will be examined in vivo by echocardiography and in vitro by single cell fluorescence microscopy/video edge detection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We are interested in understanding how transcription of protein-coding genes is regulated in vivo. Current models for transcriptional regulation are based largely on in vitro studies conducted under non-physiological conditions using highly purified general transcription factors, synthetic activator proteins, and non-chromatin DNA templates. By studying transcription of endogenous genes that are required for proper Drosophila melanogaster development, we hope to identify cis- and trans-acting factors that play regulatory roles in vivo and determine how the coordinated action of different sets of factors establish the broad transcriptional repertoire of eukaryotic cells. To this end, we are studying transcription of the Drosophila sevenless (sev) gene. The sev gene was chosen because minor alterations in the level or pattern of sev transcription cause easily observable phenotypes in the developing fly, making sev transcription amenable to genetic dissection. Knowledge gained from studies of sev will be tested on a genome wide basis to assess the generality of the Sev model.Sev-based genetic screens conducted in our laboratory have identified mutations in components of the RNA polymerase (pol) II preinitiation complex [TBP-associated factor (TAF)60 and TAF110 and RNA pol II subunits] as well as factors that modulate transcription through their affects on chromatin structure [Trithorax group (TRX-G) genes, RPD3, and SIN3]. Additional mutations have been identified that may also encode transcriptional regulators. Phenotypic and molecular characterization of mutant flies is being used to identify genes that are transcriptionally regulated by the encoded factors, and analysis of genetic interactions between mutant flies is being used to assess functional interactions between the encoded factors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have developed a protocol for accurately synchronozing Madin-Darby bovine kidney cells to the Gl/S border of the cell cycle. Using this system, we have found that bovine satellite I DNA initiates its replication 1.0 - 1.5 hours into 5 phase of synchronized MDBK cells. Concurrently, microbubbles appear in the satellite I DNA as visualized by electron microscopy; these occur at a fixed position on the satellite I restriction map. In this proposal we describe experiments which will utilize synchronized cells at specific points in the S phase of the cell cycle. We propose to hybridize Okazaki fragment probes derived from the beginning stages of S phase to restriction fragments of satellite I DNA to confirm the replicative nature of microbubbles and to map the origin of replication more precisely. We have found that bovine satellite DNA is enriched on the nuclear matrix. By restriction analysis we will define those domains of satellites I + IV are protected by the matrix, and which may correlate with the map position of the origin of replication. DNA sequence analysis of these domains will demonstrate if specific DNA sequences are recognized for attachement to the nuclear matrix. We have found that bovine satellite IV was derived from bovine satellite I which gained a DNA insertion. Despite their evolutionary relationship, bovine satellite IV initiates its replication later in S that satellite I. Comparison of nuclear matrix bound sequences (origins) of homologous regions in these two satellites will show if DNA sequence itself is directly involved in temporal control of initiation of replication.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Experimental allergic encephalitis serves as a model for the study of therapies for demyelinating disease of the central nervous system. Its similarity to multiple sclerosis lies not only in clinical features and pathology, but more importantly like MS susceptibility to EAE is linked closely to immune response genes of the major histocompatibility complex (MHC). It has been shown that treatment in vivo with monoclonal antibody to the cell surface product of the susceptibility gene (I-A antiger can prevent the induction of acute EAE, and reverse the course of chronic EAE. Similar observations have been made with other autoimmune disease wherein susceptibility was linked to IR genes of the MHC. While the precise mechanism of action of anti I-A antibody is not known, it is quite clear from a number of systems that its actions are multimodal. This proposal attempts to characterize the haplotype specific nature of suppression of EAE using F1 restricted T cell lines and clones. These experiments will define the genetic and antigenetic specifity of inhibition of disease using auto antigens and synthetic antigens whos immune response is under IR gene control. At present, the mechanism by which IR genes confer susceptibility to disease is not known. Understanding mechanisms by which antibody to IR gene products regulate auto immune disease might lead to an improved knowledge of the pathogenesis of EAE.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The principal objective of the proposed research is to gain further insight into the mechanisms of integrative recombination of viruses. Bacteriophage Mu is a virus that closely resembles mammalian tumor viruses in respect to its mode of integration and replication. Accordingly, phage Mu may prove to be a useful model for understanding the mechanism of integration of animal virus DNA. We propose to study the metabolism of Mu DNA in conditions designed to favour the detection and characterization of replicative and/or integrative forms of the viral genome. Specific objectives of this research plan include (1) visualization, by electron microscopy, of early replicative forms of Mu DNA produced in anucleate minicells after induction of prophage Mu,(2) characterization of Mu-associated heterogeneous circular DNA with respect to its mode of formation and possible role in late replication or maturation of Mu DNA, (3) examination of the products of prophage excision in minicells for possible integrative forms of Mu, and (4) investigation of Mu development in host strains that allow separation of Mu-specific DNA forms from host DNA on the basis of differences in buoyant density.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mass-analyzed ion kinetic energy spectra (MIKES) of C5H5-plus and C5H5-minus ions were used to demonstrate that the frequently proposed cyclopentyl cation is not an important structure for C5H5-plus ions. Factors affecting dynamic range in \"direct mixture\" analysis using MIKES were explored. Usable dynamic range is limited by background secondary ion currents and source defocusing to ca. 10 to the 4th power:1. Computer programs for MIKES analysis have been refined.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Applicant's Description) Project 3 will analyze how to include patient related geometric uncertainties (due primarily to patient setup variability and organ motion) in the calculation, compilation and treatment of conformal radiotherapy dose distributions. These very important and fundamental realities associated with the therapeutic treatment of cancer patients with external beam ionizing radiation are not reflected in the standard computation of dose distributions for patient treatment plans. While knowledge of the magnitude of these uncertainties, when available, can be exploited to design geometric safety margins for the treatment of tumors, these same margins often compromise the dose that can be safely delivered to the patient's target volume(s) due to the irradiation of large volumes of normal tissue. Further, even given adequate treatment of clinical target volumes, the dose distribution actually received by the patient (especially normal tissue) is not accurately represented in the single dose calculation performed prior to treatment, using a static imaging study as the underlying anatomical model. This is of importance in the continuing studies of conformal therapy techniques, as many dose escalation and optimization schemes are based on (or constrained by) the perceived probability of expressing a treatment-related complication. This project has specific aims associated with the inclusion of patient-related setup uncertainties and organ motion in the calculation of realizable dose distributions, the compilation of delivered dose distributions that reflect individual patient and organ positions over the course of treatment, and ultimately the combination of these realizable dose calculations with patient specific, realized dose compilations in the development of dynamic refinement strategies for the optimization of individual patient treatments. Stylized, realizable dose treatment plans for individual patients should result in achieving required clinical target volume coverage, with more confident descriptions of normal tissue doses. Optimal safe treatments at a given prescription dose, or further tumor dose escalation at specified levels of normal tissue risk can then be attained. It is anticipated that the investigations will permit further optimization of treatments for individual patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "After completing two years of preclinical science studies at Cornell University Medical College, I affiliated with the laboratory of Neuroendocrinology, headed by Bruce McEwen, at the Rockefeller University. There I have chosen to study the role of cytokine expression in the central nervous system (CNS), specifically, with respect to their function in response to stress and injury in the hippocampus. Cytokines have been implicated in the pathophysiology of a number of neurologic disorders such as multiple sclerosis, Alzheimer's, and AIDS-dementia complex. Moreover, immune function in the CNS remains poorly characterize. I believe the study of this interaction will lead to a better understanding of the function cytokines perform in maintaining homeostasis in the CNS, as well as role they play in inflammatory and neurodegenerative disorders. Ultimately, this will offer us an opportunity to find targets for therapeutics and further understand the basis for a number of poorly understood diseases of the nervous system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This T32 Training Grant has focused successfully over the past 30 years on providing a dynamic research training environment for pathologists, other physician-scientists and basic scientists working in the molecular pathology of cancer. Over the past two 5-year funding periods, trainees have published a considerable number of papers, including in high quality journals, and have found faculty positions in prestigious medical schools. The present proposal features 20 Harvard Medical School and Massachusetts Institute of Technology faculty who are centered in the Massachusetts General Hospital Molecular Pathology Unit and Center for Cancer Research. This interactive, overlapping community of scientists and physician-scientists is organized into thematic programs in cancer genetics;animal models of cancer;mechanisms of gene expression and protein folding;cell cycle control and DNA damage repair;stem cell biology;and in vivo imaging for cancer biology and diagnosis. As in previous cycles, the trainees will be selected from a competitive pool of MD, MD-PhD and PhD applicants on the basis of prior academic and research achievements and evidence of a strong commitment to a career in cancer biology. The period of training will be two or three years for each successful applicant. An active and program-specific recruitment and retention program to enhance diversity ensures that diverse applicants have access to, and succeed in, the training program. In summary, the proposed T32 program renewal capitalizes on a highly interactive, experienced and focused faculty;a distinguished record of training productive physicians and scientists, including underrepresented minorities;state-of-the- art facilities and educational resources;and exposure of the trainees to basic and translational aspects of cancer biology. PUBLIC HEALTH RELEVANCE: Cancer remains a major cause of morbidity and mortality in the United States, and new approaches to the diagnosis and treatment of cancer are therefore necessary. This T32 program focuses on the training of physician-scientists and scientists who plan research careers that will improve our understanding of the molecular basis of human cancers. The program is based on the translational and basic science strengths of a Pathology department and a Cancer Center, and thus creates a dynamic platform to educate physician-scientists and scientists in the study of cancer biology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The endothelial cell is topologically poised in the vasculature to sense and respond to a host of environmental signals, including reactive oxygen species (ROS). These chemically active molecules serve important roles in normal, homeostatic signaling;and their potential for oxidative injury is ameliorated by an elaborate system of antioxidant defenses. As the flux of ROS increases, the endothelial cell responds by enhancing protective mechanisms, leading to a state of compensated stress;when the flux of ROS increases further, these protective mechanisms are overwhelmed, leading to a state of uncompensated oxidant stress. In this program project application, five project leaders have come together to investigate mechanisms that underlie oxidant signaling and adaptation to oxidant stress in the endothelial cell in health and disease. Project 1 focuses on the mitochondrion as an important component of endothelial redox signaling;Project 2 addresses the role of glucose-6- phosphate dehydrogenase and its enzymatic product, NADPH, as key determinants of the thiol redox state in the endothelial cell;Project 3 examines the role of Foxo transcription factors in promoting resistance to oxidant stress in endothelial progenitor cells;Project 4 considers the effect of ROS dependent oxidative modifications of Ras on insulin signaling in the endothelial cell;and Project 5 tests the hypothesis that mitochondrial dysfunction and resulting oxidant stress contribute to endothelial dysfunction in human atherosclerosis. This conceptually cohesive program brings together five well established project leaders who have a long history of productive collaboration to focus on a key theme in endothelial cell biology. Using contemporary methods of cell and molecular biology, as well as genetic animal models and human studies, mechanisms will be dissected at the molecular and cellular levels and applied to mammalian systems. With the work proposed in this proposal, we hope to be able to identify novel approaches to understanding the biomolecular basis of oxidant signaling and the adaptive and maladaptive consequences of oxidant stress in endothelial (patho) biology and vascular disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 17,000 new cases of malignant brain cancer are diagnosed per year in the United States. Even with surgical resection, radiation, and chemotherapy, patients with glioblastoma (GBM), the most refractory form of the disease, typically succumb to the disease within 2 years of diagnosis. A major limitation in the development of effective therapies for recurrent GBM is the lack of a reliable method to predict early response to therapy. Monitoring response using conventional magnetic resonance (MR) imaging is often unreliable and are often unable to distinguish treatment effects from tumor response. In animal models of GBM, we recently found that response to temozolomide (TMZ) and PISK pathway inhibitors is associated with a tumor-specific decrease in the conversion of hyperpolarized carbon 13 (130) pyruvate to 13C lactate detectable by non invasive 130 MR spectroscopic imaging (MRSI) as a drop in the ratio of hyperpolarized lactate to pyruvate (Lac/Pyr). We therefore hypothesize that measurement of Lac/Pyr could serve as an early indicator of drug action in patients following treatment with TMZ, PI3K inhibitors, and potentially a range of other agents. This hypothesis will be tested by 1) defining in vitro sensitivity and verifying target inhibition in recurrent human GBM cells exposed to TMZ and other novel targeted agents including XL765 and SAHA 2) defining the effect of the drugs on the conversion of 130 pyruvate to 130 lactate in the drug sensitive/resistant paired GBM cells, and relating these changes to target inhibition and drug sensitivity, 3) determining if the drug-induced changes in 13C pyruvate to 130 lactate conversion that parallel target inhibition and drug sensitivity in vitro also do so in xenografts in vivo, and 4) determining if treatment-induced alterations in pyruvate metabolism detected by hyperpolarized 130 MRSI can be used alone, or in combination with other imaging markers, as a biomarker of target inhibition and early response in clinical trials of recurrent GBM patients. Close interactions with Project 1 and all the Shared Resource Cores will be critical to the successful conduct of the proposed studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The neuronal ceroid-lipofuscinoses (NCL) are possibly the most common group of progressive neurodegenerative diseases in children, with an incidence as high as one in 12,500 live births, and with about 440,000 carriers in the USA. Juvenile NCL/Batten disease is the most common of these disorders and the subject of this proposal. Individuals with the disease were found to harbor a 1 kb deletion, which introduces a frameshift that leads to a predicted translation product of 181 amino acids, of which only the first 153 residues correspond to the first 153 of the normal 438 amino acid CLN3 gene product. The yeast homolog to CLN3 was identified and designated BTN1. We had previously shown that Btn1 p may be involved in maintaining pH homeostasis. Importantly, CLN3 is able to complement the alteration in vacuolar pH in the yeast model lacking Btn1 p, indicating that they have similar, if not the same cellular functions. Our more recent studies indicated that lacking Btnlp resulted in a defect in vacuolar transport of arginine, and again CLN3 is able to complement the defect in arginine transport. Overall our studies indicate that yeast cells work to maintain pH homeostasis, and that Btn1 p is an integral part of the biology of this process. This proposal sets out to investigate bfn1-/l-mediated disruption of pH homeostasis within the single cell of yeast. By elucidating the mechanisms by which this single celled organism balances intracellular pH and by uncovering the specific function of Btnlp and other proteins in the BTN-pathway we will establish a basis for understanding pH homeostasis in mammalian cells. We propose to further characterize the biochemistry of Btn1 p-dependent regulation of vacuolar pH. Moreover by exploiting assays that correlate to Btn1 p function such as vacuolar arginine transport and vacuolar proton pumping we will further define the structural requirements of Btn1p/CLN3. Concomitant to these studies we will identify components of the BTN1- pathway through use of a variety of genetic screens such as phenotypic suppression and synthetic lethality. Finally we will characterize the pathway of trafficking Btnlp to the vacuole. Further understanding of Btnlp (and ClnSp) in yeast will provide valuable information on the pathogenesis of Batten disease. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Regulation of the number of insulin receptors on the cell surface plays a critical role in determining insulin sensitivity. In order to study the mechanism of transcriptional regulation of the insulin receptor gene, we have cloned the 5'-end of the human and mouse insulin receptor genes. Recently, we have characterized trans-acting factors that bind to a 26 base pair enhancer located 503-528 base pairs upstream from the translation start site. This enhancer accounts for a two- to threefold increase in the level of gene expression. Using electrophoretic mobility shift analysis, we have demonstrated at least two proteins that bind to overlapping sequences in the enhancer. Although the enhancer sequence does not contain a perfect match to the consensus Sp1 binding motif, there appears to be a relatively low affinity binding site for Sp1. In addition, a second protein binds to the enhancer sequence. Although the identity of this second DNA binding protein has not been determined, it appears to be a novel factor. While the insulin receptor mediates the first step in insulin action, other proteins function downstream in the insulin action pathway. We have initiated studies into the regulation of the gene encoding pp120/HA4, a membrane glycoprotein in rat liver that undergoes insulin-stimulated tyrosine phosphorylation. Consistent with the tissue-specific expression in liver, the pp120/HA4 gene has a TATA box containing promoter. In addition, the 5'-flanking region of the gene contains putative binding sites for several factors including a glucocorticoid response element and an insulin response element. We have initiated studies to investigate the regulation of expression vectors in which the promoter plus 5'-flanking region of the pp120/HA4 gene drive the expression of the luciferase gene. Using this approach, we have begun to map cis-acting elements responsible for the ability of insulin and dexamethasone to increase gene expression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Premature vascular disease in young hyperlipidemic subjects remains a major unsolved health problem in terms of pethogenesis and treatment. Recent research has made available reference values for diagnosing hyperlipidemia, markers for genetic analysis, and methods for studying lipoprotein metabolism and arteriosclerotic disease progression. With these advances, the opportunity now exists for in depth, focused studies of lipoprotein pathophysiology in genetically characterized patients toward the objective of understanding disease mechanisms and developing better treatments. This program in \"Human Lipoprotein Pathophysiology\" siezes this opportunity with 8 coordinated projects and 4 supporting core laboratories led by experienced investigators, experts in physiology, genetics, biochemistry, immunochemistry, metabolism nutrition, and cardiology. Subjects will be characterized in terms of the heritability of lipoprotein abnormalities, the metabolism of lipoproteins, including carefully characterized subpopulations of HDL isolated by immunoabsorption. The role of LCVAT and lipid transfer proteins in the genetic hyperlipidemics will be studied and related to the kinetic abnormalities of lipoprotein metabolism in vivo. The fate of lipoproteins will be studies in human cultured macrophages and placental trophoblast cells and will further the understanding the role of macrophages and steroid hormone secreting cells in lipoprotein homeostasis and atherosclerosis. The effects of diet and drug interventions will also be studied including the effects of omega-3 fatty acid feeding on lipoprotein lipid metabolism and effects of diet and drug lowering of lipoprotein on arteriosclerosis. This work will be supported by excellent Core facilities in lipoprotein measurement, apoprotein immunoassay, and biomathematical analysis and offers promise of rapid progress in understanding mechanisms and providing treatrment for the premature vascular diseade associated with genetic hyperlipidemia.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research is designed to further our understanding of why mildly delayed young children have difficulty regulating their own behavior to social expectations. My basic premise is that the children do not fully understand expectations for behavior. The aim of this 3 year study is to provide a comprehensive analysis of children's knowledge of standards, their actions in a situation where standards are expected, and the reported ways that caregivers and teachers facilitate the growth of knowledge for standards. Particular emphasis will be directed toward exploring the different kinds of knowledge children possess. This study will involve 120 mildly delayed and normally developing 4 and 5 years olds, who are matched for developmental age. Data analyses will focus on children's understanding of acceptable and unacceptable behavior, the kinds of knowledge they possess for different situations, and the coherence between child knowledge and child behavior, and caregiver expectations and child behavior. Findings from this study should have implications for teaching and training children who have developmental disabilities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The acquired immunodeficiency syndrome (AIDS) is defined by the presence of opportunistic infections or tumors that do not occur in persons with normal immune function. Monocytes and macrophages that have been \"activated\" form part of the host defense against infection and neoplasia. It has been proposed that any putative monocyte/macrophage defect simply reflects the absence of activating signals from CD4+ T cells that are depleted in AIDS. One such signal is interferon-gamma which activates macrophages for certain defense functions. T cells also induce monocytes and macrophages to (1) develop procoagulant activity, (2) release interleukin-1 and its inhibitor, (3) release tumor necrosis factor, and (4) express antimycobacterial activity. Immunolgically stimulated T cells induce these macrophage functions by means of lymphokines known to be distinct from interferon-gamma. I propose to examine these lymphokine systems in AIDS and ARC. Using blood from human immunodeficiency virus (HIV)-infected individuals, the ability of T cells to induce each of these four macrophage functions will be examined and compared to normal controls. Furthermore, since macrophages may be infected with HIV both in vivo and in vitro, I propose to study the ability of HIV to directly induce a primary monocyte/macrophage defect for each of these four functions independent of the effects of infection of T cells. Finally, the ability of lymphokines to affect the course of HIV production in macrophages will be examined. These studies should help clarify the immunological defects in AIDS and AIDS-related complex and may suggest strategies for their treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pyrene excimer fluorescence is a microviscometric probe of the hydrocarbon region of amphiphilic molecules and is based on the rate of translational diffusion of the aromatic hydrocarbon, pyrene, measured by the relative intensities of monomer and excimer fluorescence. The rate of excimer formation is diffusion controlled and depends on both the viscosity of the environment, eta, and the concentration of pyrene dissolved in the hydrocarbon region, represented by the expression: I sub E/I sub M equals (P) TK/eta, were eta is the viscosity, T is the absolute temperature, (P) is the pyrene concentration, and K is the constant which incorporates both theoretical and instrumental parameters. The overall objective is the development of pyrene excimer fluorescence as a probe of changes in the structure of the hydrocarbon region of lipids and lipoproteins. The specific, sensitive fluorescence techniques can be used to characterize the surface monolayers of plasma lipoproteins, which contain the determinants of the enzymic and non- enzymic modifications that lipoproteins undergo in the circulation. Such information appears to be essential to identify aberrant processes and disease states and to devise favorable schemes of therapy to regulate abnormal lipid metabolism associated with hyperlipoproteinemia and atherosclerosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Personality disorder (PD) diagnosis is at a crossroads: There is broad acknowledgment of the significant, negative impact that PD has on individuals and on the course and outcome of comorbid disorders, as well as relatively widespread dissatisfaction with the current categorical system of PD diagnosis. Nonetheless, there also is reluctance to implement an alternative trait dimensional system because: (1) there is no consensus on which set of traits to assess, (2) current trait assessment methods do not reflect either dysfunction or the full range of clinical variability, and (3) the clinical utility of a trait-based system is unknown. The proposed research is significant because it will draw from the full spectrum of the personality trait domain to identify a comprehensive set of specific traits for PD diagnosis, demonstrate how PD characteristics are manifest in both stable and clinical-state-dependent forms, and provide a clinically useful PD diagnostic system that integrates assessment of trait dimensions and psychosocial functioning. The long-term goal of this research is to develop a trait-plus-dysfunction-based clinical assessment system for the reliable and valid diagnosis of personality disorder on which to base the development of effective, more specifically targeted interventions. The specific aims of the current proposal are (1) to establish a set of specific traits that collectively define the trait dimensions of individuals with PD, (2) to establish a set of psychosocial functioning dimensions that collectively define the functional impairment of individuals with PD, and (3) to evaluate prospectively the clinical utility of this two-pronged PD diagnostic system. We plan to do this in three stages: (1) Assessing a large sample on a wide range of measures of both traits and psychosocial function to determine the structure of each domain and ensure that we have identified all components needed to assess personality disorder comprehensively; (2) testing the identified components on another large sample of patients and high-risk individuals, including information provided by family and/or friends, and Axis I symptomatology, and (3) testing the clinical utility of the developed system by assessing a sample of new patients, providing the information to their mental health-care providers, surveying the clinicians for the usefulness and acceptability of the diagnostic system, and examining its predictive validity over 6 months post-intake. We expect that the newly developed system will have high predictive validity and be judged clinically useful; specifically, to be comprehensive and accurate, and useful for communication purposes, for formulating general and specific treatment plans, and for predicting general and specific treatment difficulties.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overarching goal of the proposed research is to understand, from a cognitive neuroscience perspective, how information about an event is successfully encoded into episodic memory. Functional magnetic resonance imaging (fMRI) will be employed to identify neural activity elicited during the successful encoding of experimental items such as words and pictures and their associations either with other items (item-item encoding), or with one or more contextual features (item-context encoding). The theoretical perspective guiding the proposed research derives from the proposals that an episodic memory is represented in terms of the processing engaged by an event as it is experienced, and that successful episodic encoding requires that the various features constituting an event are 'bound'or integrated into a cohesive memory representation. Guided by this perspective, experiments will address the roles of the cerebral cortex and the medial temporal lobe in the 'binding'of within- versus across- modality item-context and item-item associations. Other experiments will investigate the neural correlates of the formation of within- and across-modality associations between different contextual features. Another experiment will investigate the neural correlates of the formation of temporal associations between items presented at different times. In a second strand of the research, the focus will be on neural activity that appears to play a 'permissive'role in episodic encoding, rather than supporting processes engaged by a specific study task. Experiments will be conducted to elucidate i) the functional significance of the relative attenuation of neural activity elicited in some cortical regions by study items that are remembered rather than forgotten on a later memory test, and ii) modulations of pre-stimulus activity that also are predictive of later memory performance. Disabling impairments of episodic memory are prominent in several common neurological conditions, notably Alzheimer's disease and traumatic brain injury. Episodic memory dysfunction is also a significant component of several common psychiatric disorders, including schizophrenia, depression and post-traumatic stress disorder. In addition, episodic memory declines substantially and, relative to other kinds of memory, disproportionately, with increasing age. The proposed research will contribute to the detailed understanding of the neurocognitive processes that support normally-functioning episodic memory, an understanding that is necessary for the elucidation of different kinds of episodic memory disorders and the development of effective therapeutic interventions. PUBLIC HEALTH RELEVANCE: Disabling impairments of episodic memory - memory for unique events - are found in numerous neurological and psychiatric conditions. The proposed research will contribute to the detailed understanding of normally-functioning episodic memory that is necessary if different kinds of episodic memory disorders are to be fully understood and effectively treated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The melanocortin receptors (MCRs) are G-Protein Coupled Receptors (GPCRs) that modulate and control many critical physiological processes in animals including pigmentation, response to stress, feeding behavior, energy balance, sexual function and behavior, inflammatory response, cardiovascular function, immune response, pain and others. They also are involved in many of our most common degenerative diseases including adrenal dysfunction, obesity, anorexia, pigmentary disorders, sexual dysfunction, prolonged and neuropathic pain, inflammatory disorders, cardiovascular disease and others. Yet there are few compounds that have been developed and are pharmaceuticals in use for treatment of these diseases which are our most common and difficult to treat diseases. This grant is dedicated to developing more potent, receptor selective, and specific and most efficacious ligands for the melancortin 1(MCI), melanocortin 3 (MC3), melanocortin 4 (MC4), and melanocortin 5 (MC5) receptors. These novel ligands, which are biological stable and available for in vivo applications, will be useful for studying the pharmacology, physiology and medical applications of melanotropin ligands. The specific aims we will pursue include: 1) development and use of novel protein/peptide topologies and scaffolds, in conjunction with novel constrained amino acids, to design and synthesize novel orthosteric, allosteric, and biased melanotropin peptides and peptide mimetics with enhanced stability, bioavailability, selectivity and potency for the melanocortin receptors; 2a) to examine binding affinities, cyclic AMP production, efficacies, Ca+2 assays and assays for -arrestin. The novel ligands for the hMC1R, hMC3R, hMC4R and hMC5R receptors will be examined with special attention to the development of biased ligands; and 2b) use of novel selective ligands, especially those with biased activity, to explore novel physiological functions that can lead to novel drugs with our collaborators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Core B is designed provide transgenic animals required by the four projects of the program, including mice that express key molecules/receptors and mice that have defective genes. Core B will breed and screen transgenic animals and backcross them as necessary to render them suitable for the experiments detailed in the individual projects of the program. Because this process is very labor intensive, a centralized facility specialized ot perform this task makes a great deal of sense. Core B will also oversee the collection and preparation of reagents required as key components by at least two or more individual projects. Such reagents would include Ab to potential co-stimulatory molecules and cytokines, antibodies to adhesion molecules, APC cell l lines expressing particular co-stimulatory molecules, cell lines for cytokine bioassays. The Core will obtain the various Tg or KO mice needed for the individual projects, quarantine them if necessary, develop standardized procedures for screening for the Tg/KO gene expression in each case, and will breed and backcross the mice to C57BL/6 and B10.BR inbred strains. The goals are to have healthy, genetically homogenous mice for the investigators in the project. Investigators will be able to obtain these mice for experiments but will maintain the animals separately during the course of the sometimes lengthy adoptive transfer experiments. Many of the experiments conducted so far on the Tg and KO mice have been done with original B6 x SJL founder mice or with mice backcrossed only for one or two generations. In such cases background genes from the non B6 parent may influence experimental results. Use of non-bred mice can also lead to complications due to mismatches in major or minor histocompatibility differences and also to the presence of genes other than the gene of interest due to linkage on the same chromosome. Furthermore, the facility will allow us to generate new strains of mice by inbreeding. For instance TCR Tg mice will be generated with the SCID mutation so that contributions from expression of endogenous alpha chains can be ruled out. Also double KO mice or TCR Tg/KO mice can be created which will allow a more definitive analysis. At the moment we have the following mouse strains to contribute to the core; TCR Tg mice (specific for pigeon cytochrome C (PCC) plus I-Ek (designated AND) backcrossed 5 times to B6 and B10 BR (currently being bred onto a RAG-2 KO background); TCR Tg mice specific for HY plus Dd (already on a SCID background); TCR Tg mice specific for influenza hemagglutinin (HA) peptide plus IAd (CD4 selecting) and TCR Tg specific for HA plus Dd (CD8 selecting); beta2M KO/KO mice; CD4 KO/KO mice backcrossed 8X to B6; IL-2 KO/KO backcrossed 3X to B6; IFNgammaKO/KO backcrossed 3x to B6. IL-4 KO/KO backcrossed 4x to B10.BR; and RAG-2 KO/kO. We also have mum KO/KO mice and mice constitutively expressing high levels of bcl2. Dr. Klinman will provide the Sma58 m ice, which express I=-Ek only on germinal center B cells. Dr. Bradley has already made arrangements to obtain L-, E- and P- selectin Ko mice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "When cells divide, cohesion between sister chromatids provides a physical mechanism to hold sister chromatids together from the time of their synthesis until they segregate to opposite poles. During both mitosis and meiosis, cohesion is essential for accurate chromosome segregation. For at least 5% of all clinically recognized human pregnancies, errors in meiotic segregation give rise to aneuploid zygotes. Moreover, defects in meiotic chromosome segregation are dramatically more prevalent in older women. Our long-term goal is to understand how chromosome segregation is regulated at the molecular level and why errors in segregation occur more frequently in older oocytes. We will use genetic, cytological and biochemical strategies to study the mechanisms that control meiotic sister-chromatid cohesion and chromosome segregation in the model organism Drosophila. ORD protein is essential for meiotic cohesion in Drosophila males and females. We have shown that ORD associates with meiotic chromosomes in both sexes, and controls the association of cohesin subunits with meiotic chromosomes. To elucidate the mechanism by which ORD controls cohesin localization, we will test the hypothesis that ORD is a meiotic cohesin subunit and define the requirements for arm and centromeric localization of ORD and cohesin SMCs. We will continue our characterization of gene products that interact with ORD and use genetic techniques to indentify additional ORD interactors. Analysis of these proteins will be critical in elucidating the mechanism by which ORD promotes cohesion and also will further our understanding of the overall pathway of events required for proper chromosome segregation during Drosphila meiosis. Finally, we have developed an experimental system that will allow us to test the hypothesis that deterioration of cohesion during aging causes defects in chromosome segregation in older eggs. In addition, we will will use genetic and proteomic approaches to identify gene products that influence age-dependent nondisjunction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have described two dsRNA viruses (L-A and L-BC) and two ssRNA replicons (20S RNA and 23S RNA) in the yeast Saccharomyces cerevisiae. M dsRNA is a satellite of L-A encoding the killer toxin. We discovered 7 chromosomal genes, SKI1, 2, 3, 4, 6, 7, and 8, by their ability to prevent these replicons from causing pathogenicity to yeast cells. These four RNA replicons all make uncapped mRNAs and lack a 3 poly(A)structure. SKI1 encodes an exoribonuclease specific for uncapped mRNAs, while we showed that the SKI2, SKI3, SKI6, SKI7 and SKI8 gene products block the translation of non-poly(A) mRNAs. We showed that Ski2p is an RNA helicase, Ski6p has homology to a tRNA - processing RNAse, and Ski7p is similar to translation factor EF1alpha. We showed that mutations in 20 chromosomal genes resulting in loss of M dsRNA are deficient in 60S ribosomal subunits. SLH1 is an RNA helicase homologous to SKI2. We showed that Ski2p and Slh1p have overlapping function, and together block the translation of non-poly(A) mRNA. A ski2 slh1 double mutant treats non-poly(A) mRNA the same as it treats poly(A) mRNA, with the same rate of translation and the same duration of translation. The ski2 slh1 double mutant has no detectable difference in mRNA turnover rate. Thus the 3 poly(A) structure of mRNA is only needed for translation because of the cooperating action of Ski2p and Slh1p (and other proteins that work with them). The ribosomes and translation factors are fully able to use non-poly(A) mRNAs even in the presence of a full complement of poly(A) mRNAs competing for the translation apparatus. The standard model of 3 poly(A) action in translation is that interaction of the poly(A) binding protein (Pab1p) with initiation factor 4G (eIF4G), by circularizing the mRNA, promotes 40S subunit recruitment. However, we find that elimination of the Pab1p - eIF4G interaction does not affect the requirement of translation for the 3 poly(A), showing that this model is incorrect. Although the polyA binding protein of yeast is necessary for the preference of extracts for poly(A) mRNA in vitro, we find that in spite of deletion of the PAB1 gene, electroporated cells still prefer poly(A) mRNAs by the same factor as in isogenic wild-type cells. However, elimination of Fun12p, a protein involved in 60S ribosomal subunit joining, preferentially impairs translation of poly(A) mRNA without significantly affecting translation of poly(A)- mRNA. This suggests that the 3 poly(A) structure has a role in the 60S joining reaction. We are now pursuing an understanding of the role of poly(A) in translation and the relation to the Ski proteins.[unreadable] Mak3p is an N-acetyl transferase that acetylates the N-terminus of the LA major coat protein (Gag). We find that a homologous protein from Arabidopsis is able to substitute for the yeast Mak3p. While yeast Mak10p is also necessary for this reaction in yeast, we find that the Arabidopsis Mak3p can substitute for Mak10p as well as Mak3p.[unreadable] Our collaborators, Drs. Hisashi Naitoh and John E. Johnson, determined the structure of the L-A virus at 3.4 angstroms resolution by X-ray crystallography. The L-A dsRNA virus is 400 angstroms in diameter, and contains a single protein shell of 60 asymmetric dimers of the coat protein, a feature common among the inner protein shells of dsRNA viruses, and probably related to their unique mode of transcription and replication. The two identical Gag molecules in each dimer are in non-equivalent environments, and show substantially different conformations in specific surface regions. This virus decaps cellular mRNA in order to express its own uncapped mRNA. Our structure reveals a trench at the active site of the decapping reaction and suggests a role for nearby residues in the reaction. 7methylGDP bound to viral particles shows a density in the trench near His154, the residue to which the cap is attached in this reaction. We have now shown that Tyr150, Tyr 452 and Tyr 538 are essential for the decapping reaction. Positively charged residues in this region are also important. These experiments have permitted proposal of a mechanism of the decapping reaction.[unreadable] [unreadable] S. cerevisiae can carry any of four RNA viruses, including the dsRNA viruses L-A (and its killer toxin-encoding satellites, Mi) and L-BC, and the single-stranded RNA replicons 20S RNA and 23S RNA. The copy numbers of each of these is negatively regulated by the SKI genes, homologs of which are known in all eukaryotes. We have previously shown that Ski2p and its S. cerevisiae homolog Slh1p block the expression of mRNAs which, like those of the four RNA viruses, lack a 3 polyA structure. We have published evidence that this block includes effects on both mRNA stability and translation. However, since untranslated mRNAs are inherently unstable, it is difficult to distinguish which effect is primary. Indeed, we find that deletion of the SKI2 gene and its close homolog SLH1 result in yeast cells being nearly indifferent to the presence or absence of the 3 polyA tail on an mRNA. This result has been confirmed recently by other groups, and has important implications for the mechanism of translation.[unreadable] Others have described a construct in which the 3 untranslated region of the TRP4 mRNA includes a hepatitis delta virus self-splicing intron, whose action results in the rapid elimination of the 3 polyA of the mRNA. The resulting polyA-minus mRNA is reported to be stable in yeast, so we want to use this system to investigate the mechanism of the effects of Ski2p on expression of cellular mRNAs. We find that expression of this mRNA is elevated more that five fold in a ski2 mutant compared to an isogenic wild-type strain. Our current efforts are directed at confirming the earlier report of the stability of the mRNA in wild-type strains.[unreadable] The polyA binding protein, Pab1p, is believed to interact with the translation initiation factor, eIF4G, and since the latter interacts with the cap-binding protein, eIF4E, this is believed to circularized the mRNA and to explain the importance of the 3 polyA structure for translation. However, our finding that ski2- slh1- double mutants are indifferent to the presence of the 3 polyA structure of eukaryotic mRNAs casts some doubt on this view of the role of the polyA - binding protein in translation. Indeed, mutants of PAB1 deleting the part of the protein that interacts with eIF4G have no effect on the growth of cells. [unreadable] We are re-examining the importance of the polyA binding protein in the polyA requirement for translation. We find that although loss of the polyA binding protein results in failure of 3polyA to stimulate translation in vitro (extracts of yeast cells), the absence of PAB1 has little or no effect on the requirement for polyA for translation in vivo.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "An extended family study of a discrete, isolated population with respect to a complex of developmentally and educationally important variables is being conducted. The behavioral variables of interest include spatial and verbal ability, field dependence, automatization, handedness and ear dominance for speech and non-speech sounds. Genetic and environmental contributions to the variance will be estimated by path analysis and major gene effects will be investigated by pedigree analysis. In addition, the relationship of the indices of functional specialization of the hemispheres of the brain to cognitive and motor performance will be investigated at the population level for the very first time. The proposed program of research constitutes a rare opportunity to bring strict genetic control to the field of human behavioral science.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our immediate goal is a detailed analysis of the radiation response of two established in vivo lines (Slow and S102F) of the C3H mouse mammary tumor. This analysis will be especially concerned with evaluating the radiation response as it relates to the various growth kinetic parameters in the belief that such an approach will suggest a simplified model for optimizing protocols for fractionated radiation therapy. The initial studies are concerned with determining the single-dose TCD50/120s (i.e. the dose that shows local control in 50% of the tumors at 120 days post irradiation) for 250 kVp x-rays when the tumors are under: 1) Normal physiological conditions; the mice are breathing air., 2) Anoxic conditions; a clamp is placed across the base of the tumor for 8 minutes before and then maintained in place during irradiation., and 3) Hyperbaric conditions; the mice are breathing 95% O2 at 3 atmospheres of pressure. Preliminary data for part 1 suggest that the TCD50 for the Slow line may be about 5,800 rads while that for the S102F line is about 3,500 rads. Adriamycin in combination with x-irradiation is also being studied. Host toxicity studies were done on our C3H mice and LD50 dose (single injection intraperitoneally) is about 20 milligrams/kg. The volumetric response of the tumors from both lines was also determined and a single dose of 10 milligrams/kg was chosen to study in conjunction with x- irradiation. The method for analyzing the combined effects is the TCD50 approach as described above except the adriamycin is injected two hours before x-irradiation of the tumors. These experiments are in progress.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY The number of physician-scientists in the United States has been steadily declining, and there is a greater need for both physician scientists and Ph.D.'s in gastroenterology research. Early intervention through undergraduate research opportunities (UROs) increases interest in science-related careers, and quality mentorship is critical for the development of young investigators and essential to attract more women and underrepresented minorities into science. To address these needs, in 2001 we established the Undergraduate Student Scholars Program (USSP), an innovative URO at the University of Pennsylvania which is closely linked to our NIH/NIDDK P30 Digestive Diseases Center (referred to by us as the Center for Molecular Studies in Digestive and Liver Diseases). Since the USSP is closely linked to the Center, USSP participants have access to all of the Center's superb resources and faculty. The USSP received initial funding through an R25 Education Projects (now Program) Grant from the NIDDK in 2004, and this was successfully renewed in 2009 and 2014. The cornerstone of the USSP is an intensive laboratory-based research experience that entails close interaction with and mentorship by a training program faculty member. This laboratory work is supplemented by a comprehensive educational curriculum, which includes seminars on topics in biomedical research, ethics, and laboratory safety, and a focused student research symposium in which students interact with keynote speakers who are international leaders in academic medicine and biomedical research. Over the past 5 years, we have successfully trained 54 students (a total of 150 since 2004) from leading colleges and universities throughout the country, with increasing recruitment and continued mentorship of women and underrepresented minority students. Following completion of the program, most USSP participants pursue additional biomedical research training, including through M.D., Ph.D., and M.D.-Ph.D. programs at leading institutions. The USSP tracks the performance and outcomes of students through longitudinal data and questionnaires and is guided by multi-disciplinary and experienced Internal and External Advisory Committees, both composed of thought-leaders in research education and training. The program also works closely with the Office for Diversity and Inclusion at the University of Pennsylvania Perelman School of Medicine. The overarching interrelated specific aims of the USSP are two-fold: 1. To foster a strong interest among talented undergraduates in biomedical research with a focus on digestive, pancreatic, and liver research; and 2. To establish durable mentoring relationships between talented undergraduates interested in pursuing careers in biomedical research and supportive faculty mentors. In aggregate, this innovative URO provides the superb faculty, exceptional scientific resources, and exciting intellectual environment for trainees to expand their knowledge and interest in biomedical research. The key outcome of the USSP is to motivate trainees to pursue careers in biomedical research with a focus on digestive, pancreatic, and liver related research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this phase I STTR application is to continue assessment of the relevance and feasibility of Nogo decoy receptor therapy for the treatment of glaucoma. The secondary objective is to obtain pharmacokinetic data important for designing additional preclinical efficacy studies and formulating a drug development plan for glaucoma. Our long term objective is to ameliorate the devastating consequences of glaucoma by developing a treatment option that not only slows progression but also may provide functional regeneration. Glaucoma is the second leading cause of blindness worldwide. Current therapies that lower intraocular pressure (IOP) as a means of slowing disease progression are not fully effective. Our proposal seeks to further evaluate Nogo decoy receptor as a potential neuroprotective and regenerative therapy that does not rely on lowering IOP. Preliminary studies employing rat Nogo decoy receptor (rNgR(310)Fc) and manipulation of Nogo receptor gene (ngr1) expression suggest that Nogo decoy receptor therapy has the potential to become a first-in-class therapy for glaucoma to prevent retinal ganglion cell (RGC) loss and promote optic nerve axonal regeneration. The research proposal objectives will be accomplished within a 1 year period by meeting three Specific Aims which include: 1) determining the efficacy of human Nogo decoy receptor (hNgR(310)Fc) in preventing retinal ganglion cell (RGC) loss in a model of indirect injury: rat bead model of glaucoma, 2), evaluating the effect of human Nogo decoy receptor therapy in promoting RGC survival and optic nerve regeneration in a model of direct injury: optic nerve crush injury in rats, and 3) evaluating the pharmacokinetics and tissue distribution of Nogo decoy receptor when administered into the eye. Completion of these Specific Aims will provide critical data needed to initiate development of Nogo decoy receptor therapy for the treatment of glaucoma. Specifically, the data will be used to define an appropriate formulation and delivery strategy for use in additional preclinical glaucoma models and to determine the most appropriate development strategy to advance the therapy to clinical evaluation. Accordingly, Future Studies will include preliminary formulation development for ocular delivery. This may include formulation for controlled release in the eye depending on the outcome of the pharmacokinetics and in vivo pharmacology investigations in the current proposal. Further preclinical efficacy studies will be conducted as well as investigations to elucidate the mechanism for RGC protection by Nogo decoy receptor.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "With this proposal we aim to further the development of novel and safe oral inhibitors of sEH for the treatment of peripheral neuropathy in diabetic patients. Preclinical studies with our candidate EC5026 demonstrated efficacy for diabetic neuropathy. EicOsis has selected EC5026 for its efficacy with good PK/ADME properties and stability to enter further development stages for this indication. EicOsis also tested the compound and a backup in vitro for off target effects against an array of enzymes, determined the potential inhibition of major cytochrome P450 enzymes, and tested for de-risking with a contract research laboratory. EicOsis has used these data as a basis for an application to work with Blueprint Development Teams to create a development plan and initiate studies to test the safety of EC5026 in Phase 1 clinical trials. To further develop the compounds selected from the completed Phase I application we propose here to radiolabel the candidate and backup to carry out detailed metabolism studies and develop the label as a tracer for human Phase I clinical trial. Additionally we propose to use conventional material to test for off-target toxicity in vivo and to explore the breadth of activity of the inhibitors in alternate pain models. The studies are designed to address understanding metabolism and the spectrum of activity which if lacking often prevent drugs from being developed. The results will empower scientists in the pharmaceutical industry with critical data that speaks to the wider spectrum of activity of sEH inhibitors as analgesics of the future.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Patients treated for hyperreactive airways disease (asthma) differ greatly in treatment outcome, even when no identifiable differences exist in the objective severity of their illness. One important basis for these differences, studied during the past several years, involves the level of the focused concern patients show for their breathing difficulties. Asthmatic patients who profess little anxiety during asthmatic attacks are hospitalized infrequently, particularly when this response style is paired with relatively low airways hyperreactivity. Treatment outcome appears to differ for these patients because of behaviors, including medication requests during treatment, which reflect either a vigilant stance toward breathing difficulties or a disregard for them. The basis for the behavioral differences needs to be established in order to design appropriately planned intervention strategies. At issue here is whether the symptom denial or symptom vigilance is a byproduct of faulty perception of airways obstruction when it occurs, or whether it is the result of unique behavioral strategies employed. The principal objecties of this proposed research are to investigate: (A) whether differences in perception of airway obstruction, as indexed by subjective symptom reports, occur among Low, Moderate, and High Panic-Fear patients, (B) whether these patient groups acknowledge different types of internal cues of airway obstruction (e.g., Dyspnea or Congestion symptoms), (C) whether differences in perceptions of internal cues of airway obstruction are related to frequency of PRN requests, and (D) to apply the information obtained to clinical intervention planning within an intensive treatment program for asthma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cell cultures of neuronal tissue provide many advantages and opportunities for biochemical studies of the nervous system. We seek to exploit one system-cell cultures of sympathetic neurons-from this point of view. In this proposal, we have identified and will study three major problems for which both biochemical tools and cell culture techniques are available. These are: (1) The use of the snale venom protein alpha-bungarotoxin to probe neuronal acetylcholine receptors. We propose, in cultures of sympathetic neurons, to study the turnover, migration, and development of neuronal acetylcholine receptors as well as to study the effects of synapse formation on their post-synaptic properties (eg. distribution, concentration). We shall also determine the rate of development of these receptors during embryogenesis. (2) Nerve growth factor (NGF) receptor properties and mechanism of action. NGF has profound effects on sympathetic neurons both in vivo and in vitro. We shall use (I125)-NGF to determine the cellular target for NGF in culture; the distribution, number, and concentration of NGF receptors on the target cells, and the development of NGF receptors in vivo. We shall also seek to uncover the primary mode of action of NGF by studying the ability of various chemical substances to mimic, enchance, or inhibit the effect of NGF on cultured sympathetic neurons. (3) Creation and characterization of dividing cell lines with neuronal properties. We shall use somatic cell hybridization and suitable selection techniques to fuse primary sympathetic neurons with mouse neuroblastoma cells with the aim of creating dividing lines of cells with neuronal properties. Existing and new cell lines generated in this manner will be screened for the presence of various properties of sympathetic neurons (eg. the ability to synthesize, store, or take-up catecholamines; sensitivity to NGF or chemical transmitters).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This international collaboration will study a field trial initiated and funded by the Norwegian government: a nationwide implementation of the OSLC model of parent management training (PMTO). PMTO is an efficacious theory-based intervention that teaches parents child-rearing strategies that prevent deviant child behavior and promote healthy family development. The Norwegian goal is to have trained PMTO interventionists available in every municipality (435) to intervene at the earliest stages of deviant child behavior, thus preventing later substance abuse, school failure, and delinquency. This proposal details the research describing the processes and mechanisms associated with successful PMTO implementation. The project is a collaboration among 3 centers: OSLC, the Norwegian Center for the Study of Behavioral Problems and Innovative Practice in Oslo, and the Institute for Social Research. NIDA support will enable us to map onto work in progress funded by two Norwegian Ministries (Children & Family Affairs and Social & Health Affairs). We propose to study factors contributing to the adoption, adaptation, and implementation of PMTO in Norwegian communities. Adoption will be studied across successive generations of trainees. Generation 1 (G1) consists of a cohort of Norwegian professionals recently certified by OSLC professionals as PMTO Specialists. In 2002, G1 began training the second generation of Norwegian professionals (G2). Late in 2003, Norwegian PMTO specialists will begin training G3. We will study fidelity to and adaptation of PMTO procedures through generations of training using repeated multiple method assessments and direct observation of intervention sessions. We hypothesize successful intervention for those who retain fidelity to the core PMTO parenting practices and this will prevent negative child outcomes like substance abuse. We also expect to find additions to PMTO that are relevant to Norwegian culture but do not destroy PMTO integrity. We hypothesize that such additions will enhance adoption and sustainability of the program. Factors that contribute to fidelity, adaptation, and drift from fidelity will be evaluated, hypothesizing increasing drift with time and ensuing generations of trainees, and that fidelity drift will be correlated with reduced efficacy. To better understand the adoption and rejection of PMTO by agencies, professionals, and families, we will analyze already collected field notes and retrospective accounts from the recruitment and participation of G1 agencies and trainees. These data can be compared with prospective and concurrent data collected during the recruitment and training of G2 and G3 agencies and trainees. Annual surveys will be conducted to study beliefs and attitudes within experimental and control community agencies throughout Norway.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Automobile driving is a crucial aspect of everyday life, yet vehicular crashes pose a serious public health problem. Parkinson's disease (PD) affects about one million Americans, and drivers with PD are at special risk for driving cessation or crashes due to progressive impairments of motor function, cognition, and vision. In our ongoing study on predicting driver safety in PD (R01 NS044930, PI:Uc), we are finding that drivers with PD perform worse than elderly drivers without neurological disease in driving tasks conducted on-road in an instrumented vehicle (IV) and on experimental driving scenarios presented on a high-fidelity driving simulator. Even drivers with early PD of mild severity have impairments on cognitive and visual functions, and show particular problems with multitasking while driving, performing complex driving maneuvers, and are at increased risk of collisions. Cognitive impairment and reduced visual perception contribute to poor driving performance and safety errors more than the typical motor dysfunction of PD. Based on our own and other researchers' empirical findings on the types and circumstances of common safety errors, as well as on the underlying causes and mechanisms of driving problems in PD, we have devised a new intervention for at-risk drivers with PD. The intervention consists of a systematic review of the subject's own road drive in the IV using verbal, video, and written feedback, followed by simulator training sessions for improving visual attention, performing complex maneuvers, multitasking during driving, hazard perception, and collision avoidance. The conceptual basis of our intervention is improving procedural memory and error awareness through feedback and practice in the simulator, with the expectation that this improvement would transfer to driving on the road. This is a pilot study proposal to further develop and refine our intervention, and test its short term efficacy to see if this intervention merits further testing in a Phase III trial. Development of effective rehabilitation programs for impaired drivers with PD will reduce the risk of motor vehicle crashes and help protect and preserve the mobility and quality of life of these patients. Furthermore, this driving rehabilitation program can be adapted for use in other neurological disorders such as Alzheimer's disease, stroke, and traumatic brain injury, conditions seen in both older and younger veterans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary objective of this proposal is to investigate the cellular and molecular mechanisms that regulate mammalian craniofacial development. Mice with mutations that cause craniofacial defects are important resources for determining the factors that are essential for this complex developmental process. Goosecoid (Gsc) is a homeobox gene that is expressed in the anterior region of the primitive streak and later in craniofacial structures, the limbs, and the ventrolateral body wall. We previously generated Gsc-null mice by gene targeting in embryonic stem (ES) cells. Gsc-null mice are born alive but die soon after birth with numerous craniofacial defects and rib cage abnormalities. We have initiated studies to examine the behavior of Gsc-null cells when mixed with wild-type cells in the craniofacial regions of mouse chimeras. Such studies will reveal the tissues that directly require Gsc function and whether Gsc acts cell autonomously. We have also introduced lacZ into the Gsc locus to follow Gsc-expressing cells during development. These Gsc lacZ mice should serve as valuable resources for studying the phenotypes of our Gsc mutants. Studies in Xenopus indicate that Gsc levels are important for regulating cell fates. Therefore, we have generated mice with a duplicated Gsc locus to examine the effect of Gsc dosage on craniofacial development. Recently, a Gsc-like gene (Gscl) was reported that maps to the DiGeorge and velocardiofacial syndromes (DGS/VCFS) minimal critical region. DGS/VCFS patients present with craniofacial and heart abnormalities caused by a haploinsufficiency of a gene(s) deleted in human 22q11.2. Gscl may contribute to the development of these syndromes. Therefore, we have generated Gscl-null mice that will serve as important resources to determine the role of Gscl in the DGS/VCFS. Finally, defects are not observed in all Gsc-expressing tissues of Gsc-null mice, suggesting compensation by other genes. Compensation for Gsc by the related Gscl can now be tested by generating Gsc x Gscl compound mutants. Thus, we propose molecular, genetic, and embryological experiments for a mechanistic understanding of Gsc and Gscl function during craniofacial development. This knowledge should be useful for understanding the etiology and pathogenesis of craniofacial defects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A series of approved food, drug, and cosmetic dyes are evaluated for the effects they produce on the contractile response of guinea pig ileum to administered acetylcholine and to inhibition and release of acetylcholine following electrical field stimulation. The action of food dyes in isolated cellular or neuronal systems is investigated to learn more about their possible relationship to the production of hyperkinetic behavior in children.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gene therapy vectors derived from lentiviruses offer a number of advantages over other gene transfer vectors, and they represent a promising approach for treating a variety of diseases, such as age-related macular degeneration, Parkinson's disease, and blood cell malignancies. Unfortunately, lentiviral vectors are difficult to produce in large numbers due to the lack of stable packaging cell lines and to inefficiencies associated with standard methods of transient transfection. This Phase 1 proposal describes the development of a scalable process for lentiviral vector production that uses MaxCyte's proprietary flow electroporation technology to transfect plasmid DNAs encoding components of bovine immunodeficiency virus (BIV) gene therapy vectors into mammalian cells. The workplan includes optimization of electroporation parameters for loading BIV component plasmids into adherent HEK 293T cells at both small and large scales. These experiments are intended to demonstrate that flow electroporation is superior to other methods (e.g., calcium phosphate precipitation) for transfecting cells that are commonly used in lentivector production. In addition, protocols will be developed for transfecting BIV component plasmids into suspension cells, which offer a number of advantages over adherent cells for manufacturing biological products. The goal is to generate an efficient, suspension cell-based method for lentivirus production that can be scaled up accommodate the needs of clinical-scale testing as well as commercial manufacturing of a lentivirus-based therapeutic product. The results of these studies will provide the foundation for Phase 2 studies to optimize manufacturing protocols that can by used in a GMP facility for producing lentiviral gene therapy vectors on a commercial scale. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective is to develop an automated fluorescence polarization immunoassay (FPIA) for detection of fentanyl and analogs in blood and urine. The Phase I specific aim is to synthesize and evaluate: 1) fluoresceinlabeled fentanyl analogs usable in a FPIA; 2) fentanyl (hapten)-protein conjugates used to elicit an immune response in Balb/c mice; 3) murine monoclonal antibodies to fentanyl and analogs. The Phase I objective is twofold: 1) to choose from the synthetic components a best combination for measuring fentanyl and analogs; 2) to prepare a small \"bank\" of well-characterized murine monoclonal antibodies for internal use as well as for use by other investigators. The healthcare industry and legislators are ill-prepared for the rapid and complex impact of escalating fentanyl abuse. The need for an innovative approach to detection of fentanyl in blood or urine is generally recognized. Authorities agree that an immunoassay will be required. By providing a fluorescence polarization immunoassay for fentanyl and analogs, the drawbacks associated radioimmunoassay will be overcome.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The first years of life are the most dynamic and perhaps the most critical phase of postnatal brain development. Abnormalities in early childhood brain development have been implicated in neurodevelopmental disorders, including autism and schizophrenia, though very little is known about this crucial time period. In the previous grant cycle, we studied brain development in a unique cohort of normal children longitudinally followed and scanned at birth, 1 and 2 years of age, including 386 neonates, 297 one year olds and 251 two year olds. In addition, we have developed the innovative image analysis tools necessary to study brain development in very young children. Our studies found rapid gray matter development and white matter maturation in the first 2 years of life, with marked regional differences in the cortex and in white matter tracts, consistent with temporal patterns of sensory/motor and higher integrative function development. We also found significant relationships between white matter maturation and working memory. Preliminary data indicates that regional gray matter volume and white matter tract diffusivity in neonates is predictive of subsequent gray matter volume and white matter structure at ages 1 and 2 years. These findings indicate that neonatal brain structure, reflective of prenatal brain development and the rapid growth trajectories of the first two years of life, likely play an important role in longer trm outcome. In this competitive renewal, we will extend follow-up of our cohort to 6 years of age and focus on structure/function relationships and the predictive value of early brain structure for later childhood brain structure and cognitive function. MRIs, including structural and diffusion tensor imaging, will be done at ages 1, 2, 4, and 6 years. Cognitive development, including general cognitive function and working memory will also be assessed. Developmental trajectories of cortical gray matter (including cortical thickness and surface area) and white matter (including tract-based spatial statistics and quantitative tractography) will be studied. We predict that neonatal brain structure and developmental trajectories in the first two years of life are critical for, and predictive of, subsequent structural and cognitive development. Relevance New knowledge gained in this study will provide a dramatically improved framework for understanding abnormalities of early childhood brain development in neurodevelopmental disorders such as autism and schizophrenia and will provide the fundamental information critical for developing preventative strategies for these disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Persistent pulmonary hypertension of the newborn (PPHN), previously called persistent fetal circulation, is a birth defect affecting approximately 1 in 1250 liveborn term infants; even with complex and high-risk interventions, PPHN results in substantial mortality and morbidity. This defect results from the inappropriate muscularization of fetal pulmonary vessels, and experimental and human evidence consistently suggests that maternal cigarette smoking and antenatal exposure to non-steroidal anti-inflammatory drugs (NSAIDs), particularly aspirin or ibuprofen, may play a role in the etiology of this condition. Because these exposures are quite prevalent (e.g., ibuprofen is currently taken in the first trimester or later in pregnancy by 15% and 3.2% of women, respectively), testing these hypotheses is of considerable public health importance. The investigators propose to conduct a multicenter case-control study of PPHN in relation to maternal exposure to smoking and NSAIDs. They will also assess other potential antenatal risk factors and collect and store buccal cell specimens for future analyses. There will be 560 case infants with PPHN and four controls per case (2240). All controls will be drawn from the birth hospitals of cases; half the controls will have malformations other than PPHN, and half will have normal formations. Cases and controls will be identified within 5 months of birth at 88 birth and tertiary hospitals in the areas surrounding Boston, Philadelphia, and Toronto. Mothers of subjects will be interviewed by telephone within six months of delivery; a standardized questionnaire will inquire in detail about demographic factors; reproductive, medical, and pregnancy illness histories; medication use (including a detailed focus on use of over-the-counter analgesic/antipyretic medications), smoking, and nutrition. Because of emerging genetic research suggesting an effect of NSAIDs on pathways possibly related to the etiology of PPHN, buccal swabs will also be collected and stored for future analyses. Exposure prevalences will be compared between mothers of cases and controls and relative risks will be estimated, controlling for potential confounding factors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The adverse effects of reactive intermediates derived from toxic chemicals result primarily from covalent binding to macromolecules and lipid peroxidation. Because of the central role of lipid peroxidation in the mechanisms of toxicity, many investigations of this process have been made. Fatty acid hydroperoxides are formed not only in free radical-mediated toxicities, but also physiologically as products of the peroxidation of polyunsaturated fatty acids (PUFA) by prostaglandin H synthases and lipoxygenases.One of the major reactions of PUFA- derived hydroperoxides is with hematin and various hemoproteins. Although the formation of PUFA-derived free radicals was proposed by Tappel in 1953 and free radicals were subsequently detected with ESR, the mechanism(s) of their reaction with hemoproteins has been the subject of considerable debate. The different mechanisms proposed for the reaction of hemoproteins with hydroperoxides dictate that the initial radical produced is either the peroxyl radical (as predicted by the heterolytic peroxidase mechanism), the alkoxyl radical (via the homolytic mechanism), or both the peroxyl and alkoxyl radicals (via the Haber-Weiss-type mechanism). The reactions of the initial radical that occur subsequent to its production can be suppressed by increasing the concentration of the spin trap used in the detection and identification of these reactive species. Thus, as more spin trap is added, more of the initial radical is trapped. This prevents the initial radical from undergoing reactions with other species or itself that lead to secondary radicals. By doing this, one can determine which radical is the primary or initial radical in a multiple species ESR spectrum. We used this strategy to determine whether the peroxyl or the alkoxyl radical was the initial radical produced by the reaction of hemoproteins with hydroperoxides. In every case examined thus far (i.e., cytochrome c, hematin, and cytochrome P-450) the alkoxyl radical adduct completely dominated at the highest DMPO concentrations. Thus, the ESR data provides strong evidence for the homolytic scission of the hydroperoxide O-O bond by hemoproteins initially producing alkoxyl radicals. - macromolecules, lipid peroxidation, toxicity, peroxyl radical, alkoxyl radical", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "X-ray crystallographic techniques are being used to study some acridine and benz(a)anthracene derivatives that are frameshift mutagens, carcinogens, and/or antitumor agents. The aim of this research is to find if there is a correlation of structure with biological activity for a series of chemically similar compounds. Studies will be made of polycyclic aromatic hydrocarbons and the K-region and non-K-region epoxides, dols, phenols and ketones derived from them which have been proposed as intermediates in the carcinogenic process. Molecular complexes of the hydrocarbons and acridines with more polar planar molecules, and covalent products of the action of activated carcinogenic hydrocarbons with peptides and nucleic acid components will also be studied. Work on acridine and anthracene mustards and their derivatives will continue. Studies of the mechanism of some enzymes, particularly those in the Krebs cycle, will be made by detailed studies of substrates and inhibitors, and by attempts to prepare good crystals of the enzymes themselves. Our efforts will be concentrated on aconitase and citrate synthase.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Synapse formation and maturation require signaling between the synaptic partners and signal transduction within each of them. To learn how assembly of the presynaptic neurotransmitter release apparatus is assembled, we focused on two genes, SAD-A and SAD-B. They are the mammalian orthologues of SAD-1, a gene required for presynaptic differentiation in C. elegans. Because SADs are kinases, we hope they will provide a valuable starting point for elucidating regulatory mechanisms that govern assembly of nerve terminals. Unfortunately, initial genetic tests of this idea gave complex results because the two genes play redundant roles and are involved in multiple steps in neuronal development and because SAD-A/B double mutants die at birth, before most synapses have formed. We therefore developed two genetic strategies to circumvent these limitations of pleitropy and lethality. First, we generated a conditional allele to ablate expression in selected neuronal types. Second, we generated alleles sensitive to a specific inhibitor, which allows precise temporal control of SAD-A/B activity and facilitates substrate identification. Using these new reagents, we have obtained preliminary results indicating that SADs are indeed required for complete presynaptic differentiation of several and perhaps most synaptic types. Here we propose to confirm and extend these results, and to initiate tests of our hypothesis that SAD kinases are critical components of pathways that lead from target-derived synaptic organizing molecules to assembly of nerve terminals. First, we will use the conditional allele to bypass neonatal lethality and characterize presynaptic defects in four peripheral and central excitatory synaptic types. We will also ask whether SADs are also required for development of inhibitory synapses, and whether SADs regulate post- as well as presynaptic development. Second, we will assay synaptic development and function with SAD-A and -B mutant alleles that render the kinases selectively inhibitable by an ATP analog to which unmodified kinases are insensivitive. These alleles provide us with precise and reversible temporal control over SAD kinase activity, both in vivo and in cultures generated from the mutant mice. We can therefore ask when during development SADs are required, whether their activity is required for synaptic maintenance in adults, and whether acute inhibition of the kinases affects the function of synapses that have developed normally. Finally, we will initiate studies aimed at learning how synaptogenic signals activate SADs and how SADs, in turn, coordinate presynaptic differentiation. PUBLIC HEALTH RELEVANCE: Formation of functional connections in the brain requires differentiation of the sending and receiving elements -the nerve terminal and postsynaptic membrane, respectively- and their precise apposition to each other. Much has been learned about the signals that postsynaptic cells use to organize nerve terminals, but little is known about how the nerve terminals pattern their differentiation in response to the signals. The project centers on analysis of two genes (called SAD-A and SAD-B) newly discovered to play critical roles in this process.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will develop Immunotherapeutics for Biodefense against Lassa virus (LASV), an arenavirus that is the causative agent of Lassa fever (LF). LASV is a Biosafety Level 4 and NIAID Biodefense category A agent. LF is a major public health concern causing widespread loss of life and social disruption across West Africa. The goal of this proposal is to perform critical steps in the characterization and pre-clinical development of a new class of therapeutics to treat LF. This effort combines the expertise of virologists and immunologists at Tulane University and the University of Texas Medical Branch with scientists at Zalgen, a biotechnology company specialized in the development of immunotherapeutics, rapid diagnostics, and new vaccine platforms for emerging viral diseases with global impact. Studies proposed here will leverage our recent advances, including identification of lead candidate human monoclonal antibodies (huMAbs) that neutralize LASV infectivity and protect guinea pigs against fatal disease induced by LASV. Zalgen's exclusive technology for rapid development (<3 months) of high yield antibody expressing stable cell lines ensures that costs of producing therapeutic antibodies is minimized for primary Biodefense markets and secondary Public Health markets. In studies proposed in MILESTONE 1 we will generate multi-milligram quantities of IgG1 and IgG4 LASV huMAb subtypes in serum-free medium adapted mammalian cell transient expression platforms for purification to perform in vitro testing. We will also perform a panel of characterization assays with the huMAbs focusing on identification of a huMAb or combinations of huMAbs with broad LASV neutralization potential and low IC50. Additional studies will establish cross-reactivity profiles with related Ne and Old World arenaviral proteins and reactivity of LASV huMAbs with human cell membrane phospholipids. In MILESTONE 2 we will generate CHO or NS0 stable cell lines in chemically defined serum free medium and select high producing isolates. Limiting dilution cell cloning (LDCC) will be performed for isolation of stable, production grade CD-SFM-CHO or CD-SFM-NS0 clones. We also plan to generate Accession Cell Banks (ACB) for clonal cell lines of interest, and to convert to manufacturing with QA/QC to generate gram quantities of candidate huMAbs for in vivo studies. In MILESTONE 3 we will perform evaluation of pharmacokinetics of LASV huMAb subtypes in guinea pigs (GPs). We will also perform challenge - therapeutic dose finding studies with two lead candidate LASV huMAbs or huMAb combinations in GPs, following a single dose via optimal delivery route. Next, we will determine the therapeutic efficacy with LASV huMAbs or huMAb combinations in Good Laboratory Practice (GLP) GP studies throughout the course of a lethal infection timeline, via the optimally determined route of administration. Results of the GP studies will guide an evaluation of pharmacokinetics of two single LASV huMAbs (IgG1 or IgG4) or huMAb combinations in a GLP nonhuman primate (NHP) study, following a single intravenous, intraperitoneal, or subcutaneous dose. We will also perform therapeutic efficacy studies with LASV huMAbs or huMAb combinations in GLP NHP studies throughout the course of a lethal infection timeline, following two doses via optimal delivery route, at varying times post infection. This proposed effort will result in a new class of immunotherapeutics for LF.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies were aimed at the isolation, characterization, and function of transforming genes potentially associated with the development of human mammary tumors. Analysis of primary mammary tumors and tumor cell lines of human origin for transformation activity following transfection onto NIH/3T3 cells revealed the presence of an H-ras oncogene in a mammary carcinosarcoma cell line, HS578T. The HS578T oncogene was molecularly cloned in its biologically active form. Recombinant gene constructs and nucleotide sequence analysis identified the activating lesion as a single nucleotide change from guanine to adenine at position 35 of the first exon, substituting aspartic acid for glycine at the 12th codon of the p21 gene product. The absence of restriction polymorphism for Hpa II/Msp I, induced by this single point mutation in HS578T tumor cells, from normal cells of the same patient demonstrated the somatic origin of the genetic lesion responsible for the transforming activity of HS578T. Moreover, the presence of the transforming H-ras allele as evidenced by Msp I/Hpa II restriction polymorphism in all single cell-derived clonal lines of HS578T with underrepresentation of the normal H-ras proto-oncogene suggests a selection for the transforming H-ras allele in the HS578T tumor cell population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: An integrated, behavior-genetic analysis of learning and memory processes in Drosophila is being pursued. Behavioral assays that reliably quantify several properties of non-associative and associative learning and memory formation have been developed. Initial results have revealed substantial functional similarities between fruit flies and vertebrates for these behavioral properties. Continuation of this work is organized along the following goals: 1. Behavior-genetic analyses of memory formation 2. Behavior-genetic analyses of associative learning 3. Behavior-genetic analyses of non-associative learning 4. Anatomy and development of learning and memory 5. Analyses of sensorimotor controls", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "One curious aspect of drug addiction is that a majority of people experimenting with illicit drugs do not develop the intense craving and compulsive drug use typifying addiction. Thus, identifying factors rendering a person resistant to addiction has application not only for treatment, but also for prevention of addiction. The current proposal exploits the environmental enrichment paradigm, a non-drug, non-surgical manipulation producing an addiction-resistant phenotype for intravenous drug self-administration in rats. Specifically, rats reared in an enriched condition (EC), group housed in large cages with novel children's toys, self administer less cocaine or amphetamine than rats raised in an isolated condition (IC). Importantly, the decrease in stimulant self administration is specific to self administration and not due to general anhedonia. Our preliminary data offer good evidence that the protective addiction phenotype is due, at least in part, to decreases in activity of the transcription factor cAMP response-element binding protein (CREB), with the net effect of decreasing neuronal excitability of nucleus accumbens (NAc) medium spiny neurons. The purpose of the current proposal is to further investigate neuronal excitability as it relates to addictive behavior and identify specific mechanisms mediating NAc excitability in EC and IC rats. Aim 1 will investigate whether or not decreasing neuronal excitability in the NAc can reduce the propensity for cocaine self administration in IC rats, and also to investigate if decreased excitability will be additive to the protective EC phenotype. Next, Aim 2 will examine mRNA expression of cationic ion channels as possible proximal mechanisms mediating the protective EC phenotype. For example, the study will examine basal expression of excitatory ion channels, hypothesizing decreased expression of specific ion channels in NAc of EC rats relative to IC rats, and for coordinated downregulation of groups of excitatory ion channels. The final aim will confirm mRNA regulation identified by Aim 2 and then study protein expression of these targets in EC, IC and SC rats. The results of these experiments will uncover novel gene targets mediating resistance to addiction. Further, these experiments will identify the critical \"missing links\" between CREB activity in the NAc and addiction-relevant behavior. Lastly, these experiments will provide new tools to accelerate understanding of the role of environment in mediating resistance to addiction. PUBLIC HEALTH RELEVANCE: We know that environment plays a critical role in susceptibility to drug addiction, and rearing rats in an enriched environment renders them resistant to addiction-like behavior in models of intravenous self administration. This project will identify molecular mechanisms underlying the protective effect of environmental enrichment and strengthen our understanding of how gene transcription affects behavior. The results of this project will lay the foundation for future treatment and prevention of drug addiction in humans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract The major goal of this project is to test the idea that modulation of autophagy within the hypoxic tumor microenvironment (TME) can impact circulating tumor cell (CTC) survival, metastatic potential, and therapeutic response. The TME is a complex breeding ground for selection of aggressive tumor cells with an advantage of survival or metastatic potential and confers resistance to cytotoxic as well as targeted cancer therapy. The process of tumor metastasis consists of multiple steps, including tumor cell dissemination from the primary tumor site into the vasculature or lymphatic circulation (intravasation), survival during circulation, extravasation into the secondary site, and initiation and colonization at the target organ site. Tumor cells must successfully complete each step to give rise to clinically detectable metastatic disease. It is generally accepted that autophagy is essential for tumor cell survival under conditions of nutrient or oxygen deprivation, the hallmarks of the TME. However, the relative contribution of autophagy within the TME to the pro-survival, metastasis- prone, and therapy-resistant phenotypes of CTCs is similarly unknown. There is therefore a great opportunity to unravel this biology and shed light on better therapeutic designs as well as therapy monitoring. We hypothesize that hypoxia-induced autophagy within the TME contributes to CTC survival, tumor metastasis, and chemoresistance. To test this hypothesis, we propose the following Specific Aims: 1) To determine the importance of hypoxia-regulated autophagy in tumor growth and metastasis in xenograft mouse models; 2) To evaluate autophagy and apoptosis in xenograft tumors and CTCs; 3) To investigate the effect of hypoxia- induced autophagy on therapeutic response as detected and monitored in CTCs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose to use quantitative proteomics approaches to study the interactions between simian immunodeficiency virus (SIV) and methamphetamine (METH) in the central nervous system (CNS) of rhesus monkeys. This proposal builds on our previous works for examining the SIV infected brains of rhesus monkeys for biomarker discovery using transcriptomics (gene array) and metabolomics (mass spectrometry) based technologies. For this project, we will perform hypothesis-based studies building on the wealth of data generated in functional proteomics at the University of Nebraska Medical Center (UNMC, Drs. Howard Gendelman, Project 1 and Pawel Ciborowski, Project 3). Our working hypothesis is that in the presence of METH, the harmful effects of SIV on the CNS are increased: when SIV and METH are joined there are dual and interacting untoward effects on CNS function and neuronal viability. Furthermore, since the proteome is broadly affected during disease, we believe that the use proteomics methodologies in the rhesus monkey/SIV/METH model will enable us to explore distinct and synergistic CNS disease mechanisms. We will utilize both unbiased and directed approaches to study how the effects of SIV on the primate brain are affected by METH. These studies are designed to utilize optimal samples for such discovery and follow-up validation, minimizing experimental confounds and directly studying the targets of SIV and METH, as well as accessible biofluids. In combination with the in vitro systems in H. Gendelman's Project ,1 and clinical samples in P. Ciborowski's Project 3, these nonhuman primate studies will form the necessary bridge between the projects as well as allow us to directly assess, in the most accessible and valid system, the target structures in the brain. Given the growing epidemic of METH use, and its relatively frequent abuse in the HIV infected population, this work is relevant to public health in the US. Knowledge about how METH is toxic to the brain in the setting of HIV infection will be useful not only in discouraging METH use, but in obtaining disease-specific markers that can be used for diagnosis and guiding therapeutic interventions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objectives of this research proposal are to study, by electron microscopy, 1) the localization of surface and intracellular protective antigens of the erythrocytic stages and gametocytes of Plasmodium by the use of monoclonals and cryo-ultramicrotomy, 2) the effects of histidine analogues on the structure of knobs appearing on the membrane of erythrocytes infected with P. falciparum, 3) the role of erythrocyte cytoskeleton during merozoite invasion by the use of Imposil-antibody conjugates and 4) the effects of colchicine and taxol on the formation of zygotes and ookinetes. The identification and localization of protective antigens within the parasites will facilitate further characterization of the antigen necessary for the development of malaria vaccine. The study on the effects of histidine analogues will advance our knowledge on the alteration of host cell membrane and may produce a clue enabling the prevention of such host cell membrane alterations. Understanding the role of the erythrocyte cytoskeleton during merozoite invasion may reveal a basic mechanism of host cell invasion. The investigation on the effects of colchicine and taxol on gametogenesis and zygote transformation will contribute a better understanding of these processes, particularly the role of microtubules. Thus, these investigations should provide answers to a number of basic questions on the hostmalarial parasite interaction and immunological mechanisms in malaria and may lead to the discovery of better methods for the control of malaria.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Patients with pancreatic adenocarcinoma have an extremely high mortality rate. Curative surgery would be more common if early differential diagnosis were feasible. Our group has demonstrated that the micro-leukocyte adherence inhibition assay (micro-LAI) can successfully and specifically detect the presence of pancreatic cancer. In our continuing study to determine the sensitivity of the assay for detection of early cancer, we require a stable, standardized source of pancreas tumor antigen. This will be accomplished by propagating several established low-passage pancreatic adenocarcinoma cell lines in athymic (nude) mice. A membrane extract of each of the lines will be analyzed by micro-LAI and the most consistently reactive tumor line will be grown periodically in large quantity and stable soluble antigen will be prepared for routine use in the assay. Malignant effusion fluid and patients' sera will be biochemically fractionated and selected fractions will be analyzed for reactivity in the LAI assay. Xenogeneic antisera will be prepared against the purified antigen preparations and they will be immunochemically and biochemically compared by serological assays and by 2-dimensional isoelectric-focusing electrophoresis. Radio-iodinated, purified antigen will be prepared and a radioimmunoassay (RIA) will be developed using the most avid xenogeneic antitumor antibodies. Preliminary RIA screening of selected sera from our serum bank will be carried out in a blind fashion. A portion of the pancreas cancer specific antibodies will be labeled with iodine-131 and assayed to determine their suitability for use in external computerized photoscanning for tumor localization. This project will significantly improve the micro-LAI assay and the serological and biochemical data obtained will complement the clinical results.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this project is to determine the contribution of sensory neuron mitochondrial dysfunction and TRP channel sensitization to pain in sickle cell disease (SCD), including the development of chronic pain. SCD is accompanied by crippling pain during acute sickling episodes that increases in severity and frequency with age. By adulthood, approximately 50% of SCD patients also suffer from chronic pain. SCD involves repeated ischemia-reperfusion injury and increased neutrophil numbers/activity, which results in elevated reactive oxygen species (ROS) generation and oxidative stress. This can lead to mitochondrial dysfunction and sensitization of Transient Receptor Potential (TRP) ion channels. We show that mice with severe SCD exhibit evidence for ongoing pain and pronounced pain-like behavior in response to cold, mechanical and heat stimuli. Our preliminary data also indicate that sensory neurons from sickle mice have severely dysfunctional mitochondria. We hypothesize that ROS and oxidative stress induce mitochondrial dysfunction and TRP channel sensitization in sickle sensory neurons, leading to the chronic pain and thermal hypersensitivity that is characteristic of this devastating disease. Aim 1 will determine the time course of neuronal mitochondrial dysfunction and pain sensitivity in sickle mice from age 2 to 12 weeks. Aim 2 will determine whether sensitized TRPA1 or TRPM8 channels underlie the cold hypersensitivity and primary afferent sensitization in sickle mice. Aim 3 will determine whether treatment with mitochondrial-targeted antioxidants reverses the pain behavior and primary afferent sensitization in sickle mice. These interrelated Aims provide a multifaceted approach that will define the mechanisms by which mitochondrial dysfunction and TRP channel sensitization lead to the devastating pain observed in SCD. Furthermore, this study targets specific cold-sensitive ion channels (TRPA1 and TRPM8) on sensory neurons because cold pain and cold hypersensitivity are a major complaint of patients with SCD. Data from this study will improve our understanding of the complex SCD pain syndromes and should aid in identifying novel therapeutic targets", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Brain metastases pose a significant problem for women with advanced metastatic diseases. The rate of brain metastasis has increased significantly in the last 10 years, approaching or exceeding 35% in subpopulations of metastatic breast cancer patients, particularly those with Her2+ or triple-negative tumors. After diagnosis of multiple metastatic lesions, patients typically die within one to two years. Gap: Treatment of brain metastases is primarily palliative due to limited curative effectiveness of radiation, surger, and poor delivery of chemotherapy across the blood-brain barrier (BBB). This proposal focuses on preventing metastasis seeding and initial growth in brain using preclinical models. Hypothesis: By reducing the efficiency of metastatic cancer cell penetration into brain and increasing drug delivery and efficacy in early micrometastatic lesions, we will decrease large metastases development and improve both neurological function and overall survival. Aim 1: Demonstrate that brain invasion of metastatic breast cancer cells can be inhibited at the level of the BBB: Preliminary data indicate TGF-? inhibition reduces brain invasion, at the level of the BBB, of triple negative human metastatic breast cancer cells in vivo ~70-80%. This work is extended to characterize mechanisms of how TGF-? inhibition reduces BBB cell invasion and the role Her2+ plays in BBB invasion. Aim 2: Elucidate the causal relationship between blood-tumor barrier permeability changes and chemotherapeutic uptake and effect in brain micrometastases of breast cancer: Preliminary data suggest that, contrary to common assumptions, most micrometastatic lesions (<500 ?m diameter) of breast cancer in brain show marked changes in metastatic vasculature structure and function, including vessel co- option, reduced vascular density, enhanced permeability, and elevated VEGF expression. In this Aim, work will be completed to characterize BBB changes in micrometastases, with the goal of identifying selective difference in micrometastases, such as VEGF and Notch, which can be used for targeted therapeutic benefit. Aim 3: Develop novel strategies to modulate blood-brain barrier permeability to improve therapeutic efficacy for brain micrometastases treatment: Preliminary data demonstrate that inhibition of vascular endothelial Notch signaling in both large and small brain metastases in the presence of VEGF results in targeted increases in BBB permeability. In this Aim, the targeted increases in permeability are exploited to improve chemotherapy delivery, cytotoxic effect and overall survival in three preclinical tumor models. Impact: The goal of this proposal is to develop novel approaches to reduce breast cancer cell invasion to brain, to modulate BBB permeability and improve chemotherapy uptake into CNS metastases, with an overall purpose to reduce brain metastases related death. The work requires state-of-the-art cell targeting, BBB permeability, and drug distribution methods to link barrier changes in selected small tumors with overall brain metastasis invasion and growth.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objectives of this research project are to characterize the signaling pathways that control the hepatic regenerative response in liver injury and disease. Clinical experience suggests that derangement of this regenerative response in chronic liver disease can lead to cirrhosis or neoplasia. Studies using the partial hepatectomy (PH) model system have led to the elucidation of a number of important coordinated signaling events involved in the initiation of hepatic regeneration. These include the production of cytokines and growth factors, generation of mitochondrial reactive oxygen species (ROS), activation of stress- and mitogen-activated-protein (MAP) kinases, and induction of transcription factors. We have shown that prostaglandin synthesis is also required for initiation of this regenerative response. Inhibition of prostaglandin synthesis with non-steroidal anti-inflammatory drugs (NSAIDs) augments TNFalpha-IL-6-STAT3 signaling, prolongs MAP kinase activation, and impairs activation of Jun nuclear kinase (Jnk) during liver regeneration. These changes are remarkably similar to alterations that occur in mouse models of steatohepatitis. In steatohepatitis models, altered production of ROS occurs in response to augmented cytokine signaling, and leads to subsequent alterations in MAP kinase and Jnk activation and inhibition of liver regeneration. This suggests that changes in cytokine-regulated ROS production may be involved in the mechanism by which inhibition of prostaglandin synthesis leads to impaired liver regeneration. Therefore, we hypothesize that prostaglandin synthesis is essential for appropriate regulation of cytokine-stimulated ROS production after PH and that changes in ROS metabolism are a fundamental component of the mechanism by which inhibition of prostaglandin synthesis leads to impaired liver regeneration. This could have important implications with respect to the clinical use of NSAIDs in patients with liver disease. In this grant we propose to determine the effects of prostaglandins on ROS synthesis, expression of genes regulating ROS metabolism, and expression and activity of negative regulators of cytokine signaling during liver regeneration.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Adolescence is a peak time for the emergence of the core symptoms of psychopathology. Cognitive disorganization (CD) is a key symptom dimension of psychosis that emerges most commonly in adolescence, reflects a disorganization of thought and is defined by the presence of bizarre behavior, alogia, and impaired attention. CD transcends DSM diagnostic categories, predicts the onset and severity of psychotic disorders, and is associated with neuropsychological impairment. Genetic studies have reported that CD is highly heritable, and therefore has been proposed as a promising phenotype for molecular genetic studies. Despite its critical role in heralding risk for psychosis, little is knon about the neurobiological underpinnings of CD in adolescents. It has been reported however that adolescents experiencing disorganization have significant deficits in working memory capacity (WMC) and arousal/stress regulation (ASR). These two behavioral constructs show dramatic maturational changes during adolescence, which are necessary for the transition to higher-level cognition, affect regulation and psychosocial adaptation. Despite the strong epidemiologic evidence for the role of stress in the etiology of psychosis, and the centrality of working memory impairments in psychosis, little is known about their contribution to CD in adolescence. Examining the neural and physiological systems associated with working memory and stress regulation in adolescence, and their contribution to CD severity, offers a critical step in elucidating the pathophysiological mechanisms that contribute to the onset of psychosis. This approach is consistent with the RDoC framework, which encourages using converging measurements to study the underlying neurobiology of domains (Cognitive System and Arousal Regulation in this proposal), and constructs (working memory capacity and stress regulation) that represent fundamental behaviors expressed by individuals with clinical risk symptoms for psychosis. We will use a multimodal approach integrating functional neuroimaging, electrophysiological, and behavioral measures to ascertain converging measures of working memory and arousal/stress regulation constructs across neural, physiological, and behavioral units, and to characterize the contributions of atypical ASR and impaired WMC in the severity of CD symptoms measured through clinical scales. In Aim 1, we will evaluate the contributions of working memory impairments and atypical arousal/stress regulation in 180 adolescents (ages 9-16) to the severity of CD symptom. In AIM 2 we will model the relationship between WM and ASR constructs and their impact on CD severity. In AIM 3, we will examine the longitudinal trajectory of CD symptom severity, behavioral and electrophysiological measures of WM and ASR, and their associations with baseline neural, behavioral, and physiological measures acquired in AIMs 1 and 2. For each aim, we will explore the modulatory role of sex differences and pubertal maturation on stress-regulation and working memory during adolescence, and their influence in determining functional outcomes. IMPACT: Understanding the neural and physiological systems associated with working memory capacity and stress regulation in adolescence, and their contribution to CD severity, is a crucial step for elucidating the core pathophysiological mechanisms that promote the emergence and exacerbation of psychosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The complexity of the mixed, polymicrobic, multifactorial infection known as periodontal disease continues to pose serious difficulties for the research scientist. It is commonly accepted that the polymicrobic environment of the periodontal pocket plays an important role in microbial succession as well as pathogenesis of periodontitis (25). One of the microorganisms closely associated with the severity of this type of infection is Treponema denticola. Like so many other organisms associated with periodontal disease, its actual role in pathogenesis is unknown (15). Limited laboratory data indicate that co-culture with Peptococcus can supply some of its growth requirements (39). In addition, initial observations in our laboratory indicate that medium spent by growth of T. denticola supplied factors which modulated growth of other oral microorganisms. Thus far, no systematic attempt has been made to determine the effect of mixed culture growth on T. denticola or co-cultured organisms, or critically examine mechanisms responsible for modulation. The purpose of the proposed study is to (a) identify the ability of commonly isolated periodontal organisms such as Bacteroides gingivalis, Bacteroides intermedius, Bacteroides endodontalis, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, Capnocytophaga gingivalis, Eikenella corrodens, Peptostreptococcus anaerobius, and Streptococcus mitis to modulate growth, (b) determine if T. denticola produces factors which modulate growth of other oral bacterial noted above, and (c) partially characterize modulatory factors as to molecular weight and for stability to heat, pH (2.0 - 12.0), and trypsin. Determination of mechanisms involved in selection and maintenance of specific microflora in the periodontal lesion will allow a more complete understanding of the pathogenesis and progression of periodontal diseases. Furthermore, the data obtained from this study will provide novel information concerning the specific role of spirochetes in this process.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Anisotropy in Matrix Bioblocks: A Platform for Microvascular Engineering Lung damage has devastating consequences for pulmonary and cardiovascular health, with few treatment alternatives beyond chronic mechanical ventilation and lung transplantation. The study of vascular development is essential to developing 3D engineered grafts for regenerative lung therapies. This proposal addresses two main problems in vascular tissue engineering: 1) the need for a natural, complex scaffold material and 2) the requirement for pre-vascularized tissue constructs. These two issues will be addressed in a model context by developing human lung microvascular networks in natural, tissue-specific matrices produced by human lung fibroblasts. The first aim is to examine the assembly of natural, fibroblast matrix in confined, 3D geometries and to identify the molecular mechanisms driving anisotropic cellular growth and matrix development in response to cues from environmental geometry. This will be tested using biochemical and genetic manipulation of cytoskeletal modulators. This aim will test the hypothesis that actin is the major component influencing anisotropic cellular response to 3D confinement. Biochemical inhibitors and siRNA will be used to target specific cytoskeletal functions. The spatial organization and anisotropy of cytoskeletal assembly and matrix deposition will be evaluated for each treatment group using pairs of 3D shapes that have constant volume and area, but varying anisotropies with confocal laser scanning microscopy and quantitative autocorrelation analysis. The second aim will analyze the effects of confined, geometric microenvironments on matrix remodeling and vessel formation by human lung microvascular endothelial cells. The hypothesis that 3D microvascular network formation can be controlled using a natural matrix model will be tested. Microvessel formation and alignment in wells of varying geometries will be examined quantitatively. In summary, this predoctoral training proposal will analyze the effects of confined 3D geometries on the alignment of matrix deposition and growth patterns of microvessels. This research provides an avenue for the in vitro control of vasculogenesis orientation and can potentially bridge the gap between the biology of matrix-cell interactions and translational therapy in pulmonary medicine. By bringing together multidisciplinary areas through the integration of engineering and the life sciences, this proposal addresses fundamental questions in tissue engineering to accelerate the development of functional tissue grafts for the lung, while developing a paradigm that may also be used for other vital organs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the adult, protease-activated receptors (PARs) respond to coagulation proteases to help orchestrate cellular responses involved in hemostasis, inflammation and repair. In the previous project period, we showed that PARs also play distinct and important roles in embryonic development. Indeed, PAR1 signaling in endothelial cells is required for proper remodeling and/or integrity of developing blood vessels in mouse embryos. What biochemical and physiological processes does PAR1 monitor during blood vessel formation? Which endothelial cell responses to PAR1 activation are important for proper vessel development? We hypothesize that the coagulation cascade and PARs together provide a system for monitoring and regulating the formation, remodeling, and integrity of developing blood vessels. Toward testing this hypothesis we shall first determine whether PARs sense coagulation proteases during mouse embryonic development, and whether activation of PARs account for the roles of coagulation factors in this context. Specific questions include a) Does combined deficiency of PARs phenocopy tissue factor deficiency? b) Is tissue factor expression in or around blood vessels necessary and sufficient to support embryonic development? and c) Can embryos bearing prothrombin mutations be rescued by complementary mutations in PAR1? We shall next determine which signaling pathways are important for the function of PAR1 and other GPCRs in endothelial cells during vascular development by ablating the function of specific G protein pathways in endothelial cells in the mouse embryo. We expect these studies to illuminate a novel role for the coagulation cascade and PARs and to provide new insights regarding the mechanisms governing vascular development and perhaps new blood vessel formation in other settings.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is an open label, non randomized, Phase I dose escalating study of rhIL-12 administered by intraperitoneal (IP) infusion in adult male or female patients with measurable disseminated intraperitoneal cancer that is residual, recurrent or metastatic. Neither a control or placebo group will be utilized. Up to 30 patients will be treated weekly on an outpatient basis with the study drug. Patients will be entered in cohorts of three dose tier of rhIL-12 at doses ranging from 3-500 ng/kg administered over the first two weeks of each three week cycle for three cycles. The primary objective is to assess the safety and MTD of rhIL-12 when administered by IP infusion in this patient population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The present application of configurational statistics to model the nucleic acids or polynucleotides provides an essential first step in relating the subtle features of chemical architecture to the unique physical and biological properties of these macromolecules. Through a combination of molecular modeling, potential energy calculations, and statistical mechanical analyses it is possible to elucidate details of nucleic acid conformation and to provide a rational understanding of observed experimental phenomena. The primary objectives of the program are to classify and understand the complex array of nucleotide conformers, to comprehend the effects of primary chemical sequences on secondary and tertiary structure, and to elucidate pathway of conformational transitions. The combined computations should enhance our comprehension of both polynucleotide extension and flexibility offering new structural insight into conformation and the irregularities of various forms. The chain statistics studies may also provide a molecular basis for understanding the nature and flexibility of specifically constrained structures such as small closed circles, loops, knots, hairpins, bulges, and cruciforms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Total Parenteral nutrition (TPN) is the method of intravenous nutrition delivery bypassing the gut in patients unable to receive regular enteral nutrition (EN). It is a crucial lifesaving therapy for over 30,000 individuals in the US permanently dependent on TPN. Several fold higher number of patients require TPN for a prolonged duration. Unfortunately, side effects of this critical therapy used world-wide include potentially fatal liver and gut injury from a likely multifactorial etiology. Emerging data, including results from our K08 funding suggests that a disruption of gut-derived signals in response to a lack of luminal nutrient delivery, as occurring with TPN therapy drives such injury. Our lab has been investigating the role of such signaling. Using a novel ambulatory TPN piglet model, developed at our lab which recapitulates human TPN delivery; we have published significant alterations in the gut microbiota of animals on TPN. Specifically we have shown a significant increase in the pro-inflammatory Bacteroidetes phylum and a decrease in the Firmicutes phylum in TPN animals. We have also noted significant alterations in key hepatobiliary receptors and transporters that drive gut-systemic signaling and contribute to TPN associated injury. Recent data also suggests that alteration in inflammatory cytokines secondary to microbial shifts can lead to such injury. Therefore we believe that the altered gut microbiota, during TPN, may have a prominent role in TPN associated injury. We thus hypothesize that a restoration of the altered gut microbiota in TPN animals by a transfer of fecal microbiota from control EN animals will mitigate TPN-associated injury. As detailed in the research plan; with Aim 1 we will test the impact of rigorously monitored fecal transplantation from control EN animals to those on TPN and evaluate gut injury. As part of this aim, we shall objectively identify and quantify stool microbiota using culture independent 16S ribosomal sequencing and assess serological gut injury markers, gut permeability, histology and perform gut morphometric analysis. Aim 2 relates to exploring the impact of fecal transplantation on hepatic injury and critically testing the gut-systemic cross talk. This aim is rationalized by an alteration of hepatobiliary receptors and transporters as well as cytokine mediated injury secondary to microbial shifts with TPN therapy. Liver injury serological markers, histology and key hepatobiliary receptors, transporters and signaling molecules driving the gut- systemic cross talk will be assessed to gain mechanistic insights. This proposal complements our K08 work and could provide important insights into the role of gut microbiota in TPN associated injury and help develop strategies to mitigate complication of this life saving therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dr. Sherry Timmons is currently in the Dentist Scientist program at The University of Iowa College of Dentistry. She is continuing her clinical training in Oral Pathology, Radiology and Medicine and is progressing in her research. The broad objective of her research is to examine the interactions between alcohol and tobacco carcinogenes in the etiology of oral cancer. Specifically, her research will examine the fundamental mechanisms of alcohol involvement in oral cancer. Dr. Timmons attended the Annual IADR meeting in Vancouver, BC March 9-14, 1999 where she participated in an Experimental Pathology Oral Session. Her presentation was, \"Effects of Acetaldehyde on c-jun Expression in Squamous Cell Lines.\" Dr. Timmons co-authored an article accepted for publication in Dental Maxillofacial Radiology: Timmons S, Ruprecht A, Diehl ST II. \"Frequency of Portrayal of Foramen Transversarium of the Second Cervical Vertebra on Rotational Panoramic Radiographs.\" Projected publication date is May, 1999.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Drosophila imaginal discs are able to regenerate and thus provide us with a model system to study multipotency of cells. Under certain conditions a small set of cells in the \"weak point\" of each imaginal disc change their fate. This switch, known as transdetermination (TD), occurs with high frequency when wg is overexpressed or when a disc is cut through the \"weak point\". Expression profiling of these cells identified candidate genes which in functional tests significantly modified TD. Some of these candidates are chromatin-remodeling genes. Furthermore, we identified putative regeneration genes such as air-like, a gene orthologous to the mouse gene augmenter of liver regeneration (air), Matrix metalloproteinase 1 (Mmp1), and dReg, a novel gene that contains a C-lectin domain, Although the mammalian orthologs of these three genes have been linked to regeneration of specific tissues, the mechanism by which they act is not well understood. We propose to continue and complete the functional tests, including the generation of mutations in novel candidate Drosophila genes. Dr. Moon we will test homologous zebrafish genes. In examining regeneration, the issue of stem cells must be addressed. Do TD cells divide like stem cells? Cell clones will be induced'in TD cells to assay for asymmetrical divisions, a landmark of stem cell behavior. These analyses will then be performed in the presence and absence of characterized enhancers and suppressors of TD to test whether these genes affect developmental plasticity by modifying the cell lineage pattern, the number of founder cells or the cell-doubling times. This proposal will contribute to the understanding of which molecular signals increase regeneration potential to ensure normal tissue maintenance and repair. Short summary A few specific cells in each imaginal disc of Drosophila have stem cell properties. If challenged by injury they regenerate. We will test whether these cells divide asymmetrically, a landmark of stem-cell behavior. In addition, we will identify the genes that mediate developmental plasticity and test how their gene products affect multipotent cells and their surroundings. Drosophila has been used as a model system to study human disease and we see here an opportunity to gain insight into stem-cell biology and regeneration.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract Electronic adherence monitoring (EAM) technologies are widely used to support antiretroviral adherence. Unfortunately, EAMs such as Wisepill assume, but cannot verify, actual ingestion of oral medication. In contrast, My/Treatment/Pill (MyTPill), an innovative technology that directly measures ingestion, comprises a digital pill containing a medication and tiny radio emitter in a gelatin capsule. When the digital pill is ingested, gastric contents dissolve the capsule to activate the emitter. The digital pill ?syncs? real-time ingestion data to a smartphone application to provide vivid, indisputable measures of medication ingestion. Head-to-head comparisons of EAMs, however, have yet to be performed. Identifying the superior EAM would improve virologic suppression by enabling real-time interventions to support ART adherence. In this randomized controlled trial, we will compare MyTPill to WisePill among N=80 HIV+ men/women taking prescription opioids and once-daily ART regimens containing tenofovir and emtricitabine with a viral load >200/mL. HIV+ patients on prescription opioids have difficulty adhering to ART regimens, although the reasons are not fully known. Given the high prevalence of prescription opioid misuse among HIV+ individuals?triple that of HIV-negative persons?and striking rates of suboptimal ART adherence (46% worse than those who do not misuse), strategies to improve ART adherence in this population are critically needed. Participants will be randomly assigned in a crossover trial to (1) MyTPill x 3 mos, then WisePill x 3 mos; or (2) Wisepill x 3 mos, then MyTPill x 3 mos. Adherence measured via MyTPill and WisePill will be compared to dried blood spot (DBS) concentrations of tenofovir diphosphate (for cumulative adherence) and emtricitabine triphosphate (for recent adherence). Participants will provide DBS samples on multiple random times according to a schedule that prevents anticipation of sampling but which assesses cumulative/recent ART adherence. We will also examine which aspects of prescription opioid use, pain, withdrawal, and demographic, social, structural, and other environmental contexts (measured by timeline follow back and quantitative interviews) are most closely linked to ART adherence. Furthermore, we will examine how these aspects affect MyTPill and WisePill measures of ART adherence. Primary Aim: Determine if MyTPill, as compared to Wisepill, exhibits: (1) better measures recent and cumulative ART adherence when using DBS as the ?gold standard?; and (2) better participant experience as assessed by observed and self-reported measures (e.g., study retention, fidelity to study, protocol relative subjective index, qualitative interviews). Secondary Aim: Examine which aspect(s) of prescription opioid use (e.g., prescribed use/misuse of opioids as measured by MyTPill, technology subversion, pain, demographic, social, other structural factors) are most closely linked to ART nonadherence (per DBS), and how they affect measuring ART adherence using MyTPill and Wisepill. Public health significance: If MyTPill, a non-invasive, self-contained, and nearly automated ART EAM, is superior to other strategies, it will serve as a platform for subsequent research testing real-time ART adherence interventions; 2) specific factors associated with suboptimal ART adherence can be addressed with interventions directed towards HIV+ persons receiving prescription opioids; and 3) MyTPill can directly address the crisis of opioid misuse arising from treatment of chronic pain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Topical microbicides represent an emerging strategy for the prevention of transmission of HIV and other sexually transmitted infections. A successful topical microbicide product will be applied prior to intercourse, without necessitating partner consent, and will be active against a variety of STIs, including HIV. It will be acceptable to potential users in terms of physical characteristics, availability, ease of use, safety and efficacy properties. We have utilized the macaque vaginal safety model (currently contracted by the NIH, N01-AI-95388) to provide standardized preclinical safety data for numerous topical microbicide products in development. In this model, measures of product safety include cervicovaginal colposcopy, vaginal microbiologic evaluation, and vaginal pH monitoring. This model characterizes the vaginal environment's response to repeated topical product applications in the absence of the exogenous factors of intercourse and potentially infectious ejaculate. While our preclinical evaluations of topical microbicide products have been well-rounded in many aspects, we have yet to investigate the effects of sexual intercourse on the cervicovaginal environment. Mucosal perturbation and potential micro-trauma in the form of epithelial abrasions are likely to result from sexual activity. Additionally, the effects that seminal fluid may induce on the cervicovaginal environment, as well as its effects vis-[unreadable]-vis topical microbicide product safety and efficacy, have not yet received their due attention. We propose to enhance our standardized vaginal safety evaluations conducted in the pigtailed macaque model to include evaluations after sexual activity and with the presence of seminal fluid. With continued R33 funding, we will collect baseline data from 24 female macaques assessing the cervicovaginal environment before and after mating. In addition, we will collect parallel assessments, when mating has occurred with a placebo gel (HEC universal placebo) in place. These studies will provide urgently needed data regarding topical microbicide use with coital activity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chemical Technology researchers develop small molecule and screening approaches that other scientists can use to pursue innovations in therapeutic development. Small molecule kinase inhibitors are finding utility as drugs in several cancer indications. Recently, a collaboration with the Sarczynowski lab (Cincinnati Childrens Hospital) established the potential of small molecule IRAK inhibitors in Myelodysplastic syndrome (MDS). In an extension of this work, we have established that a known IRAK inhibitor possesses unique activity versus acute myeloid leukemia and myelomonocytic leukemia cancer lines. The agent possesses strong activity versus several kinases including Flt3, PDGRF and IRAK with in vitro activity surpassing existing drugs in this domain. We currently are evaluating candidate small molecules within in vivo models of these diseases. Importantly, these agents appear to overcome selected mutations that current plague clinically active Flt3 inhibitors and, therefore, limit their utility.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The capsular polysaccharides of Streptococcus pneumoniae are essential virulence determinants for this organism. They serve as the basis for serologic classification, with ninety serotypes being distinguished to date, and they are the target for protective antibodies. In both human infections and animal models, virulence is related to the serotype of the capsule expressed. The mechanisms underlying the virulence differences between serotypes are not known but may involve both the polysaccharide structures and other factors in the genetic background in which they are expressed. Molecular genetic studies have identified a common organization among the different S. pneumoniae capsule loci. This organization permits the exchange of capsular serotypes during transformation, leading to new combinations of virulence factors and potentially impacting on the efficacy of polysaccharide-based vaccines. Knowledge of the genetic components of the loci has permitted pathways for polysaccharide synthesis to be demonstrated or proposed. Often, enzymes expected to be essential for capsule production are not encoded by genes in these loci, and the necessary products are obtained from cellular pools contributing to pathways for peptidoglycan and teichoic acid synthesis. Thus, capsule expression and basic cellular functions are intimately linked. Recent studies have demonstrated a requirement for capsule in colonization but have suggested that capsule production is reduced in this environment as compared to other host environments. Expression of other factors important for adherence and sustained colonization may be elevated during colonization and reduced at other times. Coordinate regulation of these virulence factors is thus anticipated and recent studies have identified a surface component potentially involved in adherence whose expression is altered in response to changes in capsule production. In the proposed studies, we will continue the genetic analysis of virulence and capsule expression. The specific aims are to: 1) determine the effect of specific alterations in capsule type and structure on virulence, 2) characterize the requirement for capsule in pneumococcal infections and its expression in vitro and in vivo, and 3) characterize regulatory networks and cellular pathways associated with capsule synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Different mechanisms could explain the origin and heterogeneity of CSC such as (i) differentiation arrest (stem cells), (ii) dedifferentiation (mature cells) and (iii) transdifferentiation (bone marrow stem cells). It is conceivable that all 3 mechanisms may be corrupted by oncogenic events, resulting in an assortment of CSC and explaining their heterogeneity. Defining and characterizing this heterogeneity is of vital importance for understanding CSC biology, and for effective therapeutic translation. Our most recent results in this project include: (1)Epigenetic mechanisms play critical roles in stem cell biology by maintaining pluripotency of stem cells and promoting differentiation of more mature derivatives. If similar mechanisms are relevant for the cancer stem cell (CSC) model, then epigenetic modulation might enrich the CSC population, thereby facilitating CSC isolation and rigorous evaluation. To test this hypothesis, primary human cancer cells and liver cancer cell lines were treated with zebularine (ZEB), a potent DNA methyltransferase-1 inhibitor, and putative CSCs were isolated using the side population (SP) approach. The CSC properties of ZEB-treated and untreated subpopulations were tested using standard in vitro and in vivo assays. Whole transcriptome profiling of isolated CSCs was performed to generate CSC signatures. Clinical relevance of the CSC signatures was evaluated in diverse primary human cancers. Epigenetic modulation increased frequency of cells with CSC properties in the SP fraction isolated from human cancer cells as judged by self-renewal, superior tumor-initiating capacity in serial transplantations, and direct cell tracking experiments. Integrative transcriptome analysis revealed common traits enriched for stemness-associated genes, although each individual CSC gene expression signature exhibited activation of different oncogenic pathways (e.g., EGFR, SRC, and MYC). The common CSC signature was associated with malignant progression, which is enriched in poorly differentiated tumors, and was highly predictive of prognosis in liver and other cancers. Conclusion: Epigenetic modulation may provide a tool for prospective isolation and in-depth analysis of CSC. The liver CSC gene signatures are defined by a pernicious interaction of unique oncogene-specific and common stemness traits. These data should facilitate the identifications of therapeutic tools targeting both unique and common features of CSCs; (2)Cancer heterogeneity is dynamically regulated by genetic, epigenetic and cellular microenvironmental factors. These factors also play important roles in transcriptional reprogramming and cell fate changes in cancer cells. DNA methylation constitute an important part of the epigenetic mechanism modulating cancer and stem cells. We have studied the interaction between epigenetic alterations and local microenvironment in regulation of cancer stem cell (CSC) properties using Zebularine (Zeb), a DNA-methyltransferase1 (DNMT1) inhibitor, in combination with the modulation of cell density in culture. Seven human hepatoma cell lines, including two early passage cell lines established from primary HCC, were plated at high (HD) and low density (LD), and exposed to 100 microM of Zeb for 3 days. Thereafter, cells were dissociated and plated at clonal density over several passages (generation G1 to G5) in drug-free, serum-free, and adherent-free conditions. The differences in self-renewal, gene expression, tumorigenicity and metastatic potential of cells derived from G1 to G5 spheres were examined. shDNMT1-Huh7 cell line was generated to address the role of DNMT1 in long-term self-renewal. Results: The transient exposure to Zeb produced differential cell density-depended responses in 5/7 tested HCC cell lines. In cell lines which were scored as Zeb-sensitive, a 3-day drug pre-treatment of LD cultures (LDZ) caused a remarkable increase in primary sphere formation. This effect persisted through at least five subsequent sphere generations in methylcellulose sphere-forming assays in drug- and serum-free medium. In striking contrast, untreated LD cells failed to form primary spheres while the sphere forming potential of HD and HD Zeb treated (HDZ) cells rapidly decreased over the first three generations. The depletion of DNMT1 in Huh7, a Zeb-sensitive cell line, by a stable lentiviral transduction of DNMT1 shRNA similarly increased the self-renewal potential of LD cells, providing evidence that the density-dependent effect was epigenetically regulated. Conversely, the phenotypic response of Zeb-resistant WRL68 and HepG2 was independent of drug and plating schema despite a complete depletion of DNMT1 protein.The increase in sphere formation in LDZ cells strongly correlated with a stable overexpression of CSC-related markers (CD133, CD44, EpCAM), as well as key genes involved in ESC self-renewal (Bmi1, alapha-SUZ12) and epithelial-mesenchymal transition (EMT) (beta-catenin, ZEB1, VIM, SNAL1). Likewise, when LDZ, HD and HDZ spheres were dissociated and injected s.c. into NOD/SCID mice (100 cells/injection site), the LDZ cells generated tumors more rapidly and with higher penetrance, and showed more frequent brain and lung metastasis. Both gene reactivation and tumorigenicity progressively increased G1 to G4. Tumors derived from G1-G4 LDZ cells were also increasingly more vascular. Global transcriptome analysis of LDZ spheres at G1-G4 confirmed that a common LDZ signature was characterized by genes associated with oncogenic signaling pathways including tumor initiation, metastasis, and vasculogenesis and was able to predict clinical outcome of liver cancer patients. We conclude that epigenetic reprogramming of liver cancer cell lines induced by a combined modulation of DNA methylation and cellular microenviroment can enhance and stabilize CSC properties;(3)Classification of human liver cancer into biologically distinct subgroups suggests its origin from different hepatic lineage cells. To clarify the contribution of the lineage stage in liver oncogenesis, we transduced H-Ras/SV40 large T into hepatic progenitor cells (HCP), hepatoblasts (HB) and terminally differentiated adult hepatocytes (AH). Regardless of origin, the transformed cell types acquired common cancer stem cell traits both in vitro and in vivo. However, expression analyses distinguished tumors from different lineage stages demonstrating that distinct genetic changes occur during malignant transformation. Notably, AH-derived tumors showed specific enrichment of c-Myc target genes. Our results demonstrate that any hepatic lineage cell can be a target population for transformation via activation of diverse cell-specific pathways. Primary human liver cancer (PLC) is the third most lethal cancer worldwide with incidence rising in United States and Western Europe. PLC can arise from liver epithelial cell lineages and adult hepatic progenitor cells. Identification of cells that are susceptible to oncogenic transformation is critical for both diagnosis and treatment. Employing a mosaic mouse model of PLC we demonstrated that any cell within the hepatic lineage can be a target of malignant transformation and display a cancer stem cell mode of tumorigenesis. The identification of common and cell of origin specific phenotypic and genetic changes should provide novel therapeutic targets for the treatment of PLC. In particular, our demonstration of the general role of c-myc in PLC oncogenesis offers unique therapeutic opportunities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Toll-like receptors (TLRs) play a crucial role in pathogen recognition and activation of innate and adaptive immune responses to infection. The microbial ligands recognized by TLRs are not unique to pathogens, however, and are produced by both pathogenic and commensal microorganisms. We have recently found that recognition of commensal bacteria by TLRs is critical for the maintenance of intestinal homeostasis, tissue repair and protection from injury. On the other hand, dysregulated responses to commensal bacteria can lead to inflammatory diseases, such as inflammatory bowel disease (IBD). The role of TLRs in the initiation and pathogenesis of IBD is currently unknown. The goal of this proposal is to investigate the role of commensal-TLR interactions in the maintenance of intestinal homeostasis, tissue protection and repair, and the pathogenesis of IBD. We will generate mice that are selectively sufficient or deficient in TLR signaling in different cell types present in the intestine. Using these mice we will investigate the role of different cell types and gene products in recognition of commensal bacteria, host defense, intestinal homeostasis and tissue protection and repair. We will also examine the role of TLRs in the etiology of commensal dependent, T cell mediated colitis model and examine the contribution of TLR signaling by different cell types to pathogenesis. We will analyze the TLR-dependent pathways that are negatively regulated by IL-10 in the colon, and investigate how dysregulation of these pathways leads to initiation and maintenance of colitis. These studies will provide novel and important information that will help elucidate the role of TLRs in intestinal physiology, the pathogenesis of IBD and the biology of host-commensal interactions. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research is an extension of ongoing studies of airway smooth muscle (ASM) cell development and function. The broad goals are to define mechanisms which regulate expression of functionally critical genes, including m c receptors, pp junctions, and smooth muscle actin, and to elucidate the process governing ASM cell proliferation. Specifically, cultured human airway smooth (HASM) cells will be used to define: a. The role of inflammatory cells and, more specifically, the cytokines 11 -beta, and TNF in regulating HASM cells proliferation, gene expression, and, ultimately contractility. b. The role of the autonomic nervous system in the regulation of HASM cell proliferation and phenotypic expression. Specifically, interactions between cultured HASM and cultured autonomic neurons will be examined in depth. c. Second messenger systems which mediate effects of innervation and of cytokines on HASM cell function. The long term goal of these studies is to define mechanisms underlying airway hyperreactivity in diseases such as asthma and COPD. It is hoped that these studies will indicate biochemical and molecular loci where therapeutic intervention will help to ameliorate such bronchopulmonary disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term research goal of this project is to understand how cytoskeletal motors power the transport of diverse macromolecules within eukaryotic cells, enabling them to effectively organize their contents, move, divide, and respond to signals. This proposal focuses on cytoplasmic dynein, the largest, most complex, and least understood of the cytoskeletal motors. The specific objectives are to determine how single dynein dimers move processively, how ensembles of motors efficiently move cargo, and the role of processive movement in cells. A significant obstacle to understanding these important features of motility is a lack of tools to precisely control motor-motor and motor-cargo interactions in vitro. Using DNA nanotechnology, we have developed methods to achieve this. First, we generate stable, functional dynein heterodimers through DNA base pairing. Second, using three-dimensional (3D) DNA nanotechnology, we build synthetic cargo to which DNA-linked dynein or kinesin motors can be attached with defined numbers and spacing. To determine how dynein takes consecutive steps along microtubules, single-molecule techniques, including high-precision, multi-color fluorescence microscopy and single-molecule Forster resonance energy transfer (smFRET), will be applied to track the behavior of individual moving dynein molecules. The results of these experiments will be used to construct a model for how dynein moves processively on microtubules. To determine how coordination among dynein motors or between dynein and kinesin motors affects cargo motility, varying numbers of dynein or dynein mixed with kinesin will be attached to a 3D, synthetic DNA cargo. By analyzing the behavior of both the cargo and individual, cargo-attached motors in single-molecule motility assays, the biophysical properties of multi-motor-based transport will be determined. Long distance transport is thought to require processive motility. However, we recently discovered that dynein is sub-maximally processive. Using in vivo and in vitro approaches, we will test the hypothesis that sub-maximal processivity is especially critical for cytoplasmic dynein. Because cytoplasmic dynein is encoded by only a single gene in all eukaryotes but carries out a wide range of tasks, sub-maximal processivity may allow it to be tuned to perform a variety of cellular functions. This research will provide fundamental, mechanistic insights into how the ubiquitous and essential dynein motor works. In addition, the DNA nanotechnology tools generated here will serve as general engineering principles for studying the oligomerization state of other proteins or for studying arrays of any molecular motor in a more physiologically relevant manner.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Understanding of anatomical and physiological relationships within the neural networks driving craniofacial pain is a key step in developing effective therapeutic treatments. Many studies evaluating such networks focus on peripheral mechanisms of trigeminal nociception and on homologies between trigeminal and spinal nociceptive processing. The extended goal of this work is to investigate the cellular mechanisms underlying synaptic transmission between the central terminals of the trigeminal nerve (trigeminal afferents) and neurons located in the spinal trigeminal nucleus caudalis (Vc), a brainstem region implicated in nociceptive processing for the face. We hypothesize that synaptic connectivity of individual trigeminal afferents and neurons in the Vc is mediated by glutamate acting at non-NMDA and NMDA receptors and may be modulated by select ion channels, specifically the Transient Receptor Potential Vanilloid type 1 (TRPV1). We also believe that these functional connections may have distinct anatomical features. Both NMDA and non-NMDA receptors are observed in Vc, although their roles in mediating excitatory synaptic transmission in this region are ill-defined. TRPV1 is expressed on both peripheral and central termini of trigeminal neurons and upregulation of TRPV1 is closely associated with abnormal pain. Peripherally, TRPV1 has been characterized extensively as a molecular transducer of nociceptive information;however, the roles of centrally trafficked TRPV1 channels in pain processing remain unclear. Aim 1 will determine the functional connectivity of trigeminal afferents and Vc neurons with in vitro electrophysiological techniques assessing: synaptic latency variability as a measure of synaptic order, glutamate-mediated responses at NMDA and non-NMDA channels, and if TRPV1 modulates glutamatergic responses in Vc using brainstem synaptic transmission (e.g. NTS) as a model. Preliminary data demonstrate that individual trigeminal afferents can be identified using measures of synaptic latency variability, and chemical activation of central TRPV1 induces measurable responses. Aim 2, will determine the morphological features of Vc neurons related to synaptic latency variability and evaluate the anatomical connectivity between trigeminal afferents and Vc neurons with respect to TRPV1. The results will be integrated with findings from Aim 1 to provide a more specific understanding of the relationship between trigeminal afferent and Vc neuron subpopulations and the cellular mechanisms underlying synaptic transmission. These results will provide important, complementary anatomical and functional information about the neural networks that relay sensory information from the face to the brain, including the role of TRPV1 in the trigeminal pain pathway, and identify potential mechanisms for both neural plasticity and modulation of craniofacial pains. PUBLIC HEALTH RELEVANCE: According to the National Institute of Dental and Craniofacial Research's Panel on Pain Research 2003, 22% of the American population reports having orofacial pain, the most common type of craniofacial pain. Facial tissues are innervated by neurons which comprise the trigeminal nerve. These neurons branch both peripherally to the face and centrally to the brain creating a relay network for processing of nociceptive sensory information. The current proposal seeks to understand the cellular mechanisms driving neural integration of pain sensations transmitted from the face to the brain with the potential to identify therapeutic targets for modulation of craniofacial pain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Serine/threonine phosphatase 5 (PP5) is an enzyme that has an emerging role in the regulation of signal transduction and altered PP5 activity appears to contribute to tumor development and maintenance. While protein kinase enzymes catalyze the transfer of a phosphate group from a donor to an acceptor protein in various signaling pathways, protein phosphatases, in turn, reverse the action of kinases by removing a phosphate. PP5 belongs to the PPP-family of enzymes, which also includes PPI and PP2A. Both genetic studies and studies using inhibitors of protein phosphatases (e.g. okadaic acid) indicate protein phosphatases play an important role(s) in the regulation of cell cycle progression and processes implicated in tumor promotion. PP5 has been shown to play a negative regulatory role in a p53-mediated signaling cascade leading to the induction of a cyclin-dependent kinase inhibitor protein (p21WAFI/Cipl) and Gl/S-phase growth arrest. The expression of PP5 is also responsive to 17- beta estradiol and hypoxia inducible factor-l (HIF-l), which are both positive factors in the development of human breast cancer. In addition, the constitutive over expression of PP5 promotes cell survival during oxidative stress (an issue in cardiovascular disease and in developing tumors) and converts MCF-7 breast cancer cells from an estrogen-dependent into an estrogen-independent phenotype. Recently, increased PP5 expression has been shown by tumor microarray to have a positive correlation with breast cancer. The studies described in this proposal are designed to test the hypothesis that PP5 plays an important role in the regulation of cellular responses to hypoxic stress, and that changes in the normal biological functions of PP5 may contribute to the development of cancer. These studies will expose undergraduates to research on cancer in a form that is easy to understand (the on-off type roles of signaling pathways) and also expose them to tissue culture as well as basic cell/molecular biology techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "An exciting, new investigative theme in airway physiology and pathophysiology is neurotrophins (NTs): growth factors including brain-derived neurotrophic factor (BDNF) known for their diverse roles in the nervous system. NTs and their receptors have now been found in different lung components including airway smooth muscle (ASM), with altered expression observed in asthma, allergy, and even lung cancer. While NTs may be derived from several sources, our published and preliminary data suggest that ASM is a target of NTs, and that NTs contribute not only to ASM contractility under normal circumstances, but also to increased contractility with airway inflammation (such as that induced by TNFa). The long term goal of the proposed studies is to understand the role of NTs in ASM physiology and pathophysiology. The overall hypothesis is that NTs enhance a) sarcoplasmic reticulum (SR) Ca2+ release and Ca2+ influx; and b) Ca2+ sensitivity for force generation in ASM. We propose that BDNF is a key NT influencing ASM contractility. Finally, airway inflammation enhances BDNF signaling, leading to an overall enhancement of [Ca2+]i and force. In this proposal, we will use human ASM and the ovalbumin (OVA) mouse model to examine the relative role of the BDNF receptors (high affinity TrkB vs. low affinity p75NTR) vis-vis ASM contractility. We hypothesize that TrkB is more important for [Ca2+]i regulation, while p75NTR regulates force. Using biochemistry, pharmacology, molecular biology, immunocytochemistry, fluorescence Ca2+ imaging, force measurement techniques, and lung mechanics, we will focus on specific mechanisms that may be regulated by BDNF: the second messengers IP3 (via phospholipase C PLC) and cyclic ADP ribose (via CD38) (Aim 1); SR Ca2+ release (IP3 receptor vs. ryanodine receptor (RyR) channels) (Aim 2); Ca2+ influx via store-operated Ca2+ entry (SOCE) (Aim 3) and the force regulatory mechanisms myosin light chain (MLC20) and rhoA/rho-kinase (Aim 4). These in vitro studies in human ASM will be integrated into the OVA mouse model applied in focus studies to the TrkB knockin mouse (where TrkB functionality is reversibly inhibited). We will explore the idea that inflammation induced by TNFa increases constitutive BDNF receptor expression, and alters specific [Ca2+]i and force regulatory mechanisms, thus priming ASM for enhanced response to both BDNF and bronchoconstrictor. The Specific Aims are: Aim 1: To determine mechanisms by which BDNF modulates second messenger signaling in human ASM; Aim 2: To determine mechanisms by which BDNF modulates SR Ca2+ regulation in human ASM; Aim 3: To determine mechanisms by which BDNF modulates SOCE in human ASM; Aim 4: To determine mechanisms by which BDNF modulates force regulation in human ASM; Aim 5: To determine the role of BDNF in ASM contractility in a mouse model of airway inflammation and hyperresponsiveness. PUBLIC HEALTH RELEVANCE. There is increasing recognition that abnormalities in airway smooth muscle contractility (exacerbated by inflammation) contribute to exaggerated airway narrowing and accompanying shortness of breath in clinically important diseases such as asthma and chronic bronchitis. In this regard, the potential role of growth factors called neurotrophins in regulation of airway contractility is an exciting and emerging area of research. By establishing the role of neurotrophins in airway narrowing with or without inflammation, the proposed studies will the foundation for better understanding of airway diseases, and potential development of new therapeutic targets.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The control of programmed cell division is an area that relates to a wide range of biomedical problems including tissue regeneration, development, diffrentiation, and cancer. Surprisingly little is known about the molecular basis of cell cycle control. This proposed study will employ a lower eukaryote model, Physarum pollcephalum, to study the regulation of cell division at the genetic and molecular level. This model allows a degree of analysis not possible in animal cells. Mutations defective in mitotic cycle functions will be isolated among a set of temperature conditional lethal mutants blocking DNA replication. These will be enriched by a suicide selection strategy designed to kill growing cells with a combination of bromodeoxyuridine and methotrexate. A novel method for the use of mitotic cycle mutants to assay the functions they lack is proposed. Injection of extracts from non-mutant cells will be used to rescue the mutant defects. This functional assay will eventually lead to the purification of these factors, allowing their detailed analysis. We have shown that in Physarum tubulins are syntesized in preparation for mitosis. The basis of this regulation will be investigated. The triggering of tubulin synthesis depends on the completion of prior events in the mitotic cycle. The nature of the committment to tubulin syntesis will be investigated using mitotic cycle mutants, cell fusions, and heat shocks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of the proposed project is to characterize the transduction of Escherichia coli by the virulent coliphage T1 and to use (lambda-gal) T1 hybrids to shed light on the relationship of transcription to recombination. The following types of experiments are used to understand transduction by T1: (1) The effect of ultraviolet irradiation on T1 lysates. (2) The effect of T1 mutations on the ability of T1 to transduce. (3) The use of premature lysates to determine when transducing particles are formed. (4) The use of variant prophages of lambda to characterize gene pick-up by T1. The effect of lambda initiated transcription will be studied as follows: (1) The effect of deleting DNA between the galactose operon and the attachment site of lambda. (2) The effect of nonpolar kil mutants of lambda. (3) The effect of lambda mutants which result in the creation of a new promotor between genes N and int.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Clinical Core will be responsible for all human subject-related activity for the AADRC program. This Core has four main functions: (a) recruitment and characterization of human subjects; (b) performance of bronchoscopy, along with allergen or kinin challenge, bronchial biopsy, bronchial brushing, or bronchoalveolar lavage as required; (c) collection of blood or blood cells; and (d) microdissection of bronchial biopsies and lung samples. The study of animal models of asthma or in vitro model systems has resulted in significant progress towards elucidating the immunology and cellular and molecular biology of asthma. Despite these successes, any effort to understand clinical asthma (or how asthma is initiated and propagated) must ultimately be undertaken or validated by studying human subjects with asthma. These human studies, however, are subject to considerable difficulty, including problems recruiting appropriate subjects, the clinical heterogeneity of asthma and the difficulty in obtaining suitable clinical material for study. A highly specialized and independent Clinical Core has been designed for this AADRC program to remove the burden from investigators of the consuming tasks of human subject recruitment/characterization and bronchoscope/sample acquisition while simultaneously supplying all investigators with sufficient clinical samples to efficiently pursue the mechanistic studies that define each project. By utilizing a common core to recruit, screen and study volunteer subjects, all investigators participating in this AADRC program will have full access to the subject-derived samples, independent of whether the investigators have an appropriate clinical background. In addition, expertise of the Clinical Core and standardization in sample preparation will contribute to insuring the most consistent and highest quality samples possible. Performing bronchoscopy with lavage and biopsy in asthmatic subjects has revolutionized our understanding of the pathophysiology of asthma, leading to the recognition that airway inflammation is present in even mild asthma and pointing toward the role of airway remodeling in chronic asthma. Bronchoscopy procedures have not, however, yielded information that can be used to predict asthmatic phenotype. This program will link genetic, cellular, and biochemical findings obtained from patients? samples using state-of-the-art laboratory techniques to carefully documented asthma phenotypes, potentially allowing novel insights into the determinants of asthmatic phenotype to be gleaned.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Polysaccharides are used in food and pharmaceutical applications due to their inherent abilities such as gelation, viscosity and thickening behavior. As the macroscopic properties of these biopolymers are dictated mainly by their molecular geometry and atomic level interactions, accurate molecular details of biologically and industrially important polysaccharides are very much in need for gaining better insights about their structure-function relationships. As members of the Whistler Center for Carbohydrate Research, Purdue University, our aim further constitutes a major part of the multidisciplinary research activities of the Center. We have so far successfully deciphered the roles of cations and/or side groups in the junction zone formation of gellan related polysaccharides, galactomannan and iota-carrageenan, to name a few, towards understanding their functional properties. In this regard, our long term research goals are to determine polysaccharides architectures and their interactions with solvent and solute molecules as well as the synergistic interactions in mixed polysaccharide systems. In this ongoing study, we chose a number of biologically important and industrial useful polysaccharides, such as specimens from algal (iota-, kappa- and lambda-carrageenans);bacterial (gellan analog and cepacian);fungal (glucoronoxylomannan);plan (rhamnogalacturonan) and a binary system (bacterial acetan: plan glucomannan). More recently, we ventured in to the area of utilizing polysaccharide fibers as drug carriers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We hypothesize that alterations in growth, 4 compartment of body composition (BC), regional distribution of body fat (RDBF; via MRI, anthropometry), and bone mineralization (BM; via DEXA) in boys and girls are driven by alterations in GH and gonadal and adrenal steroid hormone levels, and are modified by energy and nutrient intake (nonconsecutive, 7 day, 24 hr diet recall), energy expenditure (EE; via doubly labeled water, indirect calorimetry, 7 day physical activity recall) and physical fitness (via V02 peak), which act on the genetic background. A better understanding of the factors that control the pubertal changes in BC, RDBF and BM can lay the ground work for the comprehension, prevention and treatment of various disease processes including non insulin-dependent diabetes mellitus (NIDDM), cardiovascular diseases (CVD) and osteopenia, whose antecedents likely occur during childhood and adolescence. However, valid and precise data for BC, RDBF, and BM and the factors controlling them are lacking for children and adolescents. Girls with the Turner Syndrome (n=8) receiving GH for at least one year, prior to estrogen replacement, and boys with constitutional delay of growth (n=8) not on androgen therapy will serve as models for adolescent growth in the presence of adequate GH, but physiologically deficient sex hormones. Girts with precocious puberty (n=8) on GnRH analogue therapy for at least one year provide a model for pubertal growth, with accompanying high levels of GH and gonadal steroids, at an inappropriately early age. Control data will be provided by a cohort of 44 normally growing children at all stages of puberty followed in a parallel protocol. With these clinical models of hormonal influence and normal controls we shall determine the hormonal and nutritional mechanisms that stimulate growth and induce the alterations in BC, RDBF and BM in boys and girls at puberty. This model will also provide insights into the role of hormonal and nutritional factors in progressive diseases (CVD, NIDDM, osteopenia). Both the boys and girls with clinical growth disorders and the controls will be entered as staggered, mixed longitudinal groups. Specific hypotheses to be tested include: 1) somatic growth and maturation will be directly related to GH, IGF-1 and gonadal and adrenal steroid levels; 2) % body fat (%BF) and visceral fat will be directly related to energy and fat intake and inversely related to EE and pre- and late-pubertal GH secretion; 3) gender differences in RDBF will be related to changes in gonadal steroid levels; 4) despite increases in GH secretion during puberty, % BF and visceral fat will increase due to the over-riding effects of gonadal steroids; 5) BM will be directly related to physical activity, calcium intake, gonadal steroids and GH secretion; and 6) EE, energy and nutrient intake, and physical fitness will modulate hormonal influences on BC, RDBF, and BM. Interactions to be tested include: 1) increased aerobic fitness will increase GH secretion and reduce %BF and visceral fat but, dietary fat intake will reduce GH secretion causing an increase in % BF and visceral fat; 2) physical activity will increase BM, but inadequate energy and calcium intake and/or excessive activity will reduce gonadal steroids and reduce BM.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Infection of alveolar macrophages by Mycobacterium tuberculosis (M tb) is a fundamental step in the progression to disease. Yet little is known about the response of macrophage to M tb, especially with respect to the impact on macrophage secretion of and response to cytokines that mediate interactions with other cells and play a key role in pathogenesis. To maximize relevance to the clinical situation and thus facilitate application of the data obtained in way that will benefit public health, studies will be carried out with clinical isolates of currently prevalent M tb strains, and in part with alveolar macrophages from normal human volunteers. Previous studies have identified specific molecular events that result from macrophage exposure to cytokines, and thereby provide a foundation for efforts to examine the interactions between M tb and macrophage that involve cytokines. The overall goal of the research proposed here is to identify the molecular mechanisms that account for M tb-induced alterations in macrophage response to key cytokines that are believed to contribute to the pathology of human tuberculosis. The specific aims will thus test the hypothesis that M tb affects macrophage response to cytokines by examining whether: 1. M tb infection of macrophage alters activation or induction of transcription factors by interferons (IFNs) and tumor necrosis factor alpha (TNFalpha); 2. M tb effects on the macrophage response to IFNs and TNFalpha impinge on regulation of gene expression; and 3. expression of downstream gene products implicated in the pathogenesis of M tb or the modulation of macrophage function reflects the effects of M tb on macrophage responses to IFNs and TNFalpha. The DNA-binding activity of specific transcription factors known to be involved in responses to IFNs and TNFalpha, including ISGF-2/IRF-1 and NFkappaB, among others will be determined with the electrophoretic mobility shift assay. Quantitating the expression of marker genes such as inducible nitric oxide synthase, intercellular adhesion molecule, and interleukin-8, among others, implicated in M tb pathogenesis and/or the function of activated macrophage will be done with the run-on assay of transcription rate, and northern blot, RNase protection, primer extension or RT-PCR experiments to determine steady- state mRNA levels. The amount of protein expressed from these and other genes will be determined by assays of enzymatic activity, and with immunochemical methods to detect the intracellular protein present in an extract, or membrane proteins present at the cell surface. Elucidating the point(s) at which macrophage responses to cytokines are altered by M tb infection, and determining the consequences for expression of genes involved in pathogenesis, may provide important information for potential therapeutic interventions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The major goal of the present study is to assess the role of hyper- tension on the development of atherosclerosis in two different arterial beds, the carotid artery and aorta, and to determine similarities or differences in the responses of these two beds to intervention with the calcium blocking agent, nifedipine. The hypothesis to be tested is that carotid artery atherosclerosis in hypertensive animals develops at a significantly greater rate and is less subject to regression compared to aortic atherosclerosis. White Carneau pigeons, aged 6-12 months and of either sex, from genetically selected high (n=91) and low (n=91) blood pressure lines will be studied. The birds in each line will be divided into four groups and will be fed either an atherogenic or control diet. Two groups of pigeons will also be fed nifedipine along with the atherogenic diet. Blood pressure, body weight, plasma cholesterol, blood calcium and plasma lipoprotein concentrations will be measured initially, and at 3 and 10 months during the study. After 10 months, subgroups of pigeons will be examined to assess atherosclerosis. The remaining pigeons will be removed from the atherogenic diet and divided into three subgroups. During any atherosclerosis regression phase, a control diet with or without nifedipine will be fed for an additional 10 months. The birds will be sacrificed and atherosclerosis evaluated by visual examination, total arterial cholesterol, calcium, collagen and elastin concentrations. Histologic sections will be examined to support the biochemical studies. The significance of the project lies in a better understanding of the role of hypertension in atherosclerosis development in aorta and carotid artery and the response of these two arterial beds to dietary intervention and the effect of the calcium channel blocker, nifedipine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cholesterol absorption plays a key role in cholesterol homeostasis and understanding the luminal events that play key roles in absorption remain poorly understood. The aims of the present study are fourfold: 1) To determine whether previously observed effects on cholesterol absorption during bile acid feeding are related to changes in pool size and intestinal transit or meal stimulated gall bladder emptying or plasma cholecystokinin levels. 2) To determine the effect of dietary sphingomyelin on cholesterol absorption, micellar solubilization and synthesis in normal adults and to assess the effects of intraluminal cholesterol solubilization, absorption and synthesis in adults with heterozygous mdr 3 deficiency (a defect leading to low biliary phospholipid content). 3) To determine the mechanism of action of a non-ionic detergent, Pluronic F- 68, by evaluating its effect on cholesterol solubilization and distribution between micelles and vesicles, on cholesterol absorption and synthesis. 4) To evaluate the intraluminal solubilization and distribution within micelles and vesicles of biliary compared to dietary cholesterol in humans and assess the impact of ezetimibe treatment on absorption of endogenous or exogenous cholesterol by assessing absorption of human contents in the biliary diverted, rat lymph fistula model. For each of these aims, subjects will be studied while consuming well-controlled diets as outpatients with a combination of human and animal techniques. Techniques employed for human studies will include state-of-the art techniques utilizing stable isotopes and isotope ratio mass spectrometry, gas chromatography. Translational studies in animals will be used including novel techniques to measure fat absorption as well as the use of the lymph fistula rat model for assessment of lipid absorption and hamsters for assessment of bile acid and sterol synthesis. Integration of animal/human techniques will provide tools to characterize the role of modifications of the intraluminal environment on cholesterol solubilization and human cholesterol absorption and synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are, currently, no accepted sensitive and specific biological markers of total dietary fat intake for use in epidemiologic studies. Self-report measures of dietary fat may be seriously biased. By examining a combination of fatty acids, found in low concentrations, reflecting various components of a low-fat diet, it may be possible to develop a biomarker of fat intake. This hypothesis was tested in a controlled, dietary intervention in 66 post-menopausal women, who were randomized to consume either 17% or 34% of calories (kcal) from fat for 7 weeks. All food was provided and blood/tissue samples were taken twice at baseline and end of study. There were highly significant (P is less than 0.001) increases in relative proportions of many erythrocyte (RBC) and/or plasma phospholipid (PL) fatty acids with the low-fat diet. A linear combination of RBC C18:1(n-7) and C18:2(n-6), for example, differentiated the two groups very well at end of study (q=0.92), correlated strongly with change in fat intake (r=O.83) and moderately with baseline fat intake(r=0.51). These findings are highly promising but before these markers become widely applicable in epidemiologic settings, there are a number of issues that need to resolved. They are addressed as specific aims in this proposal, namely: (l) To test if the changes in PL and RBC fatty acids seen with the adoption of a low-fat, high carbohydrate diet are affected by weight loss: (2) To determine the time period of dietary intake reflected in PL and RBC fatty acid concentrations; (3) To ascertain if the changes in fatty acid concentrations found with post- menopausal women are generalizable to premenopausal women and men; and (4) To investigate the cross-sectional associations between adipose tissue fatty acid concentrations, RBC and PL fatty acids, and self-report measures of dietary fat intake. These aims will be addressed in a controlled, randomized dietary intervention, where 50 men (ages 21-79) and 50 women (ages 21-44) will be randomized to consume one of three possible diets for six weeks - (i) 34% of Kcalories (kcal) from fat, isocaloric; (ii) 17% kcal from fat, isocaloric; or (iii) 17%, with 33% calorie deficit, i.e. hypocaloric. Blood samples will be taken at baseline. and weeks 2, 4, and 6 after commencement of diet. Adipose tissue aspirates will be collected at baseline. Body composition will be measured at baseline and end-of-study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In heterogametic species there is an imbalance in the number of sex-linked genes between males and females. Therefore, it is essential that transcription is equalized between the sexes and between the sex chromosomes and autosomes, a process called dosage compensation. Though different dosage compensation mechanisms have evolved, they all share a common initial step of distinguishing the X-chromosome from the autosomes. The model organism Drosophila melanogaster has evolved a single-step mechanism for dosage compensation, providing a simple model for studying how a single chromosome is discriminated from the rest of the genome. In Drosophila, selective two-fold upregulation of the single male X-chromosome equalizes transcription between the sexes. The selective identification of the X-chromosome involves the recruitment of a ribonucleoprotein complex, Male-Specific Lethal (MSL) that is expressed only in males. The MSL complex initially localizes to GA-rich sequences within seed sites on the X-chromosome, termed MSL Recognition Elements (MREs). However, MSL complex cannot bind directly to MREs and these sequences are distributed throughout the genome. The Larschan laboratory has recently identified a previously unstudied zinc finger protein, Chromatin-Linked Adapter for MSL Proteins (CLAMP), that links the MSL complex to MRE sequences. Furthermore, CLAMP is highly enriched on the X-chromosome independent of MSL complex and therefore is likely to be involved in the earliest step of X-identification. Therefore, the identification of CLAMP provides the first opportunity to define the X-chromosome features that promote its identification for dosage compensation. Based on strong preliminary data, I hypothesize that identification of the X-chromosome requires contributions from primary genome sequence and higher-order nuclear organization mediated by the CLAMP protein. I will test my hypothesis by defining the role of linear and three-dimensional genomic organization in enrichment of CLAMP on the X-chromosome. First, using Chromatin Immunoprecipitation- qPCR (ChIP-qPCR), I will determine whether enrichment of CLAMP on the X-chromosome occurs cooperatively or additively as the number of tandem MRE sequences increases. Second, I will determine if putative initial seed sites cluster together in three-dimensional nuclear space using the Chromatin Interaction Analysis by Paired-End Tag Sequencing (ChIA-pet) method. The proposed experiments will use state-of-the- art approaches to provide key insight into how initial identification of the X-chromosome is achieved.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Transgenic mice containing the HTLV-I tax gene develop tumors of peripheral nerve sheaths,iris, adrenal medulla and salivary gland, myopathy and localized lymph node hyperplasias. The mechanisms leading to these abnormalities remain unclear, but may involve the deregulation of cellular genes important for cell growth and differentiation. The expression of tax in these transgenic mice has been correlated with the induction of the genes for the interleukin-2 receptor (IL-2-R), the granulocyte-macrophage, colony-stimulating factor (GM-CSF), and more recently, nerve growth factor (NGF). Cell lines from the peripheral nerve and salivary tumors appear to secrete significant quantities of IL-6 and perhaps other B-cell-related growth factors, which may account for the lymphadenopathy associated with these tumors. This transgenic mouse system provides an important in vivo model for HTLV-I-induced transformation and tax activation of growth-regulating genes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application is submitted in response to RFA DK-06-504 to continue the efforts of the Inflammatory Bowel Disease Genetics Consortium and our role as a Genetic Research (GRC) with the central goal of identifying susceptibility genes contributing to the pathogenesis of inflammatory bowel disease (IBD). This will be accomplished through the following specific aims. Specific Aim 1: Expansion, Development and Management of Consortium Resources. Our GRC will recruit additional IBD cases, controls, and parents with a focus on ulcerative colitis and early onset cases. Extensive clinical data will be collected on recruited subjects and biospecimens (EBV-transformed lymphoblastoid cell lines, DNA, serum, whole blood) will be ascertained and stored at the NIDDK Genetics Repository for use by the Consortium, individual GRCs and outside investigators. Specific Aim 2: To employ a variety of approaches to identify genetic variation that contributes to IBD susceptibility. Our GRC will be involved in the follow-up of European ancestry genome-wide association study results. A particular initial focus of our GRC will be on a) integrating the suggestive linkage data and genome-wide association data in the IBD2 linkage region followed by additional association mapping within IBD2, and b) a resequencing, detailed haplotype and additional association study of the IL23R gene region. Specific Aim 3: To build a risk model of IBD through understanding genetic influence on variations in phenotypic expressivity, gene pathway analysis, and gene-gene (G x G) and gene-environmental (G x E) interactions. A particular initial focus of our GRC will be on gene-gene interaction analyses and pathway analyses of genes along the IL23R pathway. Our GRC may also take on other projects as genes/genomic regions advance to confirmed evidence for contribution to disease through follow-up of the Consortium's genome-wide association data. IBD is a chronic inflammatory disease of the gastrointestinal tract which primarily affects young people and is characterized by long-term illness and the need for potent medical therapy and substantial surgical therapy. The work of the IBD Genetics Consortium will enable us to identify important predisposing and disease modifying genes contributing to the pathogenesis of IBD which has the promise to: (1) identify persons at risk for disease, (2) predict disease course, (3) aid in selection of treatment, (4) understand pathophysiologic mechanisms such that novel preventive and therapeutic interventions can be developed. Advances in IBD gene identification and methodologic approaches may also be applicable to other complex genetic disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT Two of the major problems in attaining euglycemia in subjects with type 1 diabetes (T1D) are hypoglycemia (including fear of severe hypoglycemia) and post-meal hyperglycemia. The latter commonly occurs in adolescents secondary to missed meal and snack insulin boluses. Both of these issues will be addressed over the five years of this grant. As over half of severe hypoglycemic events occur during sleep, we will continue our development of algorithms to predict and prevent nocturnal hypoglycemia using a closed loop system (CLS) in which the information from a continuous glucose monitor (CGM) is used to suspend delivery of insulin by an insulin pump. Initial studies will be done in the Clinical Translational Research Centers (CTRCs) to document safety of the Florence CLS (Abbott Diabetes Care) and we will then conduct a randomized clinical trial in the home environment in years 2 and 3 of the grant. Similarly, a fully portable CLS will be used to detect and correct hyperglycemia resulting from missed food boluses. The safety and efficacy of algorithms which utilize the signal form a CGM and provide output to an insulin pump will initially be studied in the CTRC's in year 3 of the grant. In years 4 and 5 of this grant we plan to assess the combined algorithms for prevention of hypoglycemia and hyperglycemia in a fully ambulatory system developed by Abbott Diabetes Care. We believe that prevention of hypoglycemia, particularly at night, and reduction of post prandial hyperglycemia will result in improvement in glycemic control and reduction in HbA1c levels.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary The overall objective of the Administration (Admin) Core efforts is to provide a robust and dynamic infrastructure for the Center of Excellence for Encephalitic Alphavirus Therapeutics that enables consistent and notable progress through the milestones. Led by the Program Director, Dr. Jonsson (UTHSC, University of Tennessee Health Science Center), the Core will be the main interface between the Scientific Advisory Committee (SAC), the Research Project PIs, and NIAID. To support the achievement of the aims and milestones of the Center, Dr. Jonsson will be supported by Jayne McKinnie (Admin Core Associate) to: (1) ensure coordination and dissemination of information across the research projects, (2) coordinate results required for filing intellectual property, (3) publish results of disclosed leads and provide novel resources to the scientific community, (4) coordinate input into the refinement of the product development plan outlined in the Overall section with our University partners and SAC, (4) attract commercial partners for future SBIR or similar proposal efforts, and (5) administratively support the Center in collaboration with our regulatory consultants to prepare for our meetings with the FDA at the end of year 3 and at the start of year 5. Specific aims of the Admin Core will be to: Aim 1. Organize, manage, and document Center communications and findings; Aim 2. Manage and ensure fiscal responsibility and regulatory compliance; and Aim 3. Manage lead prioritization and documentation, the refinement of the Product Development Plan and a path to Commercialization.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] While much progress has been made to control preventable infectious diseases, infections remain a major cause of morbidity and mortality. Many of the traditional treatments for common infections are no longer effective because of the fast-growing problem of antimicrobial resistance, first associated with hospitals but increasingly widespread in the community. Antibiotic resistance is now a global problem of major concern. The ultimate goal of this proposed Center for Interdisciplinary Research on Antimicrobial Resistance (CIRAR) is to develop strategies to implement and evaluate a long term collaborative program of interdisciplinary research on reducing antimicrobial resistance. The specific aims of the CIRAR are: (1) To plan, implement, and evaluate outcomes of a core program and curriculum to prepare medical researchers to conduct interdisciplinary research; (2) To develop the expertise of CIRAR investigators in interdisciplinary research; (3) To conduct and evaluate outcomes of interdisciplinary demonstration projects related to the prevention and control of antimicrobial resistance; (4) To implement dissemination and implementation strategies that maximize the sustainability and expansion of CIRAR's interdisciplinary research; and (5) To formalize plans for a sustainable Interdisciplinary Research Consortium for Antimicrobial Resistance. The first component of the Center is the development and implementation of formal training for conducting interdisciplinary research which will be incorporated into several ongoing programs within the institution and made available by internet. In Years 2 and 3 the interdisciplinary research curriculum will be further disseminated. The second CIRAR component is the development of interdisciplinary demonstration projects designed to rationalize antimicrobial use and reduce antimicrobial resistance. In the final year, plans for a sustainable Consortium on antimicrobial resistance will be formalized. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION:(provided by Applicant) Antibody Production by B cells employs 3 separate types of genetic alterations. The variable region exon of immunoglobulin (Ig) heavy (IgH) and light (IgL) chain genes is assembled from germline V, D, and J segments during early B cell development. V(D)J recombination is initiated by the RAG-1/2 endonuclease, which introduces DNA double strand breaks (DSBs) at participating V, D, and J segments, and is completed by non-homologous end-joining (NHEJ) proteins that seal the RAG-generated DSBs. In response to antigen, mature B cells change the IgH constant region (CH) expressed with a particular variable region via a distinct recombination process termed class switch recombination (CSR). At this stage, the variable region coding sequence may be further altered by somatic hypermutation (SM). While the nature of the enzymes that initiate CSR or SM remains elusive, both processes, like V(D)J recombination, may involve DSBs and thus, employ NHEJ. We have shown that defective NHEJ during attempted V(D)J recombination in a p53 tumor-suppressor deficient background reproducibly leads to murine pro-B cell lymphomas that harbor translocations between the IgH and c-myc loci. We now propose to elucidate further the role of NHEJ proteins in CSR and SM, with a particular focus on how defects in these processes, and in V(D)J recombination, may lead to the generation of chromosomal translocations that contribute to the generation and maintenance of mature B cell lymphomas. To focus potential oncogenic mutations on mature, as opposed to progenitor B cells, we will employ several strategies, including conditional gene targeted mutation to inactivate NHEJ or activate specific oncogenes in mature B cells. Also, we will insert V(D)J recombination signal sequences, CSR target sequences (S regions), expressed variable region gene sequences, or meganuclease restriction enzyme (I-Sce 1) target sequences adjacent to the c-myc, bcl-2 or bcl-6 genes within various NHEJ or checkpoint-deficient ES cells to test for ability to stimulate translocations in appropriate B lineage cell populations. To eliminate large-scale breedings and facilitate rapid assays, we will introduce certain compound mutations into ES cells and assay effects on translocations and lymphomagenesis via Rag-2 Deficient Blastocyst Complementation. Once desired mature B cell lymphoma models are developed, we will employ gene-targeted mutations to determine the role of IgH expression, IgH enhancers, and particular oncogenes in the generation and maintenance of the transformed phenotype.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The research outlined in this proposal is designed to provide a basic understanding of the role of persisting antigen in the regulation and maintenance of the immune response. In view of difficulties involved in analyzing interactions between persisting antigen, antibody and the various cell types involved in the immune response in vivo, in vitro model systems have been developed which are amenable to manipulation and yet accurately reproduce phenomena observed in vivo. The aggregated human serum albumin (HSA) antigen used in this system appears to persist in rabbits for at least a year, and in the absence of anti-HSA antibody, it is capable of inducing a secondary response. The data obtained to date suggest that circulating antibody levels are maintained and regulated in vivo by an antibody feedback system. The feedback system prevents persisting antigen from inducing antibody synthesis when circulating levels are high. However, when antibody levels fall below a critical level, the antigen is exposed and stimulates HSA reactive memory cells to synthesize antibody and to produce additional memory cells. The data to be obtained from the proposed research should provide a basis for critically evaluating this mechanism for maintaining circulating antibody levels and immunological memory. Persisting antigen appears to be retained on follicular dendritic cells in the draining lymph nodes of both mice and rabbits. We plan to compare the ability of mouse dendritic cells with typical mouse macrophages in their ability to stimulate B-cells and cooperate with mouse T-cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A model system has been developed to study retroviral oncogenesis, and the genetic factors regulating susceptibility to tumor induction. Avian leukosis virus (ALV) induces bursal lymphoma in chickens, after integration of viral long terminal repeat (LTR) sequences next to the c-myc proto- oncogene. Labile or short-lived proteins appear to regulate c-myc hyperexpression, because LTR-enhanced transcription is specifically decreased after inhibition of protein synthesis. This lability is observed only in pre-B cell types, suggesting that it may be important in the B cell specificity of ALV tumor induction. Moreover, lymphoma-resistant chicken strains exhibit stable LTR-enhanced transcription (unaffected by inhibition of protein synthesis), suggesting that labile regulation of LTR-enhanced c- myc transcription may be important in lymphoma susceptibility. Five LTR binding proteins from bursal lymphoma cells have been characterized using the gel shift assay. Three proteins are specifically labile in pre-B cells (decreased binding after protein synthesis inhibition) and thus may be essential for LTR enhancement. An in vivo footprinting technique will be used to determine if labile protein binding also occurs in vivo. The contribution of each protein binding sequence to LTR enhancement will be analyzed by transfection of wild type or mutant LTR-reporter gene constructs into bursal lymphoma cells. The genes encoding the major labile proteins are being cloned and sequenced, to obtain DNA and antibody probes for analysis of the role of these proteins in lymphomagenesis. Labile protein expression will be compared in different cell types, to determine why it is specifically labile in pre-B cells. Western and Northern blot analyses will determine if this lability is regulated by changes in protein half life or by post- translational modifications such as phosphorylation. Binding protein expression will be analyzed during B cell development, by Western blotting and immunofluorescent staining, to determine if the proteins are labile in ALV target pre-B cells, and stable in mature B cells no longer susceptible to lymphoma. Protein expression will be compared in lymphoma-susceptible and -resistant strains, to determine how stable expression could be involved in resistance to lymphoma. These studies will give insight to the regulation of c-myc hyperexpression during tumor induction, and the role of the labile LTR binding proteins in tumor susceptibility. An understanding of the mechanism of ALV lymphoma susceptibility can be applied to studies of oncogene activation and lymphomagenesis in humans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Microbial calcification has proven to be a useful model for studying biologic mineral formation. The mineral species formed by Bacterionema matruchotii is crystallographically indistinguishable from biologic apatite of bones and teeth. The calcification nucleator of B. matruchotii has been identified as proteolipid: basic, hydrophobic protein complexed with acidic phospholipid. Calcifiable proteolipid has been isolated from marmoset bone and human dental calculus. A within-strain comparison of B. matruchotii and a variant exhibiting impaired calcifiability has provided insight into cellular regulation of calcification. A record of the process of mineral formation in the organism provides a base line for further studies. The purpose of the proposed research is to define the mechanism of intracellular apatite deposition in B. matruchotii through continued characterization of the nucleating entity, metabolic studies and examination of the process of apatite formation. Cellular regulation of calcification will be investigated by examining the interaction of calcifiable proteolipid with cell membrane components and through comparative studies with other mircoorganisms. Methodology developed for the microbial system can then be applied to vertebrate mineral formation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "One in two hundred children is born with limb deformities. An under- standing of the teleology of these defects, whether induced by taratogens, caused from a stochastic dysfunction of the developmental program, or the result of congenital defects, will require an under- standing of the normal process of limb development. Many of the genes that are suspected of mediating the formation of the limbs are now being identified. Particularly prominent among these genes is the set of Hox genes that may be required to specify the positional information needed for the establishment of the correct set of limb cartilages and bones. Gene targeting in mouse embryo-derived stem cells provides the means for generating mice with disruptions in each of these genes. The goal of this grant is to use gene targeting to undertake a systematic analysis of this gene complex and thereby directly establish the role of each of these genes in limb development. Not only will the phenotypic consequences of disrupting any particular gene be determined, but also through epistasis and molecular genetic analysis, it is hoped that insight will be gained as to how this set of genes functions as a network to determine the overall three-dimensional structure of the limb.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposal is to study the association, if any, that exists between oral contraceptive usage and increased risk of stroke among married women of childbearing age in a developing country. The study will be undertaken with the epidemiological case-control approach. Taiwan is selected as the study site because of her excellent household registration and vital statistics system and medical care services. The association between the use of hormonal contraceptives and thromboembolic diseases of the heart, lung and brain has now been fairly well established on a number of epidemiological studies conducted mainly in the United Kingdom and United States. Similar studies to confirm, or deny, such association is much needed in the developing countries where the genetic background, life style, and dietary habits of people are different. The family planning policy implication of the proposed study is self-evident. In the coming (second) year, approximately 200 stroke patients, 400 hospital controls, and 400 matched healthy neighborhood controls will be additionally interviewed to bring up the total sample size for the entire study to 300, 600, and 600 respectively in these categories.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a proposal to investigate the function and regulation of 20-hydroxyeicosatetraenoic acid (20- HETE) synthesis in the vasculature, more specifically, its participation in endothelium dysfunction in models of increased vascular expression of cytochrome P450 (CYP) 4A. 20-HETE is a primary eicosanoid in the microcirculation where it participates in the regulation of vascular tone. In rat renal arteries, CYP4A expression and 20-HETE production increased with decreased arterial diameter. CYP4A overexpression in small arteries and arterioles increased vascular reactivity and myogenic tone. Recent studies and preliminary results suggest that the endothelium is a target for 20-HETE bioactions. Smooth muscle-specific CYP4A1 expression via Ad-SM22-4A1 induces a marked CYP4A-dependent and 20-HETE-mediated endothelial sprouting in renal arterial microvessels. In vitro, 20-HETE is a potent angiogenic factor stimulating capillary-like tube formation of endothelial cells by a mechanism that may include MAPK activation and induction of inflammatory and angiogenic proteins (IL-8 and VEGF). In vivo, intravenous injection of Adv-CYP4A2 causes hypertension and renal arteries from these rats display endothelial dysfunction, which can be reversed by inhibition of CYP4A activity. Arteries from Adv-CYP4A2-transduced rats produce more 20-HETE and less NO;they also express higher levels of inflammatory proteins (ICAM and VCAM). These findings raise the possibility that vascular 20-HETE is an important determinant of endothelial dysfunction, a condition that is characterized by decreased NO bioavailability and enhanced endothelial activation, and are the basis for the proposal's hypothesis: Vascular overexpression of CYP4A fosters prohypertensive mechanisms via increased production of 20-HETE in a manner that may include endothelial dysfunction and activation. This hypothesis will be tested by 1) determining the relationship between hypertension, endothelial dysfunction and activation, CYP4A expression and 20-HETE synthesis;2) determining whether the functional consequences of increased vascular expression of CYP4A is associated with endothelial expression and synthesis of CYP4A and 20-HETE, respectively;and 3) exploring mechanisms underlying 20-HETE mediated endothelial dysfunction and activation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We are better defining the natural history of Ataxia-Telangiectasia (A-T). 1) We have developed a clinical scoring system and applied that system to a cross-sectional cohort of A-T patients. Among patients less than 18 years of age there is a clear correlation between age and cumulative A-T score. 2) We have noted that there is a lengthy delay between agerage age at which neurologic systoms appear and average age of diagnosis. We have analyzed the reasons for the delay and its consequences. 3) We are surveying all patients for the presence of opportunistic infections and complications of immunization. 4) We have noted significant nutritional deficiencies in a sub-group of patients and are looking for correlations with neurologic dysfunction. 5) We have noted little or no correlation between severity of pulmonary disease and presence of immunodeficiency. Instead, there appears to be correlation between pulmonary disease and problems with swallowing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "SUMMARY Inhibition of intestinal bile acid absorption has been shown to reduce plasma cholesterol. Although, bile acid sequestrants are clinically shown to lower cholesterol, the development of better therapeutic modalities to directly inhibit bile acid absorption is necessary. Apical Sodium Dependent Bile Acid Transporter (ASBT) absorbs majority of the bile acids in the ileum and is essential for maintaining circulating bile acid pool. Therefore, ASBT inhibition represents an attractive therapeutic target for lowering plasma cholesterol. In this regard, recent studies from our laboratory demonstrated that ASBT is inhibited by signaling intermediates including protein tyrosine phosphatases (PTPases) via membrane recycling events. Also beneficial dietary components such as green tea catechin, (-)-epigallocatechin-3-gallate, EGCG, inhibits ASBT function in a lipid-raft dependent manner. We hypothesize that phosphorylation/dephosphorylation processes, lipid raft-dependent mechanisms and membrane trafficking events play critical roles in the inhibition of ASBT function. Hence, a comprehensive understanding of these inhibitory pathways is crucial to exploit their utilization as an effective therapy for hypercholesterolemia associated with diabetes mellitus. Our preliminary studies showed that ASBT function and expression are upregulated in rat model of streptozotocin (STZ)- induced diabetes mellitus. This in vivo diabetes model will provide an exceptional tool to investigate the underlying mechanisms of ASBT upregulation in diabetes mellitus as well as determining the impact of ASBT inhibition on associated hypercholesterolemia. Our studies are designed to systematically delineate the cellular and molecular mechanisms inhibiting ASBT utilizing in vitro models and examine their dysregulation in vivo utilizing diabetes mellitus rat model. In Specific Aim 1, we will investigate the regulation of ASBT function and phosphorylation by protein phosphatases (PPase) in cell culture models. In Specific Aim 2, our studies will focus on elucidating the inhibitory mechanisms of ASBT function by membrane recycling events and lipid rafts as well as delineating the molecular basis for EGCG-mediated inhibition. Studies designed for Specific Aim 3 will focus on investigating mechanisms underlying ASBT upregulation in rat model of STZ-induced diabetes mellitus and determine the efficacy of the beneficial dietary compound EGCG and specific ASBT inhibitors (developed by Biotechnology Company Albireo) in lowering the levels of plasma cholesterol. Our proposed studies are critical for providing novel insights into the regulation of ASBT under normal and pathophysiological conditions and may provide better strategies for the management of hypercholesterolemia associated with several disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Emory Consortium for Innovative AIDS Research in Nonhuman Primates aims to understand the B and T Cell Biology of Protection from and Eradication of SIV/SHIV Infection. The consortium brings together an interdisciplinary mix of highly collaborative, and productive investigators in a range of HIV vaccine and cure disciplines to address the overarching hypothesis that a successful prophylactic HIV vaccine will require a strong and sustained systemic and mucosal immune response, comprised of functionally targeted antibody and tissue resident CD8 T cell immunity, in concert with a modulated HIV-specific CD4 T cell response that maintains low numbers of HIV targets in mucosal tissue, while providing adequate help for a strong adaptive response. Moreover, we postulate that such a potent and balanced vaccine response will, in the context of active latency reversing agents, reduce viral reservoirs and thus maintain suppression of virus replication following cessation of highly active antiretroviral therapy. The approaches in FOCUS 1, aimed at understanding the mechanisms of vaccine protection, will utilize state of the art adjuvants coupled with native trimeric Env immunogens to induce and mechanistically dissect strong durable humoral responses. In addition, we aim to fully characterize and harness a novel population of tissue resident CD8 T cells to effectively synergize with the humoral immune response in providing protection from SHIV challenge. Because recent data suggest that SIV/HIV specific mucosal CD4 T cells could enhance infection, we will also explore the potential for modulating the susceptibility of these cells to enhance protection. In FOCUS 2, aimed at defining mechanisms of reservoir eradication, we will utilize novel latency reversing agents and immunostimulants to define an optimal viral reactivation program, and then will combine these with the optimized vaccine approaches defined in FOCUS 1 to explore the potential of this combination to yield a sustained suppression of virus replication following withdrawal of highly active antiretroviral therapy. These experimental approaches will be supported by an effective Operations and Management Support component and five state of the art Scientific Research Support Cores in order to fully characterize the magnitude, function, specificity and repertoire of the humoral response. Single cell analytics and transcriptomics will also support characterization of innate and adaptive signals at the cellular level.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Newborn and juvenile eyes have an emmetropization mechanism that uses visual signals to precisely adjust the axial length of the eyes to the optical power, so that images are in sharp focus. However, nearly 30% of the U.S. population develops axial myopia, in which the eyes are elongated relative to the focal plane. Although glasses, contact lenses and refractive surgery can provide an optical correction, the increased axial length of the eye significantly raises the risk of retinal detachment, choroidal neovascularization and glaucoma, making myopia the 7th leading cause of blindness in the U.S. The emmetropization mechanism has at least three main components: 1) the retina, which responds to the sign and amount of defocus (or other visual cues); 2) a signaling cascade that originates in the retina, travels through the retinal pigment epithelium (RPE) and choroid, and reaches the sclera (the fibrous outer coat of the eye); and 3) fibroblasts in the sclera which respond to the signaling cascade by regulating the axial length. Our working hypothesis is that retinal responses to visual stimuli control the remodeling of the scleral extracellular matrix and, in particular, the slippage of the layers (lamellae) of the sclera across each other. This, in turn, controls scleral extensibility, axial elongation, and refractive state. The proposed studies will examine the retinal responses to visual stimuli and the scleral responses to the signaling cascade. In the previous project period, we identified two novel dissociation paradigms in which similar visual stimuli produce different scleral responses due to the prior history of the eye (somewhat longer or shorter than normal, an eye-size factor). Specific Aims 1 and 2 will use these dissociative paradigms to test our hypothesis that the source of this variability is localized in the patterns of gene expression in the sclera, rather than in the retina, and to identify the specific scleral signaling pathways that are involved. Specific Aim 3 will use an association paradigm in which three different methods (darkness, form deprivation, and minus-lens wear) will be used to induce myopia to identify the pattern of retinal gene expression that is common to all three and contributes to the retina go response. The proposed studies will apply proteomic tools and our Pathway-focused PCR Assay that have been developed in this laboratory to assess retinal mRNA and protein-expression patterns that characterize retinal go and retina stop responses, and scleral mRNA and protein-expression patterns that characterize the sclera go, sclera stop and sclera ignores responses in our well-characterized animal model, tree shrew. These animals are closely related to primates and, like humans, have an all-fibrous sclera. These studies will enhance our understanding the emmetropization mechanism at the level of gene expression and may lead to successful optical or pharmacological interventions to slow or prevent myopia. PUBLIC HEALTH RELEVANCE Myopia (nearsightedness) affects over 25% of the U.S. population and perhaps one billion people worldwide. In addition to the cost (over $14 billion annually) of eye exams, glasses, contact lenses, and refractive surgery, even low amounts of myopia raise the risk of developing blinding conditions. This application will identify genes and gene products that control the axial length of the eye as a step toward finding ways to prevent or minimize myopia in children.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objective of this proposal is to determine the underlying pathophysiology that leads to remarkably divergent cellular events in alpha vs. Beta thalassemia. Accelerated apoptosis or programmed cell death (PCD) is most likely responsible for intramedullary hemolysis in both human and murine beta thalassemia. Integrally associated with PCD, is the very specific alteration in the asymmetry of the phospholipid bilayer with an outward movement of phosphatidylserine (PS). The mechanism by which PS is exposed and recognized on thalassemic red cells or red cell precursors, rendering them ~edible~ to macrophages, is not clear. The principal objectives of this proposal are to define the contribution of enhanced PCD to ineffective erythropoiesis in beta thalassemia and further our understanding of the mechanism(s) responsible for enhanced PCD. In particular, we will discover how and why PCD leads to alterations in the phospholipid bilayer, resulting in PS exposure at the surface of affected cells, and how this in turn acts either alone or in concert with other signaling systems to lead to the recognition and removal of the affected erythroid precursors. To pursue these goals, we propose the following three specific aims: 1. Investigate programmed cell death in murine thalassemia, 2. Investigate the mechanisms responsible for the movement of PS from the inner to outer monolayer and 3. Determine the factors that render a cell with PS on its outer surface recognizable and removable. The murine alpha and beta thalassemic models we have created provide us with mice with graded degrees of clinical severity necessary to test the proposed hypotheses. We will use fluorescently labeled annexin V to quantitate, isolate and study the biochemical and functional characteristics of subpopulations of cells with PS on their surface, define the PCD driven mechanism for PS exposure, and identify factors modulating macrophagic recognition. We will explore our hypothesis that the heme/hemichromes associated with the globin chains mediate oxidant attack and play a role in PCD and macrophagic attack on apoptotic erythroid precursors. The successful accomplishment of these objectives should enable us to develop a detailed mechanistic understanding of the pathophysiology of an import human disease. The insights generated are also likely to further our understanding of the loss of PS asymmetry and the role of oxidative damage in programmed cell death.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of newborn screening is to detect potentially fatal or disabling conditions in newborns, thereby providing a window of opportunity for early treatment, often while the child is still asymptomatic. Such early detection and treatment can have a profound impact on the clinical severity of the condition in the affected child. If left undiagnosed and untreated, the consequences of the targeted disorders can be dire, many causing irreversible neurological damage, intellectual, developmental and physical disabilities, and even death. In 2006, the American College of Medical Genetics (ACMG) developed newborn screening guidelines that recommend that all newborn infants be screened for 31 core conditions and that 26 secondary conditions identified during the core evaluations be reported. These recommendations have been accepted by the HHS Secretary's Advisory Committee on Heritable Disorders in Newborns and Children (ACHDNC) (authorized by the Children's Health Act of 2000), and by the Secretary of HHS. Most states now use this or very similar panels for newborn screening. Currently, there are thousands of rare disorders that have been identified and hundreds that could potentially benefit from newborn screening. MPSI is a rare, inherited disease of metabolism in which an individual lacks an enzyme called lysosomal alpha-L-iduronidase. Without the specific enzyme individuals cannot break down long chains of sugar molecules called glycosaminoglycans. The glycosaminglycans build up and can damage organs, including the heart. The severe form of MPS1 is known as Hurler syndrome in which death occurs in early childhood. Attenuated forms (Huler/Scheie and Scheie syndromes) have a later onset in which death occurs either in the teens or twenties. Hurler syndrome is the predominant form of the disease (75-80% of patients).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Anti-neutrophil cytoplasmic autoantibodies (ANCA) were first described in 1982. We reported the association of ANCA with necrotizing glomerulonephritis (GN) in 1988 and recognized myeloperoxidase (MPO)-ANCA. Since then, substantial insight into a number of issues has been gained including: 1) separation of ANCA antigens into MPO-ANCA or proteinase 3 (PR3)-ANCA; 2) clinical and pathological associations of ANCA with necrotizing GN and small vessel vasculitis (SVV); and 3) that ANCA participate in the pathogenesis of vascular inflammation. Despite these strides by the ANCA community little is known about the derivation of the human ANCA autoimmune response or the precise antigenic epitopes important in eliciting an autoimmune response during disease onset or during disease relapse. This project has three specific aims. The first aim studies the human PR3 and MPO-ANCA immunoglobulin gene expression. We hypothesize that MPO-ANCA and PR3-ANCA expression is dependent on specific light and heavy chain usage and that both MPO and PR3 are selecting antigens in the maturation of this autoimmune response. We developed a technique to amplify message from single B cells, allowing the study of somatic mutations of the amplified immunoglobulin genes. In the second aim, we will study specific epitopes on MPO and PR3 during disease onset and during relapse of ANCA-GN. Is the fine specificity of epitope usage during disease onset and relapse the same? We have clone fragments of MPO and PR3 into expression vectors to perform these studies. The third aim is the most exciting and may provide a new general understanding of autoimmunity and of the ANCA immune response in particular. It is known that peptides translated from the anti-sense strand of DNA bind to sense proteins with substantial selectivity and affinity. This concept is formulated as the molecular recognition theory. We have developed exciting preliminary data to support the novel concept that ANCA react not only with PR3, but also with complementary peptides to PR3. Is the immune response to the complementary peptide part of the autoimmune response? Are PR3-ANCA and antibodies to complementary peptides examples of an idiotypic anti-idiotypic relationship? We will have all the reagents necessary to explore this paradigm. The project will dissect human ANCA and ANCA antigens during disease onset and during disease relapse. We will ascertain whether the molecular recognition theory pertains to the human ANCA immune response in that ANCA may react, not only to PR3 and MPO, but to peptides complementary to these ANCA antigens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The principal aim of this project is to elucidate the molecular mechanism of active transport in bacterial organisms. In the simple fermentative organism, Streptococcus faecalis it is known that cellular accumulation of K ion and certain amino acids is coupled to ATP hydrolysis by an ATPase associated with the plasma membrane. In order to understand the coupling mechanism we are attempting to characterize the protein subunits in the ATPase and in the membrane sector with which the enzyme is associated. More particularly we are trying to isolate the protein components that are part of the \"proton-channel\" envisaged by the chemiosmotic theory. For this purpose we will use an isolated ATPase-membrane sector complex which is sensitive to the energy transfer inhibitor, dicyclohexylcarbodiimide. We are also trying to characterize a small peptide segment in the ATPase molecule which is involved in the attachment of the enzyme to the plasma membrane. This segment is selectively cleaved from the alpha chains by limited treatment of the isolated ATPase with chymotrypsin.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PLANNING AND EVALUATION (P&E) CORE The purpose of the Planning and Evaluation Core (P&E Core) is to monitor the progress toward the implementation of goals and objectives of the Partnership. Planning and evaluation are the ultimate responsibilities of the Pis, and Drs. Jaime Matta (PSM) and W. Jack Pledger (MCC) will lead this core and will be responsible for monitoring progress through ongoing evaluation processes, maximizing resources, identifying novel directions for the Partnership, and reporting progress to institutional leaders and the NCl. The P&E Core includes the Internal Advisory Committee (lAC, P&E Table 1) and the Program Steering Committee (PSC, P&E .Table 4). The lAC is responsible for internal planning and evaluation (P&E Core section 1) and new project selection (P&E Core section 2). The PSC provides external review and evaluation (P&E Core section 3). The decisions made by this core will be guided by data systematically accumulated and organized by the Evaluation Process Subcommittee (EPS) (membership below). This subcommittee is a unique strength of this core as it is led by an evaluation and process expert, Dr. Susan McMillan. Dr. McMillan is the Thompson Professor of Oncology Nursing at the University of South Florida, has training in evaluation and measurement, and has served as an evaluation consultant for several federal grants over the past decades. She is external to both PSM and MCC and as such will be able to objectively direct this subcommittee to produce formal systematic evaluations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The processes within the cochlea which enable it to achieve its high sensitivity and sharp frequency selectivity are still largely unknown. The primary goal of the research proposed here is to improve our understanding of cochlear function by developing a realistic mathematical model of cochlear mechanics. This model development will involve the formulation of hypotheses concerning cochlear function which will be tested by comparing numerical solutions of the model with experimental observations published by other researchers. Among the major objectives are: 1) an investigation of the possible role that active elements within the cochlea may play in boosting the sensitivity and frequency selectivity and frequency selectivity of basilar membrane displacements; and 2) a description of the changes in model parameters that correlate with common hearing impairments or normal developmental gradients. A secondary goal of the proposed research is the development of computer algorithms which simulate the frequency analysis of the cochlea and are fast enough to be considered for use in applications such as speech spectrograms or cochlear implant prostheses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objective of this proposed research is to develop a better encapsulant which not only protects lead-pigmented paint, but also detoxifies the lead. This liquid component is intended to combat lead poisoning, the leading environmental childhood disease and provide a safer workplace for lead abatement workers. This would be the first lead abatement product that actually detoxifies lead in case of ingestion and is a safe water base formula. The proposed liquid may be used in several commercial products such as: 1) coatings which would be applied as an interim step before stripping or repairs to protect workers during removal, 2) washing solutions, 3) encapsulants, 4) treatments of lead waste and 5) fogging solutions. Preliminary testing indicates that this liquid formula detoxifies lead by 50%. Phase I research is needed to improve the detoxification levels of the initial formula, analyze the feasibility of incorporating calcium compounds into a final encapsulant product and confirm preliminary results via independent products. Phase II research is necessary to confirm long term effectiveness, simulate actual extreme environmental conditions and formulate optimal mechanical characteristics to meet proposed ASTM methods for encapsulant products. PROPOSED COMMERCIAL APPLICATION: ACT-LAC has shown the potential to detoxify lead pigmented paint and its water base formula is non- -toxic. The ACT-LAC formula may be incorporated into A) a lead safe barrier B) encapsulant C) post washing solution D) lead waste treatment and E) fogging solutions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Research on medical decision-making makes use of insights from economics and statistics to build quantitative models of clinical decisions. These models rely on assumptions about the use of information by decision-makers, including the core assumption that people process information in a narrowly logical manner. Recently, the psychology of decision-makers, including their emotional states, is being incorporated into these models to augment the role of cognition. Research on older adults differs importantly from other populations in a number of ways that may impact medical decision-making, especially the role of emotions. Pursuing work begun with his dissertation, William Dale, MD, PhD, will enhance his interdisciplinary training in geriatrics and public policy by focusing on a deeper understanding of emotions in medical decision-making in older adults. This research agenda initially utilizes the clinical model of prostate cancer, a widely prevalent disease in older men, and one in which the role of the emotion of anxiety may influence decision-making. Although no consensus exists on the \"right\" clinical pathway for detecting and treating prostate cancer, there is wide agreement that patient preferences are vital for making choices. Initial investigations indicate the inclusion of anxiety in utility assessments for men with localized prostate cancer dramatically affect the cost-effectiveness ratios for treatments. Further research to assess the contribution of anxiety to utility assessments for these men is on-going. This study serves as a specific core example of the overall career goal of understanding the role of emotions in medical decision-making in older adults. Older adults care occurs in a complicated socio-medico-psychological context that demands sensitivity to the importance of issues such as functional impairments, multiple co-morbidities, polypharmacy, and care-giver burden. As a geriatrician, Dr. Dale is sensitive of including these concerns in pursuing his research. Enhancing his broad-based training in public policy studies and geriatrics with a targeted, in-depth understanding of psychology and medical decision-making under the expert guidance of his mentors, Dr. Dale will build the necessary expertise for a career researching the role of emotions in medical decisions for older adults.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The human HPRT locus will be cloned. Radioactive cDNA probe will be made on template in RNA from HPRT plus cells that does not associate with DNA from human cells that have an X-ray-induced deletion of the HPRT locus. The probe, nitrocellulose transfer techniques and autoradiography will be used to screen plaques formed by shotgun recombinant DNA phages for those with inserts complementary to probe sequences. After these clones are amplified those with functional HPRT genes will be identified by transfer of their DNA into HPRT- cells using micro-injection or calcium-phosphate precipitation techniques. If the approach outlined above is successful we will attempt to clone genes of the human major histocompatibility complex using DNA from cells having homozygous deletions in this region, which we have produced with X-rays. Microinjection experiments and hybrids produced by fusing human malignant with non-malignant cells will be used to determine if the following traits are dominantly expressed: morphological transformation, anchorage-independent growth and tumorigenicity in nude mice. For traits that are dominantly expressed a combination of DNA-mediated genetic transformation and microinjection techniques will be used to clone the responsible cancer genes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": ": Adriamycin (doxorubicin) is a potent, broad-spectrum antineoplastic agent effective in treating a variety of cancers including both solid tumors and leukemias. However, the clinical value of this drug is limited by the development of a cumulative and irreversible dilated cardiomyopathy that is manifested as a progressive loss of ventricular performance in patients receiving repeated doses of the drug. Adriamycin cardiotoxicity is also characterized by a dose dependent decline in mitochondrial oxidative phosphorylation and a decrease in high-energy phosphate pools. Our hypothesis is that this results from a cumulative and irreversible modification of regulatory factors affecting the membrane permeability transition (MPT) pore by adriamycin. The enhanced sensitivity to induction of the MPT leads to a futile, energy-depleting cycling of calcium across the mitochondrial membrane, which we suggest accounts for the dose-dependent loss of respiratory efficiency and ATP synthesis in cardiac tissue from adriamycin-treated rats. We further suggest that these mitochondrial changes contribute to the progressive inability of cardiac tissue to tolerate metabolic stress, particularly those associated with induction of the MPT such as ischemia and reperfusion. Another critical element to this adriamycin-induced mitochondrial cardiomyopathy is its irreversibility, which constitutes a very serous problem clinically in treating cases of pediatric or recurrent neoplasias. The fact that the effect of adriamycin on mitochondrial bioenergetics outlasts the persistence of drug residues in tissues suggests that adriamycin, at sufficient doses, initiates an irrevocable sequence of events that continue beyond elimination of drug. Our hypothesis is that adriamycin interacts with mitochondrial membranes to initiate a series of reactions that lead to increased rates of free radical generation. Furthermore, we suggest that this is a self-perpetuating process that increases in magnitude despite the termination of drug treatment and elimination of drug residues from the tissues. Careful and rigorous testing of these jointly related hypotheses will provide valuable insight into the pathogenesis of adriamycin cardiomyopathy and is essential to the development of mechanism-based therapeutic strategies to minimize the dose-limiting effects of this clinically important anticancer chemotherapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objective of the research proposed is to develop an understanding of the enzymatic reactions involved in the biosynthesis of terpenes through study of model reactions and investigations with biosynthetic enzymes. The synthesis of protosqualene pyrophosphate (a newly coined name for the cyclobutly isomer of presqualene) is proposed since this compound may be a new intermediate in sterol biosynthesis. Substrate specificity and mechanism in the enzymatic conversion of geranylgeranyl pyrophosphate to kaurene is to be studied through the use of structural analogues and stereospecifically labelled substrate. This project will be carried out in collaboration with Professor C.A. West at UCLA. An investigation of the carbonium ion reactions of the sesquiterpene, norcedrol is proposed as a possible model for tricyclic sesquiterpene biogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Wisconsin Clinical Cancer Center will develop a statewide cancer control program designed to: 1. Engage health care professionals in efforts to detect cancer earlier. 2. Develop a regional network of affiliated centers providing excellent diagnosis and treatment of cancer. 3. Conduct cooperative projects in cancer detection, diagnosis, therapy and rehabilitation. 4. Serve regional units with central data management resources. 5. Inform the public of advances in cancer control, ways they can improve outcome in cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The candidate seeks a K01 career development award that will encompass training in clinical research to complement his multidisciplinar training in immunology, microbiology, genetics and epidemiology and enable him to advance his scientific career by investigating the role of the gut microbiome in Crohn's diseae (CD)under the influence of major genetic risks.CD is an inflammatory bowel disease (IBD) resulting from defects in the mucosal immune response to enteric bacteria in genetically susceptible individuals. Over 70 susceptibility loci, particularly immune response genes such as NOD2/CARD15, IRGM and ATG16L1, have been associated with CD risk in individuals of European ancestry. Previous studies have established the distinct membership and abundance of the gut microbiota in CD patients compared with healthy controls, inferring a possible role of the microbiome in CD pathogenesis. Accumulating evidence supports the role of the major CD susceptibility genes, NOD2/CARD15, IRGM and ATG16L1 in the processing of microbial antigens and innate immunity. Moreover, studies showed significant differences in microbiome profiles among ethnic groups. However, it is unclear whether carriage of the major genetic risk alleles correlates with an abundance of any particular microbial species in the gut. In this study we will investigate whether bacterial profile differs between carriers and non-carriers of the major CD susceptibility alleles while focusing on a genetically homogeneous Ashkenazi Jewish (AJ) population, who has the highest prevalence of CD comparing with other ethnic/racial groups. The hypothesizes include: 1) the microbial profile of the gut is moderated by the carriage of CARD15, IRGM and ATG16L1 risk variants and 2) the CD risk in the AJ population is attributable to the unique combination of bacterial species in the gut and host genetics.To test these hypotheses, the candidate will recruit AJ CD patients enrolled in an ongoing registy of patients with IBD in the Division of Gastroenterology at The Mount Sinai School of Medicine, New York and already genotyped for the major CD risks. AJ controls without CD will come from the Mount Sinai Biobank. We will utilize 16s rRNAdeep sequencing technique followed by bioinformatics and statistical approaches to characterize the gut microbiota in study subjects with regard to disease status and genetic risks. By studying a more genetically homogeneous population (AJ), this study will attempt to gain a better understanding of the microbiome-host gene interaction associated with disease pathogenesis. This can help build comprehensive diagnostic tools to identify individuals at risk of developing CD, s well as develop novel personalized treatments for AJ CD patients. In addition, this project will help to establish a strong multidisciplinary foundation for the candidate's future career in translational research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have reported that beta-lapachone, a plant alkaloid, greatly increases the lethality of a variety of DNA damaging agents to mammalian dells. The drug was shown to increase the double strand breaks in DNA of the damaged cells, and to decrease the rate of disappearance of single strand breaks. It also-increased the unwinding activity of purified topoisomerase T. We hypothesized from these results that topoisomerase I is involved in repair of damaged DNA. Consistent with this, we showed that camptothecin, a specific inhibitor of topoisomerase I, also enhanced the lethality of DNA damage in cells. Our three specific aims are: (1) To test the hypothesis that topoisomerase I is involved in excision repair of damaged DNA. (2) To determine whether camptothecin or beta-lapachone enhance lethality of antineoplastic DNA damaging agents by altering topoisomerase I activity. (3) To correlate the above lethality with the increased conversion of single stranded DNA damage to double stranded breaks. The experimental approach, described in more detail below, depends upon three principle variations of established systems for studying topoisomerase I, namely (A) To use nicked gapped, and randomly damaged DNA's as substrates for the enzyme in comparison with intact DNA. (B) To determine the effects of the two drugs on both damaged cells and topoisomerase 1 activity in purified systems. (C) To make mutations of topoisomerase I and investigate the above phenomena with them as compared to the wild type enzyme, both in vivo and in purified preparations. The research proposed- in-this grant is intended to fill several gaps in our knowledge. The first concerns the role of topoisomerase I in DNA repair. How are DNA repair processes dependent upon the type of initial lesion? Is topoisomerase I involved in the repair of all kinds of DNA lesions? From a practical point of view, the combination of topoisomerase I inhibitors with DNA damaging agents may lead to enhanced antitumor activity with diminished host toxicity, and thereby improve the therapeutic indices of established antineoplastic agents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: In this revised competing application, the Principal Investigator seeks to define the relationship of cell cycle status in hematopoietic stem cells and their ability to engraft during transplantation. Four Specific Aims are enumerated. In the first specific aim, the Principal Investigator plans to identify the cytokines which induce an engraftment defect. During the previous tenure of this grant, Dr. Quesenberry demonstrated that hematopoietic stem cells cultured in the presence of a cocktail of cytokines including IL-1, IL-3, IL6 and stem cell factor display a substantially reduced capacity to engraft in non-marrow ablated recipients. Having developed a non- myelosuppressive model of marrow transplantation in which large numbers of male donor cells are infused for several days into female recipients, the Applicant was successful in demonstrating a very high engraftment efficiency in such animals. However, when marrow cells were first cultured in the presence of a number of cytokines which act to expand putative stem and progenitor cell numbers, the ability of these cells to contribute to long-term hematopoiesis was essentially eliminated. As this non-conditioning model of stem cell transplantation would find great use in a setting of gene therapy, the Applicant proposes to determine the cytokine or cytokines responsible for the induced effect. Initial studies will be directed towards defining the amount of time cells need to be exposed to these proliferative cytokines in order to induce the engraftment defect, and refinement in the methodology will also be addressed by studying two additional systems which potentially will eliminate any barrier to transplantation (male into female might engender an immunologic intolerance). The Investigators will study Ly 5.1 into Ly 5.2 congenic transplants, and a ROSA 26 transplant model in which beta galactosidase can be tracked. These refinements should optimize the model for the subsequent specific aims, which attempt to elucidate the relationship of cell cycle status to engraftment and any experimental manipulation which might impact favorably upon this defect. In the second Specific Aim, the Investigator plans to generate both whole marrow cells and rhodaminelo/Hoechstlo fractions and using either growth factor stimulation or isoleucine deprivation attempt to develop a synchronized population of cells to test whether the transplantation defect is restricted to a specific point in the cell cycle. The Investigator will monitor cell cycle using FACS analysis and will then perform transplantations with cells throughout the cell cycle. It is hoped but not fully expected that cells that transit an entire cell cycle might provide the most useful information. In the third Specific Aim, the Investigator plans to identify a cocktail of cytokines which are capable of maintaining primitive stem cell engraftment potential. If the Investigator's hypothesis is correct, these should be cytokines which maintain viability but fail to induce cell cycle progression. To the basic cytokine cocktail of IL-11, IL-3, IL-6 and \"steel\" factor, modulation using different concentrations of cytokines (based on the finding that low levels of cytokines can support survival, whereas high levels support proliferation), additional cytokines which appear to act in relatively primitive cells including LIF, TGF beta, IL-1 alpha, and MIP-1 alpha, and different adherent stromal layers including Dexter cultures and the cell line TCI will also be tested for effects on cell maintenance or expansion without an associated engraftment defect. Finally, in the fourth Specific Aim, the goal will be to develop in vitro techniques to reverse cytokine-induced engraftment defects. If successful, the Investigator hopes that such manipulation will allow marrow cells to be transiently stimulated to allow retroviral integration, followed by their manipulated return to quiescence, thereby repairing the engraftment defect and thus allowing for engraftment of the cells in which retroviral gene therapy had occurred. Once again, combinations of cytokines will be assessed by following their effects on short-term clonogenic cells populations (HPP- CFC and LPP-CFC) and most importantly, on the ability to engraft in the long-term setting.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mast cells (MCs), granulocytes, and lymphocytes are integral to the development of an allergic response. Allergic inflammation may also be generated through activation of receptors coupled to heterotrimeric G proteins (GPCRs). The purpose of this study is to understand mechanisms of G protein-mediated signal transduction in immune cells, with a focus on GPCR-mediated trafficking of leukocytes to sites of allergic inflammation. GPCRs activate a core pathway of heterotrimeric G proteins, which bind guanosine triphosphate (GTP) in exchange for guanosine diphosphate (GDP). The GTP-bound form of the G protein alpha subunit induces downstream signaling cascades, including intracellular calcium flux responsible for MC/basophil degranulation. This project focuses on a family of regulators of G protein signaling (RGS proteins), which inhibit the function of G alpha-i and G alpha-q, but not G alpha-s, proteins by increasing their GTPase activity. G alpha subunits oscillate between GDP- (inactive) and GTP- (active) bound forms based on ligand occupancy of the associated receptor. The GTPase accelerating (GAP) activity of RGS proteins limits the time of interaction of active G-alpha and its effectors, resulting in desensitization of GPCR signaling. Despite a growing body of knowledge concerning the biochemical mechanisms of RGS action, relatively little is known about the physiological role of these proteins in allergic inflammation. A major area of investigation is the recruitment of inflammatory leukocytes to sites of inflammation. Chemokines are a major class of compounds acting on leukocyte GPCRs, which orchestrate immune cell trafficking, and RGS proteins including RGS5, RGS13, and RGS16 inhibit chemokine signaling by desensitizing GPCR signals. In FY18, we continued our phenotypic characterization of patients with undefined immunodeficiencies and novel mutations in G proteins and/or RGS proteins are being characterized in collaborative studies with Drs. Orange and Su. 3 brothers in a single family with combined immunodeficiency were found to carry biallelic mutations in RGS10. We discovered that RGS10 is expressed in both human B and T cells. Patient-derived B and T cells from these patients exhibit profoundly impaired chemotaxis in response to chemokines despite having comparable RGS10 expression. Molecular studies, which are ongoing, have determined that the mutations may affect RGS10 phosphorylation and induce protein mislocalization, which could result in a gain of function. IN FY18, we defined a role for RGS5 in neutrophil trafficking. RGS5 was expressed in both human and murine neutrophils. We detected significantly more neutrophils in the airways of Rgs5-/- mice than wild-type counterparts following acute respiratory virus infection and also in the peritoneum in response to injection of thioglycollate, a biochemical proinflammatory stimulus. RGS5-deficient neutrophils responded with increased chemotaxis elicited by the chemokines C-X-C motif chemokine ligand 1 (CXCL1), CXCL2, and CXCL12, but not fMLP. Moreover, adhesion of these cells was increased in the presence of both CXCL2 and fMLP.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Abstract We have established the UCSD Clinical and Translational Research Institute (CTRI), with its hub located at the University of California, San Diego (UCSD). This regional center includes three universities (UCSD, San Diego State University and UC Riverside), 4 biomedical research institutes (Salk Institute, Sanford-Burnham Institute, J. Craig Venter Institute, and La Jolla Institute for Allergy and Immunology), and 3 academic hospitals (UCSD Medical Center, VA San Diego Medical Center and Rady Children's Hospital), one community health center (Palomar Health). The CTSA program encompasses 3 health sciences professional schools (Medicine, Pharmacy, and Nursing) and 2 Master's programs in Clinical Research and Drug Discovery Sciences. The overall program goals are to: 1) Develop an integrated education and training program across the translational science spectrum; 2) Empower community-based partnerships and integrate the stakeholders into translational research governance; 3) Diversify and enhance biomedical informatics infrastructure to provide a secure, confidential and inter-operative platform; 4) Provide administrative infrastructure to manage and improve clinical and translational research; and 5) Provide resources that enhance collaboration, encourage disruptive innovation and support clinical investigation. The CTRI KL2 program will provide a mentored research experience for junior faculty that ensures a diverse future workforce with the required skills for outstanding multi-disciplinary research in human health. Didactic programs will teach key skills in clinical and translational research, including Master's degrees for individuals seeking additional rigor. Mini-sabbaticals will offer experiential training to enhance the scholars' research programs. A Mentoring Core will train scholars to become mentors for the next generation of researchers. Outcomes and tracking will use new informatics tools to determine the impact of the curriculum on long term research productivity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal details a 5-year plan for the development of an academic career in Cardiovascular Medicine. The PI is a trained interventional cardiologist with a PhD, and has recently started his first faculty position at the University of California San Francisco (UCSF), Division of Cardiology. This proposal will utilize the Pi's skills in immunohistochemistry applied to the animal model of myocardial infarction (Ml) already used in the lab of the supervisor, Dr Randall Lee MD, a recognized expert in the application of new therapies for Ml. Additionally, this proposal will permit a dedicated period of time for the PI to learn the techniques and statistical analysis of high throughout microarray analysis. This will be achieved through a collaboration between the Division of Cardiology and the Gladstone Institute of Cardiovascular Disease at UCSF. Apoptosis, also called programmed cell death, of cardiomyocytes following Ml is a crucial aspect of the process of left ventricular remodeling that leads to heart failure. As the population ages and the incidence of heart failure increases, this is becoming and increasingly important clinical problem. Novel strategies to treat apoptosis following Ml are being developed, but are being tested in models of Ml in young animals. We will therefore compare the timescale of apoptosis, as well as the expression of pro- and anti-apoptotic genes, in old versus young animals. A specific inhibitor of caspases, which cause apoptosis in cardiomyocytes, will then be used to see if the anti-apoptotic effects seen in young animals are the same in older animals. This, in turn, will facilitate better planning of anti-apoptotic strategies for clinical trials in older patients. UCSF is an ideal research environment for an aspiring clinician-scientist. The Division of Cardiology and the Gladstone Institute of Cardiovascular Disease are centers of excellence in cardiac research. Specific aims 1 and 3 can be achieved in the lab of the project supervisor, Dr Lee. The microarray studies will be performed and the data analyzed at the Gladstone Institute under the supervision of Dr Bruce Conklin MD, a pioneer in microarray analysis. Additionally, an advisory committee of esteemed senior researchers will oversee the progress of this project.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Synthetic approaches to the macrolides methymycin, boromycin, and verrucarin A are described. The strategy for synthesis of the twelve-membered lactone of methymycin involves the combination of two units in chiral form, corresponding to C1-C8 and C9-C11 of the aglycone methynolide. A description of the planned synthesis of the more complex segment, using the addition of an oxoallyl cation to a furan to elaborate none of the required ten carbons at the first step, is presented. Lactonization of a fully functionalized hydroxy acid is proposed for completion of the aglycone. The plan for synthesis of the ionophore boromycin envisions construction of the two sections of the diolide by a common pathway. The \"southern half\" is to be assembled from five unitary segments in a specific sequence which permits a high degree of stereocontrol. The \"northern half\" is obtained by cyclization of the C-9 epi version of the \"southern half\", and these two chains are then fused around the borate core prior to completion of the macrolide ring. For verrucarin A, a cytostatic antibiotic from the fungus Myrothecium, the synthesis plan first envisions completion of a route to the sesquiterpenoid component verrucarol in stereoselective fashion. A key step in this scheme is expansion of a cyclobutyl carbinol to the bridged ring system characteristic of the trichothecenes. Attachment of the macrolide loop, assembled stepwise from three segments, to this terpenoid diol begins at the C15 hydroxyl with completion of the lactone by cyclization at the C4 hydroxyl.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Two canonical binding sites for the Ets family of transcription factors have been identified within the TCR-zeta promoter. These sites bind Elf-1, and play an important role in basal transcription of zeta. Ligand-mediated TCR degradation is inhibitable both by reagents that block vesicular acidification and by inhibitors of the ubiquitin/proteasome pathway, suggesting a complex pathway leading to TCR degradation. Our determination that the TCR is a substrate for ubiquitination has led us to evaluate ubiquitination in more detail. A family of human E2 enzymes (UbcH5A-C) have been cloned and characterized. These E2s are widely expressed in human tissues and function in conjunction with the only characterized human E3, E6-AP, in the ubiquitination of proteins. We have also been studying a novel yeast E2, UBC6, a novel type IV transmembrane protein found associated with membranes of the ER. The short transmembrane domain of UBC6 is responsible for its distribution within the cells, progressive lengthening the transmembrane segment re-targets UBC6 to the Golgi Apparatus and then to the cell surface. Two murine E3 enzymes have been characterized. One of these is the murine homolog of E6-AP, the other is a protein found to have homology to E6-AP, termed Nedd-4. Like E6-AP, Nedd-4 is an E3 enzyme, both of these function with the UbcH5 family of E2s in the ubiquitination of cellular proteins. These E3s exhibit distinct subcellular localizations, E6-AP is expressed predominantly in the nucleus, Nedd-4 is present exclusively in the cytosol. Nedd-4 has two distinct E2 bindings sites, one of which binds the amino terminus of E2s and the other showing affinity solely for the carboxyl component of E2s. These data suggest models for ubiquitination involving oligomerization of E2 and E3s.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of the project is to develop an immunoadsorption-immunosuppression program that will effectively remove platelet alloantibodies and impair their continued production. The goal is to obtain sufficient antibody reduction to permit the survival of incompatible platelets in refractory, thrombocytopenic patients. Either intact platelets, platelet membranes or platelet antigen fractions will be used as the immunoadosorbant. These materials will be immobilized to a solid support structure and plasma obtained by apheresis techniques will be passed over the platelet immunoadsorbant to remove alloantibodies. Alternatively, intact platelets will be placed in a reservoir system and plasma obtained by membrane or hollow fiber filtration will be pumped through the platelet reservoir to achieve alloantibody adsorption. Cyclosporin A will be used as the primary immunosuppressive agent. Other agents will be added only if Cyclosporin is not effective as a single drug. Quantitative platelet antibody measurements and radiochromium labelled donor platelet survival determinations will be used pre- and post-immunoadsorption-immunosuppression to document the value of the program in reversing platelet alloimmunization. Both animal and patient studies will be conducted.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Direct methods for functional neural imaging are critical to advancements in understanding neural behavior, plasticity, connectivity and pathology. If we can directly image active neurons we will have the ability to examine neural activity more precisely than is presently the case with fMRI. We have developed an MRI-based conductivity imaging technique, Magnetic Resonance Electrical Impedance Tomography (MREIT) that can reconstruct conductivity maps with near-MRI resolution. In MREIT, small external currents are applied to an object. The MR magnetic flux density patterns created by current flow may be converted to conductivity or current density slice images. We developed this technique and have refined it to the stage of producing electrical conductivity images of animal brains in vivo, using relatively low applied currents. The large changes in membrane conductance that occur during activity cause dynamic changes in paths taken by externally applied currents. Changes in spiking activity during external current application will cause differential phase accumulation in MR data that will increase the longer current is applied. Neural activity therefore becomes visible as an increase in apparent conductivities of voxels coincident with active intracellular areas. Because the contrast controlling MREIT signals, conductivity, may only acquire positive values, phase accumulations cannot be cancelled by the presence of opposite polarity or opposingly oriented signals. This may give MREIT an advantage compared with other MRI-based methods for imaging neural currents that are based on perturbations of phase or main magnetic fields caused principally by summed axonal current flows. Thus, MREIT has the potential to detect activity in complex structures including gray matter. In this proposal, we will investigate the ability of functional MREIT (fMREIT) to detect activity-related conductivity changes in neural tissue. We will develop fMREIT techniques to image neural activity in vitro, in a several standard neural preparations, while progressively refining our methods to detect and locate active cells at high signal to noise ratio and using main magnetic field strengths conveniently used in vivo. In isolated preparations, our method has the potential to enable detailed analyses of single cell mechanisms. The method could thus be considered as a non-invasive extension of patch clamping techniques, and could stand alone for this purpose. However, ultimately we wish to image activity in vivo and our final study in this program will include a tentative exploration of fMREIT in a live animal model as a precursor to further research in this area. In summary, this study will establish the basis for functional MREIT (fMREIT) techniques. This method could ultimately be used to visualize effects of more general neural behavior and enable more fundamental analyses of neural behavior in vivo than is available with existing techniques such as fMRI.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of the Molecular Resource Core (MRC) is to provide support for the analysis of genes and gene products that are relevant to the pathogenesis of arthritis. It will support studies of immune-mediated arthritis in both humans and mice. The MRC will take a multifaceted approach in that it will develop new assays and procedures, be a source of well-characterized Collagen, provide analysis of samples under standardized conditions with stringent quality control, and be a technical and educational resource. MRC facilities will isolate and purify collagen and peptides. This protein family has many unique characteristics that make it difficult or impossible to obtain reliably from commercial sources. The MRC will use state of the art equipment for molecular analyses to aid in the development and application of the most advanced molecular techniques to the study of rheumatic disease. It will perform highly sensitive determination of mRNA levels by quantitative real time polymerase chain reaction, including assisting investigators in the design of appropriate oligonucleotide primers and probe sets an. d will provide a standardized service for sample amplification, data collection and analysis. The MRC will aid both new and established investigators in the use of DNA microarray technology to discover new factors and genetic interactions that affect pathogenesis and modulation of disease. Since many rheumatic diseases are closely linked to particular MHC genes, it will provide HLA genotype determination to identify those haplotypes relative to disease with the capacity to perform fine specificity analysis down to the DNA sequence level. The availability of these services should facilitate research in rheumatic diseases for both clinical and basic scientists at the University of Tennessee, HSC.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Maximum blood flow acceleration, beat-to-beat stroke volume and continuous external power of the left ventricle are derived from instantaneous (pulsatile) measurements of blood flow velocity and pressure in the ascending aorta of patients with ischemic and/or valvular heart disease. These parameters are regarded as giving us more information about ventricular performance than those derived from measurements of average flow and pressure. Also, the calculation of aortic input impedance from the pulsatile records of flow and pressure give us additional information about the elastic properties of the aorta as well as the location of major reflecting sites. In the group of patients studied thus far, velocity and pressure were measured using two separate catheters, but ultimately these measurements will be obtained using a single catheter with an electromagnetic velocity transducer and a miniature pressure transducer mounted at the tip. Atrial pacing will be used as transient stress to produce myocardial ischemia and sublingual nitroglycerin will be used to reduce preload and afterload in patients with and without myocardial ischemia. Pathophysiology of atrial pacing will be studied with special attention to beat-to-beat measurements of stroke volume and acceleration following the termination of pacing. These postpacing beats are of interest because they provide an opportunity to study the response of the ventricle to a sudden increase in filling volume (\"auto volume load\").", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Understanding host-viral interactions at the organismal, cellular, and molecular levels are vital in mitigating viral pathogenesis. The goals of this project are to discover the viral microbiome and specific antiviral immunity genes in the model organism Hydra. By doing so, we will determine whether Hydra contains viruses similar to human viruses as well as being able to test Hydra specific immunity genes responsive to viral presence. Hydra provides an ideal model to test these host-viral interactions because of their simplistic composition, their ease of experimental manipulation, and their epithelial exposure to the environment without a protective barrier. Further, Hydra lack adaptive immunity features, do not contain any motile phagocytic cells, and only utilize mucous as a means to preserve its epithelium. Therefore, Hydra is uniquely suited for the study of host-viral innate immunity at the mucosal epithelium. To establish this, the project consists of two specific aims: 1.) Determine the Hydra viral composition utilizing viral metagenomics. 2.) Elucidate the Hydra antiviral innate immune response in the presence of cytosolic nucleic acids. By accomplishing these two aims we will identify viruses that cause pathogenic conditions in Hydra as well as discern viruses that similarly cause human pathogenesis. These viruses can then be tested in this innate immunity model system. Further, by identifying the Hydra antiviral genes responsive to viral infection, we will manipulate the Hydra through gene knockdown, to determine which genes are vital in limiting viral pathogenesis. PUBLIC HEALTH RELEVANCE: Establishing Hydra as an antiviral mucosal innate immunity model organism is important for human health as demonstrated by genetic variants in phylogenetically ancient innate immune genes are involved in the etiology of chronic inflammatory diseases of the epithelial barrier, such as Crohn's disease, atopic dermatitis, and asthma. These polygenic diseases are characterized by chronic relapsing inflammation of the mucosa. Such disease states and the response to them can be studied in the Hydra model system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Liver Carcinogenesis: A small electrophoretic class of soluble proteins of target organs contains the principal protein-carcinogen conjugates of three families of carcinogens. In pursuit of the possible role in carcinogenesis of one such conjugate, and of the interaction between carcinogen and target protein from which it arises, the following are broad aims of the proposed research: 1. Characterization of the principal soluble liver azoprotein of azo dye preneoplastic liver. 2. Identification of the principal protein target of azocarcinogens in liver. Cell Transformation to Malignancy: The goal is to investigate the extracellular fibrinolytic activities that may have specifically associated with cells transformed by viruses or chemicals. Extracellular activators and inhibitors of fibrinolysin will be isolated from cultured cells and characterized. Effects of the additions of the activators and inhibitors on normal and transformed cells will be studied, in order to learn whether the fibrinolysin activating activities of transformed cells are in part responsible for some of the characteristics associated with cell transformation to malignancy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Core F ? Immune Function The Immune Function Core F provides broadly ranging support for immunological studies. Driven by investigator need, and a rapidly changing science environment, the Core currently offers multiplexed cytokine measurements, standard flow cytometric analysis as well as flow imaging, sorting of cell populations via flow cytometry and other methods, and ELISA, ELISPOT, and infrared imaging. The re-structuring of the Core in 2010 included a cooperative agreement with the Cancer Center Cytometry and Imaging Microscopy Facility to pool the equipment, harmonize services, and maintain formal access to CLIA-level flow cytometry. Drs. Scott Sieg and James Jacobberger have been working closely together as Co-directors and have used their complementary expertise to greatly enhance Core services. Dr. Sieg provides immunological expertise, functional assays, and biomarkers, especially in the context of HIV research; Dr. Jacobberger provides expertise on flow and image cytometry instrumentation, single-cell assays, and biomarker discovery, especially in the areas of cell cycle, apoptosis, and cell signaling. A major function of the Core is to collaborate with users to develop novel immunological assays. For example Core F, in collaboration with the Proteomics & Systems Biology Core G, is refining new flow cytometric assays for characterizing the phospho-proteome. This unique platform should open new avenues of research relevant to HIV pathogenesis, clinical evaluation, and treatment. A parallel effort is focused on developing multiple pathway signaling measurements on sub-populations of cells in unfractionated human samples (e.g., blood). The Core also interacts closely with the Clinical Core D for sample procurement. The Immune Function Core (F) is currently the one of the most heavily used of all the CWRU/UH CFAR cores and during this grant cycle the Core generated net revenues of $384,177. Program income has been steadily reinvested in the purchase of advanced instruments, such as multicolor flow cytometers, and reagents. For example in 2012, the Immune Function Core purchased a BD Fortessa ? a next generation analytical instrument ? with blue, green, red and violet lasers and filters optimized to measure fluorescent proteins. This instrument was subsequently upgraded to add a high throughput 96 well plate feed system thanks to a CFAR supplementary award. As part of the recruitment of Dr. Skaly, the CFAR will purchase a BD Biosciences FACSAria with 4 or 5 lasers and 18-color capability. The FACSAria will be maintained under BSLII conditions to permit sorting of HIV-infected cells. This state-of-the-art instrument will bring sorting capacity in line with our analytical capacities and create further opportunities for Core-Core interactions since it will be used to prepare samples for sequencing and proteomic analyses by Cores E and G.. The specifically aims of the Core follow: ? To provide access to instruments and assays to support CFAR programs. ? To develop validated assays and platforms for CFAR investigators. ? To provide consultation and training at all levels. The development of innovative new platforms, such as phospho-protein analyses, spearheaded by the Immune Function Core creates new opportunities for scientific investigation and competitive advantages by CFAR investigators. The Core will continue to promote interactions among CFAR investigators, meeting the needs of the CFAR Working Groups, and cooperating with the other CFAR Cores to provide access to cutting edge technologies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project examines factors affecting the incidence of racial harassment and discrimination, and the multiple consequences of the stress of racial harassment and discrimination. Research suggests that Hispanic employees who experienced racial discrimination, and their White coworkers who were aware of it, showed higher levels Of stress. Further, African-Americans and Hispanics who reported racial discrimination exhibited poorer health conditions and psychological well-being. The project tests a model which distinguishes between racial harassment (i.e., unfavorable race-related comments or gestures) and racial discrimination (i.e., unfair employment- related treatment of employees of one race over another). Further, the model predicts that a negative organizational climate for racial/ethnic minorities affects the occurrence of racial harassment and discrimination, and that those who are racially harassed or discriminated against experience increased stress, and negative job-related (e.g., job satisfaction, organizational withdrawal), health (e.g., health conditions, health satisfaction), and psychological outcomes (e.g., self-esteem, satisfaction with life). In addition, the project explores how the stress of racial harassment and discrimination differentially affects those who experience such events, and their coworkers who are aware of, or witness such events. Finally, it also examines how employees of different racial/ethnic backgrounds are differentially affected by the stress of experiencing or being aware of racial harassment and discrimination. Participants are 1500 graduate students at an ethnically diverse (approximately 50% Hispanic), South-Western, university. Participants will complete a survey which assesses their perceptions of university climate, experiences of racial harassment and discrimination and job-related, health and psychological outcomes. Data will be analyzed by the LISREL VIII approach to structural equations modeling. Results can be used to design interventions and will contribute to the understanding of the role of culture in responses to the stress of racial harassment and discrimination.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our laboratory has in the past year conducted runs at CHESS A1, F1 and F2, NSLS X4A, X12 and X25, as well as APS to extend the resolution and improve the quality of diffraction data for crystalline complexes involved in chaperonin mediated protein folding, signal transduction, transcriptional and translational regulation. During those trips we have obtained numerous complete high resolution data sets resulting directly in the determination of the following new structures: (1) progesterone ligand binding domain complexed with progesterone; (2) human estrogen receptor ligand binding domain complexed with estradiol; (3) the chaperonin GroEL/GroES/ADP complex; (4) the T. thermophilus EF-Tu/EF-Ts complex; and (5) a P. woesei ternary complex of the TATA-binding protein, TFIIB, and a DNA sequence containing a consensus TATA box at 2.1[unreadable] (see publications section). Data recently collected at CHESS has been instrumental in solving all of the above structures. In addition a paper on the molecular mechanism for the role of phosphorylation in the regulation of hetereotrimeric G-Proteins by phosducin is also in preparation from data collected at CHESS on the phosphorylated form of phosducin. Other projects which are still in progress which have been significantly influenced by CHESS synchrotron data collection include the following: Chaperonin-Medicated Protein Folding: To understand the capacity of ATP to drive the GroEL-GroES reaction cycle, we wish to study relevant intermediates in the ATPase cycle. GroEL/GroES/(ADP)7/(AlFx)7 crystals diffract to 4[unreadable] at our home source but only initial tests were done at the CHESS F1 line this year. Searching for optimal freezing conditions is in progress. Bovine Visual Arrestin: The arrestin crystals diffract to 3.0[unreadable] resolution using synchrotron radiation (NSLS X-25 beamline and MacCHESS A-1 beamline). We have collected complete native data sets to 3.1[unreadable] (C2221; a = 169[unreadable], b = c = 191[unreadable]; crystal size: 0.2 x 0.2 x 0.03 mm3). Useful MAD, heavy atom and anomalous data sets for phasing have been collected at CHESS, X-25 and APS. Quaternary Complex of VP16 Acidic Activation Domain Complexed with hTBPc, hTFIIB on Promoter DNA. To date no high resolution structural information exists on a transcriptional activator domain bound to the preinitiation complex. Hence, this crystal structure will reveal the essential contacts made between a strong acidic activation domain and the basal machinery, resulting in transcriptional activation. The crystals diffract to about 4.0 [unreadable] on a home source (at 100 K). Several 2.6[unreadable] native data sets (P21; a = 118.7[unreadable], b = 122.2[unreadable], c = 140.8[unreadable], b = 113.0 ; crystal size: 0.4 x 0.1 x 0.05 mm3) were collected recently at CHESS F2 and F1 lines.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The title of this project has been changed to more accurately reflect the thrust of the science. The retina and pigment epithelium are neuroepitheli tissues that work in close cooperation. Specific growth and differentiatin factors are found in the eye that guide development and interactions of individual ocular tissues to form a functional visual system. Studies on t s project are focused on an understanding of the molecular biology and molecular genetics of the retina and discovering new genes that are candidates for hereditary retinal degenerations. For example, ocular tissu synthesize a number of growth factors. There now appear to be several systems present that could self-regulate growth and metabolic activity in the retina-pigment epithelium complex and be involved in eye diseases. In this regard, we have cloned and characterized a unique protein secreted from fetal human pigment epithelial cells, called pigment-epithelial derive factor (PEDF), that is \"neurotrophic\" to cultured human retinoblastoma cell and may affect neural retinal development in vivo. This protein also is a potent \"neuronotrophic\" agent in that it promotes neuronal cell survival of cultured cerebellar granule cells. Finally, PEDF is \"gliastatic\" in that i markedly retards glial cell growth. Along with being a candidate gene in retinal degenerations, the uses of PEDF in neuronal transplant in retina an other CNS areas are obvious.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "MAP kinases are key regulatory components acting on many cellular processes such as embryogenesis, differentiation, proliferation, cell death, and acute hormonal responses. MAP kinases are activated by a protein kinase cascade, consisting of a MAP/ERK kinase kinase (MEKK), which activates a MEK which activates a MAP kinase. ERK1 and ERK2, the first mammalian MAP kinases identified, serve as prototypes for the behavior of the many related enzymes. The functions of these MAP kinases depend on their activation, their subcellular localizations, their protein-protein interactions, and their substrate specificities. We propose to continue to investigate mechanisms controlling MAP kinase pathways as follows. The nuclear uptake of ERK2 is essential for some of its most important functions. We will define mechanisms of subcellular localization of ERKI/2 by examining import and export by reconstitution in permeabilized cells, and microinjection and transfection in intact cells. We will define mechanisms determining functional complexes of MAP kinases. To do this we will use a cDNA library of mutant ERK2 molecules from which we will isolate ERK2 mutants that lack high affinity interactions with proteins including substrates, activators, and phosphatases. We will study the functions of the loss-of-function ERK2 mutants to determine the roles of individual interactions in inducing ERK2-dependent phenotypes. The ERK2 activator MEK1 binds to molecules other than ERK1/2 and Raf that are essential for signal transmission through the cascade. We will attempt to identify proteins that bind MEK1 through its proline-rich insert and determine their mechanisms of action. We will study ERK5, another member of the MAP kinase family. ERK5 is most similar to ERK2, and cooperates with ERK2 to transform cells. We will examine the mechanisms of ERK5 regulation, and determine how its actions relate to the functions of ERK1/2. We will also study a putative ERK5 ortholog in C. elegans to use genetic studies to complement work in mammals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Triple negative breast cancer (TNBC) is one subtype of breast cancer that frequently relapses and leads to worse outcome than patients with hormone receptor-positive subtypes. PARP is currently the most promising drug target for TNBC, and multiple PARP inhibitors (PARPi) have been developed and tested in clinical trials. Although PARPi show higher response rate in patients carrying BRCA mutations, there are still high percentage of BRCA mutations carried patients does not respond to PARPi. Thus, developing strategies to make PARPi treatment more effective and to identify biomarkers to stratify patients is critical. Our recent publication in Nature Medicine (2016) showed that in response to reactive oxygen species, c-Met interacts with and phosphorylates PARP1 at Y907. Moreover, we demonstrated that c-Met-mediated phosphorylation is critical for PARPi resistance, and that c-Met inhibitors sensitize TNBC cells to PARPi. The long-term goal of our research is to develop the effective therapeutic strategies for TNBC. To this end, we will seek the novel biomarkers and treatment strategies to improve the efficacy of PARPi. We hypothesized that PARP1 protein is regulated by its phosphorylation, and that phosphorylation status of PARP1 and the expression of the corresponding kinases that phosphorylate PARP1 will serve as appropriate biomarkers for combinational treatment. Multiple kinases can phosphorylate the same substrates, resulting in signal crosstalk. Also, TNBC is a heterogeneous disease, and the distinct kinases play an important role in different TNBC. Therefore, we also hypothesize that similar to c-Met, other protein kinases also have functions to regulate PARP1 activity through phosphorylation in TNBC. Potential molecules we will study are EGFR, which also directly interacts with PARP1 and phosphorylate it. We will investigate the role of these kinases in PARP1 regulation and PARPi resistance. Thus, we propose the following three aims; Aim 1. To systematically validate the significance of c-Met?mediated phosphorylation in PARPi resistance in mouse models and TNBC patient samples; Aim 2. To determine the role of EGFR-mediated PARP phosphorylation in PARPi resistance in TNBC; Aim 3. To determine the role of c-Met and EGFR interplay in PARPi resistance in TNBC. If our proposal is successful, several phosphorylation sites in PARP1 can be used as biomarkers to guide the combinational treatment of PARPi and correlated kinase inhibitors. We will particularly focus on c-Met and EGFR because inhibitors of these kinases are currently used in the clinic or in clinical trials, allowing for faster progression of our biomarker-guided rationale combination therapy into clinical trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ultimately the primary function of all somatic cells in the testis is to ensure a microenvironment that provides adequate androgen and nutritive support for spermatogenesis and confers immunologic tolerance for the mitotic spermatogonia located outside the blood-testis barrier. It is well known that the pituitary hormones LH and PRL affect androgen secretion by Leydig cells. What is less well understood is the identity and function of the local peptidergic factors that make up the non- steroidal microenvironment of the interstitium. LACAF is an interstitial cell protein stimulated by HCG and growth factor but inhibited by prolactin. LACAF activates adenylyl cyclase in Leydig cells but this activation is not functionally directed towards steroidogenesis. Such differential regulation is by known Leydig cell trophic factors suggests that LACAF is an autocrine regulatory factor. This protein shows some sequence homology with the testicular protein of the VIP family of peptides that shares this capacity to activate cyclase without concomitant effects on steroidogenesis. We will study the function and regulation of these two proteins in parallel. Prolactin preferentially enhances the secretion of two other interstitial proteins, one of which is a known immunomodulator, suggesting the prolactin affects testicular immune elements. The second protein, PIL, may be a member of the transforming growth factor Beta family of peptides. The physiological significance of this regulated pattern of interstitial proteins is presently unknown. Our proposed studies will focus on the unique features of the regulation of LACAF, PIL and complement components by prolactin. The ability to determine the various forms of the prolactin receptor MRNAS affords the opportunity to determine the cell-specificity of these prolactin-induced changes in interstitial protein secretion. LACAF and PIL proteins will be further characterized. Cloned CDNAS for these interstitial proteins will be isolated and their structure characterized. Moreover, specific probes will be generated for use in determining both physiological and aberrant expression of these proteins in vivo and in vitro studies. Regulation of these interstitial proteins will be studied by analysis of their MRNA levels as a function of endocrine status using hybridization in situ and immunohistochemistry. The mechanism(s) of action of these peptides will be investigated by assessment of their effects on both seminiferous tubule and interstitial cell function. This research plan will lead to new, significant data on the mediators of intratesticular signalling and their physiological relevance in the testis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Intimal hyperplasia in arterialized vein bypass grafts is a significant cause of vein graft stenosis and delayed graft failure. Injury at the time of implantation or as a consequence of transplantation into the high pressure arterial system may contribute to these delayed events. Alterations in gene expression accompany implantation and arterialization injury. These alterations lead to intimal hyperplasia, including transformation of endothelial cells to an inflammatory state and initiating migration and transformation of smooth muscle cells from the contractile to secretory states thus creating the lesion of intimal hyperplasia. It is our hypothesis that silencing of genes upregulated by injury to the vein wall will diminish intimal hyperplasia. Furthermore, gene silencing can be accomplished within the constraints of operating room conditions. Our preliminary data demonstrates our ability to identify candidate genes associated with intimal hyperplasia and our ability to knockdown gene expression with siRNA. Using laser capture microscopy we have separated genetic events in the endothelium from those in smooth muscle. In the proposed study we will apply these technologies to (1) systematically identify silencing targets, (2) to silence target genes in vitro, (3) to silence target genes under surgical conditions, (4) to demonstrate inhibition of intimal hyperplasia in vein grafts in vivo. Both human tissue and canine models will be used as well as all cultures, microarrays, laser capture, RT-PCR, Western Blot, and related technology. This study will greatly strengthen the application of gene silencing to clinical problems in vascular surgery, and broaden our understanding of vascular wall biology. Public Lay Summary: Scar tissue formation due to surgical injury is a major cause for failure of heart bypass grafts and bypass grafts for peripheral vascular disease. The investigators propose to prevent formation of this scar tissue by controlling expression of the genes that cause it, using techniques that can be applied in the operating room. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "About 10% of all intracranial tumors are schwannomas, benign tumors comprised of Schwann cells with biallelelic NF2 mutations. NF2 mutation or loss is believed to cause schwannoma formation. Several lines of evidence implicate merlin, the NF2 protein, in small G-protein signaling. Biochemical evidence and our analysis of primary schwannoma cells suggest that merlin functions in Rac signaling. While normal Schwann cells are polarized, with one surface attached to axons and the other to basal lamina, schwannoma cells are unassociated with axons. This loss of polarity is likely to arise secondary to altered Rac signaling. Because the relevance of the Rac signaling pathway to Schwann cells is not known, we propose to evaluate a merlin-Rac cascade in Schwann cell growth and tumorigenesis. Our specific hypothesis is that merlin limits the duration of Rac signaling in Schwann cells. We further postulate that deregulated Rac signaling accounts for the failed axon-glial interactions characteristic of schwannomas. We plan to use a combination of gain-of-function and loss-of-function mouse models and primary cells from human schwannomas to test these hypotheses. We believe that analysis of primary schwannoma cells offers a rare opportunity to study a pure population of early stage human cancer cells. The proposed studies are expected to provide insight into regulation of normal axon-glial interactions, and to lead to strategies to understand and ultimately treat human schwannomas. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hermann Lotze was an agent in the development of philosophy and medicine into independent specialized subdisciplines. The advantages of specialization when held in check by general philosophy and general medicine turned to disadvantages in the subsequent generation. The fruitful interaction of science and philosophy in the period of the rise of the German mandarins gave way to isolation in the later period of their separation. The project deals with the contribution of Hermann Lotze (1817-1881) to the German movement, a broad reform of German medical science on the foundation of philosophical theory construction. His \"mechanical\" program for medicine was one of many attempts to break loose from vague speculative medical theories such as contagion, humors, crises, etc. He conceived the body as an equilibrium, and disease as a disturbances of this process. His \"theory of value\" for both medical and philosophical disciplines was based on Kantian critical teleology. This became a foundation stone for the New Psychology in its many ramifications, including pragmatism, phenomenology, analytic philosophy, social behaviorism, and genetic epistemology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Substance abuse continues to be a persistent epidemic that evades effective treatment. Multiple dependencies present a further challenge because of the need to treat different psychological and physiological effects of the different substances. This proposal has the potential to distinguish the biological basis of individual effects of METH and heroin abuse and reveal how they act in tandem to produce a speedball effect. Understanding the pharmacology of speedball reward will lead to the precise brain targets for which medications can be developed to counteract the rewarding effects of speedballs and consequently reduce the prevalence of speedball dependence.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Carnitine acetyltransferase (CrAT) is a freely reversible mitochondrial matrix enzyme that catalyzes the exchange of short-chain acyl groups between CoA and carnitine. Unlike their acyl-CoA counterparts, acylcarnitines can traverse cellular membranes. Accordingly, the interconversion between these molecules can potentially impact cellular, and perhaps inter-tissue carbon trafficking. The primary substrate of CrAT, acetyl-CoA, holds a prominent position in intermediary metabolism as the two-carbon universal end product of fatty acid, glucose and amino acid oxidation. As its major metabolic fate, acetyl-CoA typically enters the tricarboxylic acid (TCA) cycle where it drives production of reducing equivalents that in turn fuel the electron transport chain. CrAT purportedly acts to export excess carbon fuels from the mitochondria during conditions wherein the production of short-chain acyl-CoAs exceeds TCA cycle flux. By doing so, this reaction is thought to play a key role in regenerating free CoA, modulating mitochondrial acetyl-CoA/CoA balance and relieving acetyl-CoA-mediated inhibition of pyruvate dehydrogenase (PDH), the committed step in glucose oxidation. Mounting evidence from our laboratory suggest that CrAT plays a critical role in regulating whole body substrate selection and glucose tolerance;and that CrAT inactivity might contribute to obesity- and age-related metabolic dysfunction. This NRSA application proposes to elucidate the role of this enzyme in regulating whole body and mitochondrial energy homeostasis utilizing immunoaffinity purification techniques and physiological characterizations of genetic mouse models. First, we will test the hypothesis that one of the mechanisms through which CrAT controls energy metabolism is by regulating nuclear and/or non-nuclear protein acetylation. This is an important and timely line of investigation in light of the growing number of metabolic pathways that appear to be modulated by reversible acetylation. Second, we will use an inducible gene knockout approach to determine whether total body loss of CrAT negates the antidiabetic actions of carnitine therapy. These studies are expected to shed clinically relevant insights on the role of this enzyme in combating nutrient stress and metabolic disease. PUBLIC HEALTH RELEVANCE: The overarching goal of this project is to elucidate mechanisms through which supplemental L- carnitine improves glucose tolerance in obese and diabetic rodents. Results from the proposed studies are likely to yield new insights regarding the therapeutic properties of L-carnitine, a conditionally essential nutrient, while also advancing our understanding of why obesity increases risk of metabolic disorders such as type 2 diabetes. These are clinically relevant topics of intense scientific interest and controversy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Data Management and Statistical Analysis Core performs data management and analysis for the Clinical Core, the Neuroimaging Core, for the Neuropathology Core and for individual research projects affiliated with of the ADCC, and maintains all data collected by the ADCC in a centralized database. This facility also provides consultation in the design of experiments and statistical analysis to all collaborating investigators in the ADCC. Investigators have the option of running their own data analyses in consultation with Data Management and Statistical Analysis Core personnel, or having the core personnel provide consultation concerning interpretation of results. The centralization of these functions makes it possible for each of the research projects to benefit from more sophisticated and flexible statistical and computer tools than would normally be available for any one project and fosters sharing of data among collaborating investigators in the ADCC. The centralized database incorporates all the data on subjects who have been evaluated for participation in the ADC and is updated continuously with the results of evaluations of new subjects and longitudinal information about all subjects. Al changes in subjects' mental and physical status, as well as key outcome variables of drug trials, other clinical studies, neurobiological and postmortem studies are recorded. The core has fully achieved and exceeding its original specific aims. A new local area network (LAN) computer system using all new equipment has been installed this year, and is used to implement centralized data management. The database now encompasses not only data gathered since the inception of the ADC but also all data collected prior to establishment of the ADCC in a uniform format with consistent labeling and data codes. A data resource has been achieved that is unique in both size and diversity. Individual investigators at NYU or elsewhere are able to use with the database for exchange and sharing of data, and procedures for sharing data with the NIA-Alzheimer's Disease Data Coordinating Center (ADCC) have been established. Integrating the formation about the behavioral, physiological and psychosocial components of the disorders being studied at the ADCC in one database has made it possible to develop a rich resource for study the normal and abnormal concomitants of aging.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research has its long-term goal of the development of a quantitative theory of discrimination learning and choice behavior in pigeons. The experiments are done in a choice situation in which one of a pair of stimuli is presented on each trial, and one response is rewarded if it occurs in the presence of one of these stimuli, and a second response is rewarded if it occurs in the presence of the other stimulus. A large number of the parameters that are known to affect learning in other situations will be systematically studied in this one. The proposed theoretical analysis is based on statistical decision theory.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "CMV is a major cause of death for patients with AIDS. CMV pneumonia is often found at autopsy, but is rarely found antemortem for reasons that are not understood. As antipneumocystis prophylaxis is used more commonly, it is likely that CMV will become an increasingly important cause of morbidity and mortality. Effective anti-CMV drugs are now available. It is thus important to learn how to diagnose this entity invasively and noninvasively. This study will assess sputum, bronchoalveolar lavage, and transbronchial biopsy by a variety of conventional and novel approaches to CMV. Patients will be followed, but these data will not be used to determine therapy since their relevance is currently unknown. To date 24 asymptomatic patients with HIV infection have undergone bronchoscopy. Over 80% were culture positive for CMV. The importance of this study is that it is a comprehensive effort to determine how CMV pneumonia can be diagnosed antemortem: CMV is a major cause of death in AIDS patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This minimally invasive study is designed to explore various biologic effects of radiation on the pediatric CNS in an attempt to 1) obtain information on the pathophysiology of radiation-induced damage, 2) explore the association of neuropsychological deficits with biologic markers and neuroimaging abnormalities, 3) document changes in neurobehavioral functioning through longitudinal comprehensive neuropsychological assessments with comparison of various radiation therapy techniques, 4) describe changes in quality of life in pediatric patients who have received radiation therapy, and 5) attempt to identify children at increased risk of radiation-induced neurotoxicity. We are specifically evaluating effects on angiogenesis by measurement of angiogenesis related cytokines, proteins and cells including VEGF, bFGF, thrombospondin, TNF-alpha, IL-12, IL-8, circulating endothelial cells (CECs) and matrix metalloproteinases in blood and urine specimens. We are evaluating the changes in angiogenesis and vascularity by imaging, including dynamic susceptibility contrast perfusion and dynamic enhanced MRI scans. Neurotoxicity will be characterized by measuring biomarkers associated with neurotoxicity and documentation of changes in neurobehavioral functioning through longitudinal comprehensive assessments. In addition, we will describe changes in quality of life (QOL), assess changes in memory, define changes in ophthalmologic studies associated with radiation and detect changes in audiometry associated with radiation. Patients under 22 yrs of age who are referred to NCI for radiation therapy of a primary CNS tumor are eligible. Approximately 55 patients have enrolled and are followed longitudinally by a multidisciplinary team of specialists, including those from pediatric oncology, ophthalmology, radiology, audiology and neuropsychology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study will be conducted among a diverse group of urban and rural women between the ages of 21 and 44 years residing in eastern North Carolina. Women are studied as they are primarily shopping for and preparing family members'food, and thus have a major influence on family member's dietary practices. We propose to use a unique participant recruiting opportunity through a CDC-funded study in the local health department family planning clinic, to enroll a subset of participants from the larger study into a primary qualitative data collection effort to examine neighborhood context and \"food activity spaces\" as viewed by women of diverse demographic groups (Aim 1). In Aim 2, we will develop and implement a standard method for mapping farmers'markets. We will then use data collected in Aims 1 and 2 to conduct a secondary data analysis using a larger sample of women (n = 1000) recruited through the CDC-funded study in the local health department (Aim 3). The successful completion of this project will move the field of public health nutrition forward in the following three ways: (1) Determine the geographic boundaries and features of neighborhood and food activity spaces among diverse women by using participants to quantify and describe their activity spaces. This will allow future researchers to more accurately estimate buffer size in Geographic Information Systems. (2) Standardize the methodology for identifying farmers'markets, which will allow others to accurately quantify and disseminate information about markets to consumers and public health practitioners. (3) Provide greater clarity regarding disparities in access to farmers'markets and regarding potential associations between access to farmers'markets and obesity. PUBLIC HEALTH RELEVANCE: Obesity is an overwhelming public health problem necessitating community-level solutions. Here we propose to combine data from qualitative interviews, windshield tours, and a geographic information system to: (a) define and examine neighborhood context and \"food activity spaces\" as viewed by diverse women;(b) develop a high-quality measure of the food environment specifically related to farmers'markets;and (c) examine potential access disparities, as well as the association between access to farmers'markets and weight status among 1000 diverse women. Results will inform future research and promising environmental and policy changes to decrease obesity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary: Small regulatory RNAs regulate gene expression in most eukaryotes. By regulating gene expression, small regulatory RNAs, play key roles in many biological processes that include development, genome defense, oncogenesis, and antiviral immunity. Small regulatory RNAs act by seeking out and binding homologous (target) RNAs in cells. By recruiting accessory proteins to target RNAs, small RNAs are able to control gene expression at many levels that include; translation, mRNA stability, and transcription. The mechanistic underpinnings of small RNA biology are widely conserved in most eukaryotes. In particular, small RNAs play an important role in regulating gene expression within most eukaryote nuclei. My lab has established systems that are allowing us to study how and why small RNAs regulate genes in animal nuclei. We are using the model organism C. elegans to understand how small RNAs regulate gene expression in animal nuclei. We are using C. elegans to address this question because of the excellent genetic tools that are available, and because of the robust and facile nature of conducting RNAi experiments in this system. Using genetic approaches in C. elegans, we have identified a molecular pathway that uses small RNAs to recognize and mark nascent transcripts (and the genes that encode these transcripts) for silencing. We have identified accessory proteins (termed the nuclear RNAi defective (NRDE) factors), which are recruited by small RNAs to nascent transcripts emanating from RNAP Polymerase II. Finally, we have shown that the association of the NRDE factors with RNA transcripts allows that NRDE factors to inhibit RNA Polymerase II during the elongation phase of transcription. Some of the NRDE factors that we have identified are conserved in mammals. In summary, our work is helping us understand how small RNAs regulate gene expression in animal nuclei, and may lead to insights into how small RNAs regulate gene expression in mammals. We are also interested in understanding why small RNAs regulate gene expression in animal nuclei. Small regulatory RNAs direct the covalent modification of DNA and histones proteins in most eukaryotic cells. These small RNA-mediated chromatin modifications are epigenetic in nature: they alter gene expression without changing the underlying in DNA sequence. We have shown that endogenous nuclear small RNAs, and the nuclear RNAi pathway, regulate the epigenetic landscape at ~1000 genes during the normal course of reproduction. In animals that lack the nuclear RNAi machinery, germ cells loose their immortal character. Thus, C. elegans uses endogenous small RNAs to regulate epigenetic states at many genes during the normal course of reproduction and this gene-silencing process is required to mediate important biological processes. Many other biological processes such as development, imprinting, X-chromosome inactivation, and paramutation are directed by epigenetic modifications on DNA and histones. Interestingly, non-coding RNAs also contribute to many, if not all, of these processes. Given the widespread connections that exist between small RNAs, non-coding RNAs, and epigenetic processes in eukaryotes, we believe that our research exploring how small non-coding RNAs regulate epigenetic landscapes in C. elegans may prove to be globally applicable to diverse epigenetic processes in animals. We do not yet understand 1) how the recruitment of NRDE factors to pre-mRNA inhibits RNAP II elongation to direct nuclear RNAi, 2) how RNAi-guided chromatin modifications contribute to nuclear RNAi in animals, 3) if/how nuclear RNAi is regulated, or 4) if the NRDE nuclear RNAi pathway is functionally conserved in mammals. Our proposed experiments are designed to answer these questions. .", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Multiple sclerosis (MS) is a neurological disease of major socioeconomic importance with a suspected virus etiology. Theiler's murine encephalomyelitis virus (TMEV) provides one of the most relevant of the few available experimental animal models of virus-induced demyelination for the following reasons: a) chronic pathological involvement is limited to the white matter of the central nervous system (CNS); b) myelin breakdown leads to clinical disease; c) TMEV persists in the CNS for the life of the mouse; and, d) demyelination appears to be immune-mediated. Our studies during the initial grant period have defined genetic loci contributing to disease susceptibility, established an immune-mediated basis for the development of TMEV-induced demyelinating disease, and defined a critical role for MHC class II-restricted, virus-specific DTH responses in the demyelinating process. We have also demonstrated that chronic CNS demyelination can occur in the apparent absence of neuroantigen-specific autoimmune responses against the major myelin antigens. In addition, our studies have laid the groundwork for future studies on nonspecific (monoclonal antibody therapy) and specific (virus-specific tolerance induction) means of regulating virus and neuroantigen-specific CMI responses. Using in vivo-derived T cells and in vitro-propagated T cell clones/hybrids isolated from both peripheral lymphoid organs and CNS lesions, we propose to continue our research on defining the effector phenotype, epitope-specificity, T cell receptor usage and lymphokine-producing profile of the TMEV-specific T cell repertoire involved in the demyelinating process. We will also continue our studies examining the effects of and mechanisms responsible for both: nonspecific regulation of disease induction via therapy with monoclonal antibodies directed against MHC class I- and class II-restricted T cell subsets, T cell activation antigens, and T cell lymphokines; and, specific regulation of virus-specific immunity (both T cell and antibody levels) and disease induction via TMEV-specific tolerance/suppressor T cell induction. These studies should lead to a definitive understanding of the role of T cell- mediated immune responses in persistent virus-induced demyelinating processes and may be applicable to the understanding of the etiology and treatment of MS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are approximately 1.3 million US child care workers, most of who are hourly employees and classified as low-wage earners. In North Carolina, half of these workers are African American or Hispanic. Many have no health insurance, putting them at increased risk for many serious and chronic health conditions such as obesity, diabetes, and hypertension. Regular physical activity (PA) is known to decrease risk of many chronic diseases; unfortunately, most Americans, especially low-income individuals and minorities, fail to meet PA recommendations. A multi-level worksite-based PA intervention offered in child care is an innovative strategy to reach these low wage workers and address health disparities among this high-risk population. While the primary target of our intervention is worker PA, we expect that healthier and more physically active workers will be better role models and promoters of healthy lifestyles for children in their care. This two-arm, cluster RCT (n=104 centers, with 4 workers/center) will test the efficacy of a 6-month Care2bWell (CARE) intervention in increasing workers' PA compared to a Financial Empowerment (FE) control arm. The CARE intervention will include a kick-off workshop, where participants get tailored feedback about their health risks and set behavioral goals, followed by three 2-month long wellness campaigns, during which workers monitor PA, receive tailored feedback, and compete in teams for prizes. Center directors will take part in monthly webinars focused on improving the work environment to support PA and wellness. Within a social ecological framework, Perceptual Control, Social Support, and Diffusion of Innovation theories guide intrapersonal, interpersonal, and center- level intervention components. Participants in the FE arm will receive a parallel intervention to enhance financial health at the individual and center levels. The primary outcome will be accelerometer-measured minutes of moderate to vigorous PA. Primary and secondary outcomes (diet, anthropometrics, psychosocial variables, & center wellness environments) will be assessed at baseline, follow-up (6 mo), and maintenance (18 mo). Detailed process evaluation, cost-effectiveness, and meditational analysis will be used to assess mechanisms of behavior change at the individual and center levels.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fluid secreted by the lacrimal gland is an essential component of tears and is estimated to contribute approximately 50% of the liquid volume bathing the cornea (Walcott, 1998). Recently in situ measurements of fluid production from the lacrimal glands of mice have demonstrated stimulated flow rates of 0.2-0.6 uL/minute (Walcott et al., 2002; Paranyuk et al., 2001; Moore et al, 2000). Our preliminary data indicates that the NKCC1 Na+, K+, 2CI- co-transporter and Bkca channels play important roles in fluid production of the lacrimal gland. We propose the use of knockout mice along with NZB mice (compromised fluid flow, Paranyuk et al., 2001) to study their respective roles in fluid production by the lacrimal gland. Further we propose the use of activators and inhibitors of PKC to delineate its role in the regulation of the NKCC1 co-transporter and Bkca channel. Both systems have been shown to be influenced/moduate/regulate by PKC (Standen and Quayle, 1998;Zhou et al., 2001; Clerice etal., 1995). Our proposal is focused on addressing the following hypotheses: Aim 1 Hypothesis: Basolateral blockade of the salt co-transporter (NKCC1) will significantly reduce stimulated fluid flow in controls but will be similar to NKCC1 knockout flow rates. We will also compare and contrast control and knockouts with NZB. We propose the use of NZB because our preliminary results indicates that the NZB acinar cells have significantly less amounts of NKCC1 than controls. Aim 2: Hypothesis: Apical membrane BKca channels contribute to normal fluid production of the lacrimal gland. We will test control (C57) and Bkca Beta 1 knockout mice. Aim 3: Hypothesis: PKC activity affects fluid production via regulation of K channels and/or NKCC1 transporters. We will test controls, NZB and the two knockouts (Beta1 and NKCC1).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A high-performance liquid chromatography (HPLC) method with electrochemical (EC) detection was developed for improved quantitation of carmustine (BCNU) in patients with brain tumor. Using this method, the plasma BCNU elimination (concentration-time) profile was characterised for a male patient who received 293mg of BCNU as an intra-arterial infusion over 45 minutes. The elimination of BCNU in this patient was found to be biphasic with an initial distribution phase occurring between 0 and 30 minutes after the end of the infusion. The new method is 50 fold more sensitive than the HPLC-UV method and permits direct on-column injection of plasma samples without extraction. Continued clinical studies of BCNU pharmacokinetics in brain tumor patients are in progress.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gap junctions serve an essential role in the passage of molecules from the cytoplasm of one cell to its neighbor in both functional and homeostatic capabilities. They are defined as clusters of closely packed intercellular membrane channels embedded in the plasma membranes of two adjoining cells. The channels are composed of two hexamers of a protein (connexon) from a family of integral membrane proteins known as eonnexins. Here, we focus on the structure and function of connexin26 (Cx26), the smallest of the family. Mutations in the DNA sequence can result in hereditary sensorineural deafness and account for between one third to one half of the cases of prelingual inherited deafness in Caucasian populations. We have isolated preparations of Cx26 gap junctions in pure and sufficient amounts for biochemical and structural studies. These 2D crystals are amenable to electron microscopy (EM) structure determination and conformational dynamics as revealed with atomic force microscopy (AFM) done under hydorated conditions. In SPECIFIC AIM 1, we will determine the structure of the Cx26 hemichannel beyond 10 A using state of the art cryo-EM and improvements on image processing procedures. This involves improving specimen preparation, imaging at either liquid nitrogen or liquid helium temperature and implementation of a combined single particle/ 2D crystallographic approach to circumvent imperfect crystal lattices. In SPECIFIC AIM 2, we will construct Cx26 wild type and mutant cell lines with a tetracysteine domain genetic tag to improve isolation with FlAsH ligand affinity bead purification, stably express these in HeLa cells or in baculovirus-infected Sf9 insect cells and isolate the gap junctions or connexons for structural analysis using the methods developed in Specific Aim 1. We will construct two Cx26 mutants (P97L and T135A), each containing a single point mutation in one of the transmembrane helices that changes the effective pore properties. These mutations should be reflected in conformational changes in the 3D structure. In SPECIFIC AIM 3, we will expand coordinated AFM/EM experiments for visualizing conformational changes due to treatments known to close or alter gap junction mediated communication. Preliminary AFM images have visualized conformational changes at submolecular resolution. We have chosen five treatments that known to induce closure of Cx26 channels or hemichannels and are physiologically relevant. Conformational changes identified by AFM imaging will be further imaged using EM. Each of these goals is intended to complement the others and lead to structural and ph),siolo_ical models of Cx26 _ermane to the entire connexin family. _ERFORMANCE SITE(S) (organization, city, state) University of California San Diego La Jolla, California KEY PERSONNEL. See instructions on Page 11. Use Name SOSINSKY, GINA E. GAIETTA, GUIDO M. OSHIMA, ATSUNORI ADAMS, STEPHEN R. DOWNING, KENNETH H. FUJIYOSHI, YOSHINORI MOLLER, DANIEL J. NICHOLSON, BRUCE J. continuation pages as neededto provide the required information Organization UC SAN DIEGO UC SAN DIEGO UC SAN DIEGO UC SAN DIEGO LAWRENCE BERKEI.EY LAB KYOTO UNIVERSITY MAX-PLANCK INSTITUTE STATE UNIV. OF NEW YORK AT BUFFALO PHS 398 (Rev. 4/98) Page 2 Number pages consecutivelyat the bottom throughout the application. Do not use suffixessuch as 3a, 3b. in the format shown below. Role on Project PRINCIPAL INVEST CO-INVESTIGATOR POSTDOC (PROPOSED) CONSULTANT CONSULTANT CONSULTANT CONSULTANT CONSULTANT Principal Investigator/Program Director (Last, first, middle): The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page ......................................................................................................................................................... 1 Description,", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The three-dimensional structure of DNA is quite dependent on sequence. This should be intuitively obvious from the sequence specificity required by many proteins for recognition. The few available x-ray crystal structures and NMR solution structures attest to this structural flexibility as well. The location and orientation of potential ligand binding functions on the DNA such as charges, hydrogen bonding and hydrophobic sites can be modified substantially from that which one might expect on the basis of assuming a canonical B-DNA structure. Consequently, we intend to continue development of the methodology for determination of high-resolution nucleic acid structures in solution and apply this methodology to some oligonucleotides and oligonucleotide complexes. This entails methods designed to improve the accuracy and resolution of the structures determined. Improved structures can be obtained with more accurate and more numerous experimental distance and torsion angle constraints, as well as improvements in calculating structure from these constraints. Enhancements will result from improvements in our iterative complete relaxation matrix program MARDIGRAS, development of a more encompassing density matrix approach for analysis of spectra derived from any pulse sequences (even those not yet invented), development of tailored excitation pulses, inclusion of experimental molecular motion information, and development of alternative methods of reducing experimental structural constraint data to structures. The latter includes (a) for restrained molecular dynamics simulations, use of improved force fields, empirical development of improved force fields, and use of constraint terms permitting a more realistic picture of conformational flexibility, and (b) development of an alternative restrained Monte Carlo method in torsion angle and helical parameter space, which is quite promising especially for structure refinement directly against NOE intensities. Applications will include oligonucleotides of interest, in particular sequences recognized by transcription factors or regulators, genome targets, antisense oligonucleotides, and a DNA microcircle duplex. Structures of proteins (including nucleic acid complexes) which are important for initiation or regulation of transcription will be determined. In particular, the 72-residue protein GerE which is a regulatory protein that binds specifically to a target site in promoter DNA will be the subject of study. Other proteins will be evaluated as possible candidates for study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary objective of this proposal is to characterize an unusual bactericidal system in normal human serum. This system requires only early-acting complement components (C1, C4 and C2) and certain serum euglobulins unrelated to known components of the conventional or alternative pathways. The essential euglobulin components will be purified and characterized by means of various column chromatographic procedures, gel electrophoresis, isoelectric focussing and immunologic techniques. The purified euglobulins will be utilized to reconstitute the bactericidal system in the presence of the early complement components + antibody. An additional objective will be to determine the range of sensitivity to this \"early complement pathway\" of representative species from several medically important genera of enteric bacteria.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Johnson C. Smith University recognizes the tremendous benefits of an active research program to our overall educational atmosphere; therefore, released time is requested for those faculty who conduct supported research. The training of students in research techniques and the methodology of research is also a major goal of the program. This proposal consists of three projects which will involve ten students yearly and utilize the expertise of five faculty. Two of the proposed projects relate to opiate addiction including: (1) study of the role of brain enkephalins and adrenergic amines in the addiction process (Cain), (2) the development of new opiate-like compounds, followed by determination of their analgesic properties (Rowe-Anderson). Also proposed is (3) a biostatistical analysis of changing antimicrobial susceptibility patterns in a large community teaching medical center (Wright and Chopra).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Applicant's Description) Neuroblastoma is the most common extra-cranial solid tumor of childhood and only 40 percent of children with high-risk features at diagnosis survive despite intensive therapy. Goal. New approaches are needed to treat high-risk neuroblastoma since maximum dose intensity alone, or without hematopoietic rescue, has only modestly improved survival. Hypothesis. We hypothesize those new therapies that are not affected by resistance of tumor cells to chemotherapy, irradiation, or 13-cis retinoic acid will significantly improve treatment of these patients. Specific Aims. (1) To establish the safety, and within the confines of phase I studies, the efficacy (a) of cytotoxic agents with new mechanisms of activity or increased tumor specificity and (b) of biologic agents which may act on resistant tumors without overlapping toxicities with cytotoxic drugs. (2) To target in vivo levels of these agents using pharmacokinetic and dosimetric studies. Research Design. Cytotoxic agents to be studied include buthionine sulfoximine (BSO) with melphalan, a synergistic combination in which BSO sensitizes tumor cells by glutathione depletion. Melphalan will be tested in both non-ablative and marrow ablative doses. A tumor targeted cytotoxic therapy to be tested is 131 I-MIBG for specific concentrated radiotherapy delivery with ablative doses of carboplatin, etoposide, and melphalan and autologous stem cell transplantation (ASCT). Fenretinide, which is cytotoxic in vitro against drug and retinoic acid resistant cell lines, and which we have shown acts by increasing tumor cell ceramide levels, will be tested in combination with tamoxifen and possibly other agents that modulate ceramide levels. A humanized antibody/GM-CSF fusion protein directed against the GD2 antigen and capable of inducing high-level neutrophil cytotoxicity against neuroblastoma cells in vitro will be evaluated. Methods. These phase I studies will be performed by a consortium of nine institutions and will include (a) patients failing front-line phase ill trials, because of poor response to induction chemotherapy; (b) patients with progressive disease; and (c) those with minimal residual disease after myeloablative therapy. These studies will provide new approaches to improving outcome in children with high-risk neuroblastoma which can be tested in larger Children's Oncology Group phase III trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fetal alcohol syndrome (FAS) is a major cause of learning and sensory deficits in people. There is growing evidence that abnormalities of neocortical function and plasticity underlie these deficits. Animals exposed to alcohol during the third trimester equivalent of human gestation and examined following a prolonged alcohol- free period were characterized by disruption of neocortical function and plasticity. However, the mechanisms by which prenatal alcohol exposure disrupts neocortical development and plasticity remain elusive. Neural plasticity in the neocortex is especially interesting in the context of the learning deficits that characterize FAS since it shares basic mechanisms with learning and memory, including a requirement for activation of the N- methyl-D-aspartate (NMDAR) receptor and the transcription factor cAMP/calcium-dependent response element binding protein (CREB), which regulates expression of genes required for cortical plasticity. Chronic alcohol exposure has important effects on NMDA receptor function, CREB activation and intracortical inhibition, all of which are crucial for cortical function and plasticity. The central hypothesis of this proposal is that these effects result in abnormal transmission of synaptic signals to the nucleus, disrupting activation of transcription factors that regulate expression of plasticity genes. The primary goal of this proposal is to rescue cortical plasticity in an animal model of FAS. The proposed studies will use molecular-genetic and pharmacological approaches to enhance transmission of synaptic signals to the nucleus of cortical neurons. The second major goal is to prevent developmental problems in the neocortex. The proposed studies will restore cortical plasticity and inhibition to normal level during and after the period when the animal is exposed to alcohol. Collectively, these studies should provide a new and exciting opportunity to elucidate how early alcohol exposure impairs cortical function and plasticity. The results of these studies may one day contribute to devise therapeutic interventions that will prevent or alleviate morbidity in FAS. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our goal is to establish an innovative, world-class GEOHealth Hub in the Alto Mayo region of Northern Peru, a heavily deforested jungle region poised on the eastern edge of the Andes that exemplifies the dynamic and dangerous interface formed between remote traditional villages and emerging agricultural and population centers. New and old environmental issues are compounded in such regions, and the populace is not yet cognizant of the risks; lessons learned in the Alto Mayo will be readily extrapolated to similar interface situations in Peru and around the globe. Multiple environmental health risks of staggering magnitude are apparent there, but have received little objective scrutiny. Strong local traditional medicine remains pervasive, dating back millennia; modern medical care is developing. Infant and maternal mortality are alarmingly high. Through international collaboration, a GEOHealth Hub in this region would provide critical support needed by Peruvian health authorities to develop well-targeted, effective public health policies and educational strategies to address bona fide, quantified environmental/occupational health risks. We will build upon an on-going, uniquely advantageous partnership that has developed between the Peruvian Institute Nacional de Salud, the Medical College of Wisconsin (MCW) Division of Pediatric Pathology, the Milwaukee-based NIH/NIEHS-sponsored Children's Environmental Health Science Core Center (CEHSCC), the Yantalo Peru Foundation, and most recently the Oregon State University Superfund Research Program. Inherent strengths of this multidisciplinary group are magnified by new availability of a sophisticated clinical and scientific facility in Yantalo, Peru, located i the heart of the Alto Mayo, designed and operated by partnership members. Our research will focus on indoor/outdoor air quality, water/soil quality, and quality and safety of traditional medicinal plant preparations. Although health concerns of the broad populace will be addressed, the critical problems of infant/maternal mortality will be emphasized. PUBLIC HEALTH RELEVANCE: Through international collaboration, this partnership will identify needs and opportunities in environmental health in the remote, but heavily deforested Alto Mayo rainforest of Northern Peru. Perceived environmental hazards in this region are staggering, but poorly understood. Through innovative pilot studies and thoughtful planning for future studies and educational approaches we will help alleviate this alarming situation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cystic fibrosis (CF) is characterized as a defect of electrolyte transport of epithelial cells of exocrine tissues. Central to the pathophysiology of CF is the absence of cAMP- stimulated Cl- secretion and an enhanced rate of Na+ absorption. The cloning of the CF gene and characterization of the gene product, the cystic fibrosis transmembrane conductance regulator (CFTR), has revealed that CFTR functions as a cAMP-stimulated Cl- channel. Cloning of the CF gene led to the creation of an animal model for CF, the CFTR (-/-) knockout mouse. Interestingly, the CFTR (-/-) mouse did not exhibit a severe CF phenotype in many epithelial tissues, including the airways. Importantly, the absence of a CF phenotype led to the formal identification of the Ca2+-regulated, or \"alternative\" Cl- conduction pathway (Cl-A) that is molecularly distinct from CFTR and plays a protective role in preventing CF pathogenesis in the airways of the CFTR (-/-) mouse. This proposal focuses on Cl-A and hypothesizes that Cl-A is an important airway epithelial ion channel resident in the apical membrane and regulated by elevation of intracellular Ca2+. Cl-A will be investigated at three levels, 1) characterization of transepithelial Cl- currents, 2) regulation of Cl-A by Ca2+-mediated signal transduction pathways, 3) patch clamp identification of the Cl-A single channel properties. Confluent polarized epithelial preparations will be used to characterize the Ca2+-stimulated Cl- current and to identify the agonists and signal transducers that regulate this conductance. Patch Cl-Amp technique will be used to identify the Cl-A single channel properties that correlate with Ca2+-stimulated currents from CF murine airway epithelial cells. Importantly all single channel studies will be performed on cells grown on a permeablized support and only channels resident in the apical membrane will be studied. Finally, comparisons between the endogenous Cl-A and heterologously expressed candidate clones will permit the unequivocal identification of the Ca2+-activated airway Cl- channel. A complete characterization of Cl-A will lead to the genesis of new therapies for CF disease that would circumvent a defective CFTR.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cell and tissue imaging core (Core C). This program project focuses on the effects of locoregional expression ofcytokines, and their effect on the tumor microenvironment. This demands study of the efficiency of delivery of tranduced cells, and multiple different analyses of the effects of these cells on the tumor. Central questions in projects are: 1) how effectively are cells delivered to the tumor? 2) Is the appropriate message and the protein product expressed by these cells? 3) is any therapeutic effect seen? This latter component includes study of cellular infiltration, angiogcnesis, tumor senescence, and cellular recruitment to the tumor environment. In each case these studies will rely critically on quantitative optical methodologies. The Center for Biolo_c lmaging, in which this core service will be performed, is designed with this function in mind. It is equipped to perform a continuum of optical methods including all types of light and electron microscopy essential to this program project. Within the scope of this project the principal goals of the core will be various. For example we will quantify and characterize cellular infiltrates into the tumor micro-environment. We will determine whether exogenously transfected cells reach tumor targets, quantify their abundance, determine whether these cells are expressing the transduced cytokines, and examine structural changes resulting from the expression of these cytokines. To perform these functions we will use a diverse array of microscopy technologies. At the light microscopic level these include: histological, immuno-histological, in situ hybridization, laser confocal, 2 photon and live cell technologies. Furthermore, our considerable experience in computerized image processing and morphometry will allow quantitative analysis of observed phenomena to corroborate earlier, possibly quite subtle qualitative changes. This core will be used extensively by all projects, though the imaging tools used will vary from project to project. In previous submissions of the proposal, this core was rated as superior, well integrated into the program and offering essential and integrated services to the program. As such the changes in this application are few, and are described in the introduction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Coronary Drug Project is a national co-operative research study testing efficacy of five therapeutic regimens in preventing death from recurrent myocardial infarction in a group of men who have suffered one or more documented myocardial infarctions prior to entry into the study. The five drug regimens under study are Dextrothyroxine, Nicotinic Acid, Clofibrate and conjugated Equine Estrogens in high and low dosage. In addition, there is a placebo group. Mortality serves as the primary end point with recurrent myocardial infarction and coronary insufficiency as some of the secondary and tertiary end points. The participants are being followed in a routine and systematic manner according to a common protocol. There is continuous safety and data monitoring of the entire research effort. The study design includes a minimum of five years of follow-up for each participant. All laboratory procedures relevant to drug efficacy and possible toxic end side effects are performed in a central laboratory. All data acquistion is submitted on standard forms to a coordinating laboratory for computer and statistical analyse. The Coronary Drug Project Aspirin Study initiated in 1972 is a double- blind clinical trial testing the efficacy of acetylsalicylate acid in reducing mortality and morbidity from recurrent coronary heart disease. Patients are followed in the same manner as the other Coronary Drug Project patients, with careful monitoring of possible toxic end side effects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our aim is to develop novel antibody conjugates for use in the radioimmuno-detection of tumors. In response to the problems associated with antibody-DTPA-radiometal imaging methodologies with regard to liver deposition of the metal, a number of more specific chelators will be investigated. Based on the capability to tightly bind the metal, and because of their greater lipophilic character and lack of excess functionality, enhancement of membrane transport of the ligand-metal complex is to be expected. The overall synthetic strategy will allow a large number of alternatives to be considered in the future, while the compounds close correlation with the ubiquitous natural products would not indicate any future cytotoxic problems. Antibody conjugation will follow protocols established in these laboratories and elsewhere. Successful completion of Phase I followed by fulfillment of the compounds potential in the Phase II in vivo studies; will lead to a new generation of tumor imaging agents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Clinical observations and research data from animal models suggest that a delay of ovulation of the preovulatory folicle can result in defective embryonic development, abortion and birth defects. Recent data indicate that alterations in hormonal concentrations due to maternal age are responsible for the detrimental changes in the oocyte that lead to subsequent abnormal development. Estadiol rises early in relation to time of ovulation during an induced delay of ovulation or in old rats during a spontaneous delay of ovulation. Absorption of this early rise in estradiol with an antiserum during prolonged estrous cycles prevented the detrimental effects of delayed ovulation. Thus it appears that age associated changes in hormonal production can cause alterations in the preovulatory oocyte. The primary site of aging is not known and could be the ovary or the hypothalamus. The present project will use orthotopic ovarian transplants between young rats and old rats before they become acyclic to differentiate aging of the ovary from aging of the hypothalamus-pituitary unit in the alteration of those hormones responsible for defects of embryonic development. Also, pregnancis resulting from ovulations from reciprocal orthotopic transplants will be used to study the role of the aging ovary on developmental anomalies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Strabismus is a misalignment of the visual axes which is relatively common in the general population with estimates of 5-6%. When this disorder occurs in infants, it can lead to severe deficiencies such as loss of central vision from one eye, known as amblyopia. Current therapies for restoration of visual alignment are based on weakening the relatively too strong antagonistic eye muscle, either by surgical recession or pharmacological denervation with botulinum toxin and muscle tightening by surgical resection. There is currently no therapeutic approach that strengthens the weaker muscle. The proposed research is based on recent evidence showing that the strength of muscles is regulated by trophic factors. Experiments were designed to identify trophic factors that either directly affect muscle fibers or provide feedback signals between eye muscles and innervating motor neurons. In the long-term, this project seeks to supplement surgical treatment of strabismus with a pharmacological treatment targeted at trophic interactions. Using an advantageous chicken animal model, we will test the hypothesis that an experimentally weakened developing eye muscle can be strengthened with trophic factors. Injections of trophic factors into selected eye muscles during a critical period of development may restore balanced eye movements by mimicking intrinsic trophic mechanisms. The effectiveness and duration of acute and chronic application of trophic factors will be explored. Additional studies will characterize adaptive responses (molecular, physiological and morphological) that are induced by denervation with botulinum toxin, mechanical denervation, or muscle lesion. Trafficking of trophic factors in mutant mouse models with motor neuron degeneration will be analyzed to identify underlying causes of motor degeneration. Weak and strong human eye muscles of different ages will be examined to compare expression of trophic factors and their receptors. Our studies will focus on trophic factors with established effects in the oculomotor system: glial cell line-derived neurotrophic factor (GDNF), insulin-like growth factors (IGFs), and cardiotrophin-1 (CT-1). A combined pharmacological, molecular, physiological and morphological approach, including the ultra-structural level, will provide a meaningful assessment of the prospects for a trophic, pharmacological treatment of strabismus and other eye muscle disorders as an alternative or supplement to current surgical and denervation procedures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our main research interests reside in the intracellular events -changes in signaling and in gene expression- which direct T-lymphocyte differentiation and survival, both during intrathymic T-cell development and in mature T-cells. Our first experimental approach focuses on T-cell positive selection, an intrathymic process which ensures the generation of an efficient T-cell repertoire by rescuing 'useful' thymocytes from programmed cell death and by inducing their differentiation into mature T-cells. While it has been shown that T-cell Receptor (TCR) engagement on thymocytes is necessary to trigger positive selection, it has remained unclear whether attributes of TCR signals (including their strength or duration) also dictate subsequent differentiation or survival events required for complete intrathymic maturation. To address these issues, we have generated a recombinant mouse model in which expression of Zap70, a tyrosine kinase required for TCR signal transduction, decreases in thymocytes that have been signaled to undergo positive selection. Thus, thymocytes in such mice can only receive transient, but not persistent, TCR signals. Using this model, we have shown that interruption of intrathymic TCR signaling in vivo (i) prevents thymocytes from completing their intrathymic maturation, despite the fact that they undergo initial differentiation events that characterize positive selection and (ii) precludes the differentiation of CD4-lineage T-cells. By breeding these mice with mouse lines carrying defined genetic modifications, we are currently investigating the intracellular signaling pathways which transduce differentiation and survival signals during intrathymic selection. As a second approach, we are developing conditional versions of intracellular signaling molecules that can be turned 'on' or 'off' by small-size ligands, to interfere with T-cell signal transduction in vivo and to monitor signaling and gene expression during the course of an immune response or during intrathymic development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This work was aimed towards determining the biochemical function of calretinin. The initial results describe some of the basic characteristics of calretinin and will serve as the basis of further studies. These include the following: (1) Calretinin was identified in membrane fractions of rat cerebellum by both radioimmunoassay and immunoblot. Calcium produced an increase in the amount of calretinin associated with the synaptic membrane fraction and decreased the calretinin in cytosol relative to that found in the presence of EGTA; (2) Several approaches were used to search for calretinin target proteins. Iodinated calretinin was found to bind to proteins in several subcellular fractions on nitrocellulose blots of one dimensional SDS gels; (3) A purification procedure was developed for obtaining large amounts of homogeneous recombinant calretinin from plasmids. No differences were found in the physicochemical properties of native and recombinant calretinin (molecular weight, isoelectric point, calcium binding kinetics and behavior on ion exchange: gel filtration and hydrophobic columns); (4) A series of experiments used specific resins to study the hydrophobic properties of calretinin and the resins. A partial calcium dependence of calretinin binding observed for some resins (octylsepharose) was dependent on ionic strength and pH.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 70% of patients with Chronic Obstructive Pulmonary Disease (COPD) are obese or overweight. Tobacco smoking that causes COPD also is associated with other poor health habits that can lead to obesity and cardiovascular disease. Symptoms of COPD are often non-specific and include dyspnea and exercise intolerance. COPD treatment guidelines recommend stepped escalation of inhaled medications to improve these symptoms, but make little mention of the effect of co-existing obesity or weight loss interventions because of insufficient evidence. Cardiovascular disease is a leading cause of mortality among patients with COPD, and obesity is associated with important risk factors for cardiovascular disease including dyslipidemia, hypertension, and diabetes. Comprehensive lifestyle interventions that include calorie-controlled healthy eating, increased physical activity, and behavioral self-management strategies consistently result in modest, clinically significant weight loss and associated reductions in cardiovascular risk factors. That overweight and obese patients with COPD would not only reap similar clinical benefits from modest weight loss is an intuitive - but untested - concept. Therefore, we propose to conduct a highly valid, multicenter, patient-level randomized, pragmatic clinical trial to produce first-ever data on the effectiveness of a self-directed 12-month evidence-based lifestyle intervention targeting modest weight loss and increased physical activity among overweight and obese patients with COPD. We aim to serve as a Protocol Leadership Group (PLG) for the National Heart, Lung, and Blood Institute's Multi-Site Clinical Trials for the Pulmonary Trials Cooperative (PTC) in order to test if intervention participants have better outcomes through 24 months of follow-up compared to usual care control patients in terms of weight loss, exercise tolerance using the 6-Minute Walk Test, and dyspnea using the Borg dyspnea score. Secondary outcomes include, generic health-related quality of life using the SF-12, and major cardiovascular risk factors using Framingham risk score (including lipids), central obesity by waist circumference and blood pressure. We will oversee enrollment of 1000 patients at multiple clinical sites eventually chosen and contracted by the PTC's Network Management Core (NEMO). As the PLG for our proposed HOPE trial, we will cooperate with NEMO and will provide trial oversight, data management and reporting, and safety monitoring.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research will examine the role of prolactin, estradiol and brain catecholaminergic and LHRH neurons in the age-related transition to irregular estrous cyclicity and eventual complete acyclicity. Experiments, using the 8-16 month old female rat as an expermental model, have been designed to determine which of the following working hypotheses is (are) the most likely explanations(s) for the onset of irregular cyclicity and acyclicity: (1) Prolactin and/or estradiol may become elevated in aging females and feedback to alter the cyclic profile of catecholamine-LHRH dynamics and cyclic gonadotropin release. (2) Age-related changes at the hypothalamic level may change the synthesis, metabolism and/or release of catecholamines and/or LHRH. The spontaneous neural changes result in an acyclic pattern of gonadotropin release. (3) Alterations in the responsiveness of the preoptic area to the positive feedback of estradiol may be the primary age-related change such that steroids become incapable of stimulating cyclic gonadotropin surges. (4) Changes in pituitary lactotroph sensitivity to dopamine and/or estradiol may occur during the aging process and lead to changes in prolactin secretion and ultimately to acyclicity. It is possible to test these hypotheses because of several recent methodological advances. A microdissection technique used in conjunction with sensitive radioenzymatic and radioimmunoassays for norepinephine dopamine and LHRH permit analysis of changes in these parameters in specific brain structures which are known to be involved in the basal secretion and surge of gonadotropin and prolactin. Catecholamine turnover rates (alpha methylpara tyrosine method) will be used as an index of neurona activity. New receptor assays for estradiol and prolactin allow examination of changes in receptor binding in the brain and pituitary with minimal pooling animals for tissue samples. These technique allow for neuroendocrine analyses to proceed at a greater resolution than previously possible such that the questions posed in this proposal may be approached.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT The human Brainspan data was created to identify all transcripts involved in neural development and to help understand of how specific risk genes affect human brain development. In addition, these data will have important clinical relevance for translational medicine; these data can help discern which risk alleles associated with psychiatric and neurological disorders influence transcription and alternative splicing across different regions and developmental stages. Also, most Brainspan samples were processing for whole-genome sequencing (WGS) and/or DNA methylation analysis, which enables direct comparisons of single basepair changes, copy number variation, and RNA editing events in the developing human brain. As such, Brainspan data holds biologically and clinically important data on the genetic and molecular mechanisms underlying the development and increased disease susceptibility of the human brain. To expand upon this resource, we aim to create a matched profile of the human brains RNA modification landscape (epitranscriptome), for both methyl-6-adenosine (m6A) and 5-methyl-cytosine (5mC). We will profile the developmental trajectory of the RNA modifications and their activity in non-coding regions and impact on splicing, RNA editing, AU-rich regulation of transcripts, and association with DNA methylation changes (epigenetics). Finally, we will also test the impact of these modifications from patient-derived iPS cells that will be grown and assayed over five time points. This will be accomplished over five years, and across 1,075 samples, across the Mason and Sestan labs, with collaborators at the Broad institute available to help with assays and access to GTEx data from adult brains with m6A profiles. We will achieve these goals across three main aims. (1) Create a neuro-developmental map for epitranscriptome sites and levels, with an emphasis on m6A and m5C, for 35 brains from four time periods, and five regions of the brain, chosen based on their large differences seen in the BrainSpan data and prior implication in neurological development. (2) Detail the inter-individual variation in epitranscriptome levels and their epigenetic regulation using m6A variation with the changes in expression levels, and then link epigenetic changes to altered gene expression and m6A regulation. (3) We will delineate the epitranscriptome changes in autism brains and manifestation in patient-derived iPS cells, including an examination of epitranscriptome variation across 30 banked Autistic brain samples and testing of the impact on disruption of the readers and writers of RNA regulation (on induced pluripotent stem cells). These will represent the first-ever epitranscriptome maps from primary tissue of Autism brains and help guide future studies that examine the dysregulation of Autism gene expression networks and epitranscriptome states.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Macular Edema contributes to many of the leading causes of blindness in America including diabetes, age related macular degeneration and uveitis. Currently, a wide number of studies reveal that altered expression of cytokines, including vascular endothelial growth factor and tumor necrosis factor act to increase blood vessel permeability. Further, research from our laboratory, as well as others, reveals activation of atypical protein kinase C isoforms are required for the permeability response for these and other permeabilizing factors. By screening a commercially available library, we have already identified a class of inhibitors for this target and in this grant we propose to combine medicinal chemists, structural biologists and cellular and molecular physiologists to develop compounds that control retinal blood vessel permeability in multiple models of eye disease. Importantly, measures of retinal function will mimic clinical assessments. The successful completion of these studies will provide a robust chemical pharmacophore, pharmacokinetic analysis, mechanism of action and in vivo effectiveness for atypical protein kinase C inhibitors to treat macular edema with specific leads available for clinical trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Core C: Data Management and Statistical Core Project Summary/Abstract Efficient, clean, and organized data, together with valid, flexible, and reproducible statistical design and analysis techniques, are an integral component to achieving the scientific mission of the UCI ADRC: to discover, quantify, and validate factors that influence the risk of AD across the lifespan. The overarching goal of the Data Management and Statistical Core (DMSC) is to provide logistical and intellectual support to all ADRC investigators at all phases of scientific projects. Data management support includes comprehensive data management and dissemination of data arising from the Uniform Data Set (UDS) and all ADRC projects. DMSC personnel are responsible for the development of National Alzheimer's Coordinating Center (NACC) approved collection forms, maintenance of the ADRC database, and the timely transfer of accurate data to the NACC. DMSC faculty and staff will also be intimately involved in the conception, design, implementation, analysis, and reporting of research conducted by members of the ADRC. The DMSC emphasizes regular communication with ADRC investigators and proactive involvement in ADRC-sponsored projects. The DMSC also promotes the development of independent research programs among its members. During the proposed research period, projects will include the development and validation of novel data collection mechanisms for harmonizing heterogenous Big Data streams, as well as the development of novel statistical methodology for efficient biomarker discovery and validation. These DMSC-specific research endeavors will increase the intellectual contribution of the DMSC to the scientific community at large, and will also lead to improved methods for collecting, entering, and analyzing complex data obtained through local ADRC projects. !", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Translational research, bringing new laboratory findings quickly to improve prevention, treatment, quality of life, and survival for breast cancer patients, has been the focus of the team now forming the Baylor Breast Center for 25 years. During the first ten years of our SPORE, our tumor bank which made much of this rapid translation possible became a national resource, while basic cell and molecular biology research suggested new clinical implications for endocrine and chemo- therapy resistance, breast cancer prevention, metastasis, development of premalignant lesions, and roles for tumor suppressor genes BRCA-1 and -2 in DNA repair. Developmental projects ranged even further in seeking new translational possibilities. In this SPORE renewal, we build on the results developed in our earlier work and on new findings and new technologies, in several directions. (l) Compelling new data indicates that cross-talk from growth factor and stress pathways to the estrogen receptor and its coactivators may lie at the root of clinical resistance to tamoxifen, and suggests feasible clinical interventions. We will lay the preclinical groundwork for definitive trials and examine in clinical specimens the prognostic and predictive value of key intermediates from these pathways. (2) Our gone expression array studies offer the promise of a rational risk classification of DCIS. We will assess key markers indicated by these studies on a unique series of DCIS specimens, to evaluate their potential to predict progression to invasive cancer. (3) Tamoxifen can prevent ER-positive breast cancer in many high-risk women, but does nothing against the emergence of ER-negative disease. Based on ongoing treatment studies plus our own laboratory data, we now propose preclinical studies and an early clinical trial of the receptor tyrosine kinase inhibitor Iressa in chemoprevention of breast cancer in women at risk. (4) Our discovery that a single hypersensitive estrogen receptor mutant is found in many hyperplasias and in most node-positive breast cancers, but not in normal tissue, suggests that this mutant could be not only a powerful risk indicator but a uniquely specific treatment target. We will explore both possibilities. (5) Our preliminary expression array data in responders vs. nonresponders to neoadjuvant Taxotere implies that the gone expression profile might strongly predict clinical sensitivity or resistance. We will test in a randomized trial the ability of expression profiles to distinguish Taxotere from AC sensitivity, and confirm markers from these profiles in sets of retrospective clinical specimens. (6) Although important parts of our unique breast Tissue Resource were lost in last year's floods, much remains, and we propose important new accessions to further enhance this critical resource. (7) Pathology, Biostatistics, and Administrative Cores will also support key aspects of the proposed work. (8) Our highly successful Developmental Projects Program and Career Development Program will continue to encourage new ideas and new investigators in translational breast cancer research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the MENTORS (Model Education Networks to Optimize Rural Science) Project is to actualize an exportable model of development and dissemination of culturally relevant and innovative programs that stimulate interest in and enhance preparation for careers in science, technology, engineering and medicine. Integrated components that will provide mentoring and career preparation include: summer research internships, science exploration camp, Field Experiences (FEs), and innovative classroom activities for students, and a rigorous professional development (PD) program for K-12 teachers. Our project will build strong partnerships between biomedical, engineering, public health and education researchers at Texas A&M University and K-12 teachers, school/district administrators, students and parents. MENTORS focuses on school districts that are rural and have a high proportion of students who are underserved and/or underrepresented in science and medicine. Such districts often lack programs to optimally prepare students for STEM careers. The multi-disciplinary Project team, which includes experts in biomedical, health disparities and biomedical engineering research as well as K-12 educational theory, curriculum development and evaluation, will work with classroom teachers to develop innovative, career based, educational modules focused on 21st century skills. The Specific Aims of the MENTORS Project are: 1. To provide authentic research and field experiences (FEs) for elementary, middle and high school students, designed to enhance STEM education and stimulate interest in and pursuit of a broad variety of science, technology, engineering and health-related careers. 2. To engage a network of K-12 teachers and biomedical, public health, biomedical engineering and educational researchers to develop learner-centered, career focused educational modules that will attract and motivate students, particularly those who are traditionally underrepresented in science, medicine and technology. 3. To provide rigorous professional development for K-12 teachers in TX, through an established and impactful Summer Institute (SI). All activities and products will be extensively evaluated using a mixed methods approach, with a variety of instruments designed to collect and analyze the most pertinent data. These will assess the effectiveness of the model as a whole, and of the components, in achieving the objectives of enhancing student interest and developing the skills needed for educational and career success.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term aim of the proposed research is to define the molecular structure and function of the midkine signal transduction pathway which regulates cellular proliferation and differentiation Special emphasis in this effort is placed on the molecular cloning of the cell surface Midkine signal transduction receptor which we have recently identify from a Wilms~ tumor cell line using biochemical techniques. Recent studies have established the importance of Midkine expression in Wilms~ tumor, renal cell carcinoma and lung, colon and hepatocellular carcinoma. Our studies have also shown Midkine expression in autosomal dominant polycystic kidney disease (ADPKD) and its role as an important mediator in epithelial differentiation during kidney organogenesis. Our hypothesis is that Midkine functions as an autocrine or paracrine which regulates several biological processes including the growth regulation of epithelial cells in ADPKD as well as in oncogenic transformation in Wilms~ tumor and in differential mechanisms which regulate epithelial conversion of renal progenitor cells during renal development. We plan to clone Midkine receptor gene using either an expression cloning approach or ligand affinity chromatography to purify the MK receptor protein. Additionally, we also plan to examine Midkine signal transduction pathway, then define the basic developmental pattern in the kidney of expression of all identified components of the midkine signal transduction pathway including expression of the Midkine receptor in relation to its ligand.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Bleomycin is the name which has been given to a family of antitumor antibiotics, all of which are closely related structurally and can be derived synthetically from bleomycinic acid. The bleomycins are of interest because of their clinically useful activity against, e.g. skin, lung and testicular cancer. We propose a total synthesis of bleomycin B2, MW 1342, having 19 asymmetric centers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Aromatic hydrocarbons and their chlorinated derivatives are environmental pollutants that are, in many cases, toxic to most forms of life. Microorganisms that degrade toluene and naphthalene to carbon dioxide and water initiate metabolism by the enzymatic incorporation of molecular oxygen into the aromatic nucleus. These enzymes are called dioxygenases and are known to be multienzyme systems. The components of the toluene and naphthalene dioxygenases will be purified and characterized and studies on the mechanism of enzymatic oxygen fixation will be initiated. The products formed by the action of these dioxygenases on a variety of aromatic hydrocarbons and their substituted derivatives will be isolated and characterized by conventional chemical techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Trichotillomania (TTM) is a chronic impulse control disorder characterized by pulling out one's own hair, resulting in noticeable hair loss. Although the disorder typically begins in early adolescence, there are few data available to guide treatment choice for youth with TTM. Our research group developed and empirically evaluated a manualized BT program for pediatric TTM (R21 MH61457 - M. E. Franklin, PI), completing what is to our knowledge the only randomized controlled trial (RCT) of any treatment for pediatric TTM. Results indicated a clear advantage for BT over a Minimal Attention Control (MAC) condition at post-treatment, with gains well maintained up to six months for those randomized to BT. With this encouraging preliminary work as our starting point, we now seek to replicate the findings in a larger sample and extend them by employing a more scientifically rigorous comparison condition, Supportive Counseling (SC), to allow us to rule out non-specific factors such as psychoeducation and therapist contact time as the reasons for TTM symptom reduction. Our primary specific aims are to: 1) compare the efficacy of BT to SC for treating TTM at post-treatment (week 8); and 2) examine maintenance of BT gains through an eight week maintenance phase (week 16). We also plan to: 3) examine maintenance of BT gains through a naturalistic follow-up phase (weeks 28, 40); and 4) explore predictors of acute (week 8) and long-term (weeks 16, 28, & 40) response to BT, including comorbidity, initial severity, and family psychopathology. Public Health Relevance: Development of an efficacious and transportable treatment for youth with TTM may provide front-line clinicians with specific strategies for helping these youngsters now. Moreover, effective treatment delivered closer in time to the onset of the disorder may serve as a form of prevention for the psychiatric sequelae that appear to develop in the wake of TTM in adults. The proposed study constitutes a logical and necessary next step in developing an adequate evidence base for BT in pediatric TTM, and lays the groundwork for future transportability studies where its effectiveness can be examined in the settings where most families can access pediatric mental health services. Development of an efficacious and transportable treatment for youth with TTM may provide front-line clinicians with specific strategies for helping these youngsters now. Moreover, effective treatment delivered closer in time to the onset of the disorder may serve as a form of prevention for the psychiatric sequelae that appear to develop in the wake of TTM in adults. The proposed study constitutes a logical and necessary next step in developing an adequate evidence base for BT in pediatric TTM, and lays the groundwork for future transportability studies where its effectiveness can be examined in the settings where most families can access pediatric mental health services.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Concurrent with the increased prevalence of overweight and obesity in the United States, a significant portion of older adults are increasingly at risk for developing metabolic dysfunction. Metabolic abnormalities can include insulin resistance or impaired glucose tolerance, dyslipidemia, hypertension, and abdominal obesity. These metabolic abnormalities tend to cluster together, forming what is known as metabolic syndrome (MetSy). MetSy is widely considered a \"pre-disease\" state associated with development of Type II Diabetes as well as cardiovascular and cerebrovascular disease. Several studies have proposed links between components of MetSy and neurocognitive and brain dysfunction;however, few studies have examined the additive effect of the cluster of risk factors that comprise the syndrome! The long-term objective of this study is to uncover evidence that individuals with MetSy, who by definition do not exhibit any overt disease process, will demonstrate subtle differences in performance and abnormal FMRI brain response on cognitive tasks. The specific aims of the study are to demonstrate subtle deficits in verbal working memory (VWM) performance and abnormalities in task-related BOLD FMRI brain response in individuals with MetSy. It is hypothesized that in comparison to a well-matched control group, individuals with MetSy: (a) will be less accurate and slower in their reaction times on the 2-back VWM test;(b) will exhibit differences in 2-back performance that will increase in relation to increased task difficulty;(c) will have reduced BOLD response in brain systems involved in VWM including the dorsolateral prefrontal cortex, supplemental motor area, and the posterior parietal cortex;(d) will exhibit compensatory responses in areas outside the primary VWM system. Additionally, it is hypothesized that increases in brain response in relation to task demand will be higher in individuals with MetSy. Relevance to Public Health In the United States, 24% of adults and nearly 44% of adults over the age of 50 years meet the clinical criteria for metabolic syndrome. Thus, this project will yield significant clinical application for millions of Americans with metabolic syndrome by providing evidence that cerebrovascular and cognitive function are affected by subtle vascular and non-vascular abnormalities before clinical signs of major disease exist.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fragile X syndrome is the most common genetic cause of autism, occurring in 1 out of 6000 births. Affected patients also suffer from mental retardation and in some cases seizures. Current treatments involve the use of drugs to ameliorate mood and attention symptoms and to prevent seizures, but are not able to restore normal learning and emotional function. A molecular-level understanding of the neuronal defects in Fragile X syndrome will be necessary for the rational design of therapies to address the underlying cause of the disease. The protein mutated in the disease, the Fragile X mental retardation protein (FMRP), is required for regulating protein synthesis at activated synapses, the communication points between neurons. A large body of evidence suggests that the normal strengthening and weakening of synapses that underlies learning requires the careful regulation of protein synthesis by synaptic activity. Experiments have also suggested a role for FMRP in regulating both synaptic strengthening and weakening. However, the precise relationship between synaptic strengthening and weakening, protein synthesis, and FMRP is poorly understood. For instance, which proteins are synthesized during, utilized in, or required for synaptic strengthening and weakening, and which of these events are affected by FMRP loss, is not known. Research on the function of FMRP in activity-dependent local protein synthesis has been limited by the low sensitivity and resolution of methods for assessing and controlling protein synthesis in living neurons. We have developed new molecular tools that allow the real-time tracking and control of new protein synthesis and the visualization of kinase pathways involved in activity-induced protein synthesis. We propose to use these tools to examine the specificity of protein synthesis responses in synaptic strengthening versus weakening, and to study the effect of FMRP loss on these responses. We will also determine which new proteins are normally required for long-lasting synaptic plasticity, and how FMRP loss might alter those requirements. These studies will provide insight into the regulation and function of synaptic protein synthesis in persistent synaptic plasticity, identify potential molecular targets for therapeutic intervention, and produce new technologies that can benefit the larger neuroscience community.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Vitellogenin, the yolk protein precursor, serves as an excellent model protein to study the hormonal regulation of transcriptional, translational and posttranslational events. This proposal focuses on one such posttranslational event, the phosphorylation of over 90 serine hydroxyl groups in each vitellogenin polypeptide during its translocation through the secretary apparatus of the hepatocyte. Xenopus vitellogenin microheterogeneity will first be examined by resolving vitellogenin and yolk protein polypeptides and testing whether this heterogeneity arises from differential modification of a common polypeptide or rather from slightly different primary sequences. A variety of useful substrates, both in phosphorylated and unphosphorylated form, should be concomitantly obtained during this effort. Direct evidence will be sought that vitellogenin represents the principal endogenous substrate in liver microsomes from estrogen-treated animals, and optimum procedures will be defined for the extraction and purification of vitellogenin kinase activity from such liver microsomes. During kinase purification, the existence of multiple kinases will also be explored. Purified kinase(s) will be characterized, assigned to microsomal subfractions, and related to cytosolic \"phosvitin kinase.\" Male liver microsomes will also be examined for vitellogenin kinase activity, and vitellogenin phosphorylation and processing will be examined in Xenopus oocytes microinjected with vitellogenin mRNA. These studies should accomplish several goals, including (a) elucidation of the relationship among the vitellogenin and yolk protein polypeptides and the recently described 4 vitellogenin genes in Xenopus, (b) identification of the mechanism by which vitellogenin is phosphorylated during its secretory translocation, (c) indication of the relationship, if any, between the vitellogenin kinase(s) and the ubiquitous \"phosvitin kinases\" described by numerous laboratories and for which there is as yet no known biological function, and (d) documentation of whether vitellogenin kinase is coordinately induced in liver by estrogen or rather is a constituent enzyme of the endoplasmic reticulum/Golgi elements of all xeropus cells. In addition, these studies will allow for the first time a direct comparison between vitellogenin phosphorylation and an analogous (perhaps even homologous) system: the phosphorylation of milk protein (caseins).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Mouse Production and Ventricular Function Core (Unit C) has two major aims as dictated by Projects 1-3: 1) production of mutant mouse models of familial hypertrophic cardiomyopathy (FHC) and dilated cardiomyopathy (DCM) and 2) characterization of the phenotypes of these animals using both in vivo and in vitro methods. Production of mice will be accomplished in the laboratory of our co-investigators Drs. Jeffrey Robbins and James Gulick (Univ. of Cincinnati) using overexpression of transgenes linked to a cardiac specific myosin heavy chain promoter. Our collaborators Drs. Jon Seidman (Harvard Medical School) and James Lessard (Univ. of Cincinati) will provide additional mice with both myosin and actin mutations. In vivo phenotypic characterization will take place in the laboratory of Dr. David Kass (Johns Hopkins School of Medicine) by use of a miniaturized conductance/micromanometer catheter system for simultaneous measurement of left ventricular (LV) pressure and volume under near physiologic conditions. These measurements allow estimation of a variety of sophisticated parameters of systolic and diastolic function that have been previously characterized in larger mammals, including man. In vitro phenotypic characterization will be accomplished in the laboratory of Dr. Martin LeWinter (University of Vermont), using a buffer-perfused, isovolumically contracting, isolated heart preparation. The latter preparation provides estimates of a number of functional parameters under less physiologic but more controlled experimental conditions than are possible in vivo. It also provides a mechanoenergetic analysis of the efficiency and economy of the contractile machinery by relating its mechanical output (pressure-volume area or force-time integral) to its chemical energy input (oxygen consumption). In vivo and in vitro LV chamber properties delineated in the Core will be used to better understand how specific mutations result in a FHC versus a DCM phenotype in two ways: 1) they will be correlated with muscle strip (myofilament) and myofibril properties determined in Project 3 as a component of systematic integration of the results of all of the Projects and the Core at increasing levels of system complexity;2) by studying the temporal evolution of phenotypes in selected FHC and DCM models, we will determine which chamber properties are primary and which are secondary manifestations of specific mutations. The latter studies should lead to a better mechanistic understanding of the adaptations and maladaptations that occur in response to sarcomeric protein mutations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the administrative core of the University of Colorado Lung Cancer SPORE is to provide outstanding administrative and fiscal support for the entire program effort and to provide the scientific leadership for the program. The administrative core will oversee all administrative and scientific activities of the SPORE program, review and regulate financial expenditures, develop and prepare reports. The Administrative core consists of the two SPORE principal investigators, Drs. Bunn and Miller; the SPORE executive director. Donna Berrier, a 40% grants manager/administrative assistant and 10% financial manager. This core will also develop and circulate research conference schedules, coordinate scientific review, schedule the monthly scientific meetings and aid project investigators in the preparation and publication of manuscripts as well as maintain a record of all publications emanating from this grant. It will oversee the planning and evaluation efforts including the scheduling of visits by the external advisors, the planning and coordinating of the yearly internal retreats and yearly NCI SPORE meetings, the scheduling of meetings and scientists, Executive Committee, Developmental Research Committee and Career Development Committee and the SPORE advocacy program.. The Administrative Core works with the SPORE investigators and NCI program staff to insure compliance with all federal regulations and reporting requirements. It will coordinate activities with the Cancer Center and with other SPORES to ensure that there is no redundancy, and to ensure joint projects are conducted in the most economical way. The Administrative Core will assist in community outreach efforts particularly with respect to public relations and community activities through the established Cancer Center mechanisms. The core provides support for the development and career development programs as well as Visiting scientist program. The Administrative core oversees the functioning of the other four core resources.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Monocyte-derived dendritic cells (DC) are currently used in clinical trials as carders of anti-cancer vaccines. As it has been shown that DC developing and maturing in different conditions show strong differences in their abilities to produce cytokines and to induce Th1, Th2, and CTL responses, it is likely that DC will also differ in their ability to exert antitumor therapeutic effect. Despite extensive in vitro characterization of distinct functional subsets of DC, their ability to induce immune responses of different character and magnitude has not been tested in vivo. We propose to develop a model system for evaluating and optimizing myeloid and plasmacytoid DC function in rhesus macaques. Based on previous results, we hypothesize that polarized myeloid DC1, grown in GM-CSF and IL-4 and which in vitro produce high levels of IL-12 and preferentially induce Th1 and CTL responses, will prove to be the most potent DC for stimulating Th1 and CTL responses in vivo. However, plasmacytoid DC have also been shown to induce Th1 and CTL responses, as have DC cultured in GM-CSF and IL-15. We propose to test the efficacy of DC generated in different protocols and at different stages of maturation, polarized by different sets of cytokines, as well as exposed to multiple Forms of antigen, to induce different classes of immune responses in vitro and in vivo. We will first develop protocols for generating rhesus DC, loading them with antigens, and inducing polarized phenotypes in vitro. The phenotype and functional capacities of these cells, including cytokines produced, will be characterized extensively. We will next test the different types of DC for efficient localization in T cell areas of lymph nodes after intranodal injection. Finally, we will test in vivo the ability of different DC types to stimulate polarized CD4 T cell responses to antigens and to stimulate CD8 T cells responses. Immunization strategies that provoke strong Th1-type responses will potentially be used for clinical trials being performed in other projects within this P01, and in future studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Since parent training as compared to direct clinic treatment is potentially a more economical and effective means of providing treatment, the field in general is moving in this direction as a method of improving the treatment of autistic children. Both the review of the literature and our pilot data provide compelling evidence that parent training is a plausible means of increasing the availability, intensiveness and effectiveness of the treatment as compared to direct clinic intervention. However, many of these results were based on post hoc analyses or case studies. The necessary systematic comparison has not been conducted. Such a comparison will hopefully allow us to assess the relative strengths and weaknesses of each of these approaches. This is important because it is necessary for the development of a treatment program encompassing the strengths of both approaches. Also, no one has yet analyzed the effect of either of these approaches on family members other than the autistic child. Without such an analysis, it is impossible to know, from the point of view of the family, whether or not the effect of the treatment is worthwhile. To evaluate the relative strengths and weaknesses of these two approaches (parent training and direct clinic treatment), and to assess the impact on the family, we propose to compare two groups. One group will receive the parent-training program, and no clinic treatment. A control group will receive direct clinic treatment for the child and no training for the parents. The measures which will be analyzed include: (1) direct measures of the child's behavior; (2) measures of parent/child interactions in both home and laboratory settings; and (3) measures of parental psychological and marital adjustment, and attitudes toward their child's potential.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The present proposal examines the principles governing language comprehension in reading. The proposed studies examine the principles underlying the syntactic analysis of sentences, principles determining the semantic interpretation of sentences, and the relation between them. The experiments all involve recording subjects' eye movements as they read sentences or short texts. Average reading time per letter in various critical regions of the sentence is used as a measure of processing complexity, along with the gaze durations associated with each word, total sentence reading time, and the probability of making a regressive eye movement which returns to portions of the text which have already been read.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Stanford Proteomic Shared Resource is a state-of-the-art proteomics facility dedicated to providing rapid and low-cost support for proteomic methods to Stanford Cancer Center investigators. Services include protein purification, peptide and protein identification, and analytical characterization of modifications and other small compounds. The Shared Resource has the necessary scientific and technical foundation, as well as a clean physical plant to conduct high accuracy mass spectrometry. Substantial cost savings, efficiency, and scientific advancement is provided by this type of centralized facility. An advisory board provides recommendations on facility policies and activities, ensuring that this Shared Resource is aligned with the Cancer Center's strategic goals. In fiscal year 2005, the Proteomics Shared Resource has performed close to 600 proteomic analyses for 30 different laboratories, including 18 members of the Cancer Center, representing 6 of the 10 Cancer Center Programs. Use in fiscal year 2005 reflected an increase of 60% as compared to 2004. Future plans for the Shared Resource include expansion of capacity and capabilities with new mass spectrometers, an ongoing effort in improved and novel analysis software, and development of chromatographic strategies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Posterior tibial tendon dysfunction (PTTD) results in a debilitating flatfoot deformity which is increasingly recognized as a major problem in the treatment of foot and ankle disorders. In fact, this disorder is felt to be the number one foot and ankle problem requiring intensive scientific investigation. The deformity is caused, at least in part, by the loss of the tendon function with subsequent muscle imbalance. Bony procedures without major fusions are being used in an effort to treat deformity when possible. However, with a tendon transfer of one third the muscle mass and a similarly smaller tendon, the original problem of muscle imbalance remains. If the problem of muscle imbalance and increased strain could be resolved, patients could be treated early to eliminate those factors that promote deformity and decreased function. This would make for a successful long-term result. A cadaveric model has been developed for the posterior tibial tendon deficient foot. Physiologic tendon, bone and ground reaction loads can be applied and monitored, and the resulting strain in the arch of the foot can be measured. Using this model, the following fundamental questions in the treatment of PTTD will be addressed: 1) does loss of the posterior tibial tendon increase strain in the arch and thereby promote deformity?; 2) how well does the present flexor digitorum tendon transfer replace the posterior tibial tendon muscle unit and does it leave muscle imbalance and increased strain in the arch?; 3) does a bony procedure, like a medial calcaneal displacement osteotomy, adequately assist the flexor digitorum tendon transfer?; and, most importantly, 4) if the flexor digitorum longus tendon transfer is inadequate (as would be expected based on clinical results), are there procedures that can adequately normalize strain and joint position?", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a renewal application, and would provide funding to administer the Neurosurgeon Research Career Development Program (NRCDP) for another five years. The purpose of this award is to provide the basis for a continued national effort to support, train, and mentor junior neurosurgical faculty members at appropriate institutions nationwide. This application describes all key aspects for the mechanism of recruiting applicants and the developing them into surgeon-scientists through continuing the NRCDP. The proposal addresses topics such as the rationale for the program, the relevant history of the PI and Institution, and the ongoing need for the NRCDP. The proposal also addresses the rationale for the selection of a National Advisory Committee (NAC). It also details a well-developed system for the recruitment of applicants. In addition, there is a description of the existing and robust system for selection of scholar-applicants based on their potential for success, the track record of their mentor, and the support of their Chairpersons. In addition, there are details of an annual three-day retreat, which has been held for five years, that includes applicant interviews, lectures, and interactive grant writing sessions. Finally, there is a description of the advisory board meeting for review of applications. This program has been extremely successful. Five years ago, when this NRCDP was first awarded, a search of NINDS data revealed only 4 neurosurgeons with active K08 awards. Currently, there are a total of 12 neurosurgeons with active K08 awards. Of the currently active K08 awards, four are scholars of this NRCDP program and two were applicants. In other words, over the five-year span of the NRCDP, there has been roughly a 300% increase in neurosurgeon K08 awards. Moreover, 50% of all current NINDS K08?s, awarded to neurosurgeons, are affiliated with the NRCDP either through direct support or through participation in the program. Another three alumni obtained independent federal funding through other mechanisms. If the scholars funded in 2016 are excluded from the analysis, which is reasonable given that they have only been funded for a few months, then 7 out of 10, or 70%, of scholars have successfully obtained independent federal funding in a relatively short period of time. In our opinion, this is an excellent success rate. Perhaps as importantly, the NRCDP has fostered a healthy and burgeoning community of junior neurosurgeon-scientists that will form the basis for reinvigoration the long tradition of scientific research in neurosurgery.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent investigations have identified a new signal transduction pathway, termed the sphingomyelin pathway, that may mediate the action of tumor necrosis factor (TNF)-alpha and interleukin 1 (IL-1) Beta. This pathway is initiated by hydrolysis of plasma membrane sphingomyelin to ceramide by the action of a neutral sphingomyelinase. Ceramide functions as a second messenger stimulating a Ser/Thr proteins kinase termed ceramide- activated protein kinase to transduce the signal. Three lines of evidence support this notion. Firstly, the sphingomyelin pathway is activated within seconds to minutes by TNF in HL-60 cells and by IL-1 in human dermal fibroblasts and mouse EL4 T helper cells. Secondly, cell- permeable ceramide analogs bypass receptor activation and directly mimic cytokine action. Thirdly, the effects of TNF and IL-1 on this cascade have been reconstituted in cell-free extracts indicating tight coupling of this pathway to the respective receptors. This grant proposal focusses on the elucidation of the role of ceramide-activated protein kinase in this signaling cascade. Until now, this activity has only been defined as a Mg2+ dependent, membrane-bound activity capable of phosphorylating a peptide derived from this amino acid sequenced surrounding Thr669 of the epidermal growth factor receptor. This is the same site recognized by mitogen-activated protein (MAP) kinases (also known as extracellular signal-regulated protein kinases). Hence, ceramide-activated protein kinase may belong to an emerging family of proline-directed Ser/Thr protein kinases, which includes MAP and cdc2 kinases. These kinases recognize substrates containing the minimal motif, X-Thr/Ser-Pro-X, where the phosphoacceptor site is followed on the carboxyl-terminus by a proline residue and X can be any amino acid. This grant application has three goals: (1) To define the structural determinants for substrates recognition by ceramide-activated protein kinase and for ceramide stimulation, and determine the intracellular localization of enzyme activity; (2) to purify ceramide-activated protein kinase; and (3) to define how known antagonist of TNF action might act upon the sphingomyelin pathway and, in particular, ceramide-activated protein kinase. It is our belief that ceramide-activated protein kinase represents a new target for pharmacologic intervention in TNF and IL-1 action. Hence, the title of this application has been changed to reflect this emphasis. Hopefully, these studies will begin the process necessary for testing this hypothesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cocaine serves as a cofactor for susceptibility to HIV infection and AIDS progression. Cocaine also increases HIV-1 replication in peripheral blood mononuclear cells and enhances viral load in animal models. Furthermore, HIV positive cocaine users have lower CD4+ T cell counts and have a significant acceleration of decline of CD4+ T cells. Since CD4+ T cells are primary targets for HIV-1 infection and replication in vivo, it is imperative to understand the effects of cocaine on CD4+ T cell biology. This application proposes a potentially novel mechanism by which cocaine may increase HIV-1 replication. It has been proposed that cocaine enhances HIV-1 replication by regulating viral entry. Our preliminary data suggest modulation of HIV-1 post entry steps by cocaine. Our data also reveal that cocaine down regulates two anti-HIV cellular microRNAs (miRNAs), miR-125b and miR-328 in primary CD4+ T cells. Since these miRNAs target the 3'UTR of HIV-1 mRNA, we believe cocaine may target post-transcription steps of HIV-1 replication. Therefore, we hypothesize that enhanced HIV-1 replication and increased viral load by cocaine is mediated by down regulation of cellular anti-HIV miRNAs. Since HIV infected cocaine users have higher viral loads and increased risk of progression to AIDS, our findings will have far reaching implications in drug use and HIV biology. We will test our hypothesis by focusing on two aims. Aim 1: Determine effects of cocaine-induced down-regulation of anti-HIV cellular miRNAs on HIV-1 replication. Aim 2: Examine whether cocaine-induced down-regulation of anti-HIV cellular miRNAs activate latently infected HIV-1. PUBLIC HEALTH RELEVANCE: Drug use remains the second most common mode of exposure to HIV and illicit drugs serve as cofactors for susceptibility to HIV infection and disease progression. Although cocaine has been shown to enhance HIV infection and replication, the underlying mechanism remains unclear. Therefore, the focus of this proposal is to delineate the mechanism by which cocaine contributes to HIV/AIDS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project 1: Host Determinants of TB Susceptility Abstract: The outcomes of Mycobacterium tuberculosis (Mtb) infections are extremely diverse. Many infected individuals show no signs of disease. Among those that do become ill, the timing, location, and severity of pathology are remarkably variable. Genetic diversity in the host population is a major contributor to this variability, but the responsible alleles and mechanisms are poorly defined and have been difficult to address experimentally. In this project, we approach this inherently complex problem by modeling host diversity using a highly genetically diverse, but precisely genetically defined, panel of recombinant inbred mice known as the Collaborative Cross. The unique structure of this population allows the sequential evaluation of variable traits that reflect distinct aspects of pathogenesis. We took advantage of this iterative-phenotyping strategy to dissect the genetic basis of TB susceptibility at multiple levels. These prior studies implicated Kit Ligand (KitL) variation as an important determinant of TB disease in mice and humans, suggested that genetic interactions between pathogen and host are a sensitive metric for detecting new genes that alter disease progression, and highlighted the power of using ex vivo systems to dissect the interaction between Mtb and the macrophage. In the proposed project, we will continue to pursue these strategies to: Aim 1: Investigate the mechanism of KitL-linked susceptibility to TB disease in mice and humans. Aim 2: Use combinatorial mouse and bacterial genetics to dissect host-pathogen genetic interactions. Aim 3: Exploit the CC panel to dissect the Mtb-macrophage interaction. In combination, these new resources and approaches promise to provide fundamentally new insight into the role of host and bacterial genetic variation on the outcome of Mtb infection. !", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Thoracic aortic aneurysm (TAA) is a chronic condition that carries the risk of developing life-threatening complications such as aortic dissection (i.e., delamination of the aortic wall) and rupture (i.e., transmural tearing). The primary management objectives for TAA are to identify aortic dilation early and to surgically replace the aorta before it reaches a size that predicts future complications. It is estimated that 3 million patients in the US alone would meet criteria for aortic imaging surveillance based on current guidelines. Improved imaging surveillance strategies are needed to address the health and economic impacts of TAA. However, the reliability of standard aortic diameter measurements from CT images remains low. In this proposal, we will utilize a recently developed image analysis technique, Vascular Deformation Mapping (VDM), to perform semi-automated, three-dimensional assessment of aortic growth in patients with TAA undergoing imaging surveillance. This novel approach utilizes advanced computerized analysis techniques that make full use of the rich, high-resolution, volumetric data generated by modern CT scanners, to generate a more comprehensive and reliable assessment of aortic growth. VDM has the ability to significantly improve the speed and reliability of TAA growth analysis, facilitate the communication of aortic growth assessment between radiologists, care-providers and patients, and generate new biomarkers of disease progression that can be used to improve patient management decisions, and ultimately patient outcomes. In this grant effort, we propose to achieve the goals of this project through our close partnership between University of Michigan (UM) and Imbio, LLC investigators. Thus the Specific Aims of this Fast-Track proposal will develop a commercial VDM software application, including the product testing and data acquisition necessary for a FDA 510(k) application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fabry Disease: We have investigated the protein expression profiles in cells derived from patients with Fabry disease using microarray technology. Fabry disease is secondary to deficiency of the lysosomal enzyme alpha-galactosidase A, leading to altered glycosphingolipid metabolism and accumulation that is often associated with endothelial dysfunction. Current evidence suggests that there is impairment of the vascular nitric oxide pathway, with abnormalities evident in the cerebral circulation and in the dermal vasculature of patients with Fabry disease. Some of these findings have been confirmed in a mouse model of Fabry disease and are under investigation in the Fabry patient population. Future developments in array technology for proteins and DNA single nucleotide polymorphism analysis, together with gene expression microarray analysis, will open a new chapter in our understanding of the biology of lysosomal storage disorders. Additional studies are planned to investigate the role of rare lipids in the etiology of this disease.[unreadable] [unreadable] Gaucher Disease: We extended our basic investigations on the pathogenesis of the neuronopathic forms of Gaucher disease. Both glucocerebroside and glucosylsphingosine (GlcSph) are elevated in the brain of these patients. GlcSph is neurotoxic and contributes significantly to the dysfunction and destruction of brain cells. Our investigations have identified six compounds that inhibit the enzymatic synthesis of GlcSph and ranked them in the order of their effectiveness. Using compounds that reach the brain in an effective and non-toxic concentration, we propose to conduct clinical trials with these agents to improve the debilitating clinical course in patients with neuronopathic Gaucher disease. [unreadable] [unreadable] Our genomic investigations into the changes that occur in Gaucher disease have identified statistically significant increases in expression of 10-fold or more over normal in 203 genes and decreases in 78 genes. This information is critical to understanding the genesis of the hepatosplenomegaly and other pathophysiological changes that occur in patients with Gaucher disease and may lead to improved treatment strategies.[unreadable] [unreadable] Protein Transduction Domains: We are extending our strategies to extend the organ and tissue distribution of exogenous enzymes for the treatment of both Gaucher disease and Fabry disease to augment the effectiveness of enzyme replacement therapy. We have previously expressed a TAT-GC fusion protein in which recombinant glucocerebrosidase (GC) is fused to 11-amino acid peptide from the HIV-1 transactivator protein (TAT) which functions to facilitate the transport of the enzyme across the plasma membrane of a variety of cell-types in a receptor-independent manner. We have extended these studies to include additional GC constructs containing flexible spacers to increase enzyme activity in addition to a variety of TAT analogs in order to test the effect of sequence variation on the uptake of the enzyme. Further studies have also been undertaken to construct similar fusions with the enzyme alpha-galactosidase A (AGA) as a possible strategy for wider distribution of this enzyme for the treatment of Fabry disease. Efforts are underway to introduce these fusion proteins into animals to test their distribution and compare these findings with that of the native constructs. Recombinant GC-TAT and AGA-TAT were expressed in eukaryotic cells from which catalytically active, normally glycosylated enzyme fusion proteins were obtained and tested for receptor independent uptake into cultured cells. It is expected that GC-TAT will be more efficiently delivered than unmodified GC to cells in the bone marrow and lung, and perhaps additional cells that lack the mannose lectin and thereby enhance the clinical responses of patients with Gaucher to enzyme replacement therapy We anticipate that the AGA-TAT enzyme may be capable of entering heart and kidney more efficiently than the native enzyme.[unreadable] [unreadable] Enzyme Replacement Therapy: In collaboration with members of the Surgical Neurology Branch (SNB), we determined the safety of intracerebral injection of GC in non-human primates using the convection-enhanced delivery (CED) technique developed by SNB. This was followed by clinical administration of this enzyme to a patient with Type 2 Gaucher disease. Alterations in the enzyme aimed at improving the distribution and stability of the administered enzyme are currently under investigation. [unreadable] [unreadable] Gene Therapy: We are developing methods to improve gene therapy for patients with Gaucher disease and Fabry disease using adeno-associated viral vectors in conjunction with the abovementioned protein transduction domains.. The first of these constructs increased the level of glucocerebrosidase activity significantly in multiple organs and tissues when injected into experimental animals. Moreover, it very effectively transduces bone-marrow stem and progenitor cells ex vivo. We demonstrated long-term expression of GC in experimental animals following transplantation of bone-marrow-derived cells transformed by a GC lentivirus vector. It is anticipated that autologous cells transduced with this, or a related lentiviral vector, may be appropriate for gene therapy trials in patients with Gaucher disease since successful bone marrow transplantation can cure patients with type 1 (non-neuronopathic) Gaucher disease.[unreadable] [unreadable] Chaperone Therapy: We have identified a number of patients with Fabry disease in whom the reduced catalytic activity of alpha-galactosidase A is increased in the presence of the molecular chaperone 1-deoxy-galactonojirimycin (DGJ). This technique is based on the ability of certain compounds to interact with the catalytic site of mutated enzyme and escort it from the endoplasmic reticulum where it is produced through the Golgi apparatus to lysosomes. The low pH and the reduced concentration in lysosomes causes the chaperone to dissociate from the enzyme and allows it to carry out its catalytic function. We have examined the ability of DGJ to augment residual alpha-galactosidase A activity in cultured skin fibroblasts, T cells and peripheral blood mononuclear cells derived from patients with Fabry disease. A standardized protocol has been developed for the screening of patients for whom chaperone therapy with DGJ may be an effective way of increasing the enzyme levels. Further investigations are planned to evaluate variations in test conditions with DGJ as the chaperone and to test additional agents as potential chaperones for patients whose alpha-galactosidase A activity is not enhanceable using DGJ. A protocol has been initiated for a Phase 1 safety and dose-response trial with DGJ in patients with Fabry disease. When it is completed, we shall examine the clinical effectiveness of active site-specific chaperone therapy in patients with enhanceable alpha-galactosidase A activity (cf. lead investigator's report). [unreadable] [unreadable] Additional studies have demonstrated the potential for the use of chaperone therapy for the treatment of Gaucher disease. Using a similar test model as designed for chaperone therapy in Fabry disease, candidate active-site-directed molecules are under investigation. All surviving patients with Gaucher disease have residual glucocerebrosidase activity and appear to be excellent candidates for molecular chaperone therapy. Such studies underscore the possibility that small molecules may be used in place of or in conjunction with enzyme replacement therapy as it is currently practiced.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Melanoma is a malignant skin cancer with few effective treatment options for advanced stages of the disease. It is important to develop effective treatment strategies that will eliminate existing tumors and/or prevent the formation or recurrence of tumors. The work proposed in this application is to use an immunotherapy approach in the form of a gene delivery system. Electroporation (EP) will be used to introduce a plasmid DNA encoding for a cytokine into melanoma cells promoting an immune response to reduce cancer growth and prevent development. Interleukin 15 (IL-15) is a cytokine that activates natural killer (NK) cells and stimulates CD8+ T cells to exhibit long-term memory CTL activity against target cells. The research in this study focuses on delivering plasmid IL-15 (plL-15) by EP into a mouse model injected with B16.F10 murine melanoma cells. In Specific Aim 1, we will evaluate IL-15 expression levels when applying different EP parameters and different plasmid concentrations. An ELISA and real time RT PCR analysis will be used to detect protein levels and mRNA levels produced by the tumor cells, respectively. In Specific Aim2, the high and low expression levels of IL-15 obtained from Specific Aim 1 will be used to correlate expression levels of IL-15 with tumor regression. We will analyze the local and systemic cellular infiltrate stimulated by plL-15/EP using FACS, immunohistochemistry, Luminex bead array, TUNEL and ELISPOT to understand the reduction of tumor growth. A depletion study using monclonal antibodies will be used to demonstrate the roles that innate NK cells and adative T cells play when they are separately depleted. The reaction and duration of the response by these cells in the absence of one another will be evaluated when stimulated by the production of IL-15 from tumor cells. ELISPOT and FACS analysis will be used to demonstrate the results from the depletion studies. In Specific Aim 3, we will also determine the extent of IL-15 expression levels that provide viable treatments for metastatic disease using a two-tumor model. The cellular systemic responses to intratumoral treatments will be analyzed using immunohistochemistry, ELISPOT and ELISA. The relavance of this research is that the administration of plL-15/EP will provide promising stimulation of the immune response against tumor cells without causing toxicity or deleterious effects to the surrounding tissues. These experiments can promote the development of a potential anti-cancer therapy to block cancer metastasis as well as the recurrences of new diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A traditional view of metastasis is that it results from a process similar to Darwinian evolution involving the natural selection of tumor cells that are capable of migration and survival during treatment and at distant sites. In this model the selection of tumor cells exhibiting stable genetic changes occurs, the selected cells are very rare, local and cause metastasis late in tumor progression. The recent development of new technologies, including high-density microarray based expression profiling, multiphoton intravital imaging and the collection and characterization of migratory tumor cells from live tumors and bone marrow disseminated tumor cells (DTCs) from patients, have challenged this traditional model of metastasis. The new technologies indicate that metastatic ability is acquired at much earlier stages of tumor progression than predicted by the Darwinian model, is encoded throughout the bulk of the primary tumor, it is highly plastic and involves transient changes in gene expression. These results have led to the micro-environment model of metastasis. The micro-environment and Darwinian models can be reconciled if tumor progression resulting from the selection of stable genetic changes in the primary tumor during progression, contributes the micro-environments necessary to induce the transient changes in gene expression that support the invasive and metastatic phenotype. That is, the tumor micro-environment initiates the transient epigenetic expression of genes that induce tumor cell migration, survival and metastasis. Examples of such micro-environments in breast tumors are extracellular matrix density, inflammation, and hypoxia. To study these micro-environments and their effects on metastatic phenotype, we have assembled a multidisciplinary team who will collaborate using their special expertise to: 1. fate map tumor cells to determine if tumor cells migrating from different spontaneous, and nano-device generated soluble factor-derived micro-environments, have different migration, dissemination, dormancy and growth patterns in target organs. 2. Determine the spatial and temporal extent and functional consequences of these micro-environments at single cell resolution in vivo in primary tumors and in DTCs 3. Isolate and characterize the metabolomics, genomics and epigenomics of special populations of tumor cells such as the migratory and dormant tumor cells in disseminated locations. 4. Investigate ECM-dependent migratory/invasive, dormant and proliferative tumor cell phenotypes. 5. Extend key observations to human breast and head and neck squamous tumors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Support is requested for 9 predoctoral and 6 postdoctoral positions to establish a training program in the interdisciplinary area of Pharmacoinformatics. This program is proposed because current approaches to the discovery of new drugs have not fully capitalized on advances in bioinformatics, genomics, computing and other fields, and consequently, the rate of drug discovery has lagged behind breakthroughs in these areas. The pipeline of new drugs under development has not increased in recent years, and a very small number of new drags are thus responsible for bearing the enormous costs of an increasingly expensive process that evolved prior to development of massive amounts of new information and the ability to integrate it into usable and accessible systems. We propose a comprehensive, integrated program to train the next generation of scientists to: (1) use informatics approaches to identify biomolecules that are candidates for drug targeting, (2) use structural genomics, chemoinformatics, and structure-based design to develop families of chemicals to target selected biomolecules and structures, and (3) use ADMET (absorption, distribution, metabolism, elimination, and toxicity) profiling and computational predictions of efficacy to select viable drag candidates prior to biological testing. Training will emphasize in silico approaches to achieve these goals. Inter-institutional training will be conducted by sixty-three training faculty from the six institutions of the Gulf Coast Consortia (Baylor College of Medicine, M. D. Anderson Cancer Center, Rice University, the University of Houston, The University of Texas Health Science Center at Houston, and The University of Texas Medical Branch at Galveston) in the Houston -Galveston area and will utilize research training with co-mentors, didactic courses, seminars, retreats, and career development activities. Predoctoral trainees will matriculate and receive first year support at their home institutions, and stipends will be awarded on a competitive basis for years 2-4 of graduate training. The home institution will award the Ph.D. Postdoctoral trainees will receive support for two years maximum. The proposed program is designed to train future leaders in drag discovery research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PURPOSE OF RESEARCH: The goal of this project is the development and field-testing of a screening procedure to assess social and emotional dysfunction in populations of four- and five-year-old children, in order to allow for prompt further evaluation and early intervention. METHOD: A screening battery was developed which includes assessments of health, developmental history, and cognitive and sensory functioning. A Behavior Checklist was designed to tap behaviors considered by clinicians as indicators of social and emotional dysfunction for this age group. The battery was piloted with over 800 children in 1976. After revision it was administered to 750 subjects in 1977. A subsample of 85 children was seen for clinical interviews, and the results were compared to the screening procedure. Follow-up outcome data has been collected for the 1977 sample which will serve to evaluate the screening procedure. RESULTS AND FUTURE PLANS: A criterion validation study indicates that disturbed and normal children are significantly differentiated (p less than 04) on the Behavior Checklist. Further analysis will establish the predictive accuracy of the screening procedure as it relates to measures of the child's functioning at follow-up. Recommendations for screening policy, as well as for providing identified service needs, will be made after the data is analyzed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Howard University Graduate School proposes to continue its focus on the development of a new cadre of biomedical scientists at the institution. To this end, the MBRS SCORE program is requesting support for 18 new projects from faculty who have not previously participated in the MBRS Program. Seventeen of these new projects are from junior faculty (non-tenured), several of whom have just joined the faculty this academic year. These investigators are from five basic science an| two clinical departments in the College of Medicine, as well as from two undergraduate departments with strong graduate training programs. This application contains both regular (18) and pilot (4) projects. The mantra for the proposed program is \"ROAD MAP TO R01 SUPPORT\". Building on some of the success of our current SCORE participants (only 3 are returning in this application, as some have moved on to other individual support -one R01);the program's expectations are being raised leading to the following goals of opportunity: GOAL (1);To improve the scientific productivity of SCORE faculty. The measurable objective (A): That the publication rate of the participating faculty be increased overall and throughout the proposed project peroid. Measurable objective (B) that all of the continuing program investigators submit an R01 application by the end of the 02 year of funding and (C) that 50% of the new participating investigators will have submitted R01 applications, or submitted to an equivalent agency by the end of the proposed period. GOAL (2): To enhance the professional progression of SCORE faculty through the provision of essential research support. Measurable Objectivie (D): during the proposed period, nontenured junior faculty who are Pis on regular projects will make significant progress on each of the meaures of productivity( publications and grant submissions) which will enhance their probability of earning tenure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed Charity Hospital-Tulane-Louisiana State University (LSU) GCRC will serve as a facility for patient-based applied and basic research. The specific aims of the GCRC are: (1) to provide a well staffed facility that will allow in-hospital and outpatient care for patients who are subjects in research projects; (2) to allow these projects to be performed under controlled conditions with scientific accuracy; (3) to support a Core Laboratory which will provide assays for many of the researchers who use the facilities as a measure to increase productivity and reduce costs; (4) to improve the quality of patient care in the overall Charity Hospital- Tulane-LSU Medical Complex by applying the advances made in these clinical research endeavors to disease diagnosis and therapy; (5) to provide training in the methodology and design of good clinical research projects as a form of teaching for post-graduate research residents, interns, medical students, and fellows, nurses and other paramedical personnel in the facility. The program stresses: (1) a strong interdisciplinary effort by both medical schools and by several departments within each school; (2) a strong letter of commitment from Charity Hospital and the participating medical schools with regard to housing of the unit (Charity Hospital) plus renovation costs; and (3) quality (predominantly NIH funded or pending funded projects) by most of the leading investigators in both medical schools and at Charity Hospital. The Charity/Tulane/LSU GCRC will be predominantly an adult unit but with a small pediatrics and \"contagious\" areas within the same facilities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this proposal is to study the effect of sympathetic innervation on the formation of 21 and 22 adrenoceptor (AR) signaling complexes in the heart. We will investigate how these complexes either facilitate or restrict receptor signaling; we will identify the structural domains of the receptors that are needed for interaction with the other components of the signaling complex; and we will identify cellular proteins that define these subtype-specific signaling complexes. 21ARs and 22ARs are prototypical G protein coupled receptors (GPCRs), the largest family of hormone and neurotransmitter receptors in the human genome. These receptors are essential for the physiologic regulation of cardiac function in response to catecholamines (adrenaline and noradrenaline) released from sympathetic nerves. Recent studies suggest that 21ARs and 22ARs play distinct roles in the pathogenesis of heart failure, a growing health problem in the United States. We have developed an experimental system to study the important interface between sympathetic nerves and the heart using co-cultures of neonatal cardiac myocytes and sympathetic neurons. Our preliminary studies show that 21ARs and 22ARs have differential subcellular targeting relative to these synapses, and that signaling complexes form at the sites of synapse formation. The following aims are designed to characterize these signaling complexes. Aim 1. Determine the structural basis for subtype specific targeting and trafficking of 21ARs and 22ARs in cardiac myocytes. Aim 2. Determine the functional significance of subtype-specific localization of 21ARs and 22ARs relative to sympathetic synapses. Aim 3. Characterize protein components of the 21AR and 22AR signaling complexes in cardiac myocytes innervated by sympathetic neurons. Aim 4. Determine the functional significance of interacting proteins identified in Aim 3 on 21AR and 22AR signaling and trafficking in cardiac myocytes, and verify their existence in signaling complexes in the adult heart. The proposed studies will provide new information about how the brain regulates heart function. We will characterize the mechanism by which noradrenaline and adrenaline released from sympathetic nerves alters heart function by activation of two specific adrenergic receptors. This research will further our understanding of the development of diseases such as heart failure and sudden death.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Analytical Core (B) will be located in the Biomedical Biological Sciences Building, University of Kentucky. The Analytical Core will consist of the Director and research personnel for performing services of resolution and profiling of analytes using mass spectroscopy and for quantifying analytes using luminex platforms. An important part of Core activities will include methods development to analyze specific molecules of interest, and for profiling lipids within animal and human tissue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Significance Recombinant SHIVenv clones are a means to analyze the role of HIV-1 env genes in vivo for viral transmission across mucosal membranes and for SAIDS pathogenesis. Additionally, vaccines based on HIV-1 immunogens can be tested for efficacy by challenge with such recombinant viral clones. Objectives An animal model to study both HIV-1 infection and AIDS pathogenesis is not available. To analyze function of specific HIV-1 genes in vivo, SIV/HIV-1 recombinant viruses (designated SHIV) have been constructed by replacing genes in the pathogenic clone SIVmac239 with counterpart HIV-1 genes. We have made SHIV clones containing the envelope (env) gene of various HIV-1 subtype-B isolates, and have analyzed these recombinant viruses in vivo in juvenile and newborn macaques. Results A pathogenic SHIV (designated SHIV-33A), containing the env gene of HIV-1-SF33, was obtained by one passage in a juvenile rhesus macaque. SHIV-33A produced simian AIDS in juvenile macaques when given by either the intravenous or mucosal membrane (oral and vaginal mucosa) routes. In addition, newborn macaques (2 days of age) also developed simian AIDS after intravenous inoculation. Sequence analysis of the env gene of SHIV-33A revealed about 15 amino acid changes, relative to the input SHIV-33 clone. All animals showing immunodeficiency disease also exhibited a rapid and sustained decline in CD4 T-cells in both peripheral blood and lymph nodes. In vitro analysis of SHIV-33A revealed increased cytopathicity and increased replication in tissue culture cells. Future Directions Intragenic SHIV-33 recombinants and point mutants, involving the HIV-1SF33 env gene, are being constructed to determine which sequence change(s) accounts for viral adaptation and pathogenesis. Studies will also be performed to determine the mechanism of rapid depletion of CD4 T-cells in the acute stage of infection with the pathogenic SHIV-33A strain. Functional domains of SHIV-33A will be analyzed by constructing env genes with point and deletion mutations; these mutant viral clones will be examined in tissue culture cells and in rhesus macaques. KEY WORDS HIV-1 genes, AIDS pathogenesis, SHIV clones FUNDING NIH Grant AI41907", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Prevention strategies have been developed to take advantage of the metabolic and behavioral risk factors for type 2 (non-insulin- dependent) diabetes which have been identified in research with the Pima Indians and elsewhere. Obesity, impaired glucose tolerance, hyperinsulinemia, physical inactivity, and high fat diet have been implicated as risk factors for type 2 diabetes. Since these factors are potentially reversible with behavioral and pharmacologic therapy, strategies for prevention have been developed and will be tested. This project is a multicenter randomized clinical trial of prevention of non-insulin-dependent diabetes mellitus in overweight persons with impaired glucose tolerance known as the Diabetes Prevention Program. The coordinating center is at the George Washington University. During the project year recruitment was completed and treatment of volunteers has continued in four American Indian communities in collaboration with the Indian Health Service. - Diabetes mellitus, prevention, clinical trial - Human Subjects", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Treatment of Central Vein Thrombosis remains unsatisfactory. However, there is strong evidence from a recent study that quick, aggressive therapy with Urokinase for occlusions of recent onset, can produce marked benefits. Prognosis of these particular cases has been considered poor - therefore, all work on this subject is of considerable importance. We propose a blinded randomized study whereby our objective will be to enhance knowledge of effectiveness of fibrinolytic therapy in Central Retinal Vein Thrombosis - utilizing Urokinase - a plasminogen activator and fibrinolytic enzyme. The Urokinase is supplied by the National Heart and Lung Institute, and is derived from the stockpile of the Urokinase subcommittee which approved of this protocol and hence released the amount of urokinase sufficient for this project. We aim to compare anticoagulant therapy and the use of Urokinase therapy. All patients of less than 14 days onset of symptoms diagnosed as Central Retinal Vein Thrombosis will be considered for this study. Excluding criteria will be pregnant females and those in the child bearing age unless they have a negative pregnancy test and are on adequate contraception; all uncontrolled hypertensive patients with blood pressures exceeding 150/100; those patients with a history of bleeding diathesis, or recently active peptic ulcer; recent surgery, or history of significant complicating disease such as carcinoma or cerebral vascular accident. Intravenous Urokinase or Heparin will be given via infusion pump, with close monitoring by coagulation laboratory of fibrinogen level; prothrombin, thrombin, and partial thromboplastin times; euglobulin lysis time and level of fibrinogen degradation products (FDP). Any untoward bleeding will be treated with epsilon amino caproic acid for the Urokinase group and protamine for the heparin group.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our research is directed at epithelial differentiation in normal and pathologic states. This grant focuses on the roles of vitamins and hormones, as exemplified by retinoids and thyroid hormones, in the control of keratin synthesis and in the process of keratinization. For decades, clinicians have recognized that thyroid hormones and vitamin A had profound effects upon normal and abnormal skin and within recent years the interest in the use of vitamin A and its derivatives on diseases of keratinization has become widespread. At the same time studies of other systems have defined on cellular and molecular levels the mechanism by which these substances effect regulation of gene expression; they bind to specific receptors which in turn react with recognition elements in the non-coding regions of DNA to profoundly affect transcription. The experiments will focus upon several genes for epidermal keratins whose expression we have shown to be negatively regulated by both retinoic acid and its receptor as well as thyroid hormone and its receptor. The assays used will be based upon transfection involving regulatory regions of keratin genes linked to a reporter gene, either chloramphenicol acetyl transferase for quantitative studies or luciferase for localization studies. The recognition elements in the keratin gene for both receptor complexes will be identified through deletion studies. Specific point mutations will define their exact positions. Throughout these studies, the polymerase chain reaction (PCR) will be used to amplify appropriate areas of DNA. Parallel studies will be undertaken in cell lines and primary cultures of epithelial cells. Finally, the effects of retinoids and thyroid hormones on differentiating epidermis in tissue culture will be determined. Thus, our aims in the 5 years for which we are requesting support will be: 1. to identify and characterize the recognition elements (REs) for the nuclear receptors of retinoic acid and thyroid hormone in human keratin genes - basal cell-specific K#5 and K#14, stratum corneum-specific K#10, hyperproliferation-associated K#6 and K#16, and the simple-epithelia- specific K#8, K#17, K#18 and K#19. 2. to define the molecular mechanisms by which retinoids and thyroid hormones regulate keratin gene expression and to delineate the interactions among the two classes of mediators and their receptors; 3. to analyze the effects of retinoids and thyroid hormones on epidermal differentiation in vitro. Our proposal, which will define the mechanisms of action of retinoids and thyroid hormones in epidermis, has important implications for health care for it will ultimately lead to understanding of the normal and pathologic processes in skin and should indicate the potential of new therapeutic agents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research is designed to study the mechanism of DNA replication using T even bacteriophage infected E. coli as a model system for the investigation with emphasis on the studies of the initiation process. T4 and T6 DNA-delay proteins, which have been defined genetically and biochemically as needed for the specific initiation process of the respective phage have been purified. They form a complex which has ATP-dependent topoisomerase activity of untwisting superhelical DNA. Because of the critical role DNA topoisomerase plays in DNA replication, the mechanism of action, the subunit strucutre and the DNA sequence of this novel enzyme, which we have discovered, will be investigated in detail. On the other hand, the origin(s) of T4 DNA replication which we proposed to be the intended target of the initiation proteins will also be characterized with respect to its structure and function. We plan to carry out experiments designed to test the hypothesis that the initiation process is mediated through direct interaction of the initiation proteins with the origin of DNA replication since both components have now been purified and isolated in our laboratory. This will include the use of T2, T4 and T6 initiation proteins of the DNA-delay gene products in a comparative study to demonstrate the specificity of the initiation process.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Protein kinases mediate many cell signaling events, and their tight control is essential for regulating essential processes ranging from cell division to energy metabolism. Thus, it is not surprising that protein kinases are directly or indirectly involved in many diseases and that kinases are key drug targets. For example, Src kinase was the first identified proto-oncogene and the formation of a de-regulated Abl fusion protein (BCR-Abl) is the cause of disease in 95% of patients with chronic myeloid leukemia. X-ray crystal structures have shown that the same kinases can obtain an active and various inactive conformations, implying that kinases are inherently flexible. How the active and inactive states are stabilized and how the states interconvert are key questions in understanding kinase regulation. Because X-ray crystal structures provide only static snapshots, we will use nuclear magnetic resonance (NMR) experiments to study the time scales and amplitudes of structural interconversions in Abl and Src kinase domains. [unreadable] [unreadable] BCR-Abl is the target of the clinically highly successful drug imatinib (Gleevec(r), Novartis) in the treatment of chronic myelogenous leukemia (CML). Why does imatinib bind and inhibit c-Abl but not the structurally closely related c-Src kinase? The crystal structure of Src in complex with imatinib shows protein-drug interactions similar to that of Abl, even though the affinity of imatinib for Src is orders of magnitude lower than for Abl. Because imatinib binds only to the inactive conformation of the kinase, drug binding is intimately related to the interconversion between active and inactive states. The goal of this study is to examine whether differences in this interconversion underlie the differential sensitivities for imatinib. Therefore, we will compare the time scales and amplitudes of backbone motions between Src and Abl kinases in the presence of imatinib by NMR experiments. In preparation for these dynamics experiments, we have established expression systems and NMR conditions and will next pursue the assignment of the Src and Abl NMR spectra. [unreadable] [unreadable] Kinase inhibitory drugs such as imatinib have a great therapeutic potential because of the many signaling events that protein kinases mediate. However, these drugs have to be exceptionally specific for their target kinase and resistance mutations can render these drugs ineffective as seen in leukemia patients under imatinib treatment. The proposed experiments will clarify how inhibitors such as imatinib exploit the characteristic movements of kinase proteins, rather than just their structures, to achieve specificity. Furthermore, the results will have broader impact on the understanding of the fundamental mechanisms of kinase regulation and of drug resistance mutations that are known to arise in cancer patients undergoing kinase inhibitory treatment. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Major depressive disorder (MDD) is characterized by reductions in the density and size of neuronal and glial[unreadable] cells in prefrontal cortex. We find these cellular changes to be age-dependent as younger depressed show[unreadable] prominent reductions in the density of glial cells (astrocytes), whereas older depressed have marked[unreadable] reductions in the density of pyramidal (presumably glutamate) neurons and calbindin-immunoreactive[unreadable] interneurons (mostly co-localizing GABA). Since, astrocytes regulate concentration of glutamate, an excess[unreadable] of which is neurotoxic, we propose that an early deficit in astrocytes in MDD could lead to an increase in the[unreadable] extracellular concentration of glutamate and to a reduction in pyramidal and GABA neurons later in life.[unreadable] Hence, there may be imbalances in GABA/glutamate homeostasis that are consistent with neuroimaging[unreadable] studies revealing changes in levels of GABA and glutamate in MDD which are reversible with antidepressant[unreadable] (SSRI) treatment. Cortical neurons are regulated in complex ways by serotonin acting (at least) at serotonin-[unreadable] 1A and -2A receptors located on these neurons and astrocytes. Pathology in ascending serotoninergic[unreadable] axons and postsynaptic receptors may be related to the activity, number and size of glutamate and/or GABA[unreadable] neurons and astrocytes in MDD. To date, there have been no studies on the expression or localization of[unreadable] serotonin receptors on specific cortical cell types in depression.[unreadable] The overall hypothesis is that in depression there will be age-dependent reductions in the density of[unreadable] astrocytes, glutamate pyramidal neurons and GABA interneurons, and that the expression of regulatory[unreadable] serotonin-1A and -2A receptors on these cells will be altered. These cell reductions will also be correlated[unreadable] with an age-related loss of serotonin innervation in prefrontal layers. To test these hypotheses, we will[unreadable] directly identify and quantify the packing density of astrocytes and glutamate and GABA neurons expressing[unreadable] mRNA for specific proteins (Aim 1). Moreover, we will assess the integrity of the serotonin system regulators[unreadable] (postsynaptic receptors and presynaptic axons) of prefrontal cells by estimating the proportion of cell types[unreadable] expressing mRNA for serotonin-1 A and -2A receptors (Aim 2), and the density of serotonin axons[unreadable] expressing the serotonin transporter (Aim 3). Double in situ hybridization, immunohistochemistry and 3-D[unreadable] cell counting techniques will be used in the same postmortem tissue sampled from the prefrontal cortex of[unreadable] younger and older subjects with MDD and non-depressed controls as used in our cell counting studies.[unreadable] This project will identify the cellular substrates of glutamate, GABA and serotonin interactions in the[unreadable] cortex and their potential role in the etiology, pathophysiology and age-related progression of depression. It[unreadable] may also reveal novel targets for preventing depressive illness and better antidepressant drug treatment.[unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mathematical performance in the United States is significantly falling behind that of many other countries and failing the expectations of our nation's education leaders. This project, Number Concepts and Automaticity, is designed to target predictable factors in mathematical failure and provide the right guidance and practice to overcome deficiencies using a computer software tool. It is based on leading research in math education for elementary students. The project will focus on developing mathematical models for children, bringing hose models into working memory, and building automaticity with addition and subtraction problems. The software tool created in the project comprises a Number Concept Builder that will tie virtual manipulatives to their numeric values, an Automaticity Builder that will help students increase their ability to recall concepts from retrievable memory, and an Assessment Module that will measure student progress. The Phase I feasibility study will involve 2nd grade students. A small formative field test will guide the software tool prototype development in its earliest stages. The completed prototype will be introduced into school classrooms as part of the standard number sense curriculum. Students will participate in a controlled experiment woven into their standard classroom activities. Students with physical disabilities will engage in a single subject study to guide product accessibility. This study will seek to prove that the software can accurately measure automaticity in individual students' performance and will verify that the software can be used to increase automaticity. The study will also attempt to reveal automaticity for students with physical disabilities who have slow response times due to their impairments. Future work in Phase II and beyond will include the needs of students in throughout grades K-5 and across more areas of the math curriculum. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In rhesus macaques, a SIV/HIV-1 chimeric virus (SHIV) containing the env gene from HIV-1 89.6 is capable of initiating a systemic infection following mucosal or systemic inoculation. Previous Infection with SHIV89.6 protects 60% of rhesus monkeys from uncontrolled replication of SIVmac239 after intravaginal inoculation. Understanding the nature and anatomic location of the adaptive immune that confers protection to SHIV \"immunized\" monkeys would be significant contribution to AIDS vaccine development. The overall goal of the Primate Core is to facilitate the design and execution of the in-vivo primate studies that will be the basis of all 3 projects in the program. All the in-vivo studies will be carried out at the California National Primate Research Center (CNPRC).This Core will obtain mature, multi-parous, female rhesus monkeys, immunize the monkeys intravenously with virulence-attenuated SHIV89.6, administer immunomodulatory agents to monkeys and challenge the monkeys intravaginally with defined stocks of SIVmac239; determine vRNA levels and viral population complexity in plasma and tissues of the monkeys after SIVmac239 challenge; characterize CD4+ T cell levels in plasma and tissues of the monkeys after SIVmac239 challenge and characterize body weight levels of the monkeys before and after SIVmac239 challenge. This Core is essential for the completion of the proposed studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Obesity is a world-wide health problem that is associated with metabolic syndrome, including insulin resistance and the development of non-alcoholic fatty liver disease (NAFLD). NAFLD is characterized by hepatic steatosis and varying degrees of inflammation and fibrosis. Increasing evidence suggests that saturated free fatty acids (FFA), by promoting insulin resistance and mediating the progression from simple steatosis to steatohepatitis, are a causative link between obesity and NAFLD. However, the molecular mechanisms responsible for the effects of saturated FFA are incompletely understood. Our long-term goal is to understand the molecular mechanisms by which saturated FFA contribute to the development of NAFLD. Saturated FFA are a major source of metabolic stress that induce activation of the c-Jun NH2-terminal kinase (JNK) pathway, and gene deletion and pharmacological inhibition have established an important role for the JNK signaling pathway in obesity-induced insulin resistance and the pathogenesis of NAFLD. Our preliminary studies identified the MAP3K mixed-lineage kinase 3 (MLK3) as an important mediator of saturated FFA- induced JNK signaling. Thus, we have demonstrated that MLK3 mediates FFA-induced JNK activation, promotes obesity-induced insulin resistance and modulates FFA-induced ER stress response in specific tissues. These data indicate that MLK3 plays a critical role in the development and progression of NAFLD, most likely through metabolic stress signaling. The central hypothesis is that MLK3 contributes to the pathogenesis of NAFLD by mediating saturated FFA-induced JNK activation, thereby inducing inflammatory gene expression, promoting insulin resistance, and modulating ER stress-induced cell death in specific tissues. This hypothesis will be tested by three complementary Aims. In Aim 1 biochemical and molecular biological methods will be used to delineate the pathway(s) required for MLK3 activation by FFA. In Aim 2 biochemical and molecular biological methods will be used to determine the role of MLK3 in the FFA-induced ER stress response. In Aim 3 reciprocal bone marrow transplantation studies will be used to examine the tissue specific role of MLK3 in the development of insulin resistance. Accomplishing the goals of this proposal will increase understanding of the role of MLK3 in the pathogenesis of NAFLD. This information will provide the foundation for the potential development of novel therapeutic strategies for the treatment of this pathology. PUBLIC HEALTH RELEVANCE: Non-alcoholic fatty liver disease is the leading cause of liver dysfunction in non-alcoholic, viral hepatitis- negative populations in Europe and the USA. Saturated free fatty acids, by promoting insulin resistance and mediating the progression from simple steatosis to steatohepatitis, are a causative link between obesity and NAFLD. The proposed study will increase understanding of the molecular mechanisms by which saturated FFA contribute to the development of NAFLD. This information may suggest novel therapeutic strategies for the prevention of NAFLD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The ability of steroid hormones to influence profoundly the excitability of the CNS is well documented. The broad objective of this proposal is to explore pharmacologically the molecular mechanisms whereby steroids modulate CNS excitability. Abnormal activation of amino acid receptors has been proposed to play a role in the etiology of psychiatric disorders such as anxiety, depression and schizophrenia. Understanding the mechanisms of steroid actions on the CNS may lead to new strategies for the treatment of psychiatric disorders. During the course of our studies we came upon the unexpected finding that pregnenolone sulfate (PS), an abundant neurosteroid, acts as a positive allosteric modulator at the NMDA receptor while inhibiting the kainate, AMPA, glycine, and GABA responses. A major focus of the research plan will be to test our working hypothesis that steroids such as PS regulate the balance between excitation and inhibition on neurons derived from the vertebrate CNS by acting on excitatory and inhibitory amino acid receptors. Toward this end, whole-cell voltage-clamp and current-clamp techniques will be utilized to character electrophysiologically the effects of PS and related steroids on excitatory and inhibitory amino acid receptor-mediated responses of embryonic chick spinal cord neurons maintained in primary monolayer cell culture. The proposed study has three major parts: First, we will screen a series of steroids for activity on amino acid receptor-mediated currents. Secondly, we will determine the potency, efficacy, and mode of action for PS and each steroid identified, with the goal of elucidating the mechanism(s) of steroid action(s) through a study of their structure-activity relationships. In particular, we will focus our studies on PS and on other steroids that are active at NMDA and non-NMDA glutamate receptors. Finally, the effects of steroids on excitatory and inhibitory synaptic transmission at single synapses will be investigated. In the long run, we will determine whether chronic treatment with active steroids such as PS, progesterone, and reduced metabolites of progesterone induces functional changes of excitatory or inhibitory amino acid receptors and of synaptic transmission at identified synapses. These studies will provide a strong basis for evaluating the role of steroids as modulators of neuronal function, and a foundation for development of novel steroid-based drugs for the treatment of psychiatric disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the work described in these studies is to elucidate how critical extracellular and intracellular domains control signaling by notch receptors. The extracellular domains of all notches contain three iterated LIN12 repeats, which maintain notch receptors in their resting state prior to ligand binding. Ligand-induced proteolysis releases the intracellular portion of notch (ICN) and allow it to translocate to the nucleus, where its RAM and ankyrin-repeat domains interact with two additional factors, RBP-JK and mastermind (mami 1) to form a transcriptionally active ternary complex. The specific aims of this research plan, which will provide important new insights into the structure and function of the LIN12 and ankyrin repeat domains of notch, are to: [unreadable] [unreadable] 1. Determine how LIN12 modules prevent ligand-independent activation of notch receptors. (a) Determine high-resolution solution structures of key modules and module pairs from a Notch LIN12 domain, analyzing the structural interactions of these modules with each other and with neighboring domains. (b) Identify contacts between the LIN12 domain and the NTM extracellular stub that maintain the integrity of the Notch heterodimer, and (c) Test the functional importance of specific intermodular and LIN12-NTM contacts. [unreadable] [unreadable] 2. Determine the essential domains and residues required for assembly and function of thenotch/RBP-Jkappa/maml1 signaling complex. (a) Determine the minimal protein domains required for reconstitution of an ICN-mam11-RBP-Jkappa--DNA complex. (b) Identify contacts that contribute to energetic stabilization of ICN complexes and determine how RBP and RAMANK cooperate to recruit mamli to form a ternary complex on DNA. (c) Test the function of specific contacts responsible for stabilizing the ternary DNA-binding complex.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract The goal of this consortium is to establish tools that can be used as biomarkers and/or sensitive and reliable objective assays of social impairment in autism spectrum disorder (ASD) clinical trials. Specifically, we aim to accelerate the development of effective treatments for social impairment in ASD by validating outcome measures that will be sensitive and reliable assessments of response to treatment and EEG and eye-tracking (ET) biomarkers that can be used to reduce heterogeneity of samples via stratification, indicate early efficacy, and/or demonstrate target engagement. The consortium will conduct a naturalistic, longitudinal study of preschool (3-5 years) and school aged (6-11 years) children with ASD and typical development (TD) with IQ ranging from 50- 115. Children will be assessed across three time points (T1: Baseline, T2: 6 weeks, T3: 24 weeks) using clinician, caregiver and lab-based (LB) measures of social impairment, along with a battery of conceptually related EEG and ET tasks and independent ratings of clinical status. This battery measures key facets of social- communication in ASD using well-validated paradigms appropriate for this developmental and cognitive range. Five Collaborating Implementation Sites (?Sites?), all highly experienced in multi-site collaborative clinical research using the methodologies proposed here in both typical and atypical development, will contribute equally to recruitment, screening, diagnosis, testing, and longitudinal assessment. The Data Acquisition and Analysis Core (DAAC) will oversee consistent application of scientific standards and methodological rigor for standardized data collection, processing, and analytics. The Administrative Core will oversee the operations of the Sites, the Data Coordinating Core (DCC), and the DAAC and coordinate with federal and private partners to achieve the aims of this cooperative agreement. The will utilize informatics and technology to develop, maintain, and monitor a robust, secure, HIPAA-compliant data collection, coordination, and storage system to streamline communication and data flow throughout the consortium and ensure organized, secure data management, quality control, and reliable upload to the National Database for Autism Research and NIH/NIMH Data Repositories. The DCC benefits from the Yale Center for Clinical Investigation's quality assurance experience and the existing multi-site ASD research data systems developed by Prometheus Research. The DCC will: 1) Anticipate, respond to, and fulfill the informatics needs of the consortium in the design, harmonization, and implementation of data coordination and innovative data collection methods and tools across the Sites, DAAC, and Admin Core; 2) Provide state-of-the-art methods for: a) consortium communication; b) data management, site monitoring, privacy, and security within the consortium and across Sites; and c) dissemination of information to its stakeholders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During the last fiscal year, the following advances were made: 1) Biological activity of stem cells The Section continues to try to bring some clarity to the mesenchymal stem cell field. The concept of a post-natal mesenchymal stem cell (MSC) originated from studies focused on bone marrow stromal cells (BMSCs), which are non-hematopoietic adherent cells, a subset of which are skeletal stem cells (SSCs), able to form cartilage, bone, hematopoiesis-supportive stroma, and marrow adipocytes based on rigorous clonal and differentiation assays. Subsequently, it was speculated that BMSCs could form other mesodermal derivatives and even cell types from other germ layers. Based on BMSC surface markers, representative of fibroblastic cells, and imprecise differentiation assays, it was further imagined that MSCs are ubiquitous and equipotent. However, MSCs do not have a common embryonic origin and are not a lineage, but recent studies indicate that they are tissue-specific stem/progenitor cells. These cells share cell surface features owing to their fibroblastic nature, but they are not identical. They display different differentiation capacities based on their tissue origin, but do not trans-differentiate outside of their lineage, based on rigorous assays. For these reasons, the MSC term should be abandoned. Tissue-specific stem/progenitor cells provide the opportunity to devise methods for tissue regeneration by the cells themselves (tissue engineering). Their use in other forms of regenerative medicine based on paracrine, immunosuppressive, and immunomodulatory effects is far less clear (Robey, Faculty 1000, 2017). Human umbilical cord blood (CB) has attracted much attention as a reservoir for functional hematopoietic stem and progenitor cells, and recently, as a source of blood-borne fibroblasts (CB-BFs, also called cord blood-derived MSC). In a previous study, it was demonstrated that bone marrow stromal cell (BMSC) and CB-BF pellet cultures make cartilage in vitro. In addition, BMSC and CB-BF pellets remodeled into miniature bone/marrow organoids in vivo, suggesting that CB-BFs can support the Hematopoietic Stem Cell (HSC) niche. Compared with BM-ossicles, CB-ossicles showed a higher proportion of red marrow vs. yellow marrow. Marrow cavities from CB- and BM-ossicles included donor-derived CD146-expressing osteoprogenitors and host-derived mature hematopoietic cells, clonogenic lineage-committed progenitors, and HSCs. Furthermore, human CD34+ cells transplanted into ossicle-bearing mice engrafted and maintained human HSCs in the niche. These data indicate that CB-BFs are able to recapitulate the conditions by which the bone marrow microenvironment is formed and establish complete HSC niches, functionally supportive of hematopoietic tissue (Pievani et al, Dev, 2017). 2) Role in disease With regards to fibrous dysplasia of bone (FD), often in conjuction with the McCune-Albright Syndrome (over-active secretion of some hormones), a rare opportunity presented itself to study radiographs and tissue samples (removed during corrective orthopaedic surgery) from a single patient with FD/MAS. These radiographs and samples were collected at different times following intermittent treatment with intravenously administered pamidronate. In children, such treatment results in the formation of sclerotic bands (zebra lines) due to binding of the bisphosphonate to a mineralized surface and subsequent temporary decrease in bone resorption at the time of treatment. By way of contact microradiography, back-scattered electron microscopy and histological analysis, it was found that zebra lines formed only where the bone was normal, abruptly stopped at the interface between normal and FD bone, and were completely absent in FD bone, due to the inability of bisphosphonate to bind to unmineralized osteoid, a prominent feature of FD lesions. These results suggest that lack of zebra lines is indicative of the presence of FD bone, which may be useful in evaluating the initiation or expansion of FD lesions, and support the concept that bisphosphonates may be ineffective in FD due to their lack of binding to FD osteoid (Corsi et al, Skel Radiol, 2017). 3) Stem cells in tissue engineering and regenerative medicine Previously, a process was established for the ex vivo expansion of BMSCs under conditions that would maintain their biological properties. Human BMSCs are being manufactured around the world using many different methods (and for many different clinical applications), but little is known about the spectrum of manufacturing methods used, and their effects on BMSC characteristics and function. Eight centers (including one at the NIH CC) using or developing Good Manufacturing Practices (GMP) were surveyed as to their production methods. Among the 8 centers, all used bone marrow aspirates as the starting material, but no two centers used the same manufacturing methods. Two to four BMSC lots from each center were compared using global gene expression analysis (Agilent Chip Whole Human Genome). Among the 24 BMSC lots from the 8 centers, intra-center transcriptome variability was low and similar among centers. Principal component analysis and unsupervised hierarchical clustering analysis separated all the lots from all centers into five distinct clusters. Sufficient numbers of BMSCs from 6 of the 8 centers were available for further testing of their ability to form bone and support hematopoiesis (defining features of BMSCs) by in vivo transplantation. Cells from all 6 centers tested formed bone, but the quantity formed was highly variable, and BMSCs from only three centers supported hematopoiesis. These results show that differences in manufacturing resulted in variable BMSC characteristics, including their ability to form bone and support hematopoiesis. Although the number of centers and samples evaluated is small, the data point to a need for establishing rigorous criteria by which to establish SSCs/BMSCs in scaled-up culture conditions in order to maintain their biological properties. (Liu* and de Castro* et al, Scientific Rep, 2017, *co-first authors),", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Opsin synthesis was measured in miniature poodles affected with progressive rod-cone degeneration. Photoreceptors develop normally but begin to degenerate after the dog is fully grown. At all ages studied the rate of rod outer segment renewal was about half the normal value. Opsin synthesis, however, occurred at the normal rate until advanced stages of the disease when photoreceptor cell death was apparent. Thus, the defect may involve photoreceptor membrane assembly rather than synthesis. Peritoneal macrophages from RCS rats exhibited normal phagocytic capabilities toward bovine or rat rod outer segments as well as toward RCS retinal debris. Thus, the genetic defect is expressed in the RCS pigment epithelium but not in all phagocytic cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Program Director/Principal Investigator (Last, First, Middle): Abstract The Instrumentation Module provides design, fabrication and modification capabilities for project-specific, customized and software-flexible vision research apparatuses that are not commercially available. During the past five years, the Module fabricated many in-house, tailor-fitted and specially-designed devices that have played crucial roles in generating many high-impact and innovative vision research results. In the next grant period, this Module will continue its indispensable role in supporting vision research projects by providing new mechanical, electronic, optic and computer-based design and fabrication services and to develop highly innovative, high-impact research tools, such as the chronically implanted tetrode array recording chambers, the virtual reality visual-vestibular stimulating system, and the forced-choice, Bayesian adaptive protocol-driven optokinetic reflex machine. All these devices will be constructed and repeatedly modified, fitted and adjusted before and during experiments by the in-house machine shop and electronic/optics/computer shop. Therefore the Instrumentation Module is not only a necessity of many Vision Core labs, but also an integrated part of the research process and scientific progress. PHS 398/2590 (Rev. 11/07) Page Continuation Format Page", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Applicant's Description) This application deals with the structural biology of DNA 0-alkylation damage. As part of the applicant's long-term objectives of understanding the molecular recognition of damaged DNA, he proposes to determine the solution structure of DNA duplexes containing alkyl lesions and base analogs currently used in chemotherapy. He will establish the three-dimensional structure of the repair protein: 06-methylguanine methyltransferase (Ogt), free and after reaction with enzyme inhibitors, and of the damaged DNA/protein complex. Alkyl transferase activity has been correlated with increased sensitivity to mutations induced by methylating agents. Tumor cell lines expressing high levels of alkyl transferase activity are resistant to the action of therapeutic methylating and chloroethylating agents. In addition, alkyl transferase inhibitors used in combination with methylating chemotherapeutic agents overcome the resistance of some tumor cell lines to the drugs. The information resulting from this project will increase our knowledge of chemical mutagenesis and DNA repair and facilitate the rational design of new alkyl transferase inhibitors, which can be used in cancer therapy. Using phosphoramidite chemistry, he will synthesize large quantities of oligodeoxynucleotide duplexes containing the base analog S6-thio-2'-deoxyguanosine and S6-methyl-6-thio-2'-deoxyguanosine paired opposite to deoxycytidine and thymidine. Their three-dimensional structure will be established by NMR spectroscopy and computational methods. He will use recombinant DNA technology to obtain large quantities of highly pure Ogt protein in natural abundance as well as enriched with 15N and 13C. His solution structure will be determined by using multinuclear -- multidimensional NMR spectroscopy and computational methods. Similar methods will be used to determine the structure of protein after reaction with the enzyme inhibitors 06-methylguanine and 06-benzylguanine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A common side effect of an illness such as an infection is appetite suppression, which can quickly lead to malnutrition. Because malnutrition extends hospital stay, increases costs and exacerbates mortality, nutritional support (NS) is instituted. Unfortunately hyperglycemia is common in infected individuals and hyperglycemia negatively impacts on morbidity and mortality. We discovered that in healthy subjects the liver and muscle adapt to sustained NS by increasing their capacity to take up and oxidize glucose. As glucose is a major caloric source in NS, this adaptation is essential to minimize hyperglycemia. Infection impairs this adaptive response to NS and the current therapy administration of exogenous insulin cannot reverse this impairment. We also discovered in response to NS plasma glucagon decreases and liver derived and plasma FGF-21 increase. In addition if glucagon is increased as occurs with an infection, hepatic FGF-21 and plasma FGF-21 decrease and both hepatic and muscle carbohydrate oxidation are decreased. As FGF-21 is known to augment tissue glucose uptake our hypothesis is that, during NS, infection induced hyperglucagonemia limits FGF-21 availability and that together these changes attenuate tissue glucose uptake. We will use state of the art tracer and controlled metabolic studies to assess glucose and lipid flux in healthy and infected animals. In Aim 1 we will determine if glucagon mediated suppression of hepatic FGF-21 secretion is responsible for the decrease in muscle glucose uptake and determine whether Gs and/or Gq dependent signaling (using designer receptor technology) in the liver can explain the glucagon-mediated decrease in muscle glucose uptake. In Aim 2 we will determine if antagonizing glucagon can reverse the infection-induced impairment in tissue glucose uptake during NS. We will also determine if liver derived FGF-21 serves a protective role during infection and whether replacement of FGF-21 can sustain tissue glucose uptake during infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "All eukaryotic organisms have conserved a specific set of mechanisms for receiving and responding to extracellular signals. In most cases, these mechanisms depend on the presence of cell membrane receptors that act as signal discriminators. Such receptor systems mediate sensory transduction in the adult organism and are also employed to relay signals of developmental significance. The most ancient and diverse signalling system involves the use of a group of structurally related extracellular receptors that couple to intracellular second messenger generating systems by means of G proteins. Studies in simple organisms such as yeast point out the significance of this form of sensory transduction in mediating cell determination events during development. The aim of our studies is to investigate to role of metazoan G protein coupled receptor systems in relaying signals of developmental consequence. For these studies, we will employ the fruit fly Drosophila as an experimental system because of the wealth of genetic and molecular tools it offers as well as the easy accessibility of well--described developmental stages. We will focus specifically on the role of G protein-coupled receptors that are expressed at the earliest stages of development. A Drosophila homolog of vertebrate tachykinin receptors and a novel receptor, both expressed early in development, have been identified. These receptors will allow us to test hypotheses concerning the participation of individual receptor systems in early and cell-specific developmental responses. In addition, we will continue to identify CDNAS encoding a limited number of receptors which we predict are expressed early in embryogenesis and may therefore participate in initial determination events. The expression of each receptor within the context of well characterized developmental pathways will be analyzed using tools such as specific antibodies. These studies will allow us to predict specific roles for each receptor in mediating early developmental events or the development of specific populations of cells. Using this framework, specific predictions will be tested by the manipulation of the expression of each receptor by genetic techniques. Using this system, we will be able, for the first time, assess the potential of individual G protein coupled receptor systems to mediate the transduction of developmentally important signals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Retinal ischemia is a major cause of vision loss in common retinal disease conditions including diabetic retinopathy, glaucoma, retinopathy of prematurity, and vein occlusion. This project aims to define the mechanisms of retinal ischemic injury and identify new therapeutic targets. My long-term career goal is to pursue a distinguished career in vision research and academia. I will achieve this through establishing a strong independent research program in an academic institution that promotes interdisciplinary biomedical science and translational research. My short-term goal is to attain intensive training and supervised career development skills that are required for my career transition to become an independent investigator. Securing this award will provide me with the necessary training to achieve my short- term goals and will be the first step towards independence and achieving my long-term goals. My mentor's lab has demonstrated the involvement of the arginase enzyme in retinal neurovascular diseases. Arginase has two isoforms. Building upon the lab's finding that the mitochondrial isoform, arginase 2 (A2), has a deleterious role in retinal ischemia-reperfusion (IR) injury, I developed a project focusing on the neurovascular protective role of the cytosolic isoform arginase 1 (A1). My recently published paper shows a neuroprotective role of A1 expression in myeloid cells. Arginase competes with nitric oxide synthase (NOS) for their common substrate L-arginine. Nitric oxide (NO) produced by inducible NOS (iNOS) causes neurovascular degeneration. I predict that A1 upregulation in myeloid cells limits iNOS-derived nitrative and oxidative stress and reduces inflammation through its downstream metabolites ornithine and putrescine. Putrescine is the precursor of polyamines and it is formed from ornithine by ornithine decarboxylase (ODC, the rate-limiting enzyme in polyamine biosynthesis). These metabolites have been shown to promote reparative myeloid cells through chromatin modification. In line with this, my preliminary data show that histone deacetylase (HDAC) 3 is increased in the absence of A1 in both IR-injured retinas and stimulated macrophages. HDAC3 is essential for macrophage inflammatory gene transcription and it has been shown to suppress A1 expression. Herein, I propose a novel suppressive effect of A1 on HDAC3. My central hypothesis predicts that myeloid A1 protects against retinal IR injury through ODC-mediated suppression of HDAC3. I will be using mice with myeloid-specific deletion of A1, ODC and HDAC3, as well as the investigational drug, BCT-100 (a PEGylated form of arginase 1), together with primary macrophages isolation and treatment with inhibitors for HDAC3 or arginase downstream enzyme, ODC. My goal is to achieve the following objectives: A) Determine the effect of manipulating the arginase pathway on myeloid cells infiltration / activation in retinal IR injury and the therapeutic potential of BCT- 100. B) Describe the cross talk between the arginase pathway and HDAC3 and determine whether A1 in myeloid cells mediates its protective effect through suppression of HDAC3.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this project is two-fold. The first aim is to develop a coordinated series of instruments, primarily checklists, to elicit descriptions of the child from the several perspectives of parents, teachers and clinicians. The items on the checklists are to be weighted in terms of the relative severity of psychopathology represented as judged by a national sample of mental health professionals. Application of these weights to subscores derived from checklist content clusters will produce clinically meaningful diagnostic profiles of individual children. Treatment planning and outcome evaluation forms also are being developed. The second aim in terms of timing and the major substantive goal is to evaluate the course of psychopathology in children aged 6 to 18 years who have experienced selected treatment and no-treatment regimens. An initial, six-month and 15-month assessment of each child will be made. Separate data analyses are to be performed for children categorized in terms of a few major types of disorder and for normal school children. The independent variables will be age of child, type of treatment (or no treatment), and severity of psychopathology. The dependent variables will be checklist scores, some of which will reflect changes in total symptomatology while others will reflect changes in target complaints as identified from the diverse vantage points of parents, teachers and therapists.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research application is in response to #RFA-DA-06-004. At the heart of drug and alcohol abuse is bad decision-making, especially in response to social influences. This decision-making defect is highly reminiscent of the social decision-making impairments that characterize neurological patients with damage to ventromedial prefrontal cortex (VMPC). The VMPC patients provide a neurological model of social decision- making gone awry. There is evidence at both preclinical and clinical levels that the VMPC is a key brain region in drug abuse, and we propose that VMPC dysfunction and defective social decision-making may characterize individuals who abuse and become addicted to drugs and alcohol. In this research application, we plan to conduct studies in neurological patients in order to flesh out the neurobiological underpinnings of social decision-making, taking advantage of the unique Iowa Patient Registry that contains neuropsychological and neuroanatomical data for thousands of patients. A particular focus is on gender differences. We have intriguing preliminary findings suggesting that there might be an important sex-related difference in the relationship between the VMPC and social decision-making: in men, the right VMPC might be critical, whereas in women, the left VMPC might be critical. The experiments will test three specific aims: (1) To determine whether there is sex-related functional asymmetry of the VMPC in regard to social decision- making, using tasks such as the Ultimatum Game, Trust Game, Iowa Gambling Task, and Ellsberg Tasks; (2) To extend the investigation of sex-related functional asymmetry to other brain structures known to be critical for social and affective processes related to decision-making, namely, the amygdala and the insular cortex; (3) To investigate developmental influences on the relationship between the VMPC and social- decision making and emotional processing, by studying patients who incurred VMPC damage early in life. The experiments will furnish important new information about the neurobiological underpinnings of social decision-making and emotional processing. The link with substance abuse is direct: the decision-making deficits in VMPC patients have striking similarities with those evident in drug and alcohol abusers and addicts (\"myopia for the future\"). Thus, our research could help pinpoint sources of neural dysfunction that contribute to bad decision-making of the type that characterizes drug and alcohol abuse. Public health relevance: This research will help us understand how various parts of the brain are important for social decision-making and emotional processing, whether there are gender differences in these brain- behavior relationships, and how these relationships develop. The research could help inform treatment and prevention of drug and alcohol abuse disorders, as well as social conduct disorders, personality disorders, and anxiety and mood disorders, all of which have high comorbidity with drug and alcohol abuse. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Adapted from the application) The main goal of this core is to support and optimize the activities of the investigators performing studies with animals. This core will provide services in a. breeding and maintaining normal, congeneic, and gene-deficient mice with reproducible enteric microbiota and clinical disease in both conventional and specific pathogen-free housing, and b. generating germ-free and mono-associated infected mice. This core will provide support to Projects 2, 3 and 4.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed International Collaborations in Infectious Disease Research renewal program comprises a multidisciplinary and coordinated set of epidemiological, clinical and laboratory research activities in enteric disease, and will specifically focus on Giardia lamblia, Cryptosporidium, and E. coli infection in children. A major integrating theme will be to determine relationships between manifestations of infection in the host, and characteristics of the host and of the enteric pathogen. We will explore how infectively of each agent, and clinical expression of infection relate to the environmental, nutritional, and immunological status of the child, and to genetic and/or antigenic variability of Giardia lamblia, Cryptosporidium, and E. coli. The proposed ICIDR will be organized as five research projects and six core components and will be a collaborative thematically cohesive program among investigators from US institutions (Columbia University, Cornell University, University of Maryland, Centers for Disease Control, Univacs, ImmuCell, Merck) and institutions in Israel (Ben-Gurion University, Hebrew University, Army Health Branch Research Unit). Project 1 will prospectively follow 240 Bedouin infants from birth to age 2 years, to examine the environmental, host, and organism characteristics and the interactions between them, leading to the range of clinical manifestations associated with the three organisms. Using novel methods of pathogen identification and characterization (ELISA assays, DNA probes), genomic karyotyping, hygienic intervention protocols, and in depth analysis of nutritional/immune/environmental status, we will establish the time course and causality of these relationships, and also how asymptomatic Giardia lamblia carriage diminishes infections with other pathogens. Since Project 1 will not provide a sufficient number of cases to explore these relationships in children with severe clinical manifestations, Project 2 will identify environmental/host/organism factors associated with diarrhea in sick hospitalized children. Project 2 will also specifically assess two therapeutic interventions (azithromycin and hyperimmune anti-Cryptosporidum colostrum immunoglobulin) on the course of cryptosporidial diarrhea in a hospitalized population. Project 3 will probe the distribution of strain diversity of Giardia lamblia and Cryptosporidium using karyotype analysis and other strain markers; utilize PCR approaches for identifying and characterizing Giardia lamblia, and study in vitro cytopathogenic effects of Giardia lamblia on human intestinal epithelial cell cultures and lymphocytes. Project 4 will focus on the cellular immune response in infants in infants and young children, in relation to symptomatic and asymptomatic infections with the three groups of organisms with particular emphasis on development of different lymphocyte subpopulations e.g. 'memory cells'. Project 5 will provide an in depth understanding of the basic molecular genetic pathways underlying the high frequency of mutation and chromosome rearrangements in Giardia lamblia, and its variable gene expression. The Core components include: (a) ICIDR administration, (b) a Visiting Investigator Program, (c) Microbiology, (d) Giardia lamblia Serology, (e) Cryptosporidium/E. coli Serology, and (f) a Biostatistics center. The proposed ICIDR Program provides a unique opportunity to examine, in a Middle Eastern population with diarrheal disease patterns of developing nations, the role of Giardia lamblia, Cryptosporidium and different categories of E.coli in diarrheal diseases, and contribute knowledge essential for development of new intervention strategies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Acute lymphoblastic leukemia (ALL) is the most common cancer in children and adolescents, killing more pediatric patients every year than any other malignancy. When children with ALL relapse the survival rate is less than 50%, and this has not improved for more than 30 years. Although ALL is less common in adults, it is more deadly, reaching 5-year survival rates of less than 12% in people over 65, an age when tolerance for toxic therapies is poor. The majority of ALL cases in children and adults are the B-cell type (B-ALL) and those with the poorest outcome are characterized by overexpression of the CRLF2 gene (CRLF2 B-ALL). This type of leukemia is particularly devastating in Hispanic children where it occurs 5 times more often than in other children. In adults, CRLF2 B-ALL makes up 1/3 of all B-ALL cases. A safe and effective treatment capable of targeting CRLF2 B-ALL would have a significant health impact in the United States and in Central and South America where rates of CRLF2 B-ALL are even higher. We have discovered a novel biologic that selectively targets the signaling pathway that produces CRLF2 B-ALL while sparing normal immune cells. The proposed studies will establish the therapeutic efficacy of this novel biologic in patient-derived xenograft (PDX) models of high-risk ALL. Scientific premise: Our preliminary studies show that high physiological levels of the biologic exert an anti-leukemia effect against CRLF2 B-ALL and establish a likely mechanism for these effects. To advance the use of the biologic in clinical trial for the treatment of CRLF2 B-ALL, it is essential to establish the dose and efficacy of the biologic as a single agent and as a part of combination therapy in preclinical studies. The overall objective of the current proposal is to establish proof-of-principle for the use of the biologic and its efficacy in the treatment of high-risk CRLF2 B-ALL. The achieve this objective we will: Aim 1. Establish the dose of biologic required to achieve normal and therapeutic blood plasma levels in PDX mice and Aim 2. Evaluate the in vivo therapeutic efficacy of the biologic as a single agent and in combination therapy using PDX models of high-risk B-ALL. At the end of the proposed work we will have: 1) Determined the dosing required to generate PDX with normal and therapeutic levels of recombinant human biologic, 2) Assessed in vivo functional effects and toxicity of the biologic, 3) Evaluated the efficacy of the biologic as a single agent in the treatment of CRLF2 B-ALL using PDX models; 4) Evaluated the efficacy of the biologic to prevent relapse and restore normal B cell precursors in combination with standard of care therapy in PDX models of CRLF2 B-ALL. This work has the potential to produce a more effective and less toxic treatment strategy to improve survival in patients with high-risk CRLF2 B-ALL.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT Mossy cells (MCs) of the dentate gyrus (DG) are glutamatergic neurons that are considered to be important to normal function and their injury has been suggested to contribute to neurological and psychiatric disorders, as well as deficits after traumatic brain injury. Anatomical and slice electrophysiology studies have described MCs in detail, but there is a gap between these studies and understanding how MCs contribute to DG-dependent behavior in vivo. To address this issue, we began with a simple approach: mice were engaged in behaviors related to DG function, and MCs were examined afterwards using the neural activity marker c-fos. We quickly found that simply exploring novel objects led to a large increase in MC c-fos immunoreactivity (ir). Interestingly, most c- fos-ir did not increase in most of the other DG neurons, suggesting preferential activation of MCs by novelty. However, there was one area of the DG where c-fos-ir was consistent: a subset of GCs in dorsal DG. In contrast, the majority of MCs with c-fos-ir were ventral. Because the main projection of ventral MCs is to dorsal GCs, these data suggest that ventral MCs excite dorsal GCs. This circuitry helps explain how normally quiescent GCs become activated in dorsal DG, which is considered essential for cognitive functions of the DG. In Aim 1 we will use optogenetics to test this hypothesis, taking advantage of new mouse lines that have targeted Cre recombinase to MCs. We will also ask if dorsal MCs have effects analogous to ventral MCs, i.e., dorsal MCs contribute to ventral DG functions. In Aim 2 the underlying circuitry will be addressed. We suggest that optogenetic excitation of MCs in a normal adult mouse will recapitulate the results with c-fos: MCs excite proximal GCs weakly but distal GCs in a more robust manner. This idea has been supported by data from slices that were cut at an angle to preserve MC axons, and will be tested further in Aim 2 using voltage imaging and microelectrodes. In Aim 3 we will address the hypothesis that a large number of the distal GCs that are activated by MCs are immature. That hypothesis supports a previously published study showing that MCs are a primary source of afferent input to young GCs that are born in adulthood. This is potentially important because immature GCs are considered central to DG functions. Therefore, we will address the additional hypothesis that MCs activate adult-born GCs primarily in distal locations, leading to stronger excitation of distal GCs than proximal GCs. By providing afferent input to immature GCs, MCs could play a critical role in behaviors associated with adult DG neurogenesis. Together these experiments will significantly advance our understanding of DG circuitry and its contribution to behavior. Because MC injury is associated with several disorders, these experiments will also shed light on impairments in DG functions in those pathological conditions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pain is a common co-morbidity for HIV-infected patients. Prevalence studies suggest that, on average, half of all HIV-infected persons suffer pain. Chronic pain can lead to heavy alcohol use among HIV-infected persons, which may in turn be a barrier to treatment/control of HIV and contribute to spread of HIV. Thus there is an urgent need to address pain among persons with HIV. Opioid receptor antagonists such as naltrexone and nalmefene, which are licensed for treatment of alcohol use disorders, show promise as being effective and safe treatments for chronic pain among persons with HIV. This study will pilot test novel pharmacotherapies (opioid receptor antagonists) to improve chronic pain among HIV-infected heavy drinkers, and will explore the hypothesis that the mechanism of action for improving pain is through decreased inflammation. The specific aims of the research are: UH2/Aim 1: To assess the feasibility, tolerability and safety of using opioid receptor antagonists (low-dose naltrexone and nalmefene) to treat pain among HIV-infected persons with heavy alcohol use and chronic pain; UH3/Aim 2: to perform a 3-arm pilot randomized, double-blinded, placebo-controlled study of low-dose naltrexone and nalmefene vs. placebo among HIV-infected persons with heavy alcohol use and chronic pain to provide estimates of their effects on: 1) pain (both self-reported and experimental/cold pressor test; 2) inflammation (i.e., levels of inflammatory cytokines IL-6 and TNF-?); and 3) measures of HIV control (CD4 count and viral load). The results of this study will provide preliminary information (tolerability, effect size, etc.) to design a larger RCT of low-dose naltrexone and/or nalmefene for chronic pain among persons with heavy alcohol use. We choose to conduct this research in St. Petersburg, Russia, given that: 1) nalmefene is licensed in Russia, but not currently in the US; 2) patients are seldom on chronic opioids (which are contraindicated to use with opioid receptor antagonists) due to the unavailability of opioid agonist therapy for addiction and restricted use of opioids for pain; and 3) a high prevalence of heavy drinking and HIV exists in Russia. Addressing chronic pain is a high priority for patients with HIV, and therefore this application is highly ?patient-centered? as well as innovative. Given the US epidemic of opioid use disorders, new pharmacotherapies without addictive potential are desperately needed for HIV-infected persons with chronic pain and alcohol problems.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Principal Investigator/Program Director (Last, first, middle): Zeng, Fan-Gang Tinnitus is the perception of sound in the absence of external sound. Tinnitus is a significant public health problem that affects 50 million Americans. Severe tinnitus disrupts daily functions from sleep to work, often leading to anxiety, depression and lowered quality of life. Despite significant advances in research and development, presently there is no cure for tinnitus. The present application uses noninvasive electric stimulation in the ear and minimally-invasive electric stimulation to the round window or promontory for safe and effective treatment of tinnitus. One innovation is to evaluate stimulation sites and patterns that evoke auditory sensations while minimizing non-auditory sensations. Another innovation is to provide two novel tinnitus treatment options, especially for those who still have significant acoustic hearing and cite tinnitus, and not hearing loss, as the main indication. In the preliminary study, 10 minutes of round window stimulation completely silenced the tinnitus not only during stimulation but also for 5 hours after the stimulation in a subject who had suffered from tinnitus for 15 years. Successful completion of the present work can lead to safe and effective medical devices for tinnitus treatment. Project Summary", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Musculoskeletal symptoms, such as arthritis, tendonitis and bursitis, are very common in Systemic Lupus Erythematosus (SLE). Pain and disability from these symptoms can be significant and current treatment options for musculoskeletal pain in SLE include non-steroidal anti-inflammatory medications (NSAIDS), narcotic analgesia and corticosteroids. Each of these is limited by significant potential toxicitie including gastrointestinal bleeding and kidney toxicity in the case of NSAIDS, somnolence, constipation and nausea for narcotics and osteoporosis, osteonecrosis, diabetes, cataracts and atherosclerosis for corticosteroids. Ajulemic acid (AjA) is derived from a class of cannabinoids that retain analgesic and anti-inflammatory properties without the psychotropic effects characteristic of tetrahydrocannabinol (THC; marijuana). AjA has been shown to have potent pain- relieving effects, without psychotropic effects, in animal studies and a clinical trial in human subjects with chronic neuropathic pain. Anti-inflammatory effects of AjA have also been demonstrated in animal models as well as in human subjects and in vitro studies of human inflammatory cells. Thus far, a total of 123 subjects (healthy individuals and patients with neuropathic pain) have received AjA with no serious side effects. The overall purpose of this Phase IIa clinical trial is to evaluate the safety of AjA in SLE patients with mild to moderate musculoskeletal pain and to determine an optimum dose of AjA that will provide maximum benefit and minimal toxicity. Clinical improvement will be determined by assessment of SLE disease activity and changes in pain scales. Studies are also planned to evaluate the mechanisms of action of AjA on circulating inflammatory cells. Based on pre- clinical studies already completed in our laboratory, we expect that AjA will decrease the expression of certain pro-inflammatory proteins in peripheral blood cells and increase the expression of other proteins that enhance the resolution of inflammation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have demonstrated that Raman spectroscopy, a noninvasive technique yields significant information on both structural and chemical properties of the intact transparent crystalline lens. It is our objective to ascertain the feasibility of applying this technique to experimentally and naturally occurring cataracts. Our preliminary studies of cold-cataracts have demonstrated that Raman spectroscopy can be applied to provide information about cataractogenesis. Upon determining the Raman spectra signatures of cataracts, we will develop an in vivo method of studying the lens. Such an in vivo measurement of the Raman spectrum may suggest biochemical changes that occur in the lens before cataracts actually develop. An understanding of these biochemical alterations should produce significant information which is crucial to the development of methods for the prevention and reversal of cataract.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Attachment of bacteria to the host tissue represents the first critical step in a process that may lead to clinically manifested infections. Extracellular pathogens attach via specific surface located MSCRAMMs (Microbial Surface Components Recognizing Adhesive Matrix Molecules) to components of the extracellular matrix. We have previously demonstrated that many Gram-positive bacteria can adhere to collagen and we identified collagen binding MSCRAMMs on S. aureus and E. faecalis called CNA and ACE, respectively. We found that these MSCRAMMs can bind to multiple sites in collagen. The structure of the collagen binding domain of CNA was determined using X-ray crystallographic methods and found to contain a collagen binding extended trench on its surface. Furthermore, we found that vaccination of mice with recombinant CNA protected the animals against S. aureus induced septic death. These results form the basis for the now proposed studies where we want to identify additional collage binding MSCRAMMs and determine the structures of CNA and ACE. We hypothesize that these MSCRAMMs and other collagen adhesion receptors such as the integrins, contain trenches on their surface that represent the collagen binding sites. This hypothesis, as well as its mechanistic implication for collagen binding, will now be examined. We will identify binding sites in collagens for different adhesion receptors and characterize the interaction of synthetic triple helix collagen peptides containing these binding sites with the receptors. Finally, we will analyze in detail the established role of CNA as a virulence factor in septic arthritis and locate the protective epitopes in this MSCRAMM.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Apoptosis occurs under a variety of physiological and pathological conditions. It is essential for maintaining the homeostasis of the skin, and inappropriate apoptosis has been implicated in the pathogenesis of a multitude of skin disorders. Apoptosis mediates the decrease in cellularity during the transition between granulation to scar tissue, and aberrant apoptosis during wound healing has been attributed to the development of hypertrophic scars and keloids. Apoptosis may protect against the development of UV-induced skin cancer by selectively destroying sun-damaged keratinocytes. Mutations in the components of the surveillance machinery, such as p53 and bd-2, provide cells with an opportunity to escape from apoptosis and develop into nonmelanoma skin cancers. Early onset of apoptosis has been observed in diabetic wounds, contributing to blunted wound healing. Thus, manipulations which alter the onset or extent of apoptosis can potentially provide therapeutic interventions for the treatment of a variety of skin diseases. It is therefore important to delineate the molecular pathways and regulation of this multistep process. In this grant, I propose to use a variety of ways to manipulate the actin cytoskeleton and determine how actin remodeling contributes to UV- induced apoptosis in keratinocytes. I will examine the role of gelsolin, a Ca2+-activated actin filament severing protein which is recently identified as a direct and predominant substrate of caspase-3, in UVB-mediated apoptosis. This project is a new direction for my laboratory, but is very appropriate and timely because very little is known about the involvement of the actin cytoskeleton in apoptosis, even though the cytoskeleton is an important structural and regulatory machine. Its derangement is likely to contribute to the pathophysiological of many skin diseases. We have the tools and expertise to investigate this very important basic biological problem, and anticipate that our findings will suggest novel approaches for the treatment/or prevention of many skin diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the present project we are attempting to identify the parameters of intermediary metabolism that may be of importance for cellular radiation sensitivity, and to utilize the knowledge gained to devise improved schemes for enhancement of radiation response. Initially, we will attempt to poise cells in vitro at high and low levels of potential intracellular radical scavengers, reduced pyridine nucleotides, non- protein sulfhydryls and endogenous substrates, in the presence and absence of oxygen, and determine the radiation sensitivity of the system in terms of cell survival. Of particular value in this study are chemicals that sensitize under anoxic conditions. The use of these chemicals will enable us to control cellular electron transfer as well as the level intracellular scavengers in order to determine the importance of reductive metabolism in repair of radiation damage. Sensitizers will also be used in combination with insulin, glucose and phosphate, agents known to alter cellular radiosensitivity as well as cellular redox potential. We plan to assay radiation response with both tissue cultured cells and animal ascites tumors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposal describes the design of an adaptable computer language called AID, to be used for Assistance In Decision making for acute care of trauma and burn victims. The system should overcome the underutilization of complex data now collected by ICU information systems by providing up-to-date medical consultation to users unfamiliar with their interpretation and clinical significance. The language has been designed to provide powerful capabilities for retrieval from computer medical records so that consultation may occur without time-consuming data entry by physicians, and to provide interpretations of varying degrees of certainty when data are missing. The system will permit a useful structuring of the computer record so that significant findings are presented opportunely and concisely. The initial clinical applications are for ventilatory management, head trauma, and fluid/electrolyte balance. Development of decision algorithms in these areas should lead to a more basic understanding of the pathophysiology and treatment of trauma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The association between human leukocyte antigen (HLA) B27 and ankylosing spondylitis (AS) is among the strongest associations known between a disease and a genetic marker. Genes, both within and outside the major histocompatibility complex, are believed to account for more than 90% of the population variance in AS. While these associations clearly indicates strong genetic contributions to the susceptibility to AS, whether genes influence the severity, of AS is unknown. The severity of AS varies widely among patients, with some having severe persistent joint inflammation, functional limitation, premature work disability, and early spinal fusion. We hypothesize that the severity of -AS is genetically determined, and that genes that influence susceptibility to AS, genes that influence the severity of inflammatory responses, and genes regulating transforming growth factor-beta are among the genes that also influence its severity. We propose a model of AS severity in which genetic differences among patients influence the immune response and consequently the persistence and severity of joint inflammation. Persistent severe joint inflammation thereafter leads to spinal ankylosis and poorer long-term health outcomes. We will test this model in two stages. First, we will identify genetic markers associated with worse radiological outcomes, greater functional disability, work disability and need for total hip arthroplasty in a retrospective cohort study of 400 patients with AS of 20 years or longer. Second, we will test these genetic markers for associations with persistent active inflammation in a prospective cohort study of 200 patients with early AS who will be repeatedly assessed over 2 years. The model will be supported if the genetic markers that are associated with persistent active inflammation are also associated with poorer long-term health outcomes. In addition, the prospective cohort will enable the development and assessment of new clinical and radiographic measures of disease activity in AS for use in clinical trials. Understanding the contribution of genetic factors to differences in the severity of AS and in the health outcomes of patients would further our understanding of the pathogenesis of AS, provide important prognostic information, and could identify subgroups at high risk for poor outcomes who could then be targeted for more aggressive interventions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although there is very high interest in the demography and welfare of families headed by lesbian and gay (LG) parents, there is currently very limited longitudinal research on LG families or the transition to LG parenthood. Most of the extant literature is descriptive, comparative, or limited to distinct subgroups of LG parents. Of this research, demography studies that examined trends in LG family structures have rarely considered the family and individual processes that may account for changing trends in LG family formation; meanwhile developmental studies of LG parents have rarely considered the ecological and cultural influences on family and individual functioning, nor systematically considered how family structure may impact the family dynamic. Lastly, no studies that I am aware of have integrated family demography and biosocial approaches to understanding how stress is related to ecological and individual or family decisions and behaviors in LG families. The current research will use both secondary data analyses of longitudinal national datasets such as the U.S. Census and American Community Survey to examine within-state changes in rates of LG family structures over the past 10 years and how these changes correlate with state-level sociopolitical climates and public policies pertinent to LG family formation. This will be followed by a pilot study that will explicitly examine how sociopolitical and ecological factors ar associated with decisions regarding family formation and LG family functioning as mediated by individual mental health, relationship stability, and stress load. I have outlined 3 training goals that are critical for the success of this timely and innovative research study. The first goal is t develop conceptual and methodological expertise in the study of family demography, with an emphasis on contextual influences on family formation for lesbian and gay parents. The second goal is to increase my knowledge and expertise in using established national datasets such as the U.S. Census and the American Community survey to conduct secondary data analyses pertinent to the study of LG families. The third goal is to conduct a multi-state pilot using a biosocial model of individual and family formation and functioning to understand how contextual factors are related to LG individuals' well-being, decisions regarding the timing, method, and location for family formation, and parental functioning in preparation of submitting an R01 proposal on this topic in the later years of this K01 funding period. I have established an extremely strong and accomplished mentorship team that will guide my training and collaborate on each phase of the proposed research. In consultation with my mentors and consultants I have established a training plan that includes (1) mentor-based supervised training and directed activities, (2) formal graduate level coursework and short courses, and (3) participation in relevant research seminars, colloquia and scientific meetings. This training and pilot research will provide me with the skills, experience, and preliminary data necessary to establish an independent research program on this highly significant and innovative topic.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The efficacy of medical therapy is usually best demonstrated by randomized clinical trials but results of full-scale trials may be unavailable or may not be applicable to single patients. Single patient trials have been proposed to solve therapeutic dilemmas in dividual patients but are difficult for the clinician to perform. A single-patient clinical trial service will be established at a university hospital. The participants will publicize the role of single-patient clinical trials and teach clinicians about their applicability to certain therapeutic problems. They will assist clinicians in designing, executing, and interpreting randomized, crossover, controlled trials in appropriate patients. The value of the trials in therepeutic decision making for patient well being will be assessed on a case-by-case basis. The feasibility of the service will be assessed based on the value of single-patient clinical trials to patient care and the resources expended in providing the service. If successful, the service could become an ongoing, self-sustaining activity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal focuses on those drugs that are commonly utilized for the treatment of adult solid tumors, including both modern \"targeted\" oncology drugs, as well as cytotoxic agents. This includes studies of new candidate genes identified through recently completed genome-wide studies, both clinical and in the HapMap lymphoblastoid cell lines (LCLs). A central strategy of the proposal is the development of endophenotypes that are intermediate between clinical phenotypes, such as adverse events and measures of efficacy, and the genetic variants that affect their risk. All research projects are classified as primarily phenotype-centric, drug-centric, or gene-centric. The phenotype-centric studies will focus on markers of gene activity, such as the transcriptome, or potentially serum proteomic markers. The drug-centric studies will focus on drugs of importance for adult solid tumors. The gene-centric studies generally focus in a detailed way on candidate genes identified through these other studies. The overall goal is to identify functional relationships at the intersection of drug, gene, and phenotype, particularly those phenotypes that are affected by one or more functional polymorphisms when patients are treated with a specific anticancer drug (at a clinically relevant dose). Mathematically, this can be described by a multidimensional matrix (to be made publicly available through a PGScore database) of variation in a specific gene (or polymorphism) on the effects of a drug as measured by a specific phenotype. A consistent framework is utilized to address questions relevant to six interrelated Themes (Cytotoxicity, Transcriptome, Irinotecan, Angiogenesis, UGT, and EGFR), supported by two Platforms (Clinical Studies and Functional Studies) and four Cores (Management, Genetics-Informatics-Statistics, Liver, and LCL). Extensive collaborative clinical studies are proposed with CALGB, as well as other current PGRN Groups. Other ongoing collaborations will be continued, including studies of glucuronidation and LCLs. In addition, a new Network Resource is proposed. Cell Lines as a Resource for Pharmacogenomic Studies. PUBLIC HEALTH RELEVANCE: There is marked variability in pharmacological response to anticancer agents, which historically have been characterized by severe toxicity and inconsistent efficacy. This proposal aims to characterize the genomic basis for this variability through a series of interrelated studies addressing a breadth of modern and classical anticancer agents. This will include both laboratory and translational clinical studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are two main objectives of this research: (1) to develop the methodology - utilizing both instrumental and radiochemical neutron activation analysis - to determine selected element concentrations in cow's milk, infant formula, and human milk; and (2) to compare these element concentrations in pooled samples of cow's milk and infant formula to individual samples from human volunteers (20-30) sampled over that period of time prior to use of supplemental foods (3-6 months). The elements expected to be determined are Fe, Cu, Mn, Zn, Co, Mo, Se, Cr, As, Sb, Cs, Rb, Eu, K, Ca, Br, Na.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The current proposal, for the first time, addresses the role of neuro-immune activation within the central nervous system as a plausible mechanism underlying the visceral pain component of many common functional gastrointestinal disorders (FGID) including irritable bowel syndrome (IBS). Although increasing evidence has emerged for the role of spinal glia in the mechanisms mediating persistent pain, its implication in visceral pain has not been evaluated. We propose the general hypothesis that chronic stress, known as a key factor in the first onset or exacerbation of IBS symptoms, triggers the activation of a spinal network comprising neurons and immune cells (glia), playing a central role in the modulation of visceral nociception. Using a rat model of chronic water avoidance stress, we propose to characterize the role of stress-induced spinal glia activation in the modulation of visceral sensitivity and determine the molecular pathways engaged in the initiation and maintenance of visceral hyperalgesia. Our first specific aim is to assess the temporal profile of spinal glia activation during and after chronic stress and to test the ability of inhibitors specific of glia to reduce stress- induced visceral hyperalgesia. The second specific aim relates to the characterization of molecular mechanisms linking chronic stress and spinal glial activation. The experimental design proposed in this application includes the behavioral assessment of visceral sensitivity in response to different pharmacological treatments (antagonists, agonists, oligonucleotide antisenses), combined with in vitro analysis using Western blotting, multiplex ELISA, immunohistochemistry and quantitative RT-PCR. The long-term goal of the proposed studies is the characterization of signaling pathways underlying the observed endocrine-neural- immune interactions using selective knockout animals and glia/neurons co-cultures. Targeting specific mediators of the glial-neuronal crosstalk may provide an innovative approach for the development of novel therapeutic targets for the treatment of chronic pain conditions associated with enhanced stress responsiveness. The concept of stress-induced modulation of glial-neurons signaling and its implication in long-term alteration of the sensory system may be generalized to many other stress-sensitive pain conditions, including interstitial cystitis, non-cardiac chest pain and fibromyalgia. PUBLIC HEALTH RELEVANCE: In the current proposal, we have developed the concept that chronic stress may induce activation of the immune system in the spinal cord, initiating a cascade of events that will affect the sensory system. This new approach has considerable implication for the development of novel therapeutic targets for the treatment of chronic pain conditions associated with enhanced stress responsiveness, including functional gastrointestinal disorders, for which effective treatment remains a clinical challenge.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Schizophrenia, Parkinson's disease, Tourette's syndrome, obsessive-compulsive disorder and drug addiction, are only some of the relevant clinical conditions that have been related to dysfunction of brain dopamine systems and abnormal processing of cortical input in the striatum. The robust membrane potential depolarizations (\"up states\") during which striatal neurons become responsive to the fine structure of cortical input, are synaptically-driven events invoked by the cortex itself and probably shaped by dopamine-regulated membrane currents. The suggestion that the striatum behaves as an \"action selection\" network, raised interest in understanding how dynamic patterns of cortical activity are represented in the striatum and how this representation is modified by changes in dopamine neurotransmission. By simultaneously recording the population activity of cortical neurons and the membrane potential of striatal neurons, we have recently clarified some aspects of the temporal dynamics of up states. In this proposal, we plan to analyze the representation of spatial variations of cortical activity on striatal neurons by recording field potentials and multiunit activity from multiple cortical sites together with the membrane potential of striatal neurons in anesthetized rats. Our main goal is to understand how spatially-distributed and dynamically-changing patterns of cortical activity are reflected in the membrane potential (as an index of synaptic input) and firing pattern (as an index of neuronal output) of striatal neurons. Furthermore, we will investigate the impact of cortical activity on striatal neurons in genetically-modified mice lacking functional D1 or D2 dopamine receptors. This research will be done primarily in Argentina, at the Buenos Aires University School of Medicine, in collaboration with M. Gustavo Murer, as an extension of NIH grant # R01MH060131.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project on human immunogenetics of the major histocompatibility complex is an integrated investigation of immunological, genetic, and biochemical aspects of the HLA complex and closely linked genes. The overall objectives for this project are to obtain a better understanding of the genetic factors responsible for allograft reactions, to identify the immunological mechanisms involved in such reactions, and to understand the relationship between the HLA complex and genetic factors predisposing to diseases. The studies are divided into four projects. (1)\\Cell surface phenotype and function of human alloactivated lymphocytes. This project utilizes the techniques of T-lymphocyte cloning for the analysis of lymphocyte diversity. (2)\\In vitro production by human B lymphocytes of human monoclonal antibodies detecting cell surface antibodies is being investigated. (3)\\Serological, biochemical, and genetic analysis of the human equivalent of the thymus-leukemia (TL) antigen will be undertaken and compared with the murine counterparts. (4)\\The relationship between a disease-causing gene and HLA will be investigated using congenital adrenal hyperplasia due to 21-hydroxylase deficiency as a model. This project utilizes molecular genetic tools for the analysis of the genes in the HLA region. The major emphasis in the studies of alloreactivity during the last year has been the interactions between cell surface determinants on allocytotoxic-T cells and specific epitopes on cell surface antigens on the target cells. These studies have involved allocytotoxic, human T-cell clones. The conclusions are based upon the combined analysis of biochemical studies of MHC antigens using murine anti-HLA monoclonal antibodies, analysis of T-lymphocyte differentiation antigens with monoclonal antibodies and blocking studies of allocytotoxicity with these two sets of antibodies. The investigations have revealed a distinct but complex pattern. It has been demonstrated that multiple HLA-D-related class II antigens all carry allotypic determinants which can be recognized by cytotoxic T cells. The contribution of different T-cell differentiation antigens in the cytotoxic reaction against class I and class II antigens is also complex. While the T4 positive T cells primarily recognize class II antigens and T8 positive T cells primarily recognize class I antigens, this correlation is far from complete. The second major progress within this project on human immunogenetics has been achieved in the molecular genetic analysis of congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OH-Def.). We have during the last year cloned and characterized a cDNA for adrenal 21-hydroxylase. This cDNA has been used to map two 21-OH-genes within the murine H-2 complex. These genes are located within the S-Region of H-2 and very close to H-2D. The same cDNA probe has been used to characterize the 21-OH genes within the HLA complex. At least one of these two genes are deleted in the HLA-Bw47 positive HLA haplotypes found in some patients with the severe, salt-wasting form of 21-OH-Def. These studies indicate that CAH due to 21-OH-Def. is a defect in the structural gene for adrenal cytochrome P450 21-hydroxylase. (CS)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Continuing biological evaluation of potential anticancer and antisepsis chemotherapeutic analogues of members of the hydrolyzable tannin family of secondary plant metabolites is proposed. Characterization of the immunomodulatory properties of these species will proceed. Mechanism-of-action investigations at the molecular, cellular, and whole animal levels, coupled with structure/activity studies, are designed to reveal the basis for both up-regulation and down-regulation of cytokine (IL-lb and TNFa) secretion by certain tannins. A novel tumor targeting strategy for tannin delivery will be explored. Appreciation of the molecular-level details responsible for either triggering or suppressing cytokine release may serve as the basis for designing tannin-inspired chemotherapeutic agents for diseases as diverse as cancer and septic shock. Ongoing synthesis studies directed toward the potent marine anticancer principle diazonamide A will be brought to completion. In addition to providing a supply of this scarce material for further biological evaluation, successful execution of the plausibly biomimetic synthesis route will provide insight into the otherwise obscure biosynthesis of this complex peptide-based secondary metabolite. Finally, new studies on the synthesis of the architecturally complex 20S proteosome inhibitor TMC-95A and rationally designed analogues will be pursued. A possibly biomimetic (modified) tryptophan oxidative cyclization forms the centerpiece of the approach, and acquisition of the target and the aforementioned analogues will enable mechanism-of-action studies to move forward. Inhibition of the 20S proteosome can form the basis for chemotherapeutic intervention in a variety of disease states, including cancer, cachexia, and sepsis. The search for selective 20S proteosome inhibitors among the Ntn-type proteases will be advanced by these investigations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The present study is a continuation and further development of a novel program of research designed to reduce the onset and extent of drinking by students during their first year of college, through provision of efficacious individually oriented interventions prior to college entrance. We aim to influence the drinking behavior of college-bound high school athletes during the spring of their senior year of high school, by enhancing the influence of their parents and/or by introducing them to college student athletes who will serve as peer counselors to provide a brief motivational feedback intervention. Athletes represent a high-risk and understudied group, and existing research indicates high school athletes drink as much as or more than other students. Research also indicates that for many students, excessive drinking in college is a continuation or exacerbation of high-school drinking tendencies. However, almost all current approaches in the college-drinking domain are based on implementing interventions while the students are at college. Further, there have been no theory-driven, systematic interventions targeting high school athletes so as to prevent alcohol misuse as they transition to college. We intend to implement two early interventions for high school student athletes: (1) a Parent-Based Intervention (PBI) based on the work of Turrisi and colleagues and (2) a Brief Peer-delivered Motivational Intervention (BPMI) based on the work of Larimer and colleagues. There is sufficient empirical evidence demonstrating the efficacy of these interventions in college and older adolescent samples to warrant an examination of their unique and combined additive effects for a high-risk high school athlete sample. The integration of these two research programs will provide a unique theoretical framework and a rich database from which to assess the benefits of parent and peer-delivered intervention approaches at this late stage of adolescent development. Thus, the research will investigate, through the use of a factorial design, the unique and combined utility of the PBI and the BPMI to reduce alcohol onset, usage and negative consequences among high school athletes so as to ultimately reduce college student drinking; identify demographic and psychological characteristics of high school athletes for whom the interventions are more effective versus those for whom the interventions are relatively ineffective; and examine processes by which the interventions influence use and negative consequences for students.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although percutaneous transluminal coronary angioplasty (PTCA) appears to be a relatively safe and effective procedure, reported complications include coronary occlusion and dissection with or without acute myocardial infarction. We reported this complication of PTCA. A 77-year-old man with unstable angina pectoris by coronary angiography had focal severe stenosis of a dominant right coronary artery, a normal left coronary system, and normal left ventricular function. PTCA of the right coronary artery was performed. The tight stenosis prevented appropriate placement of a 2-mm. Gruntzig catheter; therefore, the narrowing was dilated initially by multiple inflations of a Simpson catheter advanced across a guide wire, and subsequently by a 3-mm. Gruntzig catheter at 4 atm. Post-PTCA angiograms demonstrated mild residual stenosis. Coronary angiography 3 months later disclosed significant stenosis again at the site of the previous dilatation procedure. Repeat PTCA was performed. Multiple inflations of a 3-mm. Gruntzig catheter at a maximal pressure of 9 atm reduced the stenosis to about 60% diameter reduction. A 3.7-mm. Gruntzig catheter was then positioned and inflated 5 times at a maximal pressure of 10 atm. On the fifth inflation, the proximal end of the balloon ruptured. Angiography then demonstrated extravasation of contrast material into the epicardial tissue plus run-off into the distal right coronary artery. The heart rate and arterial blood pressure quickly decreased; blood was drained from the pericardial sac, but fatal cardiac arrest ensued. At necropsy, the heart weighted 265 g. The epicardial adipose tissue surrounding the right coronary artery was hemorrhagic and an elliptical rupture site measuring 0.6 cm. in the long axis was present in the right coronary artery 3.5 cm. from the ostium. Radiographs of the major epicardial coronary arteries disclosed focal calcific deposits.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Eukaryotic cells show a high degree of macromolecular organization in which specific components are segregated into discrete structures with specialized functions. Appropriate model systems have been chosen to study the mechanisms by which proteins reach their final destination within organelles or membranes. 1. We have developed an in vitro model system using cultured epithelial cells of renal origin in which we will investigate, using biochemical, biophysical and ultrastructural methods, how tight junctions are formed and plasma membrane polarization is established during monolayer formation and after junctions are opened by experimental procedures. 2. In the same cell system production of enveloped viruses occurs in a polarized fashion. We will therefore be able to investigate the mechanism by which viral envelope glycoproteins are segregated into specific areas of the plasma membrane. 3. In vitro studies on the biosynthesis of selected membrane and organelle proteins will be carried out to identify features of the primary translation products which may be involved in directing these proteins to their functional location. The role of post-translational modifications carried out by intracellular membranes will be examined. It is hoped that these studies will contribute to a better understanding of membrane changes which may accompany cellular and organismic senescence.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary The proposed work focuses on identifying novel miRNA drivers in cancer and utilizes this and other knowledge to develop miRNA-based therapeutics. This Pathway to Independence award application includes a mentored career development plan for the transition of the candidate, Dr. Andrea Kasinski, into an independent investigator, as well an accompanying research plan describing the proposed experiments on discovering miRNAs that can potentiate KRAS-driven lung adenocarcinoma and exploring novel therapeutic strategies for re-expressing tumor-suppressive miRNAs, which includes a high-throughput screen to identify a small molecule that can restore miRNA processing and combinatorial miRNA therapies for sensitizing tumors to conventional chemotherapies. The candidate, Dr. Kasinski, is a postdoctoral fellow at Yale, in the lab of Dr. Frank Slack in the Department of Molecular, Cellular, and Developmental Biology. The work leading to her graduate degree in Genetics and Molecular Biology at Emory University was conducted in the lab of Dr. Haian Fu in the Department of Pharmacology and focused on targeting cell survival signaling for therapeutic development. In the Fu lab, Dr. Kasinski performed two very distinct projects that were interrelated based on the ultimate goal of developing targeted therapeutics: in one project Dr. Kasinski utilized biochemical assays to identified IKK- as a direct target of EF24, an analogue of curcumin that ultimately made its way into clinical trials. For the second project a series of genetic and molecular techniques were used to evaluate the transcriptional regulation of an oncogenic 14-3-3 family member, with the hope that this knowledge might spearhead subsequent studies to block 14-3-3 expression. The mentoring and career development plan will supplement her background, which is evenly split between genetic, molecular biology, whole animal studies and small molecule screening, with training and instruction in each, and in the particular areas that this project involves: murine biology, cancer biology, cell culture technique and high-throughput drug screening. Dr. Kasinski's goal is to become a faculty member in an interdisciplinary biosciences, cancer biology, or similar department at an academic, private, or government facility, in which she can research the biology of miRNAs in cancer and work to advance miRNA-based therapies. This research on miRNAs involvement in cancer requires that an innovative and selective screen be performed which Dr. Kasinski is actively pursuing first in cell culture and will advance into animal models. This assay in soft agar is selecting for miRNAs that can specifically cause normal human lung bronchial epithelial cells to become transformed. MiRNAs identified from this assay will be evaluated further in cell culture and ultimately in vivo. The proposed study involving miRNA therapeutics is two fold. The first takes into account the interaction of let-7 and LIN-28. Dr. Kasinski is in an active collaboration with Dr. Haian Fu, Co-Director of the Chemical Biology and Drug Screening Center at Emory University, to perform a high throughput screen (HTS) to identify inhibitors of this protein-RNA interaction. Hits from this HTS will be evaluated for cell permeability, kinetics and in vivo therapeutic potential, by several independent avenues of investigation. Finally building on Dr. Kasinski's current findings that miR-34 and let-7 represent valid therapeutic options for non-small cell lung cancer, she will evaluate these miRNA therapies in combination with currently used chemotherapies and targeted-therapeutics in cell culture, xenografts and in the Kras;p53 double mutant. This work is novel, timely and has clear and significant implications for human health and survival. The identification of miRNAs and targeted-therapies to perturb miRNA imbalance that occurs in cancer is invaluable to the fields of cancer and miRNA biology, and has direct clinical application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The malaria parasite Plasmodium falciparum has a complex life cycle involving two hosts. P. falciparum gametocytes are the form of the parasite which undertakes the transition from the human to the mosquito host. The developmental changes required for this transition are poorly understood. There is, however, some evidence for a shift in metabolic energy generation from glycolysis in the human host to respiration in the mosquito host. The primary goal of the proposed study is to test the hypothesis that P. falciparum mitochondria undergo developmental changes during gametocytogenesis that allow them to become functionally competent for respiration. A secondary goal is to determine if the timing of biochemical changes in developing gametocyte mitochondria correlate with changes in P. falciparum sensitivity to the antimalarial drug primaquine. The long-term goal of this study is to elucidate the mechanisms by which gametocytes regulate changes in metabolism in preparation for the shift in hosts. There are three specific aims for the proposed study. The first specific aim is to develop assays to measure the expression of marker proteins representing the three major mitochondrial activities. The marker proteins selected are cytochrome b, which is required for electron transport, mitochondrial malate dehydrogenase, which is required for the tricarboxylic acid (TCA) cycle and for the malate shuttle, and cytoplasmic malate dehydrogenase, which is required for the malate shuttle. These assays will require preparation of nucleic acid and antibody probes for each marker protein. The second specific aim is to characterize the biochemical changes in P. falciparum mitochondrial during the transition from the human to the mosquito host. To accomplish this, the expression of the marker proteins will be measured in asexual erythrocytic parasites, in parasites in early stages of gametocyte development, in parasites in late stages of gametocyte development and in zygotes. The third specific aim is to determine if there is a correlation between the biochemical changes in developing gametocyte mitochondria and the development of primaquine sensitivity. To accomplish this, a vital dye will be used to first determine how mitochondrial structure and transmembrane potential change during asexual and sexual developmental stages. Then the vital dye will be used to assay these same stages for the effects of primaquine on mitochondrial potential and cell viability. These studies will increase our understanding of the role of P. falciparum mitochondrial biogenesis in the transition from a human to a mosquito host.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cardiovascular disease remains the leading cause of death in the United States and vascular calcification and diabetes are both independent risk factors for incident cardiovascular disease. Fetuin-A is a hepatic secretory protein found in high concentrations in human serum and our preliminary studies suggest that feutin- A may simultaneously inhibit vascular calcification and promote insulin resistance and diabetes. We propose to explore the relationship of fetuin-A with the longitudinal development and progression of vascular calcification and diabetes in the Multi-Ethnic Study of Atherosclerosis (MESA) cohort. The research evaluates biological mechanisms interlinked by a single protein and will provide new insights to cardiovascular biology and diabetology simultaneously. Conducting this research within MESA has a number of advantages. MESA stored sera from the baseline visit, allowing fetuin-A measurement. MESA extensively measured subclinical cardiovascular disease;therefore, all participants already have measurements of vascular calcification, vascular function, and metabolic parameters. Additionally, MESA participants were followed longitudinally, allowing for evaluation of the associations of baseline fetuin-A concentrations with incident vascular calcification and diabetes. We propose to take advantage of this rich resource to explore the following primary aims: (1) To determine the cross-sectional and longitudinal associations of fetuin-A with vascular calcification and vascular function;and (2) To determine the associations of serum fetuin-A concentrations with adiposity, impaired fasting glucose, and diabetes mellitus. This research will take advantage of the hypothesized countervailing effects of fetuin-A on vascular calcification and diabetes to provide new insights to cardiovascular disease pathogenesis. PUBLIC HEALTH RELEVANCE: While cardiovascular disease and diabetes represent common and morbid diseases, our understanding of their pathogenesis remains incomplete. The results of this epidemiologic study will provide novel insights into their pathogenesis and may ultimately identify novel therapeutic targets for their prevention or treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Stroke is the fifth leading cause of death and the leading cause of chronic disability in the United States. Each year approximately 780,000 people suffer an acute ischemic stroke (AIS) in the United States at a cost of over $60 billion annually to the US health care system. Up to half of these strokes are due to thromboembolic occlusion of a large cerebral artery potentially accessible by catheter for endovascular treatment. Emergent large vessel occlusion (ELVO) AIS is particularly devastating and, due to the amount of thrombus burden in the arteries at the base of the brain in this disease, intravenous tPA is often insufficient to recanalize the cerebral arteries, reperfuse the brain tissue, and prevent irreversible infarction. In 2015 five randomized controlled clinical trials demonstrated the clinical superiority of catheter-based endovascular embolectomy to medical therapy alone in the first six hours after stroke onset. This has transformed the stroke community, with efforts to organize comprehensive stroke centers to treat ELVO patients rapidly along the lines of heart attack or major trauma treatment in the cardiology and trauma communities, respectively. Fewer than 10% of ischemic stroke patients receive treatment largely due to the narrow recommended time window for therapy within 3-6 hours of stroke onset. The basis for this recommendation is that the longer brain is deprived of blood flow, the more likely it is to infarct, and the more infracted brain there is, the more likely it is to hemorrhage after opening the occluded artery. Advances in neuroimaging are shifting the paradigm from ?time is brain? to ?physiology is brain?. Perfusion imaging (CT and MRI) and diffusion weighted MR imaging (DWI) now allow detection not only of infarcted brain but also of brain that is potentially salvageable if blood flow can be restored. MR DWI is the gold standard in medical imaging for identifying irreversibly infarcted brain (core infarct). Brain can progress from reversible but severe ischemia to irreversible infarction over the matter of minutes. Whereas current acute stroke diagnosis occurs by CT or MRI, treatment requires moving the patient to an X-ray angiography suite for endovascular therapy, leading to delays in care and variability in clinical outcomes. Current X-ray guided endovascular stroke treatment procedures have limitations that could be overcome using MRI guidance, including real-time tissue viability assessment and, in some cases, enhanced catheter navigation. We propose to develop the endovascular tools needed to treat patients safely and effectively whether in the strong magnetic field environment of MRI or the high ionizing radiation environment of X-ray angiography. Devices that are safe to use in the 3T MRI environment would allow patients to be treated in any standard clinical MRI scanner, thus allowing real- time assessment of brain viability during endovascular intervention. The strong magnetic field of the MRI scanner provides a unique opportunity to develop remote-controlled catheters that can navigate through the vasculature by creating magnetic forces on their tips. We have previously developed such magnetically assisted remote controlled (MARC) catheters for use in larger vessels of the body and neck; we now propose to develop smaller, more flexible versions for use in the vessels of the brain. Success in this project could transform the treatment of diseases like AIS that benefit from real-time physiologic tissue monitoring during therapy, thereby maximizing benefits and minimizing risks of treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A population of diabetics, both juvenile-onset and adult-onset, either on diet, insulin or oral hypoglycemic agents, will be screened using the photon absorption technique to detect the incidence of low bone mass. The presence of abnormally low mass will be correlated with indices of diabetic control. From the population of diabetics screened, a group of patients with low bone mass and a group with normal bone mass will be chosen for more extensive studies of calcium absorption, parathyroid hormone and vitamin D metabolism, in an effort to uncover the etiology of this abnormality. In a small population of patients with the most severe loss of bone, iliac crest biopsies will be performed for quantitative histomorphometry. One-hundred patients with diabetes who present for the first time to the clinic will be entered into a prospective study in which changes in bone mass, measured by both the photon absorption technique and radiogrammetric and radiodensitometric techniques, will be followed. Changes in bone mass will be correlated with level of diabetic control, vitamin D metabolities and parathyroid hormone levels.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposal is to research the technical and commercial feasibility of a novel wrist brace that reduces the risk for wrist fracture by resisting wrist terminal extension while otherwise allowing unconstrained movement throughout the normal range of motion. The novel wrist brace will be designed, fabricated, and validated in our laboratory under simulated impact conditions. The long-term goal of the product is to determine the efficacy of the brace on snowboarding and in-line skating populations using a prospective, randomized, longitudinal study. Wrist braces offer limited protection against distal radius fractures, yet they have achieved limited acceptance, particularly among youths, because they are bulky and limit normal range of motion. Research has also demonstrated that they do not prevent fractures under high impact energy loading conditions. We postulate that wrist guard usage will increase significantly with a device that is less constraining and which offers increased protection against distal radius fractures", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A study of the role of blood proteins and receptor-mediated endocytosis in the metamorphosis and egg formation of insects is proposed. Vitellogenin, lipophorin, and three hexameric storage proteins, which comprise most of the blood proteins of Hyalophora pupae, are to be isolated and labelled for use as physiological probes. A series of tests is designed to assess whether the probes are handled by the tissues of the insect in the same manner as before isolation and labelling. The fates of the probes when presented to developing tissues either in situ or on isolation in a culture system will be examined. It is anticipated that each protein will prove to be endocytosed by a particular tissue and at particular stages of development, and that it will either be incorporated into cellular structures intact, as in the case of vitellogenin, or hydrolyzed to release amino acids for use in synthesizing other tissue proteins. The binding selectivity of each target cell for the five probes will be determined, and since the first two to be isolated have proven to be mannose-rich glycoproteins, the role of sugars in ligand binding and endocytosis will be studied.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposal is to bring to bear state of the art theoretical methods to the study of the mechanisms of ribozyme catalysis and the factors that regulate reactivity. An overarching theme in the proposal is to facilitate active collaborations with a network of experimental groups in order to progress toward a consensus view of mechanism that may, ultimately, contribute to a deeper understanding of more complex cellular catalytic RNA systems such as the ribosome and spliceosome. The proposal focuses on application to a series of archetype ribozymes that catalyze site-specific phosphodiester bond cleavage (and ligation) but that have different active site architectures and catalytic requirements. As a baseline, the same reaction will be studied in a non-enzymatic dinucleotide model in solution, and catalyzed by the protein enzyme analog, RNase A. The tandem study of alternate mechanistic strategies and the factors that govern reactivity will provide penetrating insight into the rational design of new biomedical technology. However, these applications demand new methodological advances. Of key importance are accurate and efficient quantum mechanical/molecular mechanical (QM/MM) methods for catalysis, reliable molecular simulation force fields for RNA and metal ions, and efficient methods for sampling free energy surfaces and conformational transitions. Toward this end, we propose to: 1) improve the QM, MM and QM/MM models for ribozyme catalysis, 2) develop a new QM/MM method for prediction of pKa shifts in ribozymes;3) develop a novel free energy expansion approach to extend our QM/MM methods to the ab initio level;4) develop efficient path methods to study chemical mechanisms and conformational transitions in ribozymes. In this way we hope to greatly extend the predictive capability and range of application of state of the art theoretical methods to RNA catalysis. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to use quantum mechanical and molecular simulations methods to study the mechanisms whereby molecules of RNA catalyze important chemical reactions. The insight gained by these studies will enhance our understanding of the fundamental role RNA plays in cells and facilitate the design of new RNA-based biomedical technology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Discover new drugs with anti-Candida activity from four unique sources: antimicrobial-producing microorganisms, plant tissue cultures, \"immunoliposome\"-encapsulated anti-Candida drugs, and novel cell wall inhibitory analogs from syntheticchemical efforts of our laboratories. In vitro models for initial rapid evaluation of potentially active drugs from thesources mentioned above will be used. These in vitro models will allow us to ascertain MIC values by established methods. Since the fungal cell wall contains enzymes and components not found in human tissue, the evaluation screens will focus on compounds that inhibit fungal cell wall synthesis to develop a drug that is pathogen-specific. Animal model evaluation of potential drugs based on in vitro tests will utilize an established mouse animal of candidiasis. Parameters of the animal model systems will be varied to determine the method and route of administration, timing and dosage necessary for anti-Candida activity, and toxicity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To elucidate the mechanisms involved in the localization of neurotransmitter receptors under nerve terminals, we will analyze the interactions between acetylcholine receptors (AChRs) and cytoskeletal components in the nicotinic post-synaptic membranes isolated from Torpedo electric organ. In those isolated membranes, as at the vertebrate neuromuscular junction, AChRs are constrained and unable to diffuse freely, but upon removal of peripheral proteins from that membrane, constraints on AChR mobility are effectively removed. It is our hypothesis that interactions between AChRs and peripheral membrane proteins serve to restrict receptor mobility, and we will characterize the structures of those peripheral proteins and identify the protein interactions that account for AChR immobilization. By immunological techniques four peripheral proteins (43K, 58K, 87K, and 270K) have been identified on the cytoplasmic face of the Torpedo post-synaptic membrane and also at the vertebrate neuromuscular junction. Experiments will focus on the 43K protein, which is present in stoichiometric amount with AChRs, and the 270K protein that has been shown by immunological criteria to be related to dystrophin, the protein product of the Duchenne/Becker muscular dystrophy gene locus. Immunoelectron microscopy will be used to identify the filamentous cytoskeletal elements associated with the innervated surface of the Torpedo electrocyte and to determine the disposition of 27OK/dystrophin as well as 58K and 87K proteins relative to AChR/43K, the lipid bilayer and the cytoskeleton. Biochemical and immunochemical techniques will be used to define the mechanism of association between 43K protein and AChRs and also with lipid. The 270K protein will be isolated and characterized by protein microsequencing to determine whether that protein contains all the structural domains of muscle dystrophin. Biochemical and immunochemical techniques will be used to determine whether 27OK/dystrophin interacts directly with AChRs or other integral membrane proteins of the post-synaptic membrane and to determine whether it binds to the actin cytoskeleton. To determine whether 58K and 87K proteins are related to previously described muscle proteins, they will be isolated and characterized by protein microsequencing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies have shown that the loss of fragile X mental retardation protein (FMRP) causes a mental retardation condition known as fragile X syndrome. The protein contains two RNA binding domains known as K homology domains (KH domains). The specific function and target of these RNA binding domains are not known nor is it known why the loss of FMRP leads to mental retardation. Therefore to understand the molecular function of the FMRP, one needs to first study these domains. It is the aim of this proposal to identify RNA targets and RNA binding characteristics of the first KH domain in FMRP. The SELEX procedure (Systematic Evolution of Ligands by Exponential Enrichment) will be used to identify RNA targets. SELEX is an in vitro selection procedure that identifies high affinity nucleic acid ligands for a given protein. The sequences of the ligands obtained from SELEX are not necessary that of the in vivo target but they can be used as templates in searching genome sequences for possible targets. Once a high affinity target is identified, phage display will be used to investigate the binding of the KH domain to the target. By displaying mutant proteins on the coat of the phage, phage display enables efficient screening and selection of a large pool of mutants. The purpose of the mutation analysis is to identify the residues that participate in RNA binding. Identifying potential RNA targets and regions of the KH domain responsible for RNA binding may help to elucidate the physiological function of the FMR protein.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Biological therapy of cancer is defined as therapy directed at biological functions of the host or the tumor with change host tumor interactions in favor of host destruction of tumor cells or regulation of tumor cell proliferation. Biological response modifiers include immunotherapeutic agents, interferons, other cytokines, maturation and differentiation factors and vitamins and other nutritional factors. Augmenting agents include natural or synthetic substances which stimulate various host defense mechanisms such as macrophage tumoricidal activity, NK cells, interferon production, etc. In this project, we will investigate, in control Phase I and II studies, therapy with \"second generation\" augmenting agents of defined chemical structure. Optimal doses and schedules will be sought using a unique statistical approach and various assays of host defense activation including interferon induction, macrophage, NK cell and RES activation. Once optimal \"augmenting\" doses are defined, the agents will be introduced into Phase II trials (alone or in combination with conventional therapy) to determine 1) if the agent will increase the remission rate, duration, and survival, 2) if the degree of \"augmentation\" correlates with the therapeutic effects. Initially, MVE-2 (the 15,500 MW fraction of pyran copolymer) and ABPP (2-amino, 5-bromo, 6-phenyl pyrimidinole--an orally active interferon inducer) will be studied. These serve as models for the study of macrophage activators and interferon inducers. Each year of the project, two new augmenting agents will be introduce into Phase I trials and two will be transferred to Phase II trials. These studies should establish the therapeutic activity of a variety of augmenting agents for cancer management.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies were performed to investigate the effects of chronic, low level pre-and postnatal lead exposure on immune functions in rats. Weanling female rats were exposed to lead (as lead acetate) in their drinking water at 0, 25, and 50 ppm for 7 weeks. At the end of 7 weeks they were mated with untreated males and continued on the same dosage throughout gestation and lactation. The offspring of these females were weaned at 21 days of age and continued on the same lead exposure regimen as their mothers. These offspring were used in immune surveillance procedures between 35 and 45 days of age. Lead exposure at the levels employed had no statistically significant effect on growth and did not result in overt signs of toxicity. Thymic weights were significantly decreased in both males and females of the two lead dosage groups. Furthermore, lead exposure resulted in suppression of responsiveness of lymphocytes to mitogen stimulation and in reduced delayed hypersensitivity responsiveness. A marked depression in the antibody response to sheep red blood cells (SRBC) as well as decreased serum IgG levels were observed. Serum IgM and IgA levels were normal.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of these studies is an in-depth investigation of the thymus derived lymphocyte and its products. Lymphotoxin (LT), a product of antigen or mitogen activated T cells, is being studied as a model lymphokine. The long term goals are to study LT's cell of origin, biochemical nature, mechanism of action, relationship to other lymphokines, regulation and role in vivo. Murine lymphotoxin cDNA has been cloned with mRNA from an interleukin-2 maintained T cell clone. Its gene has been isolated and mapped to chromosome 17. The specific aims of the next 5 years are to analyze the structure of the LT gene, to precisely localize it on chromosome 17, to determine the linkage relationship of the genes for LT and tumor necrosis factor (TNF), to obtain maximal expression of LT cDNA, to analyze regulation of LT and TNF, to study expression of the transfected LT gene and to analyze the 5' regulatory sequence of the LT gene. Goals will be accomplished by sequencing the gene, by identifying LT promoter activity, and by expressing LT DNA in prokaryotic and eurkaryotic cells. Interleukin-2 maintained T cell clones will be exposed to several inducing agents and their LT and TNF mRNA analyzed by Northern blots. The murine LT gene will be ligated to strong promoters and its expression studied in human T cell tumors. Similar vectors will be used to study the effect of LT gene overproduction in transgenic mice. Such mice will be analyzed for autoimmune pathology. If the LT gene is overexpressed in the T cells of these mice, and those cells killed, a murine model for AIDS will be available. LT 5' regulatory sequences will be ligated to a neutral gene, chloramphenical acetyle transferase, in order to study regulation of the LT gene in the absence of effects attributable to the product itself. Studies with transfected T cells, B cells, macrophages and transgenic mice will provide insigt into the nature of T cell specificity of LT expression and eventually lead to an understanding of the molecular basis of LT gene activation by antigen.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this project is to elucidate the catalytic mechanism of CO dehydrogenase (a.k.a. acetyl-CoA synthase - ACS), emphasizing the structure and function of the unique Ni-Fe-S clusters contained therein. The enzyme is an alpha-2-beta-2 tetramer that catalyzes two reactions; the reversible reduction of CO2 to CO and the synthesis of acetyl-CoA. The former occurs at one Ni-Fe-S cluster in the beta subunit, while the latter reaction occurs at another Ni-Fe-S site in alpha. A tunnel through which CO migrates connects the two sites. The reaction mechanism is \"organometallic\" in nature; how does Ni promote this within an aqueous protein matrix? Mechanistic steps occurring at the two sites are synchronized, making it an attractive system to investigate details of metabolic channeling and active-site coupling. ACS is useful biotechnologically; organisms containing it reduce atmospheric levels of CO and degrade TNT. ACS is found in Clostridium difficile, a pathogenic organism responsible for the deaths of about 2000 people annually. Given the complexity of this enzyme, the general strategy used in this project will be to study each activity separately, and then compare observed properties with those of the bifunctional enzyme; such an approach may reveal some of the complexities of channeling and catalytic coupling. This approach is possible because isolated recombinant alpha subunits able to catalyze the synthesis of acetyl-CoA can be prepared, as can a homolog of the beta subunit that is able to catalyze CO/CO2 redox. Many of these developments (recombinant biosynthesis of active alpha, discovery of the tunnel and of active site coupling) arose from efforts of the previous granting period which resulted in 11 publications. The methodologies used for the project include enzyme kinetics (stopped-flow, rapid freeze-quench, and steady-state), spectroscopy (EPR, Mossbauer, and XAS), and X-ray diffraction. A rapid-freeze-quench instrument that allows samples to be prepared under reliably anaerobic conditions has been constructed, and should be an improvement over existing technology. Kinetic data will be simulated, ultimately leading to a comprehensive mechanistic model describing the general kinetic behavior of the enzyme. Crystals of alpha have been obtained, suggesting that an X-ray diffraction structure may be achievable. These studies require repetitive purification of at least 5 different proteins; funding for a preparative FPLC/HPLC is requested.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "MeCP2-duplication/triplication syndrome is a devastating neurodevelopmental disorder mostly affecting boys and is caused by duplication (or triplication) of the region in the X-chromosome containing the gene encoding MeCP2 (methyl-CpG-binding protein 2). MeCP2 binds to methylated CpG sites on DNA and recruits transcription complexes to modulate gene expression. Because MeCP2 modulates a multitude of genes, it is not surprising that pinpointing specific functions has proven difficult. In additio to MeCP2-duplication syndrome, loss-of-function mutations in MeCP2 account for about 96% of Rett Syndrome cases, another devastating neurodevelopmental disorder that is typically seen in girls. Because of the strong neurologic component seen in both Rett and MeCP2-duplication syndromes, MeCP2 was long assumed to be primarily important for neurons. However, recent studies have shown that other cells, such as astrocytes and microglia in Rett syndrome, and T cells in MeCP2-duplication syndrome, also play a significant role. This understanding opens up the possibility for treatment modalities beyond direct neuronal intervention, which has been a problematic approach due to the complexity and risk involved in any manipulation of neurons themselves. Here, we propose to test the hypothesis that MeCP2 overexpression causes defects of adaptive immunity that lead to excessive, pathologic innate inflammation, ultimately contributing to morbidity and mortality. Preliminary data in MeCP2Tg3 mice, which overexpress MeCP2 at 3-5 fold normal levels, reveals that as disease progresses, peripheral organs undergo significant weight loss associated with gross pathologic appearance and immune infiltrates. However, immune-privileged organs (brain and testes) are protected from these changes. This suggests that a peripheral destructive immune process is contributing to pathology in MeCP2Tg3 mice. In addition, MeCP2Tg3 mice have significantly increased mortality from influenza infection. In an influenza model, MeCP2Tg3 mice exhibit a deficient CD8+ T cell response and excessive innate inflammation without an apparent defect in viral clearance; we intend to test the hypothesis that CD8+ T cell defects result in a compensatory increased innate response, which causes pathology leading to increased death. The first aim is to test the hypothesis that dendritic cell malfunction contributes to defects of the adaptive immune response using both in vitro and in vivo assessment of antigen presentation and T cell activation. The second aim in this proposal will test the hypothesis that a deficient CD8+ T cell response contributes to increased mortality in MeCP2Tg3 mice via an excessive innate immune response. This will be tested by in vitro assessment of CD8+ T cell function and in vivo assessment of CD8+ T cell response to influenza. The third aim is to ameliorate death from influenza in MeCP2Tg3 mice via either bone marrow transplant or suppression of the excessive innate inflammation. Ultimately, results obtained from this work may lead to immune-based treatment strategies for patients with MeCP2-duplication syndrome.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project is part of a long-standing collaboration with the laboratory of Dr. Ettore Appella (LCB/NCI), in which the Wip1 protein was discovered. Initial characterization resulted in determining two classes of phosphorylated substrates, involving many proteins involved in cell growth regulation. The first has a diphosphorylated sequence motif (pT-X-pY), such as in p38 MAP Kinase, while the second has a mono-phosphorylated sequence motif (p(S/T)Q), such as in the p53, Chk1/2 and ATM proteins. By development of an atomic-scale computer model of the extended active site of Wip1 and a series of mutagenesis experiments, we were able to reveal the structural basis for the range of substrate specificity. This lead to the development of a cyclic peptide molecule that competitively inhibits Wip1, the first inhibitor of any kind for this family of enzymes. We then pursued development of a more drug-like, small molecule inhibitor in collaboration with Dr. Daniel Appella (LBC/NIDDK), who specializes in synthetic chemistry. The resultant small molecule is based on a pyrrole ring scaffold, with 5 different emanating sidechains to mimic the amino acids of the cyclic peptide. While successful, the final inhibition constant was still only in the low micromolar range. To further this effort, we returned to optimizing the cyclic peptide inhibitor. By multiple iterations of design and testing, we were able to drastically increase the binding affinity, resulting in an inhibition constant of 110 nM. The structural modeling involved in this process revealed both important new interactions in the extended active site, and the role of the proximal B-loop in binding substrate and regulating activity. Since the B-loop is unique to the Wip1 member of the PP2C family, its role was previously unknown. We are now applying these lessons to designing a new generation of pyrrole-based inhibitors. We are also pursuing generating sufficient Wip1 protein to determine the structure by X-ray crystallography, which will greatly aid in inhibitor optimization. Recently, we have verified the requirement of binding a 3rd magnesium ion for activity of Wip1 and the related PP2Ca homologue, and used deuterium exchange mass spectroscopy to study the functional conformational changes. We are currently determining the crystal and NMR structures of PP2Ca/cyclic peptide inhibitor complexes. Recently we used Deuterium Exchange Mass Spectroscopy to study the functional structural changes of Wip1 and the PP2Ca homologue. We have also determine the crystal structure of PP2Ca with a bound substrate.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Uncontrolled hyperbilirubinemia (jaundice) in neonates has long been known to lead to neurological dysfunction including irreversible athetoid cerebral palsy with speech, ocular and hearing impairments, and even death. Contemporary management is based upon monitoring the total serum bilirubin (TBS), taking into account other clinical parameters such as birth weight and gestational age, and administering effective treatment (phototherapy and/or the rare exchange transfusion), if dictated. Unfortunately, the trend towards discharge of apparently healthy neonates from the hospital very soon after birth has made the management of subsequent jaundice more difficult in that population. A long suggested, but not used, better predictor of the neonate's risk for neurological sequelae due to elevated bilirubin is a measure of the capacity to sequester bilirubin in the blood compartment by its binding to serum albumin. The concentration of unbound bilirubin, the driver for bilirubin escaping from the vasculature, can be calculated from the TBS and binding capacitl. Presently existing methods for assaying binding capacity and unbound bilirubin are not facile. However, all these parameters can be directly measured simply in a very small volume of whole blood with a special purpose fluorometer, the hematofluorometer, first described years ago, by making use of the natural fluorescence of bilirubin bound to albumin. This technology is amenable to point-of-care use. The aims of this project are to transform the modernized and miniaturized hematofluorometer developed in Phase I into a product suitable for operation in various point-of-care environments, including the intensive care and healthy baby nurseries, the neonatal inpatient clinic, and the pediatrician's office. The first aim is to optimize the basic optical and electronic design: redesin the electronics to support the hospital information management requirements, such as a bar code reader, printer, and interface with a computer, either directly or via the local internet. The second aim is to develop a reagent kit that is easy to use and inexpensive. Phase I work demonstrated that significantly more work is needed to design a kit meeting these requirements, and then scaling this design up into a product that can be mass produced. The third aim is to test the instrument with neonate blood samples in a clinical environment to demonstrate that it well suited to meet the needs. With this goal in mind, Stanford University's Medical School and Children's Hospital has agreed to participate as a subcontractor to evaluate the new technology. With these proposed improvements, the instrument will be ready for the next stages: releasing it to the market for immediate R&D uses and clinical studies, and eventual approval by the FDA for general use.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of the Animal Health Resources Program is to facilitate biomedical research at the Fred Hutchinson Cancer Research Center by providing quality animal care, by offering a broad range of technical (e.g., surgical) support services, and by eliminating complications of animal research due to intercurrent disease. The specific aims of this proposal are threefold. First, we plan to upgrade the general level of animal care throughout our facility and to ensure future accreditation by replacing worn and unreliable equipment (an autoclave and ten dog runs). Second, we propose the renovation of existing space for a much needed surgical facility for dog and rabbit studies, currently conducted off-site or not at all. Third, we will expand our current, disease-free microisolator rodent housing system through the purchase of 100 additional cages for rats. Support as requested will enable the Animal Health Resources to upgrade general operations, maintain accreditation standards, and meet its service goals for the coming years.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Kansas City Clinical Oncology Program (KCCOP) expects to transition to NCORP-KC. It is a very successful non-profit organization conducting clinical trials in cancer therapy, cancer symptom prevention. Quality of Life, and cancer control since 1983 with uninterrupted funding by the National Cancer Institute. The program is located in the Kansas City metropolitan area and serves the needs of both Kansas and Missouri residents spread over 16 counties and 2.5 million citizens. The program is comprised of seven component area hospitals and 52 investigators representing specialties well beyond medical, surgical, gynecologic, and radiation oncology. The other areas of expertise include outcomes research, internal medicine, healthcare administration, healthcare informatics, behavioral science, rehabilitation and survivorship, palliative care, neurology, gastroenterology, genetic counseling, pathology, cellular therapy, laboratory science, and radiology. The program has provided participation opportunities to over 4,200 patients and volunteers in cancer-related trials since its inception. Over the last five years, an average of 235 new individuals have participated per year. The program has maintained excellent quality of research as confirmed by patient satisfaction and reports from external audits. The program derives its cancer trials from SWOG, NRG, ALLIANCE, CTSU, MD Anderson, URCC, and SunCoast research bases of the National Cancer Institute. NCORP-KC wishes to thank its participants and extend its service by entering a new era of Cancer Care Delivery Research while continuing to conduct high quality trials in cancer therapy, cancer control, screening, imaging, and post-therapy surveillance. The program monitors trends and implements measures for continued improvement. It balances protecting patients, their safety, and health with providing the broadest possible access to trial activation and minority outreach participation with the help of a highly responsive and responsible support staff.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Positron emission tomography (PET) studies of neuroreceptors in vivo in man hold great promise in helping to identify the role of receptor defects in many diseases. This research is aimed at developing radiopharmaceuticals which localize in discrete regions of the brain based upon their selective affinity for the postsynaptic dopamine D2 receptor. 18F- and 11C-labeled spiperone and its analogs continue to be used for PET studies of the D2 receptor despite problems such as non-selectivity, high non-specific binding, and the difficult radiochemical synthesis of ring fluorinated spiperone. The absence of a substantially better D2 ligand is a primary reason for the use of spiperone. The work proposed here is aimed at the synthesis and evaluation of agents possessing substantially improved properties over spiperone. The goal of this work is to demonstrate the applicability of these agents for future in vivo studies in human subjects. It is anticipated that the development of these radiopharmaceuticals, their in vivo characterization, and their use will eventually aid in the study of Parkinson's disease and schizophrenia in which abnormalities in dopaminergic transmission are believed to play a role. In addition, these agents may well prove useful in studying various changes of the D2 system in aging. Non-radioactive phenyl ring fluoroalkylated benzamide compounds will be synthesized and the characterized in vitro using competitive displacement binding techniques. Those agents possessing suitable in vitro binding properties will be further assessed in vivo to determine their potential as radiopharmaceuticals for quantitative PET studies of the dopamine D2 receptor. The desired in vivo characteristics of these ligands include: good brain penetration to provide adequate counting statistics for PET studies; regional localization in D2 receptor rich areas; retention of activity in those specific areas to allow for adequate PET data acquisition; rapid clearance of non-specific uptake in non-target areas such as the cerebellum; pharmacological specificity for the dopamine D2 receptor; uptake site saturability; absence of lipophilic metabolites in the blood; and few metabolites in the brain. Those compounds which demonstrate a selective, potent binding site affinity to the D2 receptor will be radiolabeled with tritium. The tritium-labeled compound will be used in further studies to assess in vitro binding parameters of the agent. Those compounds displaying promising characteristics in vitro and in vivo will be labeled with high specific activity 18F (a short-lived positron emitter with a 110 min half-life) and utilized in PET regional localization investigations in monkeys. The dopamine D2 site densities and dissociation binding constant for the agent will be determined. PET studies in animals will determine the efficacy of the compounds as potential dopamine D2 receptor ligands for investigations in human subjects. A series of pilot experiments will be conducted in normal subjects to assess the properties of the 18F-labeled ligands in vivo to determine dopamine D2 binding parameters.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Core Component D serves three important purposes. The first purpose of this Administrative Core is to provide all Projects and Cores with appropriate administrative and secretarial support to efficiently carry out the scientific goals of the Program Project. These essential services are not provided by the Massachusetts General Hospital. The key responsibilities of this core include: (i) general administration;(ii) fiscal oversight (budgeting, purchasing);(iii) communication and coordination of data flow;(iv) seminar series coordination and travel arrangements;and (v)preparation of manuscripts and reports. The second purpose of this core component is to closely monitor the activities of the Cox-7 Animal Colony, which is the chief facility of Core C. The Surgical and Animal Core (Core C) requires additional administrative effort for the following reasons: (i) it must adhere closely to additional regulations for breeding and maintaining animals;and (ii) it is responsible for the coordination and submission of all animal protocols. The third purpose of Core D is to provide the overall scientific direction for the Program Project. This is achieved by close interaction with: (i) the Scientific Steering Committee;(ii) External and Internal Advisory Committees;and (iii) Consultants for individual projects and cores. These groups regularly evaluate the progress and agenda of the PPG. The input of these advisory groups is discussed at the monthly Scientific Steering Committee Meetings and is incorporated in specific projects and cores.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dramatic expansion of biomedical data in the post-genomic era on various organisms, including humans, has disclosed the enormous complexity of biological systems and increased our knowledge of many disease processes. This is particularly evident in reference to neoplastic disorders where molecular technologies on a genome, epigenome, transcriptome and proteomic scale have disclosed unique molecular pathways and their networks that provide significant information concerning therapeutic strategies and clinical outcomes. This in turn has affected the standard of practice of many medical disciplines including pathology. The newly developed concept of personalized and targeted therapies in cancer medicine increased and changed the traditional roles of pathology as a clinical discipline. The expanding molecular testing of tumors, with complex technologic approaches, requires a redesign of the conventional microscopic pathology services. This T32 proposal is intended to provide resources to train a new generation of pathologists via laboratory-based research training that will complement the current Accreditation Council for Graduate Medical Education-accredited clinical fellowship programs of the Department of Pathology at The University of Texas MD Anderson Cancer Center. The goal of this program is to train the next generation of physician/scientist pathologists capable of combining the conventional role of diagnostic pathology with investigative molecular approaches. To accomplish this goal, a focused 2-year research program will pair trainees with experienced educators who are also accomplished translational researchers. The trainees will work in their mentors' laboratories training in laboratory methods, processes, and team orientation needed in today's translational research. The graduates of this program will complete research projects aimed at improving patient care with patient-directed personalized treatment, concluding with the submission of a manuscript to a peer-reviewed journal. Coursework and a wide range of educational opportunities at The University of Texas MD Anderson Cancer Center will complement the laboratory research training. Combined with subspecialty clinical training, this program will produce extremely well-qualified academic, research-oriented pathologists who can, by themselves, be independent researchers and future academic mentors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Previously, Dr. Elder and his colleagues used comparative modeling of FIV and HIV aspartyl proteinases (PRs) to identify specific amino acid residues that interact with the substrate binding pocket in a unique fashion in each protease. They have discovered that HIV protease and FIV protease are very similar structurally but that each has certain unique characteristics which result in strongly preferential homologous gag substrate cleavage. The hypothesis behind the proposed work is that an in-depth understanding of both the similarities and the differences between two distantly related lentiviral proteases will promote development of protease inhibitors that may be efficacious against both FIV and HIV proteases but at that same time, relatively homologous resistance-proof. To explore this hypothesis, Dr. Elder and his colleagues will focus on substrate binding pocket interactions. They will use site-directed mutagenesis to replace FIV-specific residues with HIV-specific residues, both alone and in combinations predicted to be interactive in the three-dimensional structure. Mutant proteases will be expressed in bacteria, purified, refolded. These mutant proteases will be biochemically defined as to their substrate specificities using both natural and synthetic peptide substrates and as to the kinetics of their inhibition by a panel of previously defined HIV protease inhibitors. Interesting mutants will be placed in the context of infectious virus, and examined in cell culture for their potential to be inhibited by, or develop resistance to, known protease inhibitors during virus passage. Where appropriate, the three-dimensional structure of interesting mutant proteases can be examined by Dr. Alex Wlodawer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ivermectin has been shown in an open, in-hospital trial in South India to be effective in clearing microfilaremia in patients with bancroftian filariasis at dosages as low as 25-mcg/kg given once orally. A second, blinded, placebo-controlled trial of ivermectin and the currently used drug, diethylcarbamazine (DEC), has indicated that the efficacy of ivermectin and DEC were equivalent for 3 months, but by 6 months the ~30% microfilarial recurrence rate of the ivermectin-treated patients was significantly greater than the ~15% rate of the DEC group. Side effects were qualitatively and quantitatively identical for both groups. Thus, though ivermectin appeared somewhat less effective than DEC at 6 months, its single-oral-dose mode of administration with toxicity no greater than that of DEC means that this drug should engender greater patient compliance and, therefore, be much more effective in mass-treatment filariasis control programs than DEC. Observations on 200 Peace Corps volunteers going to Loa loa endemic areas of Africa have concluded after three years of a placebo- controlled chemoprophylaxis trial using weekly doses of DEC. In Gabon (where exposure to the parasite was heaviest) 30% of individuals in the placebo group developed overt clinical disease compared to none in the DEC-treated group (p<.02) and 50% became seropositive in the placebo group vs 13% in the DEC group (p<.02). No significant side effects were seen. These findings indicate that DEC given orally once-weekly can be an effective, acceptable chemoprophylactic for preventing loiasis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ion channels are important drug targets and unintended drug interactions with ion channels are also of critical importance, requiring the screening of all drug candidates. Conventional high throughput screening processes for soluble proteins are problematic to apply to ion channels because the channels must be incorporated into a lipid bilayer membrane and transport an ionic current to display their functionality. There are currently no high quality, high throughput assays for ion channel drug screening. Recent efforts to increase throughput have resulted in automated patch clamp systems but these are still over two orders of magnitude lower throughput than conventional drug screening technologies, requiring costly instrumentation, cells, and consumables. For existing methods of ion channel screening, there is a gap in information quality, throughput, and cost. An alternative method of ion channel measurement involves reconstituting them in artificial lipid bilayer membranes. In recent research at UCLA led by the PI, a new high-freezing point lipid membrane composition was developed enabling it to be frozen. When frozen, we showed that it was sufficiently robust to withstand shipping, a major breakthrough. These membranes, when packaged in inexpensive chips, have the potential to significantly change the way ion channel screening is done. Our company, Librede Inc., was formed by the UCLA team to further develop and explore the commercial potential of this technology, the first steps of which are proposed here in this Phase I SBIR proposal. Our ultimate goal is to create an inexpensive, disposable chip containing arrays of lipid membranes to enable low cost high throughput screening of ion channels. In the preliminary work at UCLA, the technology was demonstrated with the shipping of small numbers of membranes, with a net yield of 30%. In the proposed work, we will design and fabricate large array chips containing 48 membranes compatible with industry standard 96 well fluid handling robotics and pipetters. With these chips, we aim to demonstrate the viability of the membrane technology over a much larger scale producing over 3000 membranes. To increase their commercial viability, we will also seek to increase the yield by systematically changing the lipid concentration, solvent composition, solution volume, and thawing temperature. Although this space is potentially very large, the membrane arrays will electrically probed in an automated fashion using a custom-built electrical interface and multiplexer. We will use the materials and designs from Phase I to contract with a plastics manufacturer in Phase II to create an inexpensive, injection molded prototype. At that point, we will have demonstrated a chip able to perform industry standard ion channel screens for a significantly lower cost. These membrane array chips have the potential to increase throughput by several orders of magnitude and similarly decrease cost by several orders of magnitude-as a result, transforming the process of ion channel measurement and screening. PUBLIC HEALTH RELEVANCE: Measurement of ion channel interactions with drugs is a key process in drug discovery and drug safety screening. Due to the difficulty in working with ion channels, the existing processes used are slow, laborious, and expensive. Our team has recently developed a platform for ion channel measurement which is much less expensive and much easier to use, based on lipid membranes that can be shipped, a world first. We propose here to develop inexpensive membrane array chips and use them for large scale production, demonstrating the feasibility of this technology for ion channel screening.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Summary: One of the main reasons that promising drugs fail to live up to their potential for ameliorating disease is unforeseen toxicity. The ability to monitor, and diagnostically predict which compounds may engender toxicities from a large lead compound portfolio, will have a dramatic impact on pharmaceutical development and the FDA's ability to monitor and ensure the safety of new drugs or new combination therapies. In the past, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) has been the major technology utilized by experimentalists to separate and analyze proteomes. This technology, although a powerful method for characterizing thousands of proteins from any given sample, is low-throughput, insensitive, and has limitations with separating low molecular weight and highly basic proteins. Our laboratory accepted the challenge of developing novel methods and tools to utilize proteomic information in the discovery of protein biomarkers of toxicity. Of critical importance, our laboratory invented (patent-pending) and developed, through a collaboration with Correlogic Systems, Inc., a heuristic pattern recognition algorithm which can import the complex data streams produced by SELDI and rapidly identify (in up to 20th-dimensional space) patterns of proteins that are diagnostic for human cancer in serum or cell lysates.. A component of our research efforts focuses on the applicability of proteomic technology to identification of changes in protein patterns that could predict drug toxicity. Through a collaboration with Dr. Frank Sistare's group at the center for Drug Evaluation and Research (CDER), Dr. Petricoin obtained sera from animals treated with different drugs, with known toxicities, as well as sera from control animals. These sera were analyzed by SELDI using 1 ul of serum and each sample was analyzed in quintuplicates. Protein profiles were generated in less than 2 weeks. The SELDI profiles were then normalized and analyzed by the novel heuristic cluster algorithm, developed by the Petricoin laboratory, to identify protein patterns that were diagnostic for toxicity. The results of the analysis were remarkably clear and intriguing. Importantly, this tool was able to identify drug-induced toxicity over the vehicle alone background 100% of the time in the blinded test set. Critically, the proteomic profile proved to be more sensitive than histopathology or ELISA analysis in detecting toxicity at lower doses and at consistently earlier time points. Each of these profiles appears to be drug-specific, although the data are currently being tested to assess whether a global toxicity pattern can be uncovered.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Center for Neural Science at New York University is a premier site for research on functional aspects of vision. Eighteen investigators from the Center request support for their vision research through continuation of a Core Grant. They are bound together by their research Interests In the neural and behavioral bases of vision and visually guided behavior. The proposed Core will comprise of four modules, each of which will benefit research in the research areas represented by program faculty: 1. A Visual Displays Module which will provide support for the development of state-of-the-art software for the generation of a wide variety of visual stimuli for psychophysical, neurophysiological, and brain experiments. 2. A Design and Fabrication Module which will provide expertise to design and build novel electronic and mechanical devices such as theose required for multielectrode recording in the visual cortex, noninvasive methods of eye movement recording, and Interface devices for behavioral and blloglcal experiments. 3. A Neuroanatomy and Molecular Biology Module which will provide expert support for the processing of neuroanatomlcal material at both the light and EM level, for the analysis and localization of electrophysiological recording sites In histological material, and for the creation of transgene-containing viruses to modify neuronal funtlon experimentally. 4. A Functional Imaging Module which will provide support for hardware and software services for users of the Siemens MRI scanner and associated devices In the Center for Brain Imaging, giving access for Core researchers to functional and structural Imaging capabilities for human and animal research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed studies represent an extension of an ongoing program of research. The major objectives of this research are to describe the extant communication abilities of children and adults with severe to profound mental retardation; and to identify the factors that are associated with the attainment (or nonattainment) of specific communication and language abilities by such individuals. The proposed studies will address five specific aims related to these overall objectives that have been identified on the basis of the applicant's current and previous research. All of these aims relate to a proposed model of communication development in this population that identifies a number of critical \"cusps\" that have apparently been traversed by some individuals in the course of their communication development; but that represent the end point of that development for other individuals. The first three specific aims are: 1) To describe the extant communication abilities of a sample of young prelinguistic children with severe to profound mental retardation; 2) To describe the rate and sequence of communication skill development by these same subjects over a five year observational period; and 3) To determine whether specific child and/or environmental variables are significantly related to differential communication skill attainments by these subjects at the end of this observational period. These three aims will be addressed through a five year, repeated measures design involving 25 prelinguistic young children (ages 4 to 5 years at onset of observations) with severe to profound mental retardation. A battery of measures, including standardized assessments and direct observations of parent-child interaction, will be administered to each subject at least every six months, and more often with subjects who are arriving at the identified cusps at a faster pace. Specific aims 4 and 5 are to determine whether specific intervention procedures are effective in helping individuals with severe mental retardation make the transition from presymbolic, distal gesture communication to referential symbolic communication (Aim 4); and across one or more other critical cusps, including that between contact gesture and distal gesture communication, and/or that between single symbol and linguistic (syntactic) communication (Aim 5). Subjects in these studies will include both children and young adults; and single subject research designs will be used to determine the effectiveness of specific treatment approaches for each subject.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We conduct Monte Carlo experiments under the model of random censorship to investigate the small sample behavior of different standard deviation formulas for the Kaplan-Meier estimator. The simulation studies include the Greenwood formula, Peto's formula and an alternative modified Greenwood formula. The results of the simulation showed that the Greenwood formula underestimated the true variance more than 90% of the times. Peto's formula overestimated the true variance when the survival rates were less than or equal to 0.5. The accuracy of the three different variance estimators is investigated using the relative bias, mean square error and confidence coefficient criteria. the absolute value of the relative bias for the modified Greenwood formula was the smallest among all three different variance estimators. On the basis of the mean square error criteria, the differences were negligible between the three different variance estimators. The Greenwood formula did not approach the confidence coefficient 0.95 in any censored observation. The confidence coefficient of Peto's formula was closer to 0.95 than the other two variance estimators. The sample sizes, censorship levels and distributions of the random variable all influenced the accurancy of the variance estimators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objective of this research is to elucidate the molecular mechanism(s) by which estrogen mediates vascular lesion formation. Occlusive vascular lesions are characterized pathologically by endothelial cell hyperplasia and aberrant smooth muscle cell growth. Estrogen therapy is associated with a worsening of atherosclerosis. Until now adverse estrogenic effects have been explained by the dysregulation of the estrogen receptor (ER). Although the ER is required for the growth of cells, we consider ER actions as a late event when compared to the rapid formation of estrogen-induced oxidants that initiates the early signal for cell growth. We have shown that estrogen-induced oxidants signal endothelial cell proliferation and blood vessel formation. Since pathogenic blood vasculature formation contributes to the etiology of vascular disease, we postulate that exposure to estrogen therapy is harmful to the human cardiovascular system. This revised proposal will examine the role by which 172-estradiol (E2) activates a redox sensitive Pyk2 signaling pathway involved in the pathology of vascular lesions. Therefore, the objective of this proposal is to elucidate the molecular mechanism by which estrogen induces vascular lesion formation. The hypothesis of this proposal is that a Pyk2/GPR30/Vav1 complex mediates pathogenic blood vasculature formation in response to exposure to estrogen. We have formulated the following inter-related, focused specific aims to test our hypothesis: Aim 1: To determine whether estrogen exposure initiates Pyk2 protein-protein interactions resulting in a Pyk2/GPR30/Vav1 complex. Aim 2: To determine the mechanism by which estrogen-induced Pyk2 protein- protein interactions signal Id3 to block p21Cip1 expression. The long-term goal of this research project is to determine whether in vivo estrogen-induced Pyk2 signaling controls pathogenic blood vasculature formation. However, this long-term goal is not the purpose of this SC3 grant proposal; rather we intend to generate preliminary data from this SC3 Award that will be used to apply for successful NIH R01 funding in which we will include the determination of in vivo whether estrogen-induced Pyk2 signaling controls the formation of human vasculature in a mouse model. The PI's career goal is to become an independent investigator in the field of cardiovascular toxicology. The PI plans to achieve this objective by investigating the molecular mechanism by which estrogen induces vascular lesion formation. Florida International University offers a rich academic environment and well-respected research infrastructure that will provide a supportive setting to foster the PI's career development. The findings from the proposed research will contribute to a more profound understanding of how estrogen therapy mediates the formation of occlusive vascular lesions. This, in turn, will give new insights into the mechanisms by which estrogen increases the risk of occlusive vascular lesions and will identify novel targets for the treatment of vascular lesions in individuals exposed to these compounds.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Songbirds represent one of the few animal model systems for the study of the acquisition of human speech. Birds learn their song during development, and have a well-delineated forebrain circuitry devoted to learning and producing song. Area X is a part of the neural circuitry involved in song acquisition and modulation. Area X is also a part of the avian basal ganglia and receives a strong input from midbrain dopaminergic nuclei. Cells in Area X respond to changes in social context: their activity changes depending on whether males are singing in isolation or singing to a female. The proposed studies will investigate the role of dopamine in modulating neural activity in Area X under different social contexts, and how changes in activity in Area X may affect the responses of other nuclei in the learning pathway. Given both the homology and similarity between Area X and mammalian striatum, as well as the ecological and behavioral relevance of the bird song system, the merging of the two fields of study would provide considerable information toward understanding plasticity and song production in birds, as well as insight into striatal function in a behaviorally relevant paradigm. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The inflammasome has recently been described as an important protein complex in which a non-NFkB- mediated signaling pathway leads to up-regulation of key pro-inflammatory cytokines, including interleukin (IL)- 1b and IL-18. Members of our group recently reported that the inflammasome plays a critical role in pro- inflammatory response in murine sepsis, and this response is dependent upon mitochondrial integrity and an intact autophagy response to injury. We now present data supporting an important role for the inflammasome in predicting severity and mortality during human infection-related ALI/ARDS. Moreover, our preliminary animal studies demonstrate that statins exacerbate lung injury and inflammation via activation of the inflammasome. We therefore hypothesize that activation of the inflammasome plays a critical role in the development of infection-related ALI/ARDS and that statin administration may increase inflammasome-related downstream cytokines during lung injury. In particular, our collaboration with the ARDSnet SAILS trial investigators (a randomized trial of statins vs. placebo in infection-related ALI/ARDS) in this Ancillary Studies in Clinical Trials RFA (HL-12-012) provides a unique opportunity to obtain additional collection of key human samples to address these important processes. We therefore propose: Specific Aim 1: To determine gene expression and protein levels of the inflammasome during infection-related ALI/ARDS using prospectively collected blood (n=100) and banked plasma samples (n=600) from placebo- and statin-treated SAILS subjects. Gene expression and protein levels of the inflammasome will be correlated with 60-day mortality and additional SAILS trial secondary outcomes. We hypothesize that circulating inflammasome levels will serve as a biomarker of severity and mortality of infection-related ALI/ARDS and that inflammasome levels in statin-treated subjects will correlate with clinical outcomes. Specific Aim 2: To determine the cellular localization of expression of the inflammasome complex and role of inflammasome activation on cellular responses and function, using primary neutrophils and monocytes isolated from prospectively enrolled placebo- and statin-treated SAILS subjects (n=100), as well as primary cells isolated from control ICU subjects (n=100). We hypothesize that determining which circulating cells are the predominant source of inflammasome activation will increase sensitivity of the correlation of inflammasome levels with clinical outcomes and that localized activation of the inflammasome will result in mitochondrial dysfunction that will be enhanced in the presence of statin administration.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite aggressive surgical resections, high-dose radiation therapy, and chemotherapy delivered at toxic doses, the vast majority of patients with malignant brain tumors survive less than one year making conventional therapy for malignant brain tumors the most expensive medical therapy per quality-adjusted life-year saved currently provided in the United States. Moreover, the failure of these treatment modalities to be tumor-specific at the molecular level, results in inevitable damage to surrounding normal brain that incapacitates patients treated with these traditional modalities. The inherent specificity of immunologic recognition offers the prospect of targeting malignant cells more precisely. Within our program, direct injection of 131-I-labeled, operationally-specific, monoclonal antibodies (MAbs) into brain tumor resection cavities delivers extremely high radiation doses to tumor cells around the resection cavity and has produced promising results in Phase II clinical trials. These MAbs diffuse only short distances beyond the cavity, however. Therefore, most of the radiation extending beyond the cavity is not specifically targeted to tumor cells and the radiation dose delivered beyond the cavity declines exponentially from the cavity interface. As a result tumor cells that are known to infiltrate the brain for significant distances beyond the cavity are sub-optimally treated and lethal tumors always recur within 2 cm of the radiated re section cavity. Continuous microinfusion is a promising technique that allows homogeneous delivery of even large molecular weight molecules at high concentrations throughout large areas of the brain. Although this technique may enhance the delivery of 131-I-labeled MAbs and other therapeutic agents to diffusely infiltrating malignant brain tumors and reduce recurrence rates, the parameters that govern this technique and its limitations have not been defined. One of the major goals of this proposal is to define these parameters. In addition, this proposal is designed to investigate whether targeted radiotherapy might be improved through the use of human chimeric MAbs with increased biostability and the use of high linear energy transfer radioisotopes, such as 211-At, with greater relative biological effectiveness.The hypothesis to be tested in this proposal is that continuous microinfusion will widely deliver operationally tumor-specific monoclonal antibodies conjugated to 131-I or the alpha-emitter 211-At such that they will be specific and potent therapeutic agents against malignant brain tumors with major reductions in toxicity to normal brain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The neuronal ceroid lipofuscinoses (NCL) comprise a group of poorly understood inherited neurodegenerative diseases of children and adults. Although canine and ovine models of this disease are available, they are not economical to maintain, difficult to study genetically, and not easily manipulated experimentally. Two mouse models of NCL have been discovered. Both develop severe neurological disease; both have numerous neuronal cytoplasmic autoflourescent inclusions. Ultrastructurally these inclusions consist of stacked membranes identical to those found in the canine model of NCL and similar to inclusions considered diagnostic of human forms of the disease. One of these mouse mutants, juvenile bare (jb), has been mapped to the proximal part of Chromosome 7; the genetic linkage of the second mutant, motor neuron degeneration (Mnd), will be established, and candidate genes for both mouse diseases will be sought. These mutants will be studied to characterize the progression of lesions in affected organs, particularly the brain, during the life span. Early diagnostic lesions will be sought. The basis of the biochemical defect will be analyzed by isolating and sequencing the stored lysosomal protein(s) and searching for abnormal lysosomal enzymes. A search for other models of NCL will be conducted among the dozens of known mouse mutants with blindness, seizures, diffuse neuronal degeneration and known lysosomal disorders. Finally, the mutant models will be made available to other investigators. It is hoped that studies of these models will provide important insights into the devastating pathogenesis of Batten disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Xenotransplantation Core provides specialized, on-demand surgical services for the transplantation and evaluation of human fetal tissues in immunodeficient animals in support of the research projects carried out by the Formative Center. The core is responsible for the coordination of immunodeficient animal procurement (nude rats) with anticipated availability of fetal tissues, and sterile renal subcapsular or subcutaneous transplantation of appropriate fetal source tissue upon availability. Our group has demonstrated that fetal testis, handled and processed in the usual manner by hospital staff, is histopathologically responsive to selective toxicant exposure when implanted under the kidney capsule of nude rats. The Xenotransplantation Core facilitates the expansion of this past work into other fetal tissue types and environmental impacts by centralizing development of the xenografted animal model in support of the three primary research areas of the Formative Center: Metabolic Syndrome. Prostate Imprinting, and Lung Remodeling. The Xenotransplantation Core will function as the site of the initiation of the experiments carried out in the Formative Center's Research Projects by providing an on-demand service that coordinates with the Tissue Procurement Core to transplant fetal tissue as it becomes available into xenogeneic hosts. As such, the Core will maintain host animals for instant availability, and its personnel will have the shared responsibility of performing surgical requests within a time frame that allows confidence in graft viability upon implantation. The Xenotransplantation Core will pursue these Specific Aims: Specific Aim 1 - Provide animal surgery and model development services Specific Aim 2 - Interact with research investigators to initiate animal experiments in support of the Formative Center's research projects Specific Aim 3 - Implement current and emerging surgical techniques in xenotransplant model development and training of technical staff", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lung cancer is diagnosed late, has a 5 year survival of only 15%, and kills more people than colorectal, breast and prostate cancer combined. Less than 30% of lung cancers are resected and the majority of patients are treated with cisplatin and ionizing radiation (IR). We show that inhibition of DNA damage signaling by ATR kinase during treatment with cisplatin and IR is well-tolerated and leads to durable responses in mouse xenograft and genetic models of lung cancer. Quite unexpectedly, we show that in addition to potentiating DNA damage, two clinical ATR kinase inhibitors (ATRi?s), with unrelated structures, block expression of the immune checkpoint protein PD-L1 and increase presentation of MHC class I antigens in lung cancer cells after IR. Our finding that crosstalk exists between DNA damage signaling and immune checkpoints has not been described previously and is the focus of this proposal. Immune-inhibitory pathways, termed immune checkpoints, are coopted by tumor cells to evade cytotoxic immune cells. PD-1 is expressed on cytotoxic T cells and its ligand PD-L1 is upregulated in lung cancers. PD-L1 binding by PD-1 prevents the activation of cytotoxic T cells. Immune checkpoint blockade using anti-PD-L1 and anti-PD-1 antibodies restores anti-tumor immune responses and is emerging as an exciting lung cancer therapy. We propose that ATRi?s inhibit DNA repair and cell cycle checkpoints potentiating the DNA damage induced by cisplatin and IR while concurrently inhibiting PD-L1 expression and restoring anti-tumor immune responses. Our objective in this proposal is to define the mechanisms that connect DNA damage signaling and immune checkpoints. This objective will be accomplished by the following Specific Aims. Aim 1: To determine how ATRi?s inhibit PD-L1 expression in lung cancer cells after IR. This aim will define the contribution of ATR, ATM, IRF-1, NF-?B and p53 to PD-L1 expression after IR. Aim 2: To determine how ATRi?s increase MHC class I expression on lung cancer cells after IR. This aim will identify ATR, ATM, and p53 signaling that inhibits protein synthesis and MHC class I presentation after IR. Aim 3: To identify ATRi-induced PDL-1/PD-1 immune checkpoint blockade in lung cancer after IR. This aim will identify ATRi-induced immune checkpoint blockade in lung cancer after IR. The outcomes of these Aims will identify mechanisms that connect DNA damage signaling to immune self- tolerance. This will define a novel therapeutic opportunity to use ATRi?s to potentiate the DNA damage induced by cisplatin and IR while concurrently inducing immune checkpoint blockade.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The mammalian bombesin-like peptides, gastrin-releasing peptide (GRP) and neuromedin B (NMB), are important regulatory neuropeptides which mediate a range of biologic responses including smooth muscle contraction, stimulation of secretion, modulation of neuronal activity, and control of cell proliferation. Previous studies from this laboratory have shown that: 1) in tumor cell lines, the prepro-GRP gene is regulated at the level of primary transcription; and 2) detectable expression is restricted to a subset of neurons in the central and peripheral nervous system, neuroendocrine cells, and derivative tumor cell lines. We plan to define the molecular mechanisms important in prepro-GRP transcriptional activation in expressing cells, and suppression in nonexpressing cells. The first step in this analysis is the identification of cis-acting prepro-GRP promoter elements important for transcription. A series of fusion genes were assembled to assay the transcriptional effects of sequence elements in the prepro-GRP promoter. In these constructs, the firefly luciferase reporter gene is placed under transcriptional control of varying lengths of the human prepro-GRP promoter region. After transfection and subsequent transient expression in various host cell lines, luciferase activity is determined for each construct containing different prepro-GRP promoter elements. Using this assay system, we have made the following observations: 1) Promoter sequences between -5000 and -1600 (the origin is defined as the initiation site for transcription) exert an approximately two-fold negative effect on transcription, while sequences between -1600 and -400 exert a weak positive effect. In this positive domain, a several hundred base long 5' upstream sequence is very highly conserved between the rat and human prepro-GRP gene (-720 to -450); and 2) The constructs behave the same in host cells that do or do not normally express the prepro-GRP gene. We conclude that promoter elements between -5000 and + 100 alone do not determine cell-type specificity of transcription regulation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Laboratory of Pathology (LP) supports the clinical and research missions of the NIH and the NCI by providing anatomic pathology and tissue processing services to the Clinical Center and to all of the categorical institutes. LP is CLIA-certified with a Certificate of Accreditation and is inspected every two years for compliance with regulatory standards by the College of American Pathologists. LP's clinical services provide services in surgical pathology, autopsy pathology, hematopathology, cytopathology, chromosome pathology, pediatric pathology, flow cytometry and molecular/special diagnostics. Clinical Operations is the administrative and technical core of the clinical component of the Laboratory of Pathology. It oversees the Quality Management program, the core laboratory services, human resource management for clinical and technical staff, the Laboratory Information System (LIS) and manages the clinical diagnostic tissue archives of the laboratory. As a service to investigators throughout the NIH, scientists may request tissues for research from the archive following appropriate ethical approval (from their IRB or from the OHSR). In this fiscal year through the end of July, we processed about 250 new and ongoing requests for tissue samples. This resulted in approximately 10,500 individual biospecimens (unstained slides, special stains, tissue shavings, and tissue slides processed with RNAse precautions) provided to researchers from our tissue archive. The Clinical Operations section also processed approximately 125 medical-legal requests, which involved retrieving and processing approximately 1,000 clinical slides and/or blocks that were forwarded to other healthcare or research facilities at the request of clinicians or patients. A relatively new function within LP's Clinical Operations section has been the addition of whole slide scanning of stained tissue slides. Whole slide scanning allows the indefinite storage and electronic access of patient case material, without the risk of slide loss or deterioration of stain intensity over time. Newer technologies in slide scanning also provides a platform that employs image algorithms that can accurately and reproducibly analyze immunohistochemical stains. In fiscal year 2016, LP residents and staff scanned approximately 2,800 stained whole tissue slides of: unique and interesting pathology cases; cases submitted to LP clinical pathologists which required the slides be returned to the submitting facility; clinical cases shared with staff pathologists or clinical consultants electronically rather than risking the loss of glass slides; and cases scanned for resident study sets, research or to present at conferences on and off campus. As of July, 2017, there have been more than 4,200 while slide scans performed or supported by Clinical Operations staff. In total since inception of whole slide scanning, over 20,000 slides have been scanned. The Clinical Operations section also includes oversight of the Laboratory Information System (LIS) and collection and reporting of quality management (QM) data to LP's leadership and QM Committee monthly. The LIS is part of the SoftLab system used by the Clinical Center and it interfaces with the hospital information system so that anatomic pathology results are available online. As part of the tissue request process and to support requests for pathology reports from patients and clinicians, we conduct searches of the pathology database and provide lists of cases that match the requested criteria. We currently have reports scanned from 2008 to the present and are maintaining the report archive in real-time. The LIS is also used by Clinical Operations to generate benchmark data and quality assurance statistics for managing and continuously improving the clinical diagnostic services. The quality management program monitors LP's performance with key benchmarks, such as: cytology, surgical and biopsy turnaround times; specimen adequacy, labeling and identification compliance; and quality control of special stains. Annual QM reports are provided to LP's clinical leadership to identify areas that have demonstrated improvements, but also shows areas that need continuous improvement and monitoring. Annual QM reports are provided to the NIH Clinical Center's Office of Patient Safety and Clinical Quality as one element needed to demonstrate an ongoing quality assurance and process improvement program for clinical services that is required for Joint Commission accreditation. Clinical operations also oversees pathologist-supervised tissue division for research. Patients who undergo surgery at the NIH Clinical Center frequently have tissue divided between research and diagnostic purposes, as dictated by IRB-approved protocols.The Technical Laboratory Manager maintains the lab, and Pathologists Assistants (PAs) assist in procurements, dissection, documentation of specimen accountability, and distribution of tissues to researchers and clinical sections. The TLM also serves as contract officer technical representative for LP and assists the Clinical Manager in property management, equipment procurement, and maintenance activities for the clinical and research sections of LP.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Interactions between developing male gametes and adjacent Sertoli cells are essential for spermatogenesis. Sertoli cells express genes encoding cell adhesion molecules, growth factors, transport proteins, proteases and protease inhibitors required by male germ cells. Germ cells regulate expression by Sertoli cells of many of these genes. Consequently, expression of these genes changes as the adjacent germ progress through the stages of the cycle of the seminiferous epithelium. This application examines the regulation and biological consequences of stage-specific gene expression by Sertoli cells from the perspective of the cathepsin L gene. Transcription of this gene increases more than 10-fold as adjacent germ cells progress from stage I to stages VI and VII and then decreases to undetectable levels when germ cells reach stage X. This cycle of gene expression is a response to germ cells which causes sequential repression, stimulation and re-repression of transcription of the cathepsin L gene. We have identified 2 domains in the cathepsin L gene that potentially respond to these signals: [1] An upstream repressor domain responds to inhibitory signals from germ cells [2] A 120 bp activation domain is stimulated upon the culmination of testis maturation, which is characterized by the completion of the first wave of spermatogenesis. These domains are functional in vivo; a 3kb fragment of the cathepsin L gene confers both Sertoli cell-specific and stagespecific expression of a reporter in transgenic mice. This application also proposes to examine the function of cathepsin L in the seminiferous epithelium. Those experiments are prompted by our observation that mice which express catalytically inactive cathepsin L exhibit an increased incidence of seminiferous tubule atrophy and produce 30% fewer spermatids in nonatrophic tubules than are produced by control mice. Building on all of these data, this proposal asks four questions which are our specific aims: 1 What are the specific cis-acting elements in the activation domain and what are their functions? 2. What is the identity of the transcription factors that bind to the maturation domain? Is their expression stage-specific amd maturation-dependent? 3. What cis-acting elements and which spermatogenic cells repress cathepsin L promoter activity in Sertoli cells? 4. What is the function of cathepsin L in the seminiferous epithelium? [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Applicant's Abstract) This 5-year study, proposed by the UCLA Drug Abuse Research Center (DARC) and National Development and Research Institutes, Inc. (NDRI), is intended to increase our knowledge of the long-term outcomes and costs of a modified therapeutic community (TC) program for drug-abusing prison inmates. The Amity prison TC Program, established in 1990 at the R.J. Donovan Correctional Facility near San Diego, has been evaluated previously in a process and outcome study as one project of the NDRI Center on Therapeutic Community Research. The proposed study will conduct 5-year post-treatment follow-up interviews with the same subjects (N=715) and include an analysis of program costs. The Amity TC program provides intensive substance abuse treatment to male inmates for 9 to 12 months. In addition, participant inmates paroling from prison may voluntarily enroll in an aftercare residential program, called Vista, for up to 12 months. The project has the following specific aims: (1) To conduct secondary analyses of data previously collected on the sample. Using existing data from the previous evaluation of the Amity program, project researchers will both analyze issues of interest to the field and establish a foundation for conducting analysis with the 5-year follow-up data to be collected. (2) To assess long-term post-treatment outcomes of a prison-based TC program. Important post-treatment outcomes (drug use, criminality, employment, health status, family relationships, and psychological status) of treatment subjects will be compared with control subjects, for an approximately 5-year period after return to the community. (2A) To determine differential outcomes within selected subgroups. This aim will examine whether subgroups of the main sample have different long-term outcomes. Subgroups to be included in analysis of this aim will be defined by discharge status (dropped out, completed), race/ethnicity, primary drug problem, criminal history, criminal risk status, and psychiatric status. (2B) To examine factors that contribute to the recovery of drug-abusing offenders who did not participate in treatment. Analysis will address the need to increase our understanding of how drug-abusing offenders who do not enter formal treatment but who reduce or cease drug use are able to do so. We will use new data from the control group of the original Amity evaluation sample. (3) To provide a comprehensive economic cost analysis of the prison TC program, the aftercare TC programs, and the control group. This aim will provide needed data on the program and treatment costs for three study conditions. Accomplishment of this aim will also lay the foundation for a planned proposal to conduct a comprehensive benefit-cost analysis of prison-based TC programs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed study goals are to test whether it is possible 1) to maximize the benefits and reduce the risks of maintenance drug treatment for schizophrenia by using reduced dose strategies; 2) to employ a family management intervention to further reduce the risk of relapse, and improve social and intrafamilial functioning. The proposed study is part of a collaborative project sponsored by the NIMH. Patients will be stabilized on fluphenazine decanoate (FD) and then randomly assigned to one of the following treatment to continue for two years: continuous low dose (FD .2.5 to 10mg every two weeks); continuous standard dose (FD 12.5 to 50mg every two 'weeks); discrete targeted early intervention (placebo FD 1/2 to 2cc q two weeks). Each patient will also be randomly assigned to either applied or supportive family management. For all patients, episodes of symptom exacerbation will be treated openly with oral fluphenazine or FD, in addition to the \"blind\" study medication, without breaking the blind. Once the episode is resolved, patients will continue on study treatment only. Patients who require open medication for more than 120 days will be terminated from the double-blind medication but continued in family management for '.the full two years. Patients assigned to applied family management will receive individual family sessions according to a predetermined schedule for the first year. Families of all patients will attend a psychoeducation workshop and have the opportunity to attend monthly group meetings throughout the two year period. Outcome measures will include relapse rates, measures of psychopathology, social functioning and side effects. Additional data will be collected to compare the cost effectiveness of the six treatments. This will include the cost of study treatments, other health care and public interventions and lo t family work productivity as well as offsetting gains. Patients who fail to stabilize initially within six months on open FD or who relapse and fail to restabilize after 140 days of open medication treatment will be asked to consent to the following: an eight week double-blind randomized trial of lithium or lithium placebo in addition to open treatment with FD and continuation, of the assigned family management.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY There are two options to treat the pain and stiffness of tibiotalar OA: arthrodesis and total ankle replacement (TAR). Arthrodesis involves fusing the tibia and talus and is performed in most patients. With motion completely lost at the tibiotalar joint, 50% of arthrodesis patients are unable to return to their desired activities. Eliminating tibiotalar motion may also force the adjacent joints, including the subtalar, talonavicular, and calcaneocuboid joints to undergo compensatory hypermobility. Although not confirmed, excessive motion may cause OA at these adjacent joints. Total ankle replacement is an attractive alternative to arthrodesis as it may reduce compensatory motion by opening degrees of freedom at the tibiotalar joint. However, 33% of TAR implants fail within 10 years. Also, about 15% of TAR patients develop arthritis at the adjacent joints. With poor 10-year outcomes, TAR remains reserved for older patients. Yet, over a lifetime, younger patients may stand to benefit the most from the functionality offered by TAR. Our ultimate goal is to make TAR a viable option for restoring mobility to patients ? young and old Quantification of in-vivo kinematics (i.e. angles and translations) of the tibiotalar and adjacent joints is essential to design TAR implants that increase longevity and improve functionality. Prior studies have not measured in-vivo kinematics of the tibiotalar joint independent from the adjacent joints in TAR or arthrodesis patients. Our group has developed and validated a dual-fluoroscopy (DF) system that accurately measures in-vivo motion of each joint. DF visualizes motion relative to bony anatomy and implanted hardware, yielding ?joint arthrokinematics?. Arthrokinematic data could identify features of a TAR implant that impede articulation, motivating design changes. These data could also drive computer models that predict stresses at the bone-implant interface to identify modes of TAR failure. We propose an R21 study to measure in-vivo joint arthrokinematics during walking, stair-ascent, and balanced heelrise in: 1) post-TAR, 2) post-arthrodesis, and 3) non-pathologic controls. Compared to controls, we hypothesize: 1) motion of the adjacent joints in arthrodesis patients is increased, 2) motion of the adjacent joints in TAR patients is increased, and 3) motion of the prosthetic tibiotalar joint in TAR patients is reduced. Confirming Hypothesis 1 will provide mechanistic evidence of compensatory motion that is clinically believed to instigate OA in the joints adjacent to the fusion site. In addition, confirmation of Hypothesis 1 will provide rationale to focus future research directives on projects that advance TAR. Demonstrating that hypermobility is not eliminated (Hypothesis 2), and that TT motion is restricted (Hypothesis 3), following TAR will indicate that improvements to TAR are still required. Although an exploratory study, completion of this R21 will yield the only in-vivo measurements of articulation for the tibiotalar and adjacent joints in these cohorts. Importantly, data collected herein will support a body of future research designed to meet our ultimate goal.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abundant evidence supports the idea that progressive up-regulation of complex pro-inflammatory signaling, A42 peptide evolution, and loss of neurotrophic support underlie the initiation and propagation of Alzheimer's disease (AD). Micro RNA (miRNA)-mediated messenger RNA (mRNA) interference is a newly discovered genetic mechanism involved in the regulation of gene expression at the post-transcriptional level. The major mode of miRNA action is to target and bind to the 3' un-translated region of specific mRNAs and in doing so, quench expression of that mRNA, thereby acting as a negative regulator of gene expression. AD affects only specific areas of the brain, and mis-regulated miRNA expression is strongly linked to altered gene expression within these regions. In AD models, these same AD-enriched miRNAs induce specific pathological changes that are strikingly similar to those observed in AD brain. Accumulating evidence strongly suggests that altered miRNA-mediated processing of specific mRNA populations triggers the pathogenic expression of genes that drive the AD process. The overall goal of this project is to significantly move the miRNAin- AD field forward, and impact the scientific understanding of AD, by defining mechanistically how specific miRNAs induce molecular-genetic mechanisms that result in AD-type change. Human brains utilize only a fraction of all known miRNA species; and only a subset of these are altered in AD brain. Increases in the expression of a specific NF-?B-sensitive miRNA-146a down-regulates the abundance of at least three major mRNA targets, complement factor H (CFH), interleukin-1 receptor associated kinase (IRAK) and tetraspanin 12 (TSPAN12), that encode key modulators of the brain's innate immune system, inflammatory response and the generation of neurotrophic or neurotoxic amyloid peptides. Down-regulated TSPAN12 restricts ADAM10 (?-secretase) dependent cleavage of beta-amyloid precursor protein (APP). Stressed human brain cells up-regulate miRNA-146a in parallel with increased amyloidogenesis and shedding of A42 peptides. Hypothesis: up-regulated miRNA-146a contributes to altered innate immune and inflammatory responses, increased A42 peptide generation and decreased neurotrophic support. Three Specific Aims test predicted outcomes based on this hypothesis in AD, in stressed human brain primary cells, and in specific transgenic models of AD. These Specific Aims include (1) identification of alterations in miRNA-146a abundance in AD brain; (2) testing whether stressed human neuronal, astrocyte or microglial cells exhibit miRNA changes similar to those conducive to inflammatory neurodegeneration and A42 peptide generation in AD tissues, and testing if added exogenous miRNA-146a or anti-miRNA-146a oligonucleotides will promote or neutralize these changes; and (3) testing whether miRNA-146a, anti-miRNA-146a and modulation of CFH, IRAK or TSPAN12 expression are key regulators of the inflammatory response and amyloidogenesis in amyloid over-expressing Tg2576 mice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this set of projects is to use functional magnetic resonance imaging (fMRI) to compare the degree to which brain regions of adolescents and adults are engaged by tasks involving processing of emotionally salient stimuli. Based on developmental continuities in the relevant psychological processes, we anticipate considerable similarity across age groups in the topography of brain regions engaged by relevant tasks. However, we hypothesize that developmental differences in cortico-limbic circuits of adolescents and adults will be reflected in patterns of fMRI activation. Specifically, we hypothesize in both adults and adolescents that attention, threat perception, and memory tasks involving the processing of emotionally salient stimuli will engage the amygdala, cingulate gyrus, and association cortex of medial/inferior prefrontal cortex and temporal regions. Nevertheless, height of task-associated activation is hypothesized to differ between adolescents and adults within these regions. Moreover, prior studies distinguish puberty vs. age-related aspects of cognitive development: some aspects of attention, threat perception, or memory development relate to changes in chronological age whereas other aspects, particularly those involving emotional processes, relate to pubertal status. Therefore, we expect eventually to use emotion-evoking fMRI tasks to test hypotheses on the presence of complementary, distinguishable puberty vs. age-related components of brain development. Due to the limited fMRI database on neurodevelopmental aspects of emotional processes, one initial goal of the current project is to compare brain activation patterns in adults and adolescents, irrespective of pubertal status. A second initial goal is to generate behavioral data in adults and adolescents designed to inform the implementation of future fMRI tasks. Studies accomplishing both goals will facilitate eventual studies designed to distinguish puberty vs. age-related aspects of brain development. To meet these initial goals, the current project implemented a series of studies during the first three years in which data have been collected. For our fMRI studies, brain activation profiles were examined to four sets of emotionally evocative stimuli. This included evocative faces, standardized emotion-inducing picture sets, monetary feedback during a decision task, and threat of a mildly aversive air-puff. In total, an initial set of five fMRI studies were completed in samples of between 14 and 17 psychiatrically healthy adults. Two additional fMRI studies were completed in samples of between 14 and 17 psychiatrically healthy adolescents. During the past year, these experiments have been replicated in an additional series of 15 healthy adolescents and adults. In each of these, hypothesized brain activation patterns emerged. For one of these tasks, involving face emotion processing, we compared patterns of activation in adolescents and adults. As hypothesized, this comparison revealed signs of amygdala, orbitofrontal, and medial frontal hyperactivation in adolescents during passive viewing, as well as signs of hyperactivation in these regions in adults during attention manipulations. Subsequent studies replicated these results. These results are summarized in five recent publications from our group [1-5]. In an effort to replicate these associations, we have acquired further data sets in other samples that are larger than the initial data sets. We will be analyzing these data during the coming year, with the plan of also publishing these findings. Finally, for our behavioral studies, neuropsychological data were gathered to facilitate the development of new fMRI tasks. These included data on four additional tasks. In these tasks, between 20-60 subjects were studied, including approximately equal numbers of adolescents and adults. Three additional manuscripts using these data have been submitted and are under-review. We are also in the process of analyzing other data and preparing research reports for other tasks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The University of Alabama at Birmingham 's Center for Labor Education and Research, in its application for funding an EPA- HWWT cooperative agreement, will provide courses to four populations of workers who share the need for general and specialized training in topics related to 29 CFR 1910.120, Hazardous Waste Operations and Emergency Response. The overall goal is to improve the health and safety of members of the Communications Workers of America (CWA), Native Americans, emergency health care providers and fire fighters, by helping them reduce exposures to hazardous chemicals. Classes include Hazardous Materials Awareness, Operations, and Technician, adapted for the four different training populations; Handling Contaminated Patients; Hazardous;SCBA Fit Testing; Air Surveillance in Chemical Emergency Incidents; Health Effects of Chemical Exposure; Confined Space Entry and Rescue, Hazardous Waste Handling; and Worker Training Methods. Classes will be taught in train-the-trainer mode, and materials provided for workplace training by the participants and for outreach to their respective communities. The four target populations have in common their potential for exposures to chemicals, training budgets that are inadequate or nonexistent, and job- and finance- related restrictions on extensive travel for training. CWA workers in manufacturing, product distribution and service, health care, printing and publishing, and numerous crafts will have regional classes throughout the United States, as will members of all 557 federally-recognized Native American tribes. Indian law enforcement officers, fire fighters, highway and hospital workers, emergency planners, natural resource personnel, environmental planners, and search-and-rescue units will be trained in safe Awareness Level response and Incident Management in classes coordinated through cooperation with the Native American Fish and Wildlife Society. In the southeastern United States, emergency room personnel will be trained to handle contaminated patients; surveys show most are unprepared for this problem and are in violation of the regulations of several agencies, including OSHA 1910.120. Because fire fighters have increased risk of diseases shown to be related to the inhalation of chemicals and smoke, they will be trained in toxicology, fit testing, air monitoring, and rescue from confined spaces with hazardous atmospheres. Computer-based asynchronous training will be utilized to achieve some of the objectives, and this method formally compared with traditional training. Professional safety and health trainers will develop and deliver all training using curricula developed and piloted under previous cooperative agreements. Total number of trainees will exceed 20,000, with tribal peer trainees and community outreach participants not included in the estimate. The total proposed cost of the five- year project is 3,296,947 dollars.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Matrix metalloproteinases (MMPs) are promising targets for cancer therapy. However, recent clinical trials of MMP inhibitors have been disappointing. Certain MMPs may regulate apoptosis signaling pathways and sensitize tumor cells to apoptotic signals. Inhibition of these MMPs will promote tumor cell survival and this effect may offset the beneficial activity in inhibition of metastasis. Matrix metalloproteinase-9 (MMP-9) expression is significantly increased in malignant prostate cancers. In addition to its ability to cleave collagens and basement membrane components, MMP-9 also cleaves and activates the multifunctional cytokine TGF-[unreadable]. TGF-[unreadable] (having three isoforms in mammals: TGF-[unreadable]1, [unreadable]2, [unreadable]3) is an important regulator of normal and malignant prostate. While TGF-[unreadable]1 induces apoptosis in certain prostate cancer cells, TGF-[unreadable]2 blocks apoptosis. By activating all three isoforms of TGF-[unreadable], MMP-9 may have different net effect on prostate cancer apoptosis depending on the status of expression of TGF-[unreadable] isoforms, their receptors, and the downstream signaling molecules. There is a critical need to determine the effects of MMP-9 on apoptosis in prostate cancer. Presumable, inhibition of MMP-9 will be beneficial in patients where MMP-9-mediated TGF-[unreadable] activation has an anti-apoptotic effect in cancer cells. Selection of appropriate patients according to the status of TGF-[unreadable] signaling machinery may be critical for future clinical trials to evaluate the therapeutic efficacy of MMP-9 inhibitors. The objective of this research is to define the role of MMP-9 in prostate cancer apoptosis and how the effects of MMP-9 on apoptosis change along with prostate cancer progression. At the completion of this project, we expect to clarify the role of MMP-9 in apoptosis regulation in prostate cancer and how this role may change in relation with the changes of TGF-[unreadable] signaling at different stages of prostate cancer progression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "$ The overall objective of the research is to define the kinetics of uptake, distribution, metabolism, and excretion of inhaled lipid soluble vapors in man, by a set of differential equations, using predictable constants. The physical, chemical, and biological constants needed will be determined by simple experiments. Solution of the equations will be facilitated by a program for a large digital computer. To verify the pharmacokinetic model, the experimental conditions of suitable published studies in man will be simulated, and their results compared with the calculated data. Further verification will be done by animal experiments. The pharmacokinetic model for the rat will be elaborated, and the predicted concentrations in rats sacrificed at selected time intervals following exposure. The described pharmacokinetic model will be used to design the proper monitoring of industrial data; to predict the cumulation of inhaled compounds and their metabolites in the body; to explain the role of certain physiological factors (body build, pulmonary ventilation, tissue perfusion); and to formulate guidelines for the prediction of the uptake, distribution, metabolism, and secretion of new noxious compounds introduced in industry. Initial plans are to use toluene, trichloroethylene, carbon disulfide, and methylene chloride in the study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The chemical and biochemical properties of a number of analogues of 8-hydroxy-5-deazaflavin cofactor were studied and some substrate structure-reactivity relationships between these analogues and 8-dydroxy-5-deazaflavin dependent NADP+ reductase were established. A affinity chromatographic medium for possible use in the purification of 8-hydroxy-5-deazaflavin-dependent enzymes was synthesized.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Successful development of a vaccine against HIV will likely require the induction of both antibody and/or cellular immune responses sufficient to prevent infection or disease respectively following infectious challenge. While the induction of antibody responses for a variety of other infectious pathogens is readily achieved by a variety of vaccine formulations, live attenuated, recombinant viral vaccines or plasmid DNA vaccines only induce the induction of long-lived cellular immune responses, particularly CD8+ T cell responses. Moreover, since live attenuated HIV vaccines might be precluded from use due to safety concerns and DNA vaccines at present only induce modest CD8+ T cell responses in humans, there is an urgent need to develop ways to enhance the generation and maintenance of CD8+ T cell responses in humans in following immunization. This study focuses on how to optimize the magnitude and duration of CD8+ T cell responses following vaccination in rodents and primates using a variety of vaccine formulations. The data obtained over this past year have shown the following; 1. Prime-boost immunization with SIV Gag protein and TLR 3 or 7/8 ligands elicit potent T cell responses in non-human primates. Such responses were noted in both peripheral blood mononuclear cells and were much higher in the broncheoalveolar lavage. Upon boosting with rAd-5 SIV , CD8+ T cell responses were further enhanced. Animals were challenged with SIV Mac 251. These results show that a heterologous prime-boost immunization regimen using a protein and TLR ligand followed by rAd-5 induces potent T cell immunity. 2. Experiments in NHP have compared HIV Gag DNA versus HIV Gag protein+ Poly I:C as a prime prior to rAd-5 Gag as a boost. The data reveals that priming with DNA enhances both CD4 and CD8 responses after the rAd5 HIV Gag boost compared to rAd5 HIV Gag alone. By contrast, priming with HIV Gag protein+ Poly I:C enhanced the magnitude of CD4 responses following the rAd5 boost but not CD8 responses. Moreover, it was notable that DNA immunization but not protein + Poly I:C induced a small frequency of CD8 cells after the priming. Collectively these data suggest that priming for CD8+ T cells is critical if there is to be further expansion of such cells after the boost.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Oral infections lead to local persistent inflammation that when \"uncontrolled\" gives rise to periodontal diseases (PD), a costly public health burden. PD is not only limited to tooth loss and associated loss of self-esteem but also impacts general health. Our recent findings indicate that resolution of inflammation is an active process and reveal an urgent need to clinically navigate this essentially uncharted terrain of infammation-resolution. To address this scientific opportunity and important mission in oral health, a multidisciplinary team of experts is assembled in a new Specialized Center for Research (SCR) configuration that will use new technologies to elucidate the molecular map of oral resolution. Our team and overall project will provide systematic molecular analyses and insights into the integrated host resolution response with focus in oral inflammation and PD-related tissue injury. Recently, we found that human neutrophils (PMN), along with generating the many proinflammatory mediators during inflammation, also generate in exudates novel \"proresolving\" lipid mediators (LM) that possess potent anti-inflammatory and tissue protective actions, as well as activate novel anti-microbial mechanisms in mucosal epithelial cells. These ongoing studies give rise to an overarching hypothesis to be tested in four interrelated sub-projects in this SCR requiring a highly integrated approach. The SCR overall novel hypothesis addressed is: Periodontal disease (oral inflammation and tissue injury), certain microbes, surgical anesthesia, and other widely used drugs impact recently uncovered LM resolution pathways essential in host defense and homeostasis. They prolong oral inflammation by disrupting these uncharted resolution circuits. This SCR team consists of 4 sub-projects with complementary expertise and 3 scientific cores needed to map the molecular circuitry of oral resolution with a central theme of novel LM and their actions with PMN, newly uncovered anti-microbial mucosal defense mechanisms, and PMN-mediated destruction of engineered tissue matrices. Our mission's overall goal is to provide these essential molecular coordinates for swift translation to human oral health. Broad long-term objectives are a) to elucidate the map of oral resolution, b) provide molecular map(s) and coordinates to clinicians to improve treatment practices and our national preparedness and biodefense status, and c) provide the basis for new therapeutic approaches using agonists of resolution.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Parasitic nematodes pose major human and veterinary health problems around the world. For the past 30 years, the parasitic nematode Ascaris suum has served as a model system for medically important helminths, such as filaria and hookworms. The early development of A. suum from embryo to 2nd-stage larvae takes place bathed in perivitelline fluid (PF) within an eggshell. However, little is known about the secretion of the PF, the regulation of its composition during development, or the molecular aspects of eggshell maintenance, even though these processes are critical for survival. The present study is designed to identify the major components of the PF and to characterize the pathway of their secretion from the larvae by exploiting the advantages inherent in the A. suum and Caenorhabditis elegans model systems. Changes in composition of the A. suum PF during development will be examined, and the roles of individual components in the hatching process assessed. Recently, the investigators have identified a developmentally regulated A. suum PF protein, As-p18, and have demonstrated that it exhibited almost identical patterns of expression with C. elegans LBP-1. Most significantly, LBP-1 appears to be secreted from hypodermal cells prior to the formation of the cuticle and the classical nematode excretory/secretory apparatus which only appears later in development. The present studies are designed to 1) characterize individual components of the PF, 2) clone, sequence and functionally express specific PF proteins to examine their potential roles in hatching of \"eggshell\" maintenance, 3) identify the cells responsible for the secretion of PF proteins either by immunolocalization with specific antisera or the identification of putative homologues in the C. elegans database and the use of the C. elegans transfection system, and 4) attempt to identify promoter elements responsible for the timing and specificity of expression. On a broad scale, the investigators intend to determine whether nematodes can be engineered to secrete specific proteins during specific stages in their life cycles.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Biostatistics Core provides statistical expertise in study design, data analysis, and interpretation of the results of analysis of cancer research data. Core biostatisticians collaborate with Cancer Center investigators in framing testable questions, devising experimental plans that make efficient use of experimental materials, patient populations and community participants, and ensuring that there is sufficient statistical power to answer the research questions. They are responsible for ensuring that due consideration is given to issues Iof unbiased and efficient sample selection, accuracy and precision of measurements, unbiased assessment of outcomes, completeness and adequacy of follow-up and other aspects of research study implementation. Core biostatisticians collaborate with investigators in each of the Cancer Center Programs on research, publications and grant proposals for external funding. They support the Clinical Trials Office as active, Imembers of the Cancer Center Protocol Review Committee, Data Safety and Monitoring Committee and as Chair of the Quality Assurance Committee. A core biostatistician serves on the Internal Advisory Committee of the Bioinformatics Core. This Core has a significant role ensuring scientific rigor of cancer research at the Karmanos Cancer Institute.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dr. Lyuba Varticovski headed this initiative to adapt genetically engineered mouse tumors for preclinical testing using transplantation of cell suspension into syngeneic or immunodeficient hosts. Multiple mammary tumors have been investigated to date. A manuscript describing the model system was recently revised to include microarray analysis as requested by the reviewers. (Varticovski, L., Hollingshead, M.G., Robles, A.I., et al., Transplantation strategy for generation of genetically engineered mammary mouse models suitable for preclinical testing. Clin Cancer Res., in review). Recommendations from Dr. Bob Wiltrout and Dr. Glenn Merlino were addressed by performing microarray analysis of MMTV-PyMT and wnt1 mammary tumors, and to complete a repository for tissues on each model. Gene expression analysis of other mammary models used in the study is in progress. Additional validation of the MMTV-PyMT model using Cyclophosphamide and Paclitaxel was performed as recommended by breast cancer clinicians at CCR. Based on success in using transplantation to generate synchronous mammary tumors for preclinical testing, we began harvesting and validation of transplantation protocol for selected lung cancer tumors from genetically engineered mice. In collaboration with Dr. Charles Vinson, we examined tumor development in diabetic A ZIP/F 1 fatless transgenic mice bred with several genetically engineered mouse models. This work was a cover article in Cancer Research, 2006. The inflammatory mechanisms responsible for accelerated carcinogenesis observed in this mouse model are not known and a proposal to follow up on this work was submitted. In collaboration with the Preclinical Pharmacokinetics Section of MTP (Dr. W. Doug Figg) we are investigating differences in pharmacokinetics of Paclitaxel between man and mouse. These studies are critical for development of appropriate schedule of drug administration for preclinical testing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The application proposes to investigate the hypothesis that noradrenaline containing neurons in the caudal ventrolateral medulla (VLM) of the cat are excited by acute increases in arterial blood pressure and that adrenaline containing neurons in the rostral VLM are inhibited by acute increases in arterial blood pressure. Further it is proposed that caudal VLM neurons influence the activity of rostral VLM neurons either through monosynaptic connections or through indirect pathways. Thus, interactions between caudal and rostral VLM neurons constitute neuronal mechanisms in the VLM of the cat that are part of baroreflex control of sympathetic nerve activity. The concept that VLM neurons are part of the baroreflex will be investigated in anesthetized cats prepared with a single innervated isolated carotid sinus (other baroreceptor regions denervated) and prepared for the recording of single unit activity of caudal and rostral VLM neurons. The behavior of VLM neurons to baroreceptor input will be examined by systematically forcing carotid sinus pressure and recording neuronal activity in response to this baroreceptor input. After completion of each experiment the brain stem of these cats will be processed for immunocytochemical identification of noradrenaline and adrenaline containing neurons. Noradrenaline containing neurons will be identified as those neurons that stain only with dopamine-beta-hydroxylase antibodies (rabbit) and adrenaline containing neurons as those that stain with phenylethanolamine-N-methyltransferase antibodies (rabbit). The response properties of individual neurons will be correlated to their location within populations of noradrenaline and adrenaline containing neurons in the VLM. Additional experiments are proposed to investigate the probable neural pathways through which caudal VLM neurons can influence the activity of rostral VLM neurons. First, the projection of caudal VLM neurons to the ipsilateral rostral VLM will be determined through the use of antidromic microstimulation and/or spike-triggered averaging of recorded field potentials. Second, monosynaptic connections between caudal and rostral VLM neurons will be investigated by recording simultaneous activity from pairs of these neurons and application of cross correlation analysis. Third, the possibility of an indirect pathway from the caudal VLM to rostral VLM neurons via neurons in the NTS will be investigated by electrophysiologic determination of axonal projections from the caudal VLM to the ipsilateral NTS in combination with the electrophysiologic determination of axonal projections from the NTS to the ipsilateral rostral VLM. The proposed experiments should contribute to the understanding of the role of VLM in baroreflex control of the circulation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies on the mechanisms of radiation resistance in Salmonella typhimurium are being conducted. First a series of radiation resistant mutants of S. typhimurium were developed. Secondly, the genes controlling radiation resistance are being identified. Thirdly, the phenotypic characteristics of the radio resistant genes are being established. Bibliographic references: Silverman, G.J. and A.J. Sinskey. The Destruction of Microorganisms by Ionizing Radiation. In \"Disinfection, Sterilization and Preservation\". (Lawrence, C.A. and Block, S.S., eds.) Lea & Febiger, Philadelphia. Gomez, R.F.&A.J. Sinskey. (1975) Effect of aeration on Minimal medium Recovery of Heated Salmonella typhimurium. J. Bacteriol. 122: 106-109.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The relationship of capillary basement membrane thickening to duration of glucose intolerance and the complications of diabetes mellitus are being investigated in the Pima Indians of Arizona. A comparison of the Siperstein and Williamson techniques of analysis is being carried out on these subjects to determine if one method is more sensitive for the detection of diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research will examine long-term effects of an innovative partnership-based intervention delivery model called PROSPER (PROmoting School/community-university Partnerships to Enhance Resilience). Primary effects examined will be reductions in emerging adult substance misuse, antisocial behaviors, and sexual risk behaviors for AIDS and other STIs, along with improvements in healthy adult functioning, among project participants who received evidence-based interventions during middle school. This proposed work extends a randomized control study of the PROSPER model, involving two sequential cohorts of youth and families from an understudied rural population. A total of 28 rural school districts in IA and PA, ranging in size from approximately 1,300 to 5,200 students, were selected and randomly assigned to either the PROSPER partnership condition or to a usual programming control condition. Local teams in the PROSPER partnership sites selected and implemented family-focused and school-based evidence-based interventions (EBIs) for middle school students. The evaluation of intervention outcomes began with approximately 12,000 students (90% of those eligible) in the two cohorts who participated in a 6th grade pretest assessment; follow-up assessments with the students in these cohorts were conducted each spring, through the 12th grade. Following 12th grade, 2,000 randomly selected students from that sample were recruited for continued follow-up assessments. The proposed study will extend those follow-ups to ages 22 and 24. Aims of the proposed study are to: (1) evaluate long-term substance misuse behaviors, anti-social behaviors and sexual risk behavior outcomes of the PROSPER partnership-supported middle-school EBIs; and (2) evaluate mediators and moderators of long-term EBI effects, informed by an extension of the outcome model specified in the original study. Positive results from the first two grant periods have established PROSPER effectiveness, with evidence of: effective functioning of community teams; strong participation in EBIs; long-term, high quality EBI implementation; sustainability of teams and EBIs; and economic analyses indicating PROSPER efficiency. Most importantly, results show strong EBI outcomes through middle and high school, with positive effects on: young adolescent competencies (e.g., peer refusal skills); parenting effectiveness and family functioning; reduced adolescent conduct problems; and reduced misuse of a wide range of substances through the end of high school. These results suggest a unique opportunity for demonstrating early preventive intervention impacts on healthy adult functioning; the prospect of establishing such impacts would constitute a compelling case for national diffusion of the PROSPER model. Literature reviews failed to reveal another study with a comparably large rural sample that allows assessment of intervention effects on general health, reduced problem behaviors, reduced sexual risk behaviors for AIDS/STIs, and other benefits for participants as they transition into adulthood.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The total chemical synthesis of calyciphylline A, an important alkaloid with anticancer activity, will be developed. The focus of this highly atom-economical strategy will be a ruthenium-catalyzed cascade of isomerization reactions to construct the [6.7.5] ring system of the Calyciphylline alkaloids in a single step. At this point i time, the development of a cascade of transition-metal-catalyzed isomerization reactions has never been done. Furthermore, 4 of the rings 6 rings of calyciphylline A are formed simply by isomerization reactions, highlighting an extremely efficient and highly atom-economical approach to this complex molecule with the incorporation of only two protecting groups. Additionally, this research program is designed to stimulate development of pre-existing methods in main group and transition-metal catalysis. These studies will explore novel zinc catalyzed ProPhenol alkynylations to saturated aliphatic carbamate-protected imines, as well as additions of highly functionalized alkynes to aliphatic saturated aldehydes. In addition, a general method for transition-metal-catalyzed [5+2] cycloadditions of 1,7-enynes will be explored. The methods developed within will undoubtedly attain widespread application amongst the general synthetic community. Successful completion of the proposed project would achieve the following: 1) Provide material for further biological studies in order to investigate the molecule s a potential therapeutic agent, 2) Outline a conceptual foundation for a synthetic, and highly atom-economical approach to other molecules of this type, 3) Develop new catalytic reactions involving both main group and transition-metal catalysis, 4) Culminate in the first total synthesis from this subclass of potent, structurally unique natural products.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. A complex problem for every eukaryotic organism is how to regulate the process of chromatin remodeling, a critical function to mediate DNA accessibility throughout the genome. Using the UCSF Mass Spec Facility, we have found that Rsc4, an essential sub unit of the RSC remodeling complex, is acetylated, by Gcn5, on K25 at its N-terminus. Purification of RSC under standard growth conditions suggests that most of Rsc4 is acetylated. In vitro comparisons of WT and mutant (K25R) RSC complexes reveal similar capabilities to remodel nucleosomes, bind DNA, and recognize acetylation. Since acetylation does not appear to alter the enzymatic properties of RSC, we are taking several in vivo approaches to understand the biological processes in which acetylation is important and to understand the functional consequences of the acetylation. Specifically, we will use mass spectrometry to identify conditions where acetylation is regulated and, combined with genetic mutations, identify the genes responsible for this regulation. These studies are the first to characterize the function of acetylation in the context of a chromatin remodeling complex.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We conducted a comprehensive analysis of transcription of two candidate histone methyltransferases in human and mouse tissues, showing tissue-specific expression of alternatively spliced and read-through transcripts. We conducted several yeast two-hybrid screens to identify protein-binding partners for the putative histone methyltransferases. These screens yielded several intriguing candidates that are being characterized further by biochemical and genetic methods. We have constructed a conditional knockout plasmid to generate a knockout mouse model lacking one of the putative histone methyltransferases located on a frequently deleted chromosomal region.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT While the role of stress in alcohol use disorder (AUD) is well established, none of the current FDA-approved medications in the U.S target the stress system. One potential pharmacological target for the stress component of AUD is norepinephrine. The goal of this application is to replicate findings previously conducted in a pilot trial and to understand, mechanistically, the role of stress in the development of AUD pharmacotherapies that target noradrenergic blockade. To achieve these goals, this study proposes a 12 week, between-subject, double-blind, randomized clinical trial (RCT) with doxazosin (16 mg, or maximum tolerated dose, MTD) compared to placebo in 184 treatment seeking individuals with AUD. We will: (1) examine alcohol drinking and clinical outcomes at baseline, throughout treatment, and at posttreatment; (2) conduct a stress-induced alcohol cue-reactivity in a bar- laboratory; and (3) test moderators of doxazosin response to inform a personalized treatment approach. There are three aims in this research plan. In Aim 1, we will test if doxazosin decreases alcohol consumption (primary outcome) and alcohol craving (secondary outcome) in naturalistic conditions throughout the study. Then in Aim 2, in the bar laboratory, we will measure acute craving after a single oral dose of 32.4 mg yohimbine (to initiate the neuroendocrine process associated with stress induction and to enhance exposure therapy), combined with an alcohol cue reactivity protocol (to selectively target alcohol cues). Finally, in the exploratory aims, to further shed light on potential personalized medicine approaches with noradrenergic pharmacotherapies for AUD, we will test the hypotheses that baseline family history density of alcoholism, blood pressure and rs1611115 polymorphism moderate doxazosin's effect on alcohol consumption in individuals with AUD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The studies investigate the neural processes underlying the initiation and guidance of voluntary limb movement and address two general questions. First, what parameters of movement are specified by central programs and how are these parameters expressed in the neuronal activities of the major motor systems. Second, what role does afferent information play in the initiation and control of movement and how can this information be used by the nervous system to alter existing programs to better achieve the behavioral objective. Chronic experiments are performed on cats trained to make precisely controlled tracking movements with their forelimb muscles using a compensatory display. In previous studies it was shown that the central programs controlling rapid voluntary adjustments in isometric force as well as in limb position could be described by a pulse-step model. Separate processes were shown to control the movement trajectory and the final position of the limb. The present studies will assess the role of the sensory cortex in mediating somatosensory information used for rapid reaction time movements, and the role of proprioceptive information in providing the central programs with initial position information. In a separate series of experiments we will evaluate the role of segmental mechanisms in determining the configuration of the initial pulsatile output controlling movement trajectory. Lesions of the rubrospinal tract will indicate the role of this structure in the control of limb trajectory. Single unit recordings of corticospinal and corticorubral neurons will indicate the role of these systems in controlling motor output parameter.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Data collection and survival analyses for patients undergoing various breast cancer treatment procedures were performed using CDMAS. Treat- ment evaluation and identification of prognostic indicators are end-points.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall aim of this research application is to investigate the relationship of early problem identification and linkage to behavioral health care with later development of substance abuse and psychological injury outcomes and attrition from the armed services. This observational study is based on a quasi-experimental design with longitudinal data files and statistical models of deployment-related factors and military health services on the odds of long-term poor substance abuse outcomes, poor psychological outcomes (e.g. post-traumatic stress disorder, depression), thoughts of harmful behavior, and attrition from the Army, associated with service in Operation Enduring Freedom (OEF) or Operation Iraqi Freedom (OIF). We will form a cohort of Army members (active-duty and Reserve component) with deployments ending in FY2008-2010. Outcomes will be tracked, starting at 12 months through 24 months, for all cohort members and 36 months for early cohort entrants. We will compare the outcomes of a group that receives early treatment initiation and engagement among the OEF/OIF cohort to a comparison group from the cohort (weighted for trauma exposure and self-report problems using propensity scores) that does not receive early treatment initiation and engagement. We also will compare the outcomes for the OEF/OIF returnee group that a military health system (MHS) professional identifies and refers early for follow-up assessment to a group from the cohort with self-report of potential problems (weighted using propensity scores) without early identification and referral. The role of this early identification and referral (within two months of return), and this early linkage to treatment (within 5 months of return) will be the primary focus of the analysis. We will merge MHS and Veterans Administration (VA) data for the cohort members to form complete longitudinal files previously unavailable for analyses, with observations of veterans'delayed problem presentation and service utilization in both health systems, and long-term adverse substance abuse and psychological outcomes and attrition from the Army. We also will be able to describe and better understand which combat veterans seek care from the VA among all that are eligible, as VA statistics suggest that 35% or fewer of OEF/OIF are using VA services to which they are entitled. The findings will provide operationally actionable data useful to quality improvement programs in the MHS and VA on urgent issues requiring clinical and policy attention. PUBLIC HEALTH RELEVANCE: This application will provide operationally actionable data useful to quality improvement programs in the military health system (MHS) and Veterans Administration (VA) on urgent issues requiring clinical and policy attention. We will investigate the substance abuse and psychological outcomes of receiving early identification and linkage to behavioral health care in a cohort of Army members returning from OEF/OIF, merging MHS and VA data to create a previously unavailable data files.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: The critical step in porphyrin biosynthesis is the condensation of two molecules of amino-levulinic acid (ALA) into the monopyrrole, porphobilinogen. This reaction is catalyzed by porphorbilinogen synthase (PBGS). All PBGS contain at least 8 metal ions; the mammalian enzyme is a Zn(4)Zn(4) species, with two Zn(2+) per dimeric unit in a homooctameric structure. In each homodimer, one Zn is bound to what is known as the \"A\" site, one Zn(2+) is bound at the \"B\" site. This proposal has four specific aims focussed on determination of the structure and catalytic mechanism of this essential enzyme. The first aim is to determine the chemical structure of the PBGS-catalyzed reaction intermediates. Three approaches will be used. First, site-directed mutagenesis will be used to make substitutions at residues thought to play a role as a covalent catalyst, a metal ligand, or Bronsted base/acid. Second, fluorinated substrate(s) will be used that cannot undergo turnover. Third, rapid quench approaches will be tried. The second aim is to determine the manner by which PBGS from various species use metal ions. This will involve the preparation of a number of PBGSs from various sources, with various combinations of Zn(2+), Mg(2+), Mn(2+), followed by analysis of metal ligation by EXAFS and ESR, and function. The inhibition of these several forms of PBGS by Pb(2+) is the focus of the third aim which is to determine whether the mechanism of lead inhibition is common to all of these enzyme forms. The last aim is to complete the x-ray structural determination of PBGS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The broad objectives of our program have been the characterization of polypeptides which may have physiological and immunological importance for the establishment and survival of malaria parasites in their hosts. The know protein (KP) of Plasmodium falciparum interacts with the host erythrocyte membrane and induces membrane protrusions called knobs. These protrusions are essential for the survival of this parasite in the host. The objectives of this project are: (1) To identify the structural domain(s) of the KP of one isolate of P. falciparum which are involved in the induction of knobs. (2) To determine whether these domains are conserved in other geographical isolates as well as in a different species of malaria which induces knobs and (3) To determine whether these domains are immunogenic in man. To achieve these objectives, KP and its peptides will be expressed in yeast and E. coli. The expressed polypeptides will be tested for reactivity with specific cytoskeletal proteins of the host erythrocyte membrane and with immune human sera. The conserved sequences of KP will be identified by analysis of the KP gene of different geographical isolates of P. falciparum and that of Plasmodium brasilianum.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The microenvironment of the periodontal pocket contain an abundance of sialylated glycoproteins. P. gingivalis a key stone pathogen has been shown to enable the emergence of dysbiotic oral bacterial communities, hence play an important role in periodontal pathogenesis. They also express sialoglycorptoeases which break down the terminal sialic acids in the sialoglycoproteins. This breakdown contributes to many important biological functions by either exposing or masking the interacting receptors. Studies have shown the importance of these enzymes in virulence and pathogenesis of P. gingivalis and also in other bacteria. Our preliminary in vitro data showed the involvement of P. gingivalis sialoglycoproteases (PG-Sgps) (PG0778 & PG1724) in virulence modulation and show binding specificity to common glycoprotein subtypes found in eukaryotic cells. However, there is still a significant gap in knowledge of how PG-Sgps orchestrate host- pathogen interaction and virulence leading to dysbiosis. Our central hypothesis is that PG-Sgps are involved in virulence regulation and contribute to host interaction through surface glycan modifications of specific virulence glycoproteins and mediate immune evasion through interactions with cell surface sialylated ligands in neutrophils. We will test our hypothesis by completion of the following specific aims: (1). Identify the functional role of P. gingivalis sialoglycoproteases and surface glycan changes influencing pathogenic potential of the organism. (2).Study the in vivo virulence of P. gingivalis sialoglycoproteases and identify PG-Sgp mediated neutrophil interactions involved in dysregulation of innate immunity. This work will advance our understanding of P. gingivalis virulence and pathogenicity by studying the functional role of PG-Sgps through identification of specific glycan modification in two important virulence determinants of P. gingivalis. And to study, how PG-Sgps interact with neutrophils influencing the pathogenic potential of P. gingivalis. This will eventually lead us to expand our scope for understanding specific sialic acid ligand interactions and their role in immune evasion and subversion of P. gingivalis. This study will lead us to design potential therapeutic targets against the sialic acid specific ligands. Hence, will pave the way for future pharmacological prevention of P. gingivalis infection. .", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Thyroid hormones (T3,T4) play a critical but only partially understood role in the development and maintenance of normal peripheral and central nervous system function. The clinical, psychiatric, and neurological effects of hypothyroidism and hyperthyroidism include: mental retardation, learning disability, deafness, optic neuropathy, delirium, dementia, psychosis, paranoid ideation, and depression. The clinical manifestations may be related to thyroid mediated changes in neurotransmitter regulation and brain gene expression. Deficiency of thyroid hormone during intrauterine life and subsequent to birth underlies the syndrome of cretinism or congenital hypothyroidism, which is a cause of mental retardation, impaired learning and motor disability. The syndrome occurs in 1 in 5000 births. For our understanding of the pathophysioloby and treatment of the human condition, an animal model of congenital hypothyoidism is essential. The purpose of the present proposed series of studies is to utilize an interdisciplinary approach to determine the utility of an inbred mouse animal model of congenital hypothyroidism, the hyt/hyt mouse. This model has particular relevance to human fetal thyroid disorders and may help us learn more about the treatment, etiology, and substrate of mental retardation as well as the initial and pervasive effects of thyroid hormones on the developing brain. The specific aims are: 1) To characterize the behavioral, neuroanatomical (particularly in cerebral cortex layers I-IV) and thyroid gland and hormone differences that exist in the hyt/hyt hypothyroid mouse fetus and neonate relative to normal mouse offspring; and 2) To choose and isolate, several mouse brain mRNAs which are regulated by thyroid hormone(s) during the intrauterine period following the onset of fetal mouse thyroid function.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Translational research in head and neck cancer is critically needed to enhance applications of the knowledge gained through the rapid expansion of our understanding of the molecular basis of cell growth and development. A comprehensive faculty career development program is proposed to meet the urgent need for increased number of clinicians and basic scientists devoted to head and neck cancer research. Although some of this need has been addressed through NIH T32 training grants for post doctoral fellows, transition to an independent, funded research career in head and neck cancer for full time faculty has been difficult. Direct funding of Assistant Professors with outstanding new translational research programs relevant to head and neck cancer is proposed to address this limitation. Focus on young faculty members will ensure attraction of new, energetic investigators, who when successful, will likely maintain a long-term research interest in head and neck oncology. A rigorous selection process is outlined that includes University-wide recruitment, strong emphasis on recruitment of women and minorities and comprehensive basic and clinical science mentoring. Close evaluation and monitoring is achieved through monthly meetings and annual internal and external review for continued funding. A major emphasis is placed on salary support for clinician scientists to ensure adequate time commitment to their SPORE research program.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The principal objective of this study is to increase our understanding of the primary causes of cataract formation in the human. Animal lenses will be used initially; when and where appropriate, clear and cataractous human lenses removed at the time of cataract extraction, enucleation, or autopsy, will be used. Attention will be focused on the factors regulating anaerobi glycolysis, the principal source of biological energy in the lens. Hexokinase, the pacemaker of this metabolic pathway and phosphofructokinase, an equally important point of metabolic control, will be studied in detail. Isolation and characterization of these enzymes will be attempted. Changes in overall glycolytic activity will be correlated with changes in hexokinase and phosphofructokinase activity. A well developed lens-culture system will be used to study an experimental analogue of the \"hypoglycemic\" cataract; it is in this type of cataract that a link between decreased hexokinase activity, disordered glycolysis and cataract formation may be substantiated. Well established biochemical and physiological techniques will be employed in all these studies. A continuing effort will be made to extend and compare results with the animal lens to the human lens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Staff in residential facilities and family caregivers are challenged by disruptive behaviors, disengagement from the environment, and other problems often displayed by people with dementia. Memory interventions show promise as a behavioral technique to help staff and family caregivers improve the quality of life for people with dementia. Market potential exists for an engaging group activity for persons with dementia that provides appropriate levels of cognitive and physical involvement, accesses long-term memories, focuses attention and concentration, provides a high probability of success for people at different levels of dementia and in different cohorts, and is water-proof and washable. Memory Magic is an activity designed to meet these criteria. Phase I feasibility testing revealed that Memory Magic elicited both higher quality and amount of engagement than standard programming, produced more positive and less negative affect, and was related to a reduction in inappropriate behaviors compared to levels seen in regular activity programming. Specific aims of Phase II research are to continue research and development efforts by improving the design, developing mass manufacturing capabilities, and testing refined versions across multiple sites in the Midwest, South, East and West resulting in a market-ready Memory Magic game for Phase II commercialization. PROPOSED COMMERCIAL APPLICATIONS: Long-term care, assisted living and adult day care facilities have expressed interest in purchasing Memory MagicTM to increase the quality of life and care for residents. An estimated 1.8 million persons with dementia reside in 25,000 nursing homes alone. Another 2.8 million live at home. Facilities such as Menorah Park Center for the Aging, Montefiore Home, Judson Park, and Rockynol Presbyterian Homes have expressed strong interest in Memory MagicTM.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A neuronal projection has been neuroanatomically identified to originate from the median raphe (MR) nucleus and to terminate in the hypothalamic suprachiasmatic (SCh) nuclei. These neurons have been further identified neurochemically to contain serotonin and to release this monoamine at their suprachiasmatic terminals. Stimulation and lesion studies at the level of the MR nucleus have suggested that this MR-SCh projection inhibited luteinizing hormone (LH) release and ovulation and may be involved in regulating other anterior and/or posterior pituitary hormones. The overall objective of the proposed research is to establish the nature of MR-SCh inputs to hypothalamic-pituitary function. We will manipulate the activity of this system by stimulating or lesioning the MR nucleus using stereotaxic procedures. Responses in various endocrine situations will be monitored by direct measurement of hypothalamic serotonin or (LH, follicle stimulating hormone and prolactin).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Virologic and immunologic factors in the pathogenesis of subacute and chronic measles encephalitis are being investigated. Strains of measles virus and cell cultures containing defective measles virus genome have been isolated in our laboratory from brains of patients with subacute sclerosing panencephalitis (SSPE). These viruses and cells are being characterized by a combined biochemical-immunologic-genetic approach. Attempts are being made to detect virus or viral products in brains of patients with subacute and chronic nervous system diseases other than SSPE and multiple sclerosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent studies in our laboratory have established for the first time that the major efferent outflow from the arm area of the primate motor cortex is derived not from its widely studied population of fast PT cells, but instead from: (1) A much larger population of very small PT cells, the majority of which send their axons to the contralateral cord; and (2) A separate population of small corticorubral neurons, that exert excitatory effects upon rubrospinal tract neurons. Additional studies of the motor functions of these small cell systems are therefore badly needed. We are therefore proposing three sets of experiments, that are designed to provide new information about the voluntary 'transient' and steady or 'tonic' motor functions of the monkey's slow pyramidal tract and corticorubrospinal pathways. In the first set, microelectrode recording techniques will be used to observe and compare the behavior of large samples of these small cells with that of fast PT neurons, in monkeys that have been trained to generate both transient and maintained, steady outputs in forelimb muscle tension. In the second set, additional information will be obtained about the functions of the rubrospinal pathway, by recording the activity of cells within the red nucleus in animals that have been trained to perform precisely the same set of forearm motor tasks. Finally, a careful, quantitative electromyographic study will also be carried out, in order to determine the specific motor deficits that are produced in the performance of these same tasks by surgical interruption of pyramidal and corticorubrospinal pathways. BIBLIOGRAPHIC REFERENCES: Humphrey, D.R. Neuronal size and systematic sampling errors in cortical single-unit recording. Fed. Proc., 34 (1975) 445. Humphrey, D.R. Statistical features of neuronal ouput populations in the arm area of primate motor cortex. Physiologist, 18 (1975) 255.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary objective of the multicenter study is to compare the safety, tolerability and efficacy of SB204269 with placebo as add on therapy in patients with refractory simple or complex partial seizures. Another objective is to gather population pharmacokinetic for SB204269 as adjunct therapy in patients. The study consists of 3 phases: Baseline, Double blind, and long term extension phases. Throughout the study physical and neurological exams will be performed. SB204269 can cause mild side effects reversible by lowering dosage.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Genome-wide association studies have been very successful in identifying hundreds of variants associated to complex diseases and phenotypes. In contrast, due to high levels of linkage disequilibrium at any given locus, only a handful of causal variants have been identified so far. In an attempt to bridge this gap, several fine- mapping studies involving dense genotyping or sequencing are currently being performed in multiple populations such as Europeans, Asians, African Americans or Latinos. Fine mapping studies over multiple populations can leverage different genetic variation across populations to increase the accuracy for localizing the causal variant in a joint analysis of multiple populations as compared to studies in which only one population is analyzed at a time. Surprisingly, despite the large potential of multi ethnic fine mapping studies, current multi population fine mapping studies employ standard statistical techniques within locus specific ad- hoc frameworks. In this application we will introduce novel metrics and automated frameworks for quantifying the performance of fine mapping methods as well as novel statistical methods that leverage multi ethnic genetic variation to increase the localization accuracy for fine mapping. PUBLIC HEALTH RELEVANCE: Resistance to a wide range of cancers, including breast cancer and various other diseases, is known to include a substantial genetically heritable component. Genome wide association studies have been very successful in identifying loci associated to various diseases including breast cancer. In contrast, the underlying genetic causal variants have yet to be identified for large number of phenotypes including most cancers. In this application, we will develop novel methods and metrics for multi-ethnic fine mapping studies and apply them to real fine mapping breast cancer data sets.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lens proteins undergo disulfide and non-disulfide crosslinking during aging. Such crosslinkings are associated with protein aggregation, insolubilization, light scattering and loss of lens accommodation. While disulfide crosslinking is well studied, the biochemical nature and the mechanism of formation of non-disulfide crosslinking are poorly understood. We propose to test a novel hypothesis that non-disulfide covalent crosslinking of proteins in the lens could arise from glycation-mediated crosslinking of the complexes that are formed between ?-crystallin and its chaperoned proteins, which leads to the formation of high molecular weight proteins and protein insolubilization during lens aging. Our preliminary studies strongly support this hypothesis. In the proposed project, we will systematically investigate this hypothesis via three specific aims. In Aim 1, we will perform experiments to establish the long-term stability of ?-crystallin-client protein complexes under the conditions of the lens by employing fluorescence resonance energy transfer (FRET)- based assays. We will then determine whether ?-crystallin-client protein complexes undergo more covalent crosslinking by glycation than their individual protein components by quantifying protein-crosslinking advanced glycation end products (AGEs). In Aim 2, we will extend our studies to intact human and mouse lenses to determine whether oxidative or thermal stress (to promote ?-crystallin-client protein complex formation) would promote glycation-mediated protein crosslinking in the lens. We will then determine the collective effects of stress and glycation on light transmittance and stiffness (resilience) in lenses. In Aim 3, we will first determine whether crosslinking by ?-crystallin-client protein glycation has a direct relationship with non-disulfide crosslinked high-molecular-weight proteins in aging lenses; we will then use a novel inhibitor that we developed during the previous funding period to inhibit protein crosslinking in human lenses. Finally, we will determine whether the inhibitor prevents losses in light transmittance and losses in resilience due to AGE-mediated protein crosslinking. Together, the three aims will test an innovative concept of protein crosslinking in the lens and test a novel chemical inhibitor against such crosslinking. The findings in this study could lead to innovative therapies against presbyopia and cataracts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "National surveys report that over 13% of nursing homes receive deficiency ratings related to the preservation of residents' dignity and care respectful of them as unique individuals. Nurse aides (NAs), who provide over 60% of personal care, learn few skills that help them address these deficiencies. NAs are often unprepared to offer empathic care that would foster meaningful and respectful caring relationships. Story Sharing: Enriching Nurse Aide-Resident Relations, a longitudinal mixed methodology study guided by Watson's theory of transpersonal caring, aims to improve caring behaviors toward residents by increasing NA levels of empathy, job attitude, mutuality, and self-efficacy. The effects of a three-session Story Sharing (StS) intervention will be compared to a Communication Skills (CS) condition. Interventions will be randomly assigned to six nursing homes. A convenience sample of 20 NAs in each of the six homes (N=120 NAs) will participate in the study. Based on the outcomes of a feasibility pilot study, a pre- and posttest for the outcome measures (empathy, job attitude, mutuality, and self-efficacy) will be administered with follow-up at 3- and 6- month intervals. NA turnover rates will be monitored at baseline and at 6-month intervals for 12 months post intervention. NAs will participate in small-group interviews. Interpretive analysis of open-ended interview data will reveal the nature of caring relationships among the sample of NAs and residents. A resident matched to each participating NA will complete the Caring Behaviors Assessment and Mutuality Scales and participate in an interview. The study will also include monthly support forums. Statistical analysis of the instrumentation responses and interpretive analysis of interview texts will be used. Story Sharing promises to offer a new caring intervention for NAs as they develop empathic and mutual care relationships that recognize the uniqueness and dignity of each resident.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Juvenile and adult organisms may be more susceptible to lead neurotoxicity than commonly recognized. Recent data indicate that juvenile rats chronically exposed to lead in drinking water at concentrations as low as 50 ppm (with corresponding blood lead values of 20-30 Mug/dl) manifest distinctive behavioral phenomena. These appear as markedly elevated rates of responding in operant (schedule-controlled) behavior testing. Four tissues require clarification: (1) How low an exposure level is able to produce such an effect? (2) Is the effect specific to rats and fixed-interval reinforcement schedules? (3) Is there a critical age period for its appearance? (4) What are the biological correlates, such as tissue lead concentrations and morphologic findings? These issues are addressed by a series of related experiments. Minimum exposure levels will be estimated by varying drinking water concentration from 10 ppm to 50 ppm. Critical periods will be assessed by comparing juvenile rats (21 days old at the start of exposure) and adult rats (90 days old). Behavioral specificity will be explored in an experiment comparing two different reinforcement schedules, and species specificity assessed by studies in mice and guinea pigs. Finally, the biological correlates will be examined by extensive assays of tissue levels, by accelerating excretion of lead with complexing agents, and by extensive histopathology. The ultimate aim is a comprehensive evaluation of the previous findings and their possible biological mechanisms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Marijuana is the most commonly used illegal drug of abuse in the United States. Chronic use of marijuana produces tolerance or dependence, and cannabis-related disorders. These effects are mainly mediated by ?9-tetrahydrocannabinol (?9-THC), the major psychoactive constituent in marijuana, which activates cannabinoid receptors in the CNS. However, the mechanisms by which ?9-THC alters synaptic function and develops tolerance are largely unknown. Accumulated information suggests that astroglial cells, an important element of tripartite synapses, actively participate in neuronal synaptic transmission and plasticity. The presence of bidirectional communication between astroglial cells and neurons suggests that alterations in astroglial behaviors by abused drugs may influence neuronal synaptic transmission and neuroadaptation and produce tolerance and addiction. Indeed, astroglial cells express cannabinoid receptors, which are targets of ?9-THC. It has been shown that ?9-THC exposure during development significantly decreases expression of glutamate transporters in cerebellar astroglial cells, indicating that glutamate uptake is reduced in astroglial cells exposed to ?9-THC. In our preliminary studies, we observed that chronic exposure to ?9-THC significantly attenuated hippocampal long-term potentiation (LTP) and reduced expression of the glutamate receptor. We also found that astroglial cells are able to undergo plasticity in response to presynaptic high-frequency stimulation. The plasticity of astroglial-neuronal synapses may be an important and integrated component of neuronal synaptic plasticity. Thus, we hypothesize that ?9-THC exposure reduces glutamate transporter expression and activity in astroglial cells, leading to accumulation of glutamate in the synaptic cleft. This results in functional desensitization and down-regulation of the glutamate receptors on both neurons and astroglial cells, which contributes to the ?9-THC exposure-altered synaptic plasticity. To test this hypothesis, we will accomplish the following two specific aims: 1) To test the hypothesis that chronic exposure to ?9-THC reduces hippocampal glutamate transporter expression and activity in astroglial cells;2) To test the hypothesis that chronic exposure to ?9-THC reduces hippocampal glutamate receptor expression and function on both neurons and astroglial cells, resulting in impairments in synaptic plasticity. PUBLIC HEALTH RELEVANCE: The proposed research will provide important information on the involvement of astroglial cells in marijuana-induced neuroadaptive changes in synaptic circuits, and is expected to further our understanding of the molecular and cellular mechanisms underlying marijuana abuse-induced tolerance and alterations in synaptic plasticity and cognitive function.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of the proposed study are to participate as a cooperative member of the Gynecologic Oncology Group, performing clinical cooperative trials in solid tumors. Investigations will include a variety of Phase I, Phase II, and Phase III studies in patients with gynecologic malignancies using chemotherapy, radiation therapy, surgical therapy and combinations of these modalities. Adjuvant therapeutic trials are specifically planned for: 1) adenocarcinoma of the endometrium, 2) adeno-carcinoma of the ovary, 3) uterine sarcoma, 4) squamous cell carcinoma of the vulva, 5) advanced, recurrent pelvic malignancy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In excitable cells, voltage-gated potassium channels play an important role in regulating the duration of the action potential. We hypothesize that the targeting of channels to particular membrane microdomains and their organization in macromolecular complexes allow cells to increase the efficiency of their response to extracellular signals. The work proposed in this application is directed towards understanding the molecular mechanisms underlying the trafficking and turnover of Kv1.5 and KV2.1 potassium channels. We propose to characterize the macromolecular complex (channelosome) that modulates the function of a delayed rectifier potassium channel, Kv1.5. We will use a combination of biochemical and molecular tools, patch-clamp analyses, and in vivo mouse models to pursue the following specific aims: (1) To determine the mechanisms that regulate trafficking of Kv1.5 polypeptide to caveolae, specifically to characterize the interactions between Kv1.5 and caveolin-3 (Cav-3) and SAP97. (2) To examine the role of posttranslational modifications on the trafficking, assembly, and stability on Kv1.5/Cav-3/SAP97 macromolecular complex. To map the site of interactions between SAP97 and Cav-3 and prove that this interaction contributes to the targeting of Kv1.5 to caveolae. (3) To characterize the role of N-terminal cysteines of Kv2.1 and their posttranslational modifications in regulating the assembly, trafficking, and gating of Kv2.1. The work outlined in this application should greatly increase our understanding of the regulation of expression of Kv1.5 and Kv2.1. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ProjectAbstract PXEInternationalResearchMeeting2016willsetaresearchagendafor pseudoxanthomaelasticum(PXE).PXEcauseslegalblindness,cardiovasculardiseaseand severalotherdebilitatingsymptoms.Thismeetingwillfocusonsomepromisingpotential interventionsandtreatments.Itwillalsoprovideanopportunitytospeedupthesciencetothe pointofmeaningfuloutcomesforpeoplelivingwiththedisease.Thesediscussionswillcreatea roadmapfordevelopingtherapy,whichistheoverallgoalofthisforum.Theorganizing committeeconsistsofadiversegroupofscientists,clinicians,andadvocateswhowillbringtheir expertiseinseveralareastotheworkshoptoroundoutacompetentandcreativeorganizing committee. Pseudoxanthomaelasticum(PXE)iscausedbymutationsintheABCC6gene.Thegeneis primarilyexpressedintheliver.RecentfindingssuggestthatABCC6regulatesthecellular releaseofnucleosidetriphosphates,predominantlyadenosinetriphosphate(ATP).Outsidethe cellATPisrapidlyconvertedintoadenosinemonophosphate(AMP)andinorganic pyrophosphate(PPi).MeasurementsofPPilevelsinasamplingof12individualswith confirmedABCC6mutationsrevealedsignificantlylowerconcentrationsofPPiintheirbloodas comparedtohealthycontrols.Thus,itismostlikelythatPPiisakeyinhibitorofmineralization foundinPXE. Severalnewideashaveresultedfromtheserecentdiscoveries,someleadingtopotential therapeuticareas.Thesepotentialtargetsmayprovetomitigatesomeofthemorbidity associatedwiththedisease.Drugrepurposing,useofalreadyapproveddrugsandnovel therapeuticsareallbeingconsidered.Arecentmeetingofaffectedindividualsresultedin researchprioritiessetthroughapatientcenteredoutcomesprocess.Thesewillalsobe consideredatthismeeting.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Histrelin is an LHRH agonist with which there is considerable human experience, although not when delivered by implant. It has recently been approved by the US FDA for inhibition of early puberty in children. Potent LHRH agonists when administered frequently, or even more effectively, continuously, cause desensitization of the pituitary and markedly reduced circulating levels of LH. This, in turn, suppresses testosterone synthesis. Based on experience with other analogs given by injection, depression to essentially castrate levels can be attained. This is a desired end in treatment of symptoms associated with prostate cancer. A subdermal implant would constitute a convenient way of continuously dosing with an LHRH agonist. Such an implant has been developed and shown in vitro and in animal studies to deliver an essentially constant dose of a period of 6 months or longer. The rate-controlling membrane is a hydrogel. Specific goals of the study are: (a) determine whether these histrelin-delivering implants can be used in humans; (b) ascertain the dose of histrelin necessary to suppress gonadotropin and testosterone secretions, and (c) determine the blood levels of histrelin during effective treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The RNA sequencing technique will be adapted to the analysis of polyribonucleotides at low molar concentrations by investigatng the incorporation of radioactive label into the nucleoside fragments that are successively removed by the periodate-Beta-elimination procedure. The study of the synthesis and properties of polynucleotides containing ribo-deoxyribo hybrid sequences will be undertaken. A new approach to the chemical synthesis of polynucleotides on solid supports will be investigated. BIBLIOGRAPHIC REFERENCES: Guanosine Tetraphosphate and Its Analogues. Chemical Synthesis of Guanosine 3',5'-Dipyrophosphate, Deoxyguanosine 3',5'-Dipyrophosphate, Guanosine 2',5'-Bis(methylenediphosphonate), and Guanosine 3',5'-Bis(methylenediphosphonate) G.N. Bennett, G.R. Gough, and P.T. Gilham (1976), Biochemistry 15:4623-4628. Separation of Oligonucleotides, Nucleotides, and Nucleosides on Columns of Polystyrene Anion-Exchangers with Solvent Systems Containing Ethanol. G.T. Asteriadis, M.A. Armbruster, and P.T. Gilham (1976), Anal. Biochem. 70:64-74.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed program, entitled BASE (Broadening Access to Science Education), will offer an annual two-week residential summer science enrichment program on the Fairfield University campus for 24 young women who are rising juniors and seniors from our neighboring city of Bridgeport, CT, a city comprised of many health disparity populations and some of the worst poverty in the nation. The program will include three key components. The first component is the Research Immersion Experience, a weeklong scientific research experience that will engage students in exciting faculty-led research projects assisted by current undergraduate science majors. Projects will be based on faculty research expertise, covering topics in biology, chemistry, biochemistry, neuroscience, psychology, mathematics, and engineering. Students will be exposed to science content, methodology, data collection and analysis, and quantitative reasoning, while receiving individual mentorship by female scientist role models at both the faculty and undergraduate level. Students will continue exploring their research topics in the second week through writing, reading, and discussion, with the guidance of their undergraduate counselors, culminating in formal student research presentations on the last day of the camp. The second component will be the Science and Health Careers Exploration, which will take place in the first two days of the second camp week. This will expose students to various careers in science, technology, and healthcare, and the academic paths required to get there. It will include presentations by faculty and professionals in STEM and healthcare careers, and mentorship on academic majors, courses, skill sets, and organizations that can support them in these pursuits. The final component of the program is the College Admissions Counseling, which will be a two-day set of activities to follow the career exploration component. In collaboration with our Office of Undergraduate Admissions, we will educate students about the process and requirements for admission to college, financial aid opportunities, as well as engaging students in mock interviews and essay writing. Also included will be a student/parent information session, to inform parents and integrate them into the process. Continued mentorship and access will be provided to students after the camp through a resource website with contacts and information on organizations, programs, and career paths introduced in the camp. Together, BASE camp will provide a significant experience that excites and informs students about the process and promise of science, and inspires them to pursue science-based careers to address the growing challenges facing health disparity populations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The hepatitis B virus (HBV) is a small, enveloped hepatotropic virus with a double stranded DNA genome that causes acute and chronic necroinflammatory liver disease (hepatitis) and hepatocellular carcinoma. Since HBV is not directly cytopathic for the infected hepatocyte, the cellular immune response to HBV encoded antigens is thought to be responsible for the liver disease caused by HBV, and to play an important role in viral clearance as well. Because HLA class I restricted, CD8- positive cytotoxic T lymphocytes (CTL) are known to interact directly with processed viral peptides at the cell surface, we have proposed that they play a direct role in viral clearance by destroying infected cells, and possibly by non-cytolytic anti-viral mechanisms as well. During the past several years we established the necessary models and technology to test this hypothesis. In a recent series of studies, we demonstrated that patients with acute viral hepatitis who eventually clear the virus develop a polyclonal, HLA class I restricted, CD-8 positive CTL response to multiple HBV encoded antigens, but that this response is not detectable in patients with chronic HBV infection. We now propose to define the molecular and cellular characteristics of the HBV specific CTL response during acute and chronic viral hepatitis, and to determine the role it plays in viral clearance and liver cell injury. A desired by- produce of this research may be the rational design of CTL-based therapeutic strategies to terminate persistent HBV infection, thereby reducing the human and economic toll of chronic liver disease and hepatocellular carcinoma in the 300 million chronic HBV carriers alive throughout the world today. Because of the power of the technology we have developed, the same strategy can be applied to study the CTL response to any human pathogen whose genome has been cloned and sequenced.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "My long term goal is to understand the physiological mechanisms associated with human color vision. The cynomolgus monkey, which has color vision identical to that of normal human trichromats, will be the experimental animal. Micropipette electrodes filled with the marker enzyme horseradish peroxidase (HRP) will be used to record intracellularly from lateral geniculate axons terminating in the striate cortex. After physiologically characterizing a fiber, it will be injected iontophoretically with HRP to reveal its cortical terminations. Distinctive axonal arborization patterns will be correlated with specific function properties to better understand the significance of both the geniculate and the cortical layers in color processing. Intracellular recordings will be made from monkey striate cortical cells with HRP-filled micropipettes. The cells' physiological properties will be determined and then they will be injected with HRP. The study of the laminar positions, dendritic arborizations and axonal projections of functionally characterized color-sensitive cortical cells provides a most promising way to understand the intracortical circuitry for color processing, the synaptic connections and interactions involved, the integrative mechanisms leading to their receptive field construction and the development of their spectral sensitivities. Tungsten electrodes will be used to record extracellularly from single cells for long periods of time, as well as from many cells in a single experiment. The functional organization of the color cells in the monkey's striate cortex will be examined by making long, oblique electrode tracks through single cortical layers. The double opponent color cells in the supragranular blobs will be compared with those in layer 4, those in 4A will be correlated with those in 4Cb, upper versus lower 4Cb will be examined, and the more complicated color cells in the supragranular and infragranular layers will be contrasted. This approach will provide much useful information about the integrative steps involved in cortical color processing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY The human complement system is a collection of ~30 membrane-bound or serum proteins which form a tightly regulated proteolytic cascade with antimicrobial and homeostatic effector functions. Complement dysregulation contributes significantly to a wide array of human diseases including arthritis, age-related macular degeneration, and transplant rejection. Although only two complement-directed drugs are currently FDA approved, therapeutic intervention of complement has gained traction with the advent of several new drugs in the preclinical and clinical pipeline. Nature has provided an unexpected source of novel complement inhibitory mechanisms in the form of microbial immune evasion proteins, and strides have recently been made in translating these evolutionarily-optimized inhibitory templates into novel therapeutics. However, relatively few examples of classical pathway specific immune evasion strategies have been described. Given the causal link of the classical pathway in neurodegenerative conditions such as Alzheimer?s disease and schizophrenia, this represents a critical deficit in our knowledge which stands to be addressed by the scientific community. The etiological agent of Lyme disease, Borrelia burgdorferi, is transmitted to humans by the bite of infected Ixodes ticks where it disseminates and colonizes remote tissues. Untreated B. burgdorferi infection causes disease in immune competent hosts, in part, by evading innate immune systems such as complement. Recently we reported that the lipoprotein B. burgdorferi BBK32 specifically prevents activation of the complement classical pathway by acting as a nanomolar inhibitor of the first component of complement, C1. We hypothesize that in addition to BBK32, B. burgdorferi expresses additional factors which also recognize and interfere with C1, and that one of these immunomodulators is the lipoprotein CspA. The overarching goal of this project is to shed light on the interaction of B. burgdorferi with the classical pathway of complement as outlined by three Specific Aims. In the first Aim, we will investigate the structural and molecular basis for C1 inhibition by BBK32 and follow-up on interesting preliminary data which suggests that other Lyme disease associated Borrelia BBK32?s have differential activity. In the second aim, we will use a multi-platform approach to quantitatively measure the novel interaction between CspA and the pattern recognition protein of the classical pathway, C1q. In Aim 3, we will use in vivo imaging in mouse models of Lyme borreliosis to explore the hypothesis that the presence of multiple overlapping classical pathway evasion proteins in B. burgdorferi contributes significantly to its ability to cause infection. By completing this research plan, we will greatly improve our understanding of how an important human pathogen causes disease while simultaneously laying the groundwork for the development of novel classical pathway complement-directed therapeutics for the treatment of devastating human autoimmune, inflammatory, and neurodegenerative diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Restoration of muscle function after injury requires the coordinate restoration of muscle fibers and microcirculation, as well as muscle and vascular innervation. Hypertension may have a significant effect on muscle regeneration, particularly on the regeneration of muscle blood vessels and the restoration of normal vascular control mechanisms. Applicants' preliminary studies, in which muscle injury was induced by tourniquet ischemia, Marcaine injection, or nerve-intact graft, indicate that experimental muscle injury in the SHR is associated with significantly slower and less complete regenerative recovery than occurs in the normotensive Wistar Kyoto rat (WKY). Moreover, the rate and extent of regenerative recovery in SHR is more severely limited when both muscle fibers and blood vessels are involved than when only the fibers are affected. In addition to being hypertensive, SHR is hyperinsulinemic, a state in which insulin receptors and IGF-I receptors are down-regulated. Down-regulation of receptors would impede the action of these growth factors during the repair process after injury. The investigators propose to determine: whether the impaired regenerative recovery of SHR muscle is linked to impaired regeneration of muscle blood vessels and lack of restoration of normal mechanisms controlling blood flow; whether the degree of vascular injury is related to the level and duration of hypertension; and whether the delayed recovery is related to decreased effectiveness of insulin and insulin-like growth factors, IGF-I and IGF-II. There are four AIMS to investigate various aspects of regeneration in extensor digitorum longus (EDL) and soleus (SOL) in hypertension, addressing: I) the effects of age and duration of hypertension on recovery; II) the effect of blood pressure, per se , on the recovery process, by studying muscle injury in SHR with pharmacologically normalized blood pressure (prazocin, propranolol and captopril), WKY with Golblatt hypertension, and cross-grafted EDL muscles between WKY and SHR; III) the functional state of the vessels in recovering muscle, by in vitro and in vivo assessments of arteriolar reactivity, capillary function, and blood flow; IV) the effect of the hyperinsulinemic state in SHR on effectiveness of IGF-I in promoting regenerative growth, by studying IGF-I receptor number and affinity, and concentration and binding characteristics of IGF-I plasma binding proteins. Hypertension and hyperinsulinemia are important risk factors in the progressive development of cardiovascular disease in humans, and these studies may elucidate mechanisms by which they interact, particularly with regard to regenerating vasculature in injured tissue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this application is to determine whether a peripherally acting mu-opioid receptor antagonist, methylnaltrexone (MTNX), attenuates certain subjective effects of a mu opioid analgesic, morphine, and to determine if MTNX changes the abuse liability-related profile of morphine to be more positive in healthy volunteers. It is currently used in patients on chronic opioid therapy for the relief of opioid-induced constipation, a peripherally-mediated side effect. Because it does not cross the blood-brain barrier, it does not antagonize the analgesic effect of opioids that is centrally mediated. Results from two small-N studies conducted in healthy volunteers indicate that aversive subjective effects that are induced by opioids, such as nausea, might be attenuated by pretreatment with MTNX. The study results were not definitive for several reasons, and we feel a more systematic laboratory analysis would clearly be of value. We intend to use MTNX as a probe to determine whether, and to what extent, certain subjective effects of opioids are mediated by peripheral opiate receptors. Healthy volunteers with no history of opioid abuse or dependence will participate in a placebo- controlled, double-blind, randomized, crossover trial in which they will be exposed to placebo, morphine alone (0.14 mg/kg, iv), two doses of MTNX (0.225, 0.45 mg/kg, sc) combined with morphine, and the higher dose of MTNX tested alone. The primary dependent measure in this study is subjective effects. Secondary measures include physiological and psychomotor effects. The study is significant from a basic psychopharmacology and pharmacology standpoint in that it will address the extent to which prototypic subjective effects of mu opioids are mediated at peripheral opiate receptors. It is significant from a clinical standpoint in that will address whether MTNX might have potential therapeutic implications beyond its ability to relieve constipation. It is significant and innovative from an abuse liability standpoint. Abuse liability of a drug is putatively influenced not only by positive effects of a drug but also negative effects, so if negative effects of morphine are attenuated by MTNX, the abuse liability-related profile of morphine may shift to where morphine is predominantly liked by the majority of healthy volunteers, a finding which would run counter to a number of studies in the extant literature with healthy volunteers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Surgical techniques for the treatment of stress incontinence (SUI) have significantly evolved over the last 100 years. The gold standard Burch urethropexy and pubovaginal sling procedures are now being performed less frequently, with the increased use of the newer minimally invasive mid-urethral sling procedures, the most common being the tension-free vaginal tape procedure (TVT). The TVT procedure is comparable in efficacy to the open Burch procedure with low morbidity and fewer complications. Because the sling is placed at the level of the mid-urethra under no tension, it was thought that the TVT would yield fewer postoperative lower urinary tract symptoms. However, a review of the literature has not borne this out, with postoperative storage symptoms reported in up to 42% of women. The primary purpose of the proposed randomized clinical trial is to test whether a perioperative behavioral/pelvic floor muscle training program can reduce the occurrence of these postoperative storage symptoms and voiding dysfunction in women undergoing a TVT procedure for SUI. Behavioral interventions are known to be effective for treating urge incontinence and voiding dysfunction unrelated to surgery, but have not been tested as a preventive adjunctive strategy. Approximately 400 subjects will be randomized to a perioperative behavioral program or usual care. The intervention will be implemented 2 weeks preoperatively, and reinforced before leaving the hospital and two weeks postoperatively. The primary outcome will be complaints of urgency, frequency, nocturia and urge incontinence using the overactive bladder questionnaire (OABq). Evaluations will be performed at 2 and 6 weeks, 3, 6, and 12 months postop, and will include the OABq, questionnaire for urinary diagnosis (QUID), urogenital distress inventory (UDI), pelvic organ prolapse/urinary incontinence sexual function questionnaire (PISQ), patient global impression of severity (PGI-S) and SF-36. Subjects will also complete a 7-day bladder diary to assess frequency of storage symptoms. Secondary aims are to determine whether this intervention reduces time to voiding and symptoms of voiding dysfunction, whether it impacts on patient satisfaction and quality of life, and to identify predictors of postoperative storage symptoms and voiding dysfunction symptoms. This type of information will allow physicians to more effectively counsel and treat their incontinent female patients to further enhance long-term quality of life.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Acquired Immunodeficiency Syndrome (AIDS) is no longer restricted to gay white men. Women, especially women of color, represent the most rapidly- growing segment of persons infected with HIV (human immunodeficiency virus). Despite the increasing frequency of HIV infection in women, virtually all the information available regarding the natural history of HIV infection and its effects on the lung is derived from studies of infected men. Recent reports suggest that important gender differences exist in the type and frequency of complications, particularly in regard to respiratory disease. In both genders in the general population, cigarette smoking represents the major preventable cause of respiratory morbidity and mortality, accelerating loss of lung function and predisposing to bacterial and other infections. In HIV-infected individuals, preliminary reports (again mainly in men) suggest that cigarette smoking may predispose to bacterial pneumonia and may worsen the pulmonary dysfunction which occurs after PCP. Based on these data, we hypothesize that cigarette smoking among HIV-infected women alters the spectrum, frequency, and severity of respiratory complications (both non- infectious and infectious) of HIV infection. Specifically, we surmise that in HIV-seropositive women, cigarette smoking increases airway hyperresponsiveness, accelerates the loss of lung function, and predisposes to the development of obstructive lung disease. Furthermore, we conjecture that cigarette smoking affects the type and extent of acute and chronic respiratory infections that these women experience. To test this hypothesis, we will assess respiratory health by lung function testing, by questionnaire and by surveillance for infectious complications over the course of a 3 year observation period in a cohort of HIV- seropositive women - representatively diverse in terms of race/ethnicity, HIV risk factors, socioeconomic status, and smoking exposure. For both non-infectious and infectious outcomes multiple regression analysis will be used to discern the relative importance of cigarette smoking as an independent risk for loss of lung function and infectious complications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application responds to AHRQ FOA PAR-09-071. We propose to implement improvements in a regional health-information exchange (HIE) designed to enable clinicians who manage transitions of care, to work more effectively and efficiently and to utilize the HIE more fully. The Keystone Health Information ExchangeTM (KeyHIETM), created with support from AHRQ Implementation Grant UC1HS016162, enables 8 hospitals and emergency departments and 38 ambulatory clinics in 31 counties of central and northeastern Pennsylvania to exchange test results and clinical summaries. KeyHIE contains administrative information for a population of 2.6 million over 1 million visits and 40,000 hospital discharges annually. Patients who authorize the sharing of their records (as 300,000 have already), will enable the clinicians caring for them to view their clinical information from across the region. KeyHIE uses technology and standards published by the Health Information Technology Standards Panel (HITSP), and will become part of the National Health Information Network (NHIN) when it is implemented. The proposed project, the next phase of KeyHIE's development, is designed to support safer, more effective transitions of care. To achieve this, we will add critical transitions-of-care clinicians as exchange users-case managers, home-health nurses, emergency responders, and long-term care facility clinicians. We will also expand the exchange's clinical content by adding additional laboratory and radiology reports; medication lists from pharmacy benefit management companies (PBMs) and retail pharmacies; and problem lists, allergy lists, electrocardiogram (ECG) traces, and consult reports. Finally, we will automate the distribution of patient information from participants' EHRs to KeyHIE and from KeyHIE to the EHRs of other participants who care for the patient. We will test the hypothesis that when KeyHIE contains more comprehensive clinical information, use of the exchange will increase, care will become more efficient and of higher quality, and patient outcomes will improve. Using de-identified data, we will monitor the use of the exchange over five years. We will conduct baseline and follow-up surveys with clinicians to understand their motivations for using the exchange, and with clinicians and patients to understand their perceptions of the exchange's usefulness. We will collect detailed data from a subset of high-use clinicians about whether and how their diagnostic and care plans are (or would be) affected by information available in the exchange and how their patients' outcomes change with exchange use. Finally, we will conduct a pre/post sub-study of care efficiency at selected provider facilities. PUBLIC HEALTH RELEVANCE: Project Narrative - This project will study whether and how the addition of specific types of clinician users and clinical information to a regional health-information exchange increases acceptance and use of the exchange, improves care-process quality and efficiency, and improves patient outcomes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] The goal of this PPG is to test specific hypotheses of how genes within four discrete regions of Chr 13 initiate the cascade of events determining blood presure salt-sensitivity and renal dysfunction in the Dahl S (SS) rat; and to identify specific genetic polymorphisms in two of these regions that determine blood pressure, salt-sensitivity, renal damage, and vascular angiogenesis. We have demonstrated that introgression of small regions of Chr 13 from the inbred Brown Norway (BN) strain of rat into the genomic background of the SS rat strain (consomic SS-13BN) substantially reduced salt-induced hypertension and restored angiogenic capacity to the SS rat. Four of 26 overlapping SS.BN congenic strains within Chr 13 containing BN substitutions (congenic strains 1,5,9, and 26) have been selected based on their protective actions on salt-induced hypertension and vascular angiogenesis effects. We propose three closely inter-related scientific projects that bring together a team of uniquely qualified geneticists and physiologists. Project by Cowley will determine the sequential physiological and gene expression changes in response to salt intake upon the kidney, adrenal gland, and vasculature. Since sex differences affected the degree of protection from salt-induced hypertension in several of the congenic strains, one of these strains (strain 9) has been selected to determine variations of genomic and physiological pathways that may explain these differences. Project by Roman focuses on a region of Chr 13 congenic strain 5 to identify the specific gene that \"protects\" from salt-induced hypertension in both male and female rats. Project by Greene focuses on Chr 13 congenic strain 9, to characterize the mutation that regulates the renin gene and thereby impacts upon angiotensin II formation and angiogenesis. Validation of differentially expressed and/or candidate genes will be tested using rat transgenic apporaches for Projects by Roman and Greene. The four Cores that will provide support for the PPG include: A) Administrative Core; B) Genomics Core; C) Transgenics Core; D) Research Services Core. [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are 2 purposes to the intended work. The first is to design, synthesize and test new radiopharmaceuticals which may have value in quantifying tumors and growth. The second is to use the newer agents, and more conventional radiopharmaceuticals, in the serial study of increasing and decreasing tissue masses. There are therapeutic implications to the work, since a radiolabeled compound which concentrated at a given site could be used to deliver a significant amount of localized irradiation. A principal lead has been the observation that arsenic can be substituted for phosphorus in a number of compounds of biological interest. While phosphorus does not have readily available radionuclides which emit gamma or positron emissions, arsenic does. Hence an active synthetic program is underway.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Statistical mechanics of cross-bridge action is considered in order to develop constitutive equations that express fiber tension as a function of degree of activation and time history of speed of contraction. The kinetic equation of A.F. Huxley is generalized to apply to the partially activated state. The rate parameters of attachment and detachment, and cross-bridge compliance are assumed to be step functions of extension, x, with a finite number of discontinuities. This assumption enables integration of the kinetic equation and its moments with respect to x analytically resulting in equations where x has been eliminated. When the constants in the rate parameters and the force function are chosen such that Hill's force-velocity relation and features of the transient kinetic and tension data can be fitted, the resulting cross-bridge mechanism is quite similar to the one proposed by Podolsky and co-workers. Because the derived constitutive equations simplify mathematical analysis, they enable the evaluation of the influence of various cross-bridge parameters on the mechanical behavior of muscle fibers. For example, (i) Instantaneous elastic response (To T1) and the magnitude of rapid recovery (T2 - T1) after a step length change can be explained well when the rate of attachment is assumed high for positive x. In that case T2 corresponds to the force generated by cross-bridges in the region of negative x. (ii) Kinetic transients occur as a result of the jumps that exist in the distribution of attached cross-bridges during the isometric state. Because of the hyperbolic nature of the kinetic equation, these jumps propagate in the -x direction causing rapid changes in the speed of contraction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Genetic and developmental studies of the correlations of the high uric acid levels of excretion in the urine and characteristics of the peculiar spotting pattern of Dalmatian dogs are being determined for backcross progeny of hybrid animals x Dalmatian. Hybrids are obtained from crosses of Dalmatians to Collies and to English Pointers. Crosses to Collies are repetitions of the experiments of Trimble and Keeler (1938 J. Heredity 29:280). Not so many variables are involved in the crosses to English Pointers. The presence or absence of pigment cells in the skin and hair follicles of pigmented and non-pigmented areas are being determined by electron microscopy of tissues from Dalmatians and 3 phenotypes of Shetland Sheepdogs (harlequin, merle and piebald).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project's goal is to use a well-characterized model system to define processes governing the formation and activity of CD4+CD25+Foxp3+ regulatory T (Treg) cells. The proposal will make use of existing lineages of transgenic mice expressing the influenza virus hemagglutinin (HA) under the control of a variety of promoters (HA Tg mice) and/or HA-specific T cell receptors with varying affinities and specificities for the HA (TCR Tg mice). By analyzing double transgenic mice co-expressing HA-specific TCRs and neo-self HA peptides (TCRxHA Tg mice), we have shown that highly specific interactions with HA-derived peptides can induce thymocytes to undergo selection to become Treg cells that suppress conventional CD4+ T (Tconv) cell responses in vitro, and can modify immune responses in vivo. We have shown that the formation of these Treg cells is highly sensitive to variations in the amount of the HA that is expressed in different HA Tg lineages, and that expression of HA selectively by antigen presenting cells in one of these lineages (designated HACII mice) induces the spontaneous development of inflammatory arthritis in TCRxHACII mice despite the presence of antigen-specific Treg cells. We will use this unique model system to examine how TCR specificity directs Treg cell formation and determines the capacity of Treg cells to modulate anti-self and anti-viral immune responses. In Aim 1 we will examine how TCR recognition of self-peptides shapes Treg repertoire formation. We will modulate the reactivity of the TCR for self-peptides and determine the effects on thymic Treg cell development in TCRxHA Tg mice, and will use adoptive transfer approaches to examine the development of Treg cells in the periphery of HA Tg mice under various conditions. In Aim 2 we will examine how Treg specificity for self-peptides shapes anti-viral immunity. We will analyze the ability of Treg cells from TCRxHA Tg mice to modulate antibody responses to influenza viruses with which they possess varying degrees of crossreactivity, and examine HA Tg mice that do not co-express TCR transgenes for their frequencies of virus- specific CD4+ Treg and Tconv cells. In Aim 3 we will determine how TCR specificity impacts the ability of Treg cells to prevent autoimmune arthritis. We will either deplete Treg cells, or add Treg cells with varying degrees of reactivity with the HA to pre-arthritic TCRxHACII mice, and determine the effects on cellular processes that accompany arthritis development. We will also analyze the expansion and differentiation of Treg cells with varying degrees of reactivity for the HA following transfer into HACII or TS1xHACII mice. These studies will provide fundamental insights into the mechanisms of immune repertoire formation and tolerance. They will enhance our understanding of the role immune regulation plays in anti-viral immunity and how its failure can contribute to autoimmunity, and will enhance the ability of Treg cells to b exploit in diagnostic therapeutic settings. PUBLIC HEALTH RELEVANCE: Regulatory T cells play an vital role in preventing the immune system from mounting harmful responses to the body's cells and tissues, such as can occur in autoimmune diseases. These cells can also modify the activity of the immune system in settings such as infection, transplantation and cancer. This proposal uses genetically-modified mice to analyze mechanisms and cellular processes than govern the development and activity of regulatory T cells, and to develop novel insights into regulatory T cell biology that will facilitate their application for diagnosis and therapy of human diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "According to research into the social causes of mental illness, migration tends to raise prevalence and incidence rates of mental illness in European immigrant populations. Lower socio-economic status in a community also effects the rates of mental illness of a population. A great number of effects that, in theory, have been proposed as causes for increases in incidence rates of mental illness in a population appear to fit the situation of Asian Americans. Yet, they are one of the smallest groups in the American mentally ill population. The present study proposes to use the San Diego Asian American population as a case study in order to develop a working hypothesis that may account for the extremely low usage of mental health facilities by Asian Americans. The expectations of the study are that Asian Americans under-utilize the available facilities and that only a very select group are seen at all. This group is expected to consist of military wives and Asian Americans under the age of 55. The San Diego Asian American population will be used as an indicator of these expectations but these results will not serve as an end in themselves. In order for this study to be of any positive value, it must be seen as the basis of a more comprehensive study dealing with Asian American mental health. The Asian American population that is represented by San Diego must be seen as a single case and not as being representative of all Asian American populations. The data obtained from this study can be used as a guideline for future reference and become part of a descriptive analysis of the Asian American population that does utilize mental health facilities. A twenty-nine point questionaire has been developed to serve as a data base for the study. The questionaire has three broad areas in which data can be organized for analysis: 1) basic demographic data, 2) psychiatric background of patient and family and 3) symptom review for the treatment period.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The study proposed to determine if physiological variables can be identified in Asian, African and Native Americans which might act as risk or protective factors for the development of alcoholism. Measures include neuronal response by resting EEG and event-related potentials, subjective response, and autonomic response by blood pressure and heart rate, genotyping and neuropsychological evaluation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the Opportunities in Genomics Research Program at The Genome Institute at Washington University is to increase the retention of underrepresented students in STEM and to increase their rate of matriculation into highly competitive Ph.D. programs. OGR consists of two programs: an eight-week summer program (Undergraduate Scholars) and a one-year post-baccalaureate program (Extensive Study). Prior data for the OGR programs indicate that over 45% of the graduates have matriculated into Ph.D. and other graduate and professional programs. OGR participants will engage in activities that will increase the percent of our students moving into Ph.D. programs to at least 65% overall. These activities include hypothesis-driven, mentored research, sessions focused on critical thinking, virtual academic year activities and graduate preparation courses and workshops. These activities will build on the work we began with OGR in 2007, leading to greater outcomes as the program moves forward. In addition to student outreach, we remain committed to the community at large and we will provide seminars focused on health disparities in genomics that will be available to the public and health advocacy groups. Our efforts with the OGR program and community outreach are helping to enhance diversity in the scientific workforce as well as enhancing the public's knowledge of genomics and its relevance to human health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cancer patients treated by radiation usually suffer organ damages in addition to other side effects (hair loss, nausea, and diarrhea). For example, patients with liver cancer, when treated by X-ray, suffer from liver damages and liver function reduction. Unfortunately, there is only one approved drug, Amifostine, to reduce head & neck damages caused by radiation, but not for liver damage. The direct effect of radiation therapy is to cause DNA damages (whether it be cancer cells or normal cells). In addition to the direct effect, there are also some indirect effects which can also cause liver damages. For example, the elevation of TNF-alpha levels in liver by radiation is an indirectly effect of radiation treatment; high levels of TNF-alpha appeared to cause liver damages. The applicant organization has a series of patented small-molecules TNF-alpha modulators. Out of these compounds, UTL-5b, -5d and -5g have been selected due to (a) their ability to modulate TNF-alpha and (b) they are associated with very high LD50 values and satisfactory results on a 14-day repeat dose toxicity study in mice. Preliminary study showed that when an animal was pre-treated with a small molecule TNF-alpha modulator (UTL-5d) before an X-ray treatment, liver damages appeared to be reduced. This preliminary study indicates that (1) TNF-alpha plays an important role in damaging the liver, and (2) a UTL-compound lowered TNF-alpha levels and may be a potential radiation protector for liver. The goal of this SBIR Phase I study is to validate the feasibility of using our TNF-alpha modulators to reduce liver damages on mice treated by radiation. Upon the approval of this Phase I study, the following specific aims of this Phase I study will be conducted: (1) Develop at least two injectable formulations employing these compounds; (2) Develop and validate an HPLC method for each of these formulations; (3) Conduct animal studies to confirm the radiation protection effects of UTL compounds; (4) Analyze TNF-alpha levels and attempt to correlate the reduction of TNF-alpha levels reduction with liver damages; (5) Conduct a radiation dose escalation study. The ultimate goal is to develop a radioprotective drug to be used in conjunction with radio therapy. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study is testing the hypothesis that a change in aldosterone and renal blood flow response to AII, induced by converting enzyme inhibitors, is specific for a single subset of hypertensive patients and also for one form of therapy. Thus, non-specific vasodilators will not produce an increase aldosterone or renal blood flow response to AII, similar to what has been previously documented for converting enzyme inhibitors. Used CDMAS only this year.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Moyamoya disease is an obliterative vasculopathy of the large arteries at the base of the brain. In the United States, it most commonly affects women in their third and fourth decades of life, causing ischemic stroke. The natural history of this disorder is not defined. It is very likely that hemodynamic factors play an important role in the risk of future stroke. Some patients are treated surgically with a variety of procedures intended to improve collateral blood flow to the brain. The effect of these procedures on cerebral blood flow and on subsequent stroke risk reduction is unknown. This blinded, prospective study will investigate the role of cerebral hemodynamic factors in the pathogenesis of stroke in patients with moyamoya disease. We will test the hypothesis that increased oxygen extraction in the cerebral hemisphere distal to the occlusion is a predictor of the subsequent risk of ipsilateral ischemic stroke in medically treated patients (Specific Aim 1). The secondary objectives of this project are (1) to determine other predictive factors for stroke in this population; (2) to obtain preliminary data on the effects of different medical treatment regimens in this population; (3) to determine the temporal changes in hemodynamic impairment in medically treated patients; (4) to determine the effects of surgery on hemodynamic impairment in the subset of patients that undergo surgical revascularization; and (5) to obtain estimates of surgical complication rates for patients with and without hemodynamic impairment. These objectives will be accomplished by the use of positron emission tomography (PET) to measure regional cerebral oxygen extraction fraction (OEF) in patients with moyamoya disease. Information on clinical, epidemiologic and angiographic risk factors will be obtained on enrollment. Patients are subsequently followed every six months by telephone to determine the subsequent occurrence of stroke and the interim treatment for stroke prevention. Results of the PET study are kept confidential from the patient, treating physician and investigator monitoring the occurrence of stroke. We will determine if a regional increase in OEF is associated with an increased risk of ipsilateral ischemic stroke. We will also do a multivariate analysis to determine the independent predictors of stroke in this sample. We will perform a post-hoc longitudinal analysis of the effect of different medical treatment regimens. Patients will return annually for follow up PET studies. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "NIAAA has designated underage drinking as a priority research area. Of note, the highest prevalence of problem alcohol use is among young adults ages 18-25. Heavy drinking that occurs during this period can have important immediate and lifelong adverse consequences. Behavioral interventions, notably BASICS (Brief Alcohol Screening and Intervention for College Students), have been developed to help young adults reduce their drinking. Although these interventions are effective, including with college students mandated to treatment and others with minimal motivation to stop drinking, the effect sizes are modest, suggesting that new approaches are needed to enhance these interventions. A promising strategy yet to be tested in young adults is the use of the opiate antagonist naltrexone. In other research, naltrexone has been shown to reduce the amount of alcohol consumed, even in the absence of strong internal motivation to change, and to [unreadable] reduce the frequency of any and heavy drinking in problem drinkers seeking treatment. [unreadable] [unreadable] Thus we propose to conduct an 8 week double-blind placebo-controlled trial to test the combined efficacy of BASICS + naltrexone in 132 young adults aged 18-25 who drink heavily. A novel strategy will be used for administering low-dose naltrexone, in which daily dosing will be combined with targeted dosing in anticipation of high-risk situations. The main hypotheses are that daily + targeted naltrexone will result in greater reductions in frequency of heavy and any drinking compared with daily + targeted placebo. In order to enhance the sensitivity with which we are able to assess naltrexone's effects on drinking, daily ratings will be obtained during treatment. These will permit us to examine alternative measures of alcohol involvement (e.g., reports of subjective intoxication, estimated blood alcohol levels) in addition to the traditional measures based on number of drinks consumed. These data will also be used to examine potential mediators (e.g., craving, subjective effects of alcohol) of treatment response in order to better understand the effects of naltrexone. The durability of treatment effects will be examined at 3, 6 and 12 months after randomization. Demonstration of the efficacy of naltrexone in this population will provide the essential information needed for its adoption by college counseling centers and other health care settings committed to reducing the risk of heavy drinking in young adults. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is proposed to study the physiological and biochemical mechanism of membrane transport using a comparative and evolutionary approach. We postulate that the earliest cell utilized a \"proton economy\" (primary proton pumps generated a proton gradient which was used for proton-substrate cotransport and other processes). Later in evolution a \"sodium economy\" developed and found its most sophisticated expression in animal cells. It is proposed to compare cotransport systems utilizing protons (lactose carrier of E. coli) with cotransport utilizing sodium ion (proline carrier of E. coli). In addition we plan to study a representative of the intermediate class of carriers that utilize either protons or sodium for cotransport (melibiose carrier of E. coli). Mutants will be isolated which show changes in sugar specificity or catiom recognition. We intend to clone the mutant gene for each of the transport proteins and determine the DNA sequence by the Sanger dideoxynucleotide method. A comparison between the altered amino acid sequence of the carrier protein and the altered physiology of transport will help to correlate three dimensional structure with function. A second project involves a study of the comparative physiology of cell volume regulation. As with animal cells microorganisms without cell walls (such as Mycoplasma gallisepticum) must continuously pump out the NaCl and water that diffuse into the cell as a result of colloid osmotic and Donnan forces. It is proposed to investigate the ionic mechanism for NaCl extrusion from M. gallisepticum and compare it with that used by animal cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project has two primary aims. The first aim is to understand how proteins that are destined to travel through the secretory pathway are targeted to transport sites in the endoplasmic reticulum (ER) or the bacterial inner membrane (IM). For the last decade we have been investigating the role of a ribonucleoprotein called the signal recognition particle (SRP) and its membrane-bound receptor in this process. Although SRP was initially believed to exist only in eukaryotic cells, the sequencing of a large number of microbial genomes has demonstrated that the particle is found in most (if not all) organisms. Previous studies have shown that in mammalian cells SRP recognizes the \"signal sequences\" found on virtually all secreted and membrane proteins as they emerge during translation and then catalyzes their translocation across the ER membrane upon interaction with the SRP receptor. A few years ago we demonstrated that bacterial SRP has a somewhat more restricted function in that it only targets integral membrane proteins to the IM. Consistent with the work of other laboratories, we found that most secreted proteins, by contrast, are targeted to the IM by molecular chaperones. In recent studies we have continued to analyze protein targeting mechanisms and protein targeting signals in bacteria. In the past year we have focused on the targeting of a class of very large (~100-400 kD) secreted toxins called autotransporters. These proteins are produced by a wide range of pathogenic Gram negative bacteria and often contain exceptionally long signal peptides that are distinguished by a unique N-terminal sequence motif. Although work by others has suggested that the unusual signal peptides are recognized by SRP, we have found that a model autotransporter produced by E. coli O157:H7 called EspP is targeted to the IM by an SRP-independent pathway. Surprisingly, we found that the unusual signal peptide was not required for translocation of EspP across the IM, but instead was essential for late stages of protein biogenesis that occur after the protein is translocated across the IM. These results not only provide insight into the biogenesis of a class of proteins associated with bacterial pathogenesis, but also provide evidence for a novel signal peptide function. The second aim of the project is to elucidate the function of factors that facilitate the transport of proteins across or insertion into the ER or bacterial IM. In the last year we have continued to study the structure and membrane distribution of YidC, a highly conserved bacterial protein that has been shown to play an important but still poorly defined role in membrane protein biogenesis. Using a variety of different biochemical methods we have obtained evidence that YidC exists in a homooligomeric form similar to that observed for its mitochondrial homolog (a protein called Oxa1p). The results suggest that YidC is a multisubunit complex that could function as a protein conducting channel. In addition, recent results indicate that YidC is freely mobile within the IM and call into question results published by another group suggesting that YidC has a strictly polar localization.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pathogenic bacteria are often capable of killing cells within the hosts they infect. The molecular mechanisms underlying the cytotoxic actions of bacteria on human cells are only partly understood, but generally they involve secretion of killer proteins from bacteria, and often these proteins are injected into host cells by Type III or IV secretion mechanisms. Examples of apoptosis-inducing virulence factors have been found in Yersinia (YopJ), Shigella (ipa), Salmonella (sip), and Anthrax (Lethal Factor) species. The mechanisms by which these bacterial gene products induce apoptosis of human cells are diverse. Knowledge of apoptosis-inducing virulence factors and information about their mechanisms of action can provide the basis for strategies designed to treat or prevent bacterial disease. Using bioinformatics approaches, we have identified several candidate apoptosis-inducing virulence factors encoded in the genomes of pathogenic bacteria. We propose to characterize the apoptosis-inducing activity of these bacterial proteins and to determine their mechanisms of action in human cells. It is hypothesized that some of these bacterial proteins will prove to be virulence factors, which are important for causation of disease. A combination of cellular, molecular, and biochemical methods will be applied to characterize the mechanisms of these candidate virulence factors, with the validated factors then progressing into more in-depth structural analysis through collaborations with other scientists participating in the project. Where warranted, bacterial apoptosis-inducing proteins will become targets for chemical compound library screening, and the resulting chemical entities will be used for further studies of the pathobiology of the target proteins. Altogether, the information derived from these studies may reveal strategies for preventing or ameliorating cell death induced by pathogenic bacteria, thus complementing traditional antibiotics in the prevention and treatment of bacterial diseases and agents of biological warfare.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A large but inconclusive body of evidence implicates the role of biogenic amines in the biology of depression and in the mechanism of action of antidepressant drugs. Neurochemical and pharmacological data derived from acute drug effects constitute the bulk of evidence. However, in clinical practice, the efficacy of these agents is slow in developing and requires prolonged periods of treatment. This consideration focuses on the relevance of the study of chronic effects of antidepressants. We propose to study the effects of long-term administration of MAO inhibitors, tricyclic antidepressants, chlorpromazine, and some antihistamines that have been shown to have antidepressant potential on the basis of preclinical pharmacology. The effects of these drugs on the synthesis, uptake, release and catabolism, of the biogenic amines (norepinephrine, dopamine, serotonin, and acetylcholine) and the enzymes involved in their syntheses and catabolism as well as their effects on the availability of tryptophan for brain serotonin synthesis, will be investigated. The evaluation of neurochemical and pharmacological effects of long-term administration of these psychoactive agents will aid (1) in finding a unifying concept for the antidepressant action of these agents and, (2) in postulating on the underlying defect in pathological depression and, (3) in improving the basis of prediction of clinical activity of antidepressant drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The vast majority of current therapeutic agents function by binding to disease-associated macromolecules and modulating their activity. Recent developments, however, have made increasingly realistic the possibility of developing next-generation therapeutics that do not simply bind targets implicated in disease, but instead alter the covalent structure of genes and gene products in ways that can more effectively treat-or even cure-many diseases. While the possibility of precisely manipulating genes and proteins in mammalian cells and, eventually, in humans, has enormous potential, several major challenges must be overcome to fully realize this vision. Perhaps the most significant of these challenges is the efficient creation of the macromolecules that are needed to alter genomes or proteomes with a high degree of selectivity and potency. To realize a vision in which arbitrary genes or proteins can be manipulated in mammalian cells to treat disease thus requires new approaches to rapidly generating macromolecules with precise, tailor-made properties. During the last granting period, we developed a system that enables proteins to evolve continuously in the laboratory, requiring virtually no researcher intervention. The resulting system, phage-assisted continuous evolution (PACE), allows proteins to undergo directed evolution at a rate ~100-fold faster than conventional methods. In the first applications of PACE, we rapidly evolved RNA polymerases with dramatically different DNA promoter specificities. We also identified the vulnerabilities of drug candidates to the evolution of drug resistance by using PACE to evolve proteases that are resistant to HCV protease inhibitors currently used in human clinical trials. In addition, we developed important PACE capabilities beyond basic positive selection, including small- molecule modulation of selection stringency and negative selection against undesired activities. These initial studies established PACE as a robust and general platform to evolve proteins with tailor-made properties at an unprecedented speed. In the next granting period, we propose to apply these developments to continuously evolve four classes of proteins or RNAs, each with the ability to manipulate the covalent structure of genes or gene products, and each with potential relevance to the development of next-generation human therapeutics: recombinase enzymes that insert DNA of interest into safe-harbor loci in the human genome, proteases that specifically cleave disease- associated proteins, orthogonal Cas9 (CRISPR) nucleases with altered PAM specificities and enhanced activities, and smart Cas9 guide RNAs that mediate genome engineering only in those cells that are in specific disease-associated cell states. Success would establish the novel therapeutic potential of these proteins and RNAs to address a wide range of human diseases, including many human genetic disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will be the first large-scale investigation to evaluate changes in visual enhancement across the adult life span. Visual enhancement is the advantage afforded by seeing as well as hearing the talker, as compared to listening alone, and is an issue of critical significance to the audiological rehabilitation of older persons. The unique contributions of the present proposal are threefold. First, we will assess how vision-only speech recognition and auditory-visual integration skills contribute to changes in visual enhancement, and determine if changes relate to more global declines in older persons' abilities to integrate two sensory signals, be they speech or non-speech. Performance will be related to perceived communication handicap. Second, two quantitative models of auditory- visual integration, one developed by Blamey, Alcantara, Whitford, & Clark (1989) and the other by Braida (1991), will allow us to determine whether age affects how individuals integrate specific features of the speech signal, and also, how it affects their ability to achieve optimum integration of the auditory and visual signals. Third, we will determine whether or not the presence of hearing loss may affect integration performance for both older and younger adults, and whether the type of speech material interacts with one's ability to integrate. The results may affect several stages of the geriatric audiological rehabilitation plan, including how we assess speech recognition skills and the kinds of follow-up, support, and counseling we provide to clients and their families after an individual receives appropriate amplification. In addition, the results may have important theoretical implications for models of aging and auditory-visual speech recognition, and may elucidate variables that contribute to the communication handicap experienced by persons who have hearing loss. Over a five-year period 200 subjects, selected on the basis of age and hearing status, will be enrolled in a set of cross-sectional experiments. The experimental methodologies have been developed and tested through a series of extensive pilot studies. The experiments are designed to assess subjects' levels of visual enhancement, vision-only speech recognition, auditory-visual integration, auditory-auditory speech integration, and auditory-auditory non- speech integration. It is hypothesized that visual enhancement will be shown to decline with age, and that this decline will be associated with a global decline in the ability to integrate sensory information, not to a change in vision-only speech recognition abilities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To combine the disciplines of Surgical Oncology, Medical Oncology, Radiation Oncology, Pathology, and Immunology in a truly synergistic attack on the clinical cancer problem. To participate in Southwest Oncology Group phase I, II, and III, and adjuvant studies. To conduct preclinical and clinical pilot studies for possible use by the Southwest Oncology Group. To provide core laboratory support and suitable administrative facilities for the clinical evaluation of various techniques to predict response to endocrine ablation in breast and other cancers. To utilize Southwest Oncology Group studies to link basic science to modern clinical care within the area as an educational mechanism. To undertake a retrospective study of the critical management variables of metastatic breast cancer. To study acute pulmonary distress associated with antiestrogen therapy in metastatic breast cancer patients. The undersigned agrees to accept responsibility for the scientific and technical conduct of the project and for provision of required progress reports if a grant is awarded as the result of this application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fragile X syndrome (FXS) is the most common form of inherited cognitive impairment, and children with this disorder often have additional behavioral and neurological problems, including increased anxiety, autistic tendencies, hyperactivity and epilepsy. Since the GABA system plays an important role in regulation of the neural systems involved in these behavioral phenotypes, GABA system deficits could be present in FXS. While alterations in GABAA receptors (GABAARs) have been identified, knowledge of the regional and cellular localization of changes in GABAAR subunits remains very limited. Thus, the broad goal of this project is to test the hypothesis that expression and localization of specific subunits of the GABAAR are altered during postnatal development in a mouse model of FXS and that these changes are associated with functional deficits, including increased neuronal excitability and altered anxiety- related behavior. Specific Aim 1 will identify changes in the 12 subunit of the GABAAR in Fmr1 knockout mice that lack the Fragile X mental retardation protein (FMRP), using immunohistochemical methods. Importantly, the patterns of expression of the 12 subunit will be followed throughout early postnatal development in wild-type and Fmr1 knockout mice in order to identify early changes that could be associated with loss of FMRP function and precede potential compensatory changes that may become apparent later in life. Specific Aim 2 will identify functional deficits that could be associated with altered expression of the 12 subunit in Fmr1 knockout mice. In vitro electrophysiological studies will be used to assess GABAergic function at the axon initial segment of dentate granule cells where the 12 subunit is normally prominent and where a loss of 12 subunit-containing GABAARs could influence axon potential generation and increase granule cell excitability. Behavioral tests will be used to evaluate the anxiolytic and sedative effects of a classical benzodiazepine that could be altered in response to GABAAR subunit changes in Fmr1-deficient mice. Specific Aim 3 will identify changes in the expression and localization of the 4 subunit of the GABAAR in the Fmr1 knockout mouse and determine if such changes are reflected in alterations in tonic inhibition and its modulation by neurosteroids in the dentate gyrus. Specific Aim 4 will determine if GABAAR-related pharmacological treatment will ameliorate some of the behavioral changes and GABAAR subunit deficits in Fmr1 knockout mice. These studies will provide unique information about GABAAR subunit alterations during postnatal development in a mouse model of FXS and could provide a framework for new GABAAR subunit-targeted treatments for this disorder.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a one-year, investigator-initiated, phase II study in 16 patients with Huntington's disease. The protocol is designed to test the safety, tolerance, bioavailability, and efficacy of OPC-14117, a new oxygen free radical scavenger with excellent central nervous system penetration. It will also evaluate the potential benefits of this drug on the movement disorders and mental features of the disease. This study is part of a multi-center trial initiated by the Huntington Study Group, a consortium of clinical investigators and basic scientists with the aim of developing therapeutic agents for HD. The study has been completed and the results have been submitted for publication.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to generate evidence to inform non-pharmaceutical interventions (NPIs) for pandemic influenza. We are proposing three parts to our research: 1. Preventing pandemic entry into island countries - This component would review historic evidence about successes and failures with control of the 1918, 1957 and 1968 influenza pandemics entry into island countries and lessons that can be learned from this. It would assess the feasibility of using known NPI to exclude pandemic influenza from island countries of varying size, from continental islands (Australia) to medium sized islands (NZ, Fiji), to small islands (e.g. American Samoa, Marshall Islands). This information would be incorporated into a 'threshold analysis' tool for use by health and border authorities. 2. Seasonal influenza prevalence in airline passengers - This component would measure the prevalence of influenza like illness (ILI) and infection in arriving passengers according to origins and travel times and estimate the proportion who are incubating disease. It would also assess the positive predictive value of a short screening questionnaire. 3. Seasonal influenza hospitalizations and housing conditions - This component would measure rates of hospitalization with influenza in an established cohort of 225,000 people living in public housing in New Zealand. It would specifically measure the risk of influenza in relation to age, ethnicity, socio-economic status, household crowding levels and tobacco smoke exposure. Results of these related projects would inform pandemic influenza prevention in a number of ways: [unreadable] -Results would support decision-making about the feasibility of preventing pandemic influenza entry into island countries and the combination of NPIs that are likely to be most effective at achieving this. [unreadable] - Data on the prevalence of influenza in passengers from various origins and flying times would provide an important input for decisions about the efficacy of traveler exclusion, quarantine and screening. Results could also be contrasted with the patterns seen in Northern Hemisphere sites. [unreadable] - Findings from the housing cohort would be used to guide advice about reducing influenza transmission in households, particularly in terms of crowding levels and exposure to active and passive smoking. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cooperative binding systems are being studied taking into account site or subunit interactions, ligand interactions, aggregation and redistribution in proteins, and model systems. Methods are being developed to evaluate reasonable values for the parameters describing these systems, particularly those involving the action of fibrinogen and the processes of protein-DNA interactions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Cardiovascular Health Study (CHS) was initiated by the NHLBI in 1987 to determine the risk factors for development and progression of cardiovascular disease (CVD) in older adults. The study recruited and examined 5,888 adults aged 65 and older at entry in four communities. Exam components emphasized measures of subclinical disease such as carotid ultrasound, cranial magnetic resonance imaging, echocardiography, ankle-brachial index, spirometry, and retinal photography, as well as more traditional questionnaire, physiologic, and biochemical measures of CVD burden and risk. Nine annual follow-up exams were conducted through 1999, followed by semi-annual phone calls to ascertain new CVD events (including incident myocardial infarction, angina, congestive heart failure, stroke, transient ischemic attack, peripheral arterial disease, and death). The data and specimens collected in CHS represent a major national resource for the study of development and progression of cardiovascular and several other chronic diseases in older adults. The current Transition Phase contract provides partial support to the study's Coordinating Center for maintenance of ongoing operations to assist researchers evaluating CHS'extensive longitudinal data and samples, promote participation by new investigators, and expand access to the data and sample repositories for the research community. Additional funding for infrastructure activities and for scientific research is acquired through investigator-initiated grants and other outside sources. This contract provides a mechanism to preserve key study resources and potential access to the cohort while sharing responsibility for study design, management, and support with interested investigators independent of contract support. It assists the study in maintaining core functions while assuring open, efficient, and active use of the CHS data and specimens for the scientific community.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The efficacy of antiretroviral therapy (ART) in HIV-1 infected individuals is determined by restoration of peripheral blood CD4+ T cell numbers and viral suppression. However, peripheral blood represents only 2% of the total lymphocytes in the body. In contrast, gut associated lymphoid tissue (GALT) harbors 80% of the lymphocytes in the body. Our previous studies showed that severe CD4+ T cell depletion occurs in GALT during primary HIV infection and that CD4+ T cell restoration in GALT is modest and slow compared to peripheral blood during ART. These changes in GALT are not adequately reflected in peripheral blood analysis. The kinetics and mechanisms of CD4+ T cell restoration and function in GALT following ART have not been fully determined. Simian immunodeficiency virus (SIV) infected rhesus macaques provide an excellent animal model to study the gut mucosal immune system in comparison to peripheral blood compartment. The overall objective of this research proposal is to examine the suppression of viral replication and kinetics and mechanisms of restoration of gut mucosal immune system and function in comparison to mucosal and peripheral lymph nodes and peripheral blood in rhesus macaques starting ART (combination of PMPA and FTC) during primary or chronic SIV infection. Our hypothesis is that slow restoration of CD4+ T cells in GALT during therapy can be attributed to the disruption of the functional organization of the gut mucosal tissue occurring very early in SIV infection and this may not adequately support survival and maintenance of the CD4+ T cells homing to gut mucosa. Longitudinal evaluation in the SIV model will lead to characterization of the mechanisms and relationship between CD4+ T cell restoration in GALT, peripheral blood, and lymph node compartments (peripheral, and those draining mucosal sites). There are 3 specific aims. In rhesus macaques initiating ART during primary or chronic SIV infection, (1) to determine suppression of SIV replication and genomic diversity, and the kinetics of CD4+ T cell restoration in GALT in comparison to peripheral blood and lymph nodes;(2) to determine the homing and survival of CD4+ T cells in the GALT microenvironment and (3) to investigate the molecular processes involved in the restoration of gut mucosal immune system. The proposal capitalizes on our experience in enteropathogenic studies in the SIV model, expertise in multi-color flow cytometry, in vivo molecular imaging, autologous T cell transfer and gene expression methodologies. The proposed studies promise to provide valuable insights into the impact of impaired gut microenvironment on the viral suppression and restoration of gut mucosal immune system compared to mucosal and peripheral lymph node compartments, and molecular basis of pathophysiologic processes in GALT.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sphingolipids are important mediators and regulators of cell signaling pathways. Our studies have focused on the actions of two classes of sphingolipids represented by glycosphingolipids (GSLs) and sphingosine-1-phosphate. Our work is aimed at defining the normal functions of these sphingolipids and understanding their roles in disease processes. GSLs are found in the outer leaflet of the plasma membrane and are concentrated in specialized signaling structures. They are particularly abundant in neuronal cells in the form of gangliosides (sialic acid containing GSLs). Through genetic disruption of genes that encode synthetic enzymes for GSLs, we have created a series of mice that express limited glycosphingolipid structures. We are using these mice to discover the functions of GSLs. When the cellular machinery responsible for GSL degradation is defective, GSL storage diseases result in which profound neurodegeneration occurs. Examples are Tay-Sachs, Sandhoff and Gaucher diseases. We are attempting to understand how the accumulation of GSLs cause neurodegeneration through the construction of animal models of the diseases. Our major accomplishments this year include a demonstration the expanded macrophage/microglial population in the brain of Sandhoff disease mice is compounded by the infiltration of cells from the periphery. Coincident with the cellular infiltration was an increased expression of macrophage-inflammatory protein 1alpha (MIP-1alpha), a leukocyte chemokine, in astrocytes. Deletion of MIP-1alpha expression resulted in a substantial decrease in infiltration and macrophage/microglial-associated pathology together with neuronal apoptosis in Sandhoff disease mice. These mice without MIP-1alpha showed improved neurologic status and a longer lifespan. The results indicate that the pathogenesis of Sandhoff disease involves an increase in MIP-1alpha that induces monocytes to infiltrate the brain, expand the activated macrophage/microglial population, and trigger apoptosis of neurons, resulting in a rapid neurodegenerative course We also continued our studies on the G-protein coupled receptor for sphingosine-1-phosphate, S1P1. To determine the precise role of the S1P1 receptor on T-cells, we established a T-cell-specific S1P1 knock-out mouse. The mutant mice showed a block in the egress of mature T-cells into the periphery. The expression of the S1P1 receptor was up-regulated in mature thymocytes, and its deletion altered the chemotactic responses of thymocytes to sphingosine 1-phosphate. The results indicated that the expression of the S1P1 receptor on T-cells controls their exit from the thymus and entry into the blood and, thus, has a central role in regulating the numbers of peripheral T-cells..", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronic inflammation promotes DNA damage to overlying epithelium and is frequently associated with neoplastic disease. In addition, DNA damage to epithelial tissues results in the release of a number of cytokines and chemokines that promote an inflammatory response. In particular, a persistent DNA damage response (PDDR) leading to cellular senescence is particularly potent in producing a sustained source of pro- inflammatory mediators; this response has been called the senescence associated secretory phenotype (SASP). It would therefore seem reasonable that a self-perpetuating cycle of DNA damage/SASP and inflammation could occur that would lead to continuing mutation pressure in overlying epithelium. Our proposed studies will examine this linkage. This is possible due to a novel bioimaging approach that monitors dermal microvascular hemoglobin (Hgb) content. We show that focal areas of hyperemia occur following the application of chemical carcinogens or ultraviolet (UV) light to mouse skin. Hyperemia preceded tumor formation, tumors were found to invariably occur in these areas of hyperemia, and these areas were seen to persist and expand following the cessation of an initial carcinogenic UV exposure. Only areas with increased hyperemia were associated with epidermal hyperplasia and dermal inflammation. Moreover, celecoxib, a known anti-inflammatory agent, was shown to suppress these areas of hyperemia along with subsequent tumor formation. We hypothesize that initial carcinogenic exposures elicit a PDDR coupled SASP in epithelial or dermal cells that then drive a self- perpetuating cycle of DNA damage/dermal inflammatory angiogenesis that precedes neoplastic development. We will utilize our unique bioimaging strategy to examine this idea in two aims. In the first aim, we will expose mice to a carcinogenic dose of UV of limited duration, then map out early changes in microvascular blood supply. We will then verify that hyperemic areas correspond to zones of inflammatory angiogenesis that precede microscopic or macroscopic neoplastic disease. We will also determine whether these hyperemic zones exhibit enrichment for characteristic SASP inflammatory mediators. In the second aim, we will examine whether the epidermis overlying early hyperemic areas exhibit increased mutations of the key DNA damage regulator, p53, which is known to augment the SASP response. Moreover, we will determine whether dermal fibroblasts or epidermal keratinocytes in early hyperemic foci exhibit a senescence marker, heterochromatin protein 1g (HP1g), and markers of double stranded DNA breaks, gH2AX and p53BP1. In addition, using a transgenic mouse model (Big Blue mice), we will determine whether hyperemic areas are associated with increased epidermal mutation frequency and whether this mutation pressure persists in the absence of further UV exposures. In both aims, we will examine the capacity of two anti-inflammatory agents (celecoxib and a CXCR2 receptor antagonist) to suppress the formation of hyperemic foci and the associated features of inflammation, senescence and increased epidermal mutation pressure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is clear evidence that nutritional factors alter the toxic response of the lung to oxidants although the mechanisms for these effects are not known. We plan to study the influence of 1) protein deficiency, 2) calorie deprivation, 3) vitamin E deficiency, 4) copper deficiency and 5) selenium deficiency on O2-protective enzymes of the lung Superoxide dismutase, glutathione peroxidase and reductase, glucose-6-phosphate dehydrogenase and catalase), lung reduced and oxidized glutathione levels, lung fatty acid composition and lung morphology before and during exposure to normobaric hyperoxia. In addition, we plan to assess freshly isolated lung cells (alveolar Type II, endothelial, macrophage and fibroblasts) to determine whether any differences exist in cellular distribution of protective enzymes normally or with the various nutritional alterations before or during exposure to hyperoxia.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The cerebral microvessels isolated from brains of gerbils subjected to 15 minutes of complete cerebral ischemia and various periods of recovery had shown an increased monoamine uptake which mirrored the increased uptake of these substances across the blood-brain barrier observed in in vivo studies. Thus, the results suggest that the capillaries are the site responsible for the altered passage of the amine from blood to brain.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Human herpesvirus-6 (HHV-6), the newly identified lymphotropic herpesvirus, is the etiological agent of exanthem subitum and emerging studies also suggest it's involvement in fatal hepatitis, acute and chronic hepatitis, persistent lymphadenopathy, acute mononucleosis and encephalitis. HHV-6 infects productively CD4+ human T lymphocytes, which may potentially have several important implications in human diseases. Our overall objective is to define the role of HHV-6 glycoproteins in its basic biology including in its interactions with human T cells. Our preliminary studies reveal several unique biological properties of HHV-6. HHV-6 strains show variations in their in vitro tropism, DNA restriction sites and in their reactivities with our MAbs against HHV-6 strain GS and Z-29. These properties segregated 20 HHV-6 strains into two groups, tentatively designated as HHV-6 group I and group II. Reactivities of human sera also demonstrated antigenic differences among these two groups and suggest that individuals may be infected with HHV-6 from one or both groups. Glycoproteins, gp82-gp105 complex of group I and gp185-gp210 of group II show major antigenic variations and unique structural features. Our immediate specific goals outlined in this proposal are designed to define in detail the immunochemical, antigenic, genetic and functional relationships among the two variable envelope glycoproteins of group I and group II strains and to determine the potential significance of these variations in the biology of HHV-6. 1. Monoclonal antibodies and mono-specific antibodies will be used to determine the identity, precursor-product, synthesis, structural and antigenic relationships among the polypeptides in gp82-gp105 complex of group I strain HHV-6(GS) and gp185-gp210 of group II strain HHV-6(Z-29) and their counterparts. 2. A 624bp genomic fragment encoding a part of gp82-gp105 characterized by us will be used to identify the genes encoding gp82-gp105 of HHV-6(GS) and (Z-29). Genomic and cDNA libraries of HHV-6(Z-29) constructed in lambda gt11 will be used to identify and sequence gene(s) encoding gp185-gp210 and the counterpart(s) in HHV- 6(GS. Identified genes will be sequenced, characterized and the genetic basis of variations among these glycoproteins will be determined. 3. Monoclonal antibodies, mono-specific antibodies and cloned genes will be used to examine the functions and functional relationships among these glycoproteins, their association with the inner and outer surfaces of infected cell membranes at different time post-infection and to evaluate the potential significance of variations in the biology of HHV-6. The studies outlined in this proposal are significant because these studies will lead into important insight in to the biology of HHV-6.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] The need for a reliable and safe field anesthesia machine that is portable, lightweight, tippable, does not require compressed gas, easy to setup and maintain, insensitive to shock, and cost effective, has been expressed repeatedly by physicians devoted to civilian and military healthcare in the field. This is amplified by the growing trend towards ambulatory surgery, the global war against terrorism and requirements for appropriate medical devices to facilitate military and civilian casualty care, as well as healthcare needs in remote environments where physicians are constrained by the absence of suitable anesthesia equipment. Studies of combat and civilian surgical care shows that existing devices have not evolved to address the unique requirements of surgery outside of the traditional operating room. Evidence suggests that hemorrhage is the biggest cause of death in the battlefield, and that it can be minimized by stabilizing the patient prior to evacuation. The situations encountered in the field are austere and require anesthesia systems that will function reliably in extreme environmental conditions. In addition, since the last 32 years, there has been over a 50% increase in the number of surgeries performed on an outpatient basis, yet there are no anesthesia machines specifically designed for these surgical settings. Studies of patient trauma cases show that vaporizers are the second biggest contributor to patient trauma, second only to breathing circuits. They are unreliable if tipped or agitated and can deliver unsafe concentration of anesthetic gas to patients. We have invented a vaporizer that overcomes these drawbacks and propose to optimize it for potential commercialization through further development. Preliminary results are promising. Tests will be conducted on optimized prototypes under simulated physiologic conditions that mimic field operational environments. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The YMCA's out-of-school time (OST) programs (afterschool programs ~3-6pm and all-day summer camp programs) serve millions of children nationally and represent one of the few settings outside the school where youth can be physically active and eat nutritious foods (e.g., fruits/vegetables [FV]). In November 2011, the YMCA of the USA adopted Healthy Eating and Physical Activity Standards for its OST programs that focus on the amount of activity children accumulate while attending YMCA OST programs and require that children are served FV and water each day at the programs. While these standards represent an important step towards creating healthier OST programs, they fail to describe the strategies YMCA OST programs can use to meet them. This results in a sizable gap between standards and changes in routine practice. Over the past 2 years, working with YMCA OST programs, we have pilot tested and refined a coordinated and comprehensive evidence-based set of strategies, the Healthy Eating and Physical Activity (HEPA) Strategies, to optimize the implementation of the Standards. Our objective here is to evaluate the impact of HEPA Strategies on achieving the Healthy Eating and Physical Activity Standards and the associated costs with using the HEPA Strategies. To accomplish this, we will use an immediate intervention vs. wait-listed standard practice comparison group design (total of 20 programs across the state) and will work with South Carolina YMCA leadership to implement the Healthy Eating and Physical Activity Standards and HEPA Strategies in YMCA programs across the state. We developed an innovative model to achieve the objectives and have drawn extensively from the restaurant/franchisee industry to create a diffusion-support framework for statewide implementation that utilizes existing YMCA OST programs as demonstration/training sites and develops regional training sites where program leaders and staff from across the state will receive hands-on experience implementing the Standards and Strategies in real-world settings. We will use direct observation and accelerometry to measure all primary outcomes. The following Aims are proposed: Aim 1. Evaluate the uptake of and adherence to the Physical Activity and Healthy Eating Standards and identify organizational, staff, and setting characteristics that influence the implementation process; Aim 2. Evaluate the impact of the Physical Activity and Healthy Eating Standards on children's physical activity levels and fruits/vegetables and water served in YMCA OST programs; Aim 3. Evaluate the cost-effectiveness of meeting the Physical Activity and Healthy Eating Standards. This work is significant and represents an important step in identifying processes by which effective, low- and no-cost strategies can be integrated into the existing YMCA organizational framework to ensure national YMCA Standards are achieved. This application is innovative because it will establish the effectiveness of the Standards using a diffusion-support framework in real-world settings to increase children's physical activity and healthy eating in YMCA OST programs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Angiogenesis, the formation of new blood vessels, is a key element of a number of normal and pathological processes. Whereas vascularization is a primary event associated with normal embryogenesis and wound healing, neovascularization is of central importance also n a variety of disease states. Neovascularization in the eye is an almost universal aspect of ocular disease and injury, new blood vessels are associated with immunological reactions and the inflammatory processes, and angiogenesis plays a pivotal role in permitting the growth and differentiation of autochthonous tumors and their metastases. The proposed research is directed at a greater understanding of the process of neovascularization, with special emphasis on lymphocyte-induced angiogenesis. Specifically, the aims of this research include: 1. Identification and characterization of angiogenic lymphokines released by lymphocytes following immune stimulation; 2. Delineation of site-dependent selectivity in the neovascular response as this may be reflected by heterogeneity in the microvascular endothelial cell phenotype; 3. Development and validation of in vitro correlates of angiogenesis and of more quantitative and readily reproducible in vivo tests of neovascular reactions, using the mouse as an animal model. 4. Study of mouse mutants with specific vascular or immunological deficiencies and with cells obtained from those mice, as a means of developing specific disease models applicable to human diseases. Research methods include cell culture analysis of migration and proliferation; flow cytometry and cell sorting; production and testing of monoclonal antibodies against angiogenic lymphokines; biochemical procedures designed for purification of specific angiokines, and image analysis for assessment of new blood vessel formation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Administrative Core (Core B) will consist of the Program Director (10% effort) and two assistants each with partial effort. One is Ms. Lisa Russell, administrative assistant, who will perform duties related to grant preparation, grant reporting, manuscript preparation, human and animal subjects review and arranging meetings and conference calls among the PPG investigators, the internal advisory committee and the external advisory committee, and other non-financial issues mentioned below. She will need to be appointed at 50% effort to perform all of these duties. The other is Ms. Cher Herh, a grants management specialist who will manage all financial components of the grant including a) prior to submission, preparing financial elements of the application, b) performing all postfunding accounting on a continuing daily basis with monthly reports to the Program Director, project and core leaders, and c) supporting annual reporting for all four projects and two cores. She will be appointed at 25% effort. In addition, our Senior Electronics/Computer Technician Mr. Jeff Struthers will provide 10% effort within the Core to support maintenance of computers and peripheral equipment used in research in all projects and cores. The remainder of his time is distributed among the projects to support each project's unique research equipment electronics and hardware needs. This is both in maintenance of existing equipment and in the development of new equipment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During the past 25 years, studies in epithelial cancers have defined genetic changes in the tumor cells themselves. However, it is clear that cancer progression and metastasis requires complex interactions between tumor cells and the surrounding stromal. Our work has studied the role of cytosolic phospholipase A2 (cPLA2) in the development of lung cancer, focusing on tumor cells themselves. Activation of cPLA2 represents the rare limiting step in prostaglandins(PG) production. Constitutive increases in PG production are associated with many cancers, including non-small cell lung cancer (NSCLC). We have shown that NSCLC cell lines expressing oncogenic K-Ras have elevated levels of PGE2 production, mediated through induction of cPLA2 and COX-2. Blocking this pathway inhibits transformed growth in vitro and in xeno graft models, supporting a role for cPLA2 in lung cancer progression. However, less is known regarding the role of cPLA2 in the tumor microenvironment. During the past funding period, we showed that cPLA2 knockout (KO) mice are protected against chemicallyinduced lung tumorigenesis. However, these studies do not discriminate between the action of cPLA2 in the tumor cells versus the surrounding stromal. To address the function of cPLA2 in the tumor microenvironment, we have developed a model in which mouse tumor cells are directly injected into the lungs of syngeneic, immune competent mice. Using bioluminescence imaging we have shown that these cells form well-defined primary tumors which metastasize to other lobes of the lungs and into the mediastinal lymph nodes. When identical cells are injected into cPLA2 KO mice, while primary tumor growth is not significantly altered, there is a profound inhibition of metastasis. Tumors growing in cPLA2 KO mice exhibited alterations in the pattern of macrophages surrounding the tumor, supporting a role for cPLA2 in macrophage recruitment/function. Transplanting cPLA2 KO bone marrow into wild-type mice was sufficient to inhibit tumor metastasis and promote survival. Based on these results, we hypothesize that cPLA2 is required for mobilization and/or function of tumor associated macrophages which play a critical role in lung cancer progression and metastasis. This project will investigate the role of cPLA2 in both tumor cells and stromal, and define molecular events mediated by cPLA2 in both compartments. Two Specific Aims are proposed. Aim 1 will assess the contribution of cPLA2 in stromal cells using murine lung cancer cells injected into either WT or cPLA2 KO mice. Changes in stromal cells will be defined and in vitro co-cultures will examine cross-talk between cancer cells and macrophages. Aim 2 will use genetic mice to define the downstream effectors of cPLA2 in the tumor microenvironment. These studies will provide a better understanding of tumor-stromal interactions critical for metastasis and define therapeutic targets to block progression of lung cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract Child maltreatment is among the most devastating of childhood adversities and increases vulnerability for deleterious outcomes across a number of domains of functioning. However, not all who experience child maltreatment develop problems and those with similar maltreatment experiences may show very different outcomes. Differential biological embedding of maltreatment likely explains this heterogeneity in outcomes. Epigenetics, or the processes that influence gene expression, are crucial to our understanding of how child maltreatment influences later development. DNA methylation (DNAm) is the most well-studied epigenetic mechanism and has been proposed to link early life experiences to alterations in gene expression and mental health symptoms. Only a few studies have used a genome-wide approach to examine these associations and importantly have shown different DNAm patterns for maltreated youth within genes linked with mental health, but did not actually assess outcomes. No longitudinal investigations have tested genome-wide differences in DNAm that emerge after maltreatment and predict the emergence of, or change in, mental health symptoms. A relatively new measure of biological vulnerability, DNAm age, has emerged as an indicator of biological aging that is associated with life stress and has initial support for predicting risk for poor mental health functioning. Although maltreatment is linked with advanced biological aging using other indices (e.g., telomere length), there have been no longitudinal studies testing the effect of maltreatment on DNAm age, nor testing DNAm age as a mediator between maltreatment and later mental health symptoms. To address these gaps, we will use an existing longitudinal sample of maltreated and comparison children (n=454) followed for 10 years with 4 waves of archived samples and a myriad of psychosocial data. This data is optimal to assess the timing of maltreatment effects on genome-wide DNAm, DNAm patterns associated with subsequent mental health, and the potential resolution of problematic DNAm patterns across adolescence (ages 9-23). This is an unprecedented opportunity to capitalize on existing data from a well-designed longitudinal study of maltreated youth with a matched comparison group, which allows us to separate maltreatment effects from other aspects of early life stress and adversity, including neighborhood and socioeconomic factors. This study will contribute important new evidence crucial to our understanding of how maltreatment affects genome-wide epigenetic patterns of vulnerability that result in mental health problems. This will further the development of clinical risk assessment and prevention approaches for maltreated youth which will be of substantial benefit to population health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The research to be conducted involves the study of the preparation and chemistry of penta and hexacoordinated phosphorus compounds. Detailed studies of the structures and reactivities of these substances will be conducted. In particular extensive nuclear magnetic resonance studies of the various nuclei present in these molecules will be carried out. Pentacoordinated phosphorus compounds have been implicated as intermediates in important biological processes. The role of hexacoordinated phosphorus compounds has not been established. Recent evidence from several sources indicates that these materials are often intermediates in a number of reactions which occur in living systems. Because of this, extensive studies of the formation and chemistry of these materials are contemplated. Studies of intra and intermolecular ligand reorganization of P(V) and P(VI) compounds will be conducted as will ligand reorganization in selected P(III) compounds.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pseudomonas aeruginosa produces a variety of extracellular products which may play a significant role in the pathogenesis of Pseudomonas infections in experimental animal models and in humans. Some of these factors such as exotoxin A, protease and phospholipase C (hemolysin) are likely candidates to be major virulence factors of P. aeruginosa; however, the role and interaction of these factors in the pathogenesis of Pseudomonas infections has not yet been clarified. The ultimate objective of this research is to employ genetical techniques completed with other experimental methods to obtain valuable information concerning the role of toxins as virulence factor of P. aeruginosa. As further insight into the role of specific virulence factors in the pathogenesis of Pseudomonas infections is gained, it is likely that the principal of genetics, including genetic engineering problems of developing prophylactic and therapeutic agents for patients with Pseudomonas infections. Immediate objectives are : (1) to elucidate the genetic mechanisms which regulate the production of exotoxin A, proteases and hemolysin. The well characterized conjugal mating system of P. aeruginosa will be used to further map and analyze genetic determinats controlling toxin production, protease production and hemolysin in production. The transducing system of P. aeruginosa will be potentially useful for fine structure mapping of virulence determinant genes and to determine the role, if any, that bacteriophage play in the production of virulence factors. (2) To isolate and fully characterize spontaneous or chemically induced mutants of P. aeruginosa with altered ability to produce virulence determinants. Such mutants will be subjected to further extensive biochemical and genetical analyses. (3) To examine and compare the virulence of mutants and wild-type strains in appropriate experimental animal models. Ultimately, preliminary studies could be conducted with experimental animals to determine if a mutant strain or a CRM product has potential to be an effective immunoprophylactic agent Pseudomonas infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent studies on the mechanism of ionization during thermospray liquid chromatography-mass spectrometry have extended earlier work and substantiated the conclusion that, for most analytes, ion formation is predominantly a gas phase process akin to chemical ionization. The finding has important implications for the prediction of suitable experimental conditions for new analytes. Tandem mass spectrometry of acylcarnitines and steroid glucuronides has indicated appropriate conditions for the analyses, with a particular view to the differentiation of isomers. These studies have additionally improved understanding of the processes of ion production following fast atom bombardment and of collisionally activated decomposition.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Current estimates suggest that approximately half of arteriovenous fistulas (AVF) fail to functionally mature and become regularly usable for hemodialysis by six months of surgical creation. The process of maturation (and its failure) is poorly understood, and no effective therapies to promote maturation are currently known. An NIDDK-sponsored Hemodialysis Fistula Maturation (HFM)(U01 DK-082189-01) cohort study is underway to examine whether a broad range of clinical, demographic, physiological and process-of-care variables in patients undergoing AVF placement influence maturation and usability of the AVF for dialysis. While the parent study will correlate pre-operative vascular function and venous histological features with post-operative ultrasound data, its largely global assessments will not furnish a detailed picture of the local and specific biological mechanisms that determine AVF maturation. Hence, the proposed ancillary clinical study examine the complex interplay between pre-existing endothelial and smooth muscle cell dysfunction, endothelial repair, and systemic inflammation. The specific objectives are (1) to delineate genome-wide expression patterns from endothelial and smooth muscle cells within the donor vein wall at the time of AVF creation, and to identify unique genomic patterns (i.e. signatures) that are associated with AVF outward remodeling and changes in blood flow rate, (2) to determine the initial and ongoing balance between endothelial injury and repair following vein graft implantation, as defined by circulating endothelial cells (CECs) and endothelial progenitor cells (EPCs), and obtain novel genomic signatures of EPCs associated with a successful or unsuccessful AVF maturation, and, (3) to evaluate the role of systemic inflammation, as determined by genome-wide expression patterns of circulating monocytes, as they relate to AVF maturation and can be used to predict outcomes and identify novel pathways. We propose to enter 100 patients (50 patient test set, 50 patient validation set) at the University of Florida who will participate in the parent study. We propose to employ laser microdissection for endothelial and smooth muscle cell capture within the vein wall, as well as macro-scale and novel microfluidic capture technologies for blood EPCs and monocytes obtained preoperatively and on post-op day 14 for subsequent genome-wide expression analysis using a novel proprietary Affymetrix HH/2 exon array. Using these novel technologies, we will accomplish two major goals: (1) develop novel genomic signatures from vascular tissue cell populations and enriched blood leukocytes that can be used to predict a successful outcome, and (2) develop new insights into novel signaling pathways and potential mechanisms for therapeutic intervention. The proposed studies will effectively exploit the unique data collection effort in the parent HFM study and add both predictive genomic modeling of outcome, and significant mechanistic insight into the physiology and pathophysiology of AVF development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The first developmental choice faced by germ cells in the majority of higher eukaryotes is oogenesis versus spermatogenesis, but very little is known about how this basic event is genetically encoded in any organism. We have chosen to study the developmental genetics of this germline sex determination process in the fruit fly, Drosophila melanogaster. Our long-term goals are to: 1) identify the major germline sex determination regulatory genes, 2) determine how they are related in terms of a developmental hierarchy and 3) understand in detail how these regulatory gene products function. We are question oriented and take advantage of genetic, molecular, and biochemical techniques as needed. Current molecular studies are focused on OVO, a highly conserved zinc finger domain protein functioning at a key intersection in the hierarchy. We have found that the ovo+ locus has alternate promotors and 5' exons (ovo-a and ovo-b), only one of which has an in frame AUG (ovo-a). OVO-B mRNA is translated from an AUG in exon 2. We found that OVO-B mRNA is produced only in females and is controlled by the sex-karyotype and somatic inductive signals, while OVO-A mRNA is produced in both males and females. To test the function of OVO-A and OVO-B, we constructed multiple transgenes. Analysis of these constructs in an ovo minus genetic background indicate that the female-specific OVO-B isoform is sufficient for zygotic female fertility (female sterility is fully rescued), but is insufficient for full germline viability in the next generation (there is incompletely penetrant maternal effect sterility). This suggests that maternally deposited OVO-A activity is important in the early embryo, while positive OVO-B activity is required for female germline development in the zygote. Flies bearing a ovo-b promoter knockouts or with new in-frame AUG initiation codons in the ovo-B 5'-UTR show dominant negative female sterility suggesting that excessive OVO-A activity is detrimental to the female germline. Interestingly, the ovo-b promoter knockouts also show significant rescuing activity in an ovo minus genetic background. We are investigating whether this simultaneous dominant negative and wild-type allele results in the production of both OVO-A and OVO-B proteins (the later a result of either ectopic ovo-b promoter activity or OVO-A AUG initiation codon suppression). We are also looking for biochemical function by assaying for the binding of bacterially expressed OVO to its own promotor and to the ovarian tumor promoter (we have shown that both are regulated by ovo+ dose). Interestingly, OVO binds at the transcription initiation site of both these TATA-less promoters, in vitro, as well as to upstream sites. We have shown that the upstream sites are functional by using reporter genes. Deletions of the upstream OVO binding sites abolish transcriptional activity and transcription can be reconstituted by adding back any of several slightly different OVO binding sites. The core promoter OVO binding sites also appear to be important. Point mutations and core promoter swapping experiments suggest that only a subset of core promoters can respond to the enhancers at the ovarian tumor locus. For example the ovo-b and ovarian tumor promoters, both of which have strong OVO binding sites at the core promoter, support good transcription using the ovarian tumor enhancers, while the ovo-a promoter, or the ovo-b promoter with a mutated OVO binding site do not.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROVIDED. During development of the central nervous system (CMS) post-migratory cells of both neuronal and oligodendroglial origin send out processes that navigate through the parenchyma in search for a target signal that induces their maturation, i.e synapse formation for neuronal and myelin sheath formation for oligodendroglial cells. The regulation of this process navigation has been well characterized for neuronal cells, where a sensorimotor structure, the neuronal growth cone, located at the process tip is the \"organelle\" that senses environmental cues. In contrast, surprisingly little is known about oligodendroglial process pathfinding and targeting. Our preliminary data demonstrate that oligodendrocyte processes possess at their distal ends \"organelles\" that structurally and functionally resemble neuronal growth cones and that we will refer to as oligodendroglial growth cone-like structures (OLG-growth cones). In neurons, mRNA transport and locally restricted protein synthesis within the growth cone are considered main regulators of axon/neurite pathfinding and targeting. Our preliminary data suggest that similar regulatory mechanisms may be important for the pathfinding and targeting of oligodendrocyte processes. Thus, we formulate the central hypothesis that premyelinating, post-migratory oligodendrocytes possess OLG-growth cones that containmRNAs, the locally restricted translation of which is critical for the regulation of oligodendroglial process pathfinding and targeting, i.e. myelination. This hypothesis will be tested in two specific aims. 1) We will determine the role of locally restricted protein synthesis for OLG-growth cone pathfinding and in particular for chemotropic turning in response to non-permissive cues. In this set of experiments the extent to which locally restricted protein synthesis occurs in pathfinding OLG-growth cones will be determined. In addition, chemotropic turning of separated processes will be analyzed after inhibition of protein synthesis. It is expected that inhibition of locally restricted protein synthesis will perturb OLG- growth cone chemotropic turning responses. 2) Using a microarray approach, we will identify mRNAs that are present in OLG-growth cones specifically in response to axonal targeting signals. This set of experiments is expected to reveal candidate proteins that are important for OLG-growth cone targeting, i.e. the initiation of myelin sheath formation. In continuing studies based on the findings of the present proposal, we are planning to further characterize these proteins. The proposed studies thus represent a first step into a set of experiments designed to better understand the regulation of oligodendrocyte process pathfinding and targeting. Such advancement in knowledge is of particular interest since deficiencies in process outgrowth and axon recognition appear to be among the main limiting factors for repair of lesions present in the CMS of patients suffering from the major demyelinating disease in humans, Multiple Sclerosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract We propose to develop an advanced rapid point of care diagnostic for HIV. This approach will leverage a novel D4 diagnostic assay platform: The D4 is so named because of the chain of events that occur upon addition of blood: (1) Dispense blood onto chip; (2) Dissolution of ?soluble? detection reagent spots (3) Diffusion of coated virion across a surface and binding to specific capture Ab spots; (4) Detection of binding with fluorescence imaging. We will enhance HIV virion capture using a lectin, Griffithsin, which can bind to the glycan shield of HIV virions spikes and induce exposure of CD4 binding sites which can be captured by antibodies. Individual virions can be captured, labelled and imaged using next generation cell phone based microscopy. This diagnostic test enables not only rapid ?window independent? diagnosis independent of patient antibodies, but also potentially for routine viral load monitoring.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies will focus on the potential role of extracellular proteases, namely the plasminogen activator/plasminogen (PA/pign) system in synaptic remodeling and axonal regeneration in the injured spinal cord of rodents. The PA/plgn system has been suggested to play a role both in axonal outgrowth and synaptic plasticity. The PA genes are induced following peripheral nerve injury and are required for timely PNS regeneration. Studies will be directed at assessing the role of these PA/plgn genes in the ability of dorsal root ganglia sensory axons to regenerate their CNS collaterals in the dorsal column of the spinal cord. Most importantly studies will be performed to assess whether the interesting crossed phrenic nerve response, an example of synaptic remodeling following spinal cord injury, requires the induction of the PA/pign system to promote these synaptic changes that permit a restoration of lung function. Mice deficient in various PA/plgn genes will be compared for their plasticity and regenerative abilities to demonstrate any requirement for these protease activities. Furthermore, expression of the PA/pign system [tissue plasminogen activator (tPA), urokinase plasminogen activator(uPA), plasminogen, plasminogen activator inhibitor-i (PAIl), and protease nexin(PN)] in the lesioned or injured spinal cord will be studied by in situ hybridization with 35S-CRNA probes and antibodies to these PA system proteins, to determine whether this gene system is also induced in the CNS and whether it plays a role in axonal regeneration and synaptic plasticity. An understanding of this protease system in the injured spinal cord may allow us to effectively enhance axonal regeneration by supplying either PA/plgn or their inhibitors to the site of injury. Similarly, PA/pign expression may be important for synaptic remodeling at sites below a spinal cord lesion, such that induction of the PA/pign genes may lead to partial restoration of function.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is well-established that a brief period of global brain ischemia causes delayed cell death in hippocampal CA1 pyramidal neurons days after reperfusion in animals and humans. Although numerous factors have been indicated in this phenomenon, the mechanisms underlying this delayed neuronal cell death are still poorly understood. We have demonstrated that cerebral infarction and neurological deficits are significantly reduced in transgenic mice overexpressing CuZn-superoxide dismutase (Sod1) activity after acute focal stroke, whereas vasogenic edema, infarction and neurological deficits are exacerbated in mutant mice deficient in Sod1 or in mitochondrial manganese SOD (Sod2) activities. But the role of these antioxidant enzymes on the delayed hippocampal neuronal injury after global ischemia is still unknown. Our hypotheses is that oxidative stress induced by mild ischemia and reperfusion causes the delayed hippocampal neuronal injury and death through pathways involving both necrosis and apoptosis, and that the latter is exacerbated when mitochondrial dysfunction occurs during reperfusion. It is our aim to test our hypothesis using transgenic mice overexpressing Sod1 and Sod2 activities and knockout mutant mice that contain no Sod1 -/- (homozygous), half (heterozygous, Sod1 +/-) or Sod2 +/- activities. In order to dissect out the role of neuronal-specific expression of Sod1 in ischemic neuronal protection, we will generate mice that contain only neurons expressing Sod1 activity using neuronal- specific enolase (NSE) promoter with Sod1 genomic DNA construct. In order to elucidate the oxidative role of subcellular compartmentation (i.e., cytosolic vs. mitochondria) in necrosis and apoptosis, we will generate mice that contain genotypes with combinations of increased Sod1 expression and Sod2 +/- knockout mutants. We believe these are unique and fresh approaches that will provide insights into the oxidative mechanism in mitochondria that underlies apoptosis in delayed hippocampal cell death after global cerebral ischemia and reperfusion.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Proper functioning of the immune system, in detecting and mounting an immune response against invading pathogens, is dependent on lymphocyte recirculation through secondary lymphoid organs. Factors that affect lymphocyte entry and exit into these lymphoid tissues are not completely understood. Furthermore, persistent pathological disruption of lymphocyte recirculation can lead to sequestration of lymphocytes in lymphoid organs, as seen in HIV infection, and may contribute to immunosuppression of the host. The long-term goal of our research program is to gain a detailed molecular understanding of regulation of lymphocyte trafficking in acute and chronic inflammation using viral infections as a model system. We have found that expression of a phospholipid receptor, sphingosine-1-phosphate receptor-1 (S1P1), within lymphocytes is required for their ability to exit from secondary lymphoid organs. Both innate and adaptive immune responses alter the expression and function of this receptor leading to lymphocyte sequestration. A variety of cytokines, including interferon alpha/beta (IFN), up-regulate expression of the transmembrane C-type lectin, CD69. CD69 interacts with and down-modulates surface expression and function of S1P1, a G protein coupled receptor, leading to transient lymphocyte sequestration. In addition, engagement of the antigen receptor on T cells leads to transcriptional down-regulation of S1P1 mRNA expression and a prolonged inhibition of lymphocyte egress. Thus, S1P1 function is tightly regulated by two distinct mechanisms during the early stages of an immune response. The goal of this proposal is to further understand the regulation of lymphocyte trafficking by S1P1 and CD69 and its impact on generation of T cell responses. Cellular and biochemical approaches will be utilized to determine: (1) the molecular requirements for S1P1-CD69 interaction; (2) the molecular mechanism of inhibition of S1P1 function by CD69; and (3) the effects of genetic perturbation of regulation of S1P1 expression and function on in vivo T cell responses to a viral infection. Our hope is that a detailed understanding of factors that regulate lymphocyte trafficking can contribute not only to vaccine development and improved immunotherapies, but also provide targets for therapeutic disruption of these processes in autoimmune and chronic inflammatory diseases. PUBLIC HEALTH RELEVENCE: Circulation of white blood cells through lymphoid tissues is essential for initiation of immune responses to infectious microorganisms such as viruses but could also have detrimental consequences in chronic inflammatory and autoimmune diseases. The goal of this proposal is to gain a detailed understanding of how two specific molecules known to affect white blood cell trafficking regulate this process. The knowledge gained from these studies will facilitate development of therapies that target white blood cell trafficking for treatment of inflammatory diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project aims at determining if the administration of Betaseron affects the progression of secondary-progressive multiple sclerosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Bone metastasis occurs in 49% of patients with cancer of the breast. Although the diagnosis is currently made by x-rays and scintiscanning, there are no accurate indicators for the evaluation of the early response to therapy. Since bone matrix is the largest store of collagen, the invasion of bone by metastatic cancer, is accompanied by an elevated excretion of hydroxyproline (HYPRO) in urine. HYPRO is excreted as dialyzable and non-dialyzable forms; the former represents breakdown of mature collagen, the latter is an index by synthesis of new collagen. It is therefore likely that serial determinations of dialyzable and non-dialyzable HYPRO can be an important tool in the evaluation of therapeutic response in patients with cancer of the breast with skeletal metastasis. In continuation of studies conducted during the first stage of this project (CA 20939-01, Jan.-Dec. 1977), we proposed to determine in patients with cancer of the breast: 1) If the higly simplified procedure of HYPRO/creatinine ratio in morning urine \"SPOT-HYPRO\" currently used in our laboratories, is an accurate index of the presence and response to therapy of bone metastasis. 2) If the elevation of non-dialyzable HYPRO excretion, observed 24-48h after chemotherapy is an index of immediate response to therapy and if the tests have prognostic value for long-term treatment. 3) If there is correlation between the metastasis/normal bone ratio of densitometry in bone scans with the urinary excretion of HYPRO, as preliminary data from our laboratories appear to indicate. 4) The validity of HYPRO measurements as an index of bone metastasis by determining the glucosylgalactosylhydroxylysne/galactosylhydroxylysine ratio in urine. This test differentiates the tissue of origin of the increased collagen turnover as bone or soft tissues.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Co-infection with multiple parasites is common in sub-Saharan Africa. Malaria and schistosomiasis are co-endemic diseases that affect nearly a billion people worldwide; 89% of whom reside in Africa. These diseases impose an enormous public health burden individually, but little is known about the interactive effect that occurs with dual infection. Vaccines are urgently needed to eradicate these parasites yet vaccine development depends upon reproducing predictable host immune responses. Evidence suggests that patent helminth infection can alter host immune responses to acute protozoal infection. Our central hypothesis is that underlying helminthiasis, such as schistosomiasis, modulates the human immune response, affecting the incidence and severity of concomitant falciparum malaria. Prospective cohort field studies that we conducted in Mali, West Africa demonstrated age-specific protective effects in children with S. haematobium infection upon the time to acquisition of falciparum malaria, burden of malaria disease and geometric mean parasite density as compared to matched controls without schistosomiasis. Immunologic assays in sera and PBMC demonstrate alterations in Th-2 balance in children with helminth infection at the time of a malaria infection. Flow cytometry studies demonstrate marked increases in IL-4 production, predominantly by CD4+ central memory T cell (TCM) subsets, in children aged 4-8 years with schistosomiasis compared to matched controls. IL-4 is critical to memory induction in both the humoral arm the development of memory T cell responses. Using valuable samples that were collected during this clinical study, we will study the complex humoral and cell mediated immune responses that occur in response to malaria and assess differences in children with or without underlying schistosomiasis. We expect these studies to 1) demonstrate the importance of co-parasitic infections in the developing world and elucidate immunologic mechanisms that result in these clinical interactions, 2) provide insight into altered immune responses induced by polyparasitism that will be critical to malaria vaccine trial design, and 3) influence parasite treatment policies and control programs throughout much of the tropics. Our central aim is to examine the immunomodulatory effect that patent Schistosoma haematobium has upon the humoral and cellular response elicited to acute Plasmodium falciparum infection and how this immune response affects the incidence and severity of clinical malarial disease. We expect these studies to 1) demonstrate the importance of co-parasitic infections in the developing world and elucidate immunologic mechanisms that result in these clinical interactions, 2) provide insight into altered immune responses induced by polyparasitism that will be critical to malaria vaccine trial design, and 3) influence parasite treatment programs and preventative health policies throughout much of the tropics. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Collaborative Cross (CC) is a panel of recombinant inbred lines derived from eight founder mouse strains. We have recently genotyped 450 CC lines to provide the first accurate description of the genetic architecture of the CC population. We are now focused on making the largest number of lines accessible to the researchers as soon as possible through genotyping and re-derivation of CC lines nearing the thresholds for distribution (>90% inbred) and completion (>98% inbred). There are currently 27 CC lines in the distribution list and we expect to have a minimum of 50 CC lines by the end of 2012 and 100 CC lines by 2103. The CC is a Genetic Reference Population and, therefore, the number of lines available has a direct impact in the power of detecting loci with small to moderate effects and on the ability to select lines that are uniquely suited to specific research projects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is concerned with skeletal muscle function, energy metabolism and biochemical adaptations induced by exercise training. Particular attention is given to the response of the different skeletal muscle fiber types, since each fiber is characterized by distinct biochemical and physiological properties. Further, these distinctions probably have a direct influence on the adaptive responses induced within each fiber type by exercise training. We will evaluate aspects of adenylate metabolism and the purine nucleotide cycle in working muscle and during recovery. This will include an assessment of the factors that are important in the activation of AMP deaminase in vivo and the amine source for adenylate resynthesis following intense muscle use in rats. The involvement of certain amino acids in energy metabolism will be evaluated during steady-state muscle use with an isolated perfused hindlimb preparation. We will determine the energy contribution of branched-chain amino acid oxidation, the extent of oxidative deamination of glutarate, and the mode of amine nitrogen elimination from the working muscle. A major adaptation induced in skeletal muscle by training is an increase in the capacity for ATP provision via oxidative metabolism. The impact of this adaptation on the above processes will be made. In addition, investigations with muscle stimulated in vitro will be performed to explore the physiological significance of the adaptive changes induced by training.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The median survival for patients diagnosed with glioblastoma multiforme (GBM) is one year. Although surgery, chemotherapy and radioactive therapy have continuously improved, long-term survival from GBM has not improved since 1950. One of the methods that have been hypothesized to provide more effective treatment for GBM is through immunomodulation. To investigate this hypothesis, we previously determined that immunosuppressive regulatory T cells (Tregs) are found in human GBM, which is recapitulated in experimental mouse models of glioma. Using an experimental mouse model of glioma, we also determined that Treg depletion increases long-term survival. Based on those discoveries, this research proposal aims to extend those observations by investigating the immunomodulatory enzyme, indoleamine 2, 3 dioxygenase (IDO), which is currently being targeted in Phase 1 clinical trials for breast, lung, and pancreatic cancers, as well as melanoma. IDO has been shown to regulate the conversion of Treg into IL-17A-producing pro- inflammatory CD4+ T helper 17 cells (Th17s). However, no previous investigation has determined the roles of IDO and Th17s in GBM. Therefore, this study aims to identify: 1) the role of IDO, 2) the role of Th17s and 3) the mechanism by which IDO regulates Treg/Th17 levels in orthotopic and transgenic mouse models of glioma. Aim 1 will provide an analysis of in vivo transgenic and orthotopic mouse models of glioma in combination with select IDO deficiency using wild-type (WT), IDO-/- and CD4-/- mice, IDO inhibitors, IDO knockdown with shRNA and CD4+ T- and dendritic cell-adoptive transfers. The changes in IDO mRNA levels, IDO enzyme activity, IDO protein localization, Treg/Th17 levels and glioma invasiveness will be analyzed and correlated to mouse survival. Aim 2 will critically test in vivo orthotopic mouse models of glioma with selective IL-17A deficiency using WT-, IL-17A-/--, ROR3t-/-- and CD4-/--mice, in conjunction with CD4+ T cell adoptive transfers. The changes in Treg/Th17 frequency, Treg/Th17-related cell surface and cytokine expression, IL- 17A protein expression, IDO enzyme activity and glioma invasiveness will be analyzed and correlated to mouse survival. Aim 3 will investigate the mechanism by which IDO regulates Treg/Th17 conversion in vivo using IL-17 reporter-, CD4-/--, CD4+pSTAT5-/--, and CD4+pSTAT3-/--orthotopic mouse models of glioma 1IDO inhibition with CD4+ T cell adoptive transfers. The changes in Treg/Th17 frequency, pSTAT3 and pSTAT5 levels in CD4+ T cells, conversion of adoptively transferred Tregs and Th17s, absolute Treg/Th17 numbers, and CD4+CD25+-, CD4+CD25+IL-17+-, CD4+IL-17+-T cell suppressor capacity will be analyzed. Human GBM is a fatal type of glioma, which results in the infiltration of Tregs and Th17s, as well as the selective expression of IDO. This proposal aims to understand the roles that IDO plays in maintaining the balance between Tregs and Th17s using pre-clinical experimental mouse models of malignant glioma. PUBLIC HEALTH RELEVANCE: Glioblastoma multiforme (GBM) is a highly malignant primary tumor of the central nervous system of glial origin with a median survival time of 15 months. We and others have shown that CD4+ T lymphocytes infiltrate human GBM, as well as mouse models that recapitulate GBM, leading to studies demonstrating that, immunomodulation is a potential target for future treatment of GBM. To further these studies, this proposal aims to investigate the role of indoleamine 2, 3 dioxygenase (IDO), the role of t helper 17 cells (Th17s) and the molecular mechanism by which IDO regulates Treg/Th17 levels using intracranially-injected and spontaneously-developing mouse models of GBM.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Covalent modification by isoprenoid lipids (prenylation) is a critical post-translational event for many proteins involved in cellular signaling and cancer. The primary goal of this research program is to design and test prenyl function inhibitors and to identify and characterize proteins that are farnesylated in vivo. The studies outlined in this proposal will result in the preparation of new tools to probe the specificity of the prenyltransferases protein- farnesyltransferase (FTase) and protein-geranylgeranyltransferase (GGTase-I), enable further development of prenyl function inhibitors and identify new prenylated proteins as potential targets for therapeutic intervention. By varying the chemical structure of the prenyl lipid, we are developing reagents to probe the biological function of the posttranslational modification. Taking advantage of the fact that the prenyl group forms a substantial part of the peptide substrate binding site in the prenyltransferases has allowed us to develop peptide selective inhibitors of prenyl function. These unnatural analogues may allow for the selective interference with specific prenylation targets and may provide lead compounds to alleviate the potential toxicity associated with complete inhibition of protein prenylation. In particular, the unnatural analogues may be useful to obtain a more complete understanding of the role that alternative prenylation plays in Farnesyl transferase inhibitor (FTI) evasion by oncogenes such as K-Ras. Critical to understanding the clinical effects of existing and future FTIs and geranylgeranyl transferase inhibitors (GGTIs), is the identification of in vivo substrates of FTase and GGTase-I. Our innovative strategy of using unnatural, transferable prenyl analogues and analogue specific monoclonal antibodies to identify prenylated cellular proteins will provide valuable information on the cellular targets of inhibitors of prenylation. We expect these studies to result in the identification of previously unknown prenylated proteins. The specific aims of this project are: 1) Synthesis of farnesyl and geranylgeranyl diphosphate analogues to study protein prenylation. 2) Screening these compounds for substrate specificity of FTase and GGTase-I and development of prenyl function inhibitors. 3) Identification of prenylated proteins in cells. The results from these studies may provide leads to new molecules to treat cancer and also identify new molecular targets to develop anti-cancer therapies. PUBLIC HEALTH RELEVANCE Substantial evidence points to the central role of proteins normally modified by a prenyl group in cancer progression. We propose to design and test prenyl function inhibitors and to identify and characterize proteins that are modified with prenyl groups. The results from these studies may provide leads to new molecules to treat cancer and also identify new molecular targets to develop future anti-cancer therapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although stannous fluoride has been used as a proven anti-caries agent for over twenty years, its mechanism of action is still not clearly understood. Only recently has the compound Sn3F3PO4 been identified as one of the main reaction products of in vitro experiments involving the treatment of enamel surfaces with 10 percent stannous fluoride solutions. It is possible that Sn3F3PO4 plays an important role in the anti-caries mechanism of stannous fluoride in vivo. This investigation will study the reaction of stannous fluoride with sound and carious dental enamel as it exists in the human mouth. Both permanent and primary teeth scheduled for extractions will be treated with a freshly prepared 10 percent stannous fluoride solution for varying time intervals. Contralateral teeth in the same patients will be treated with distilled water and will serve as controls. Up to fourteen days later the teeth will be extracted and subjected to analysis. Surface reaction in the first 10 to 25 micron layer will be identified using X-ray powder diffraction methods. The products in the carious dentin will be analyzed similarly. The enamel fluoride biopsy technic will be used to remove the first two to three microns of enamel surface for chemical assay using atomic absorption, fluoride electrode, and other chemical methods. The reaction products of the hydroxyapatite stannous fluoride system will be isolated in pure form. Acid solubility studies will be done on these compounds using conventional technics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "I have been committed to a career in basic science since my first exposure to research as an undergraduate student. After completing an MD/PhD program, I went on to pursue surgical residency and then a fellowship in abdominal transplant surgery. To achieve my goal of becoming a successful independent researcher, I recently entered the laboratory of Dr. John Engelhardt, a national authority on the molecular pathophysiology of cystic fibrosis (CF) and strategies to utilize recombinant adenovirus-associated viruses (rAAV) as gene therapy vectors. Dr. Engelhardt has proven track records in both mentoring physician- scientists and securing research funding, and his guidance will be a critical determinant of my success in developing my independent research laboratory. Under Dr. Engelhardt's mentorship, I am focused on combining my basic science training with my medical background to tackle an important clinical question that directly impacts my patients, namely studying the pathophysiology of cystic fibrosis-related diabetes (CFRD). This is a clinically important question, as CF is the most common lethal autosomal recessive disease in Caucasians, resulting from defects in the cystic fibrosis transmembrane conductance regulator (CFTR) channel. This K-award will allow critical protected time for me to fully develop my independent research career. During the period of this award, I intend to: 1) expand my skills in molecular genetics, 2) master the techniques relevant to CF research and learn to generate pathophysiologically important hypotheses about CFRD, 3) develop skill in the implementation of rAAV gene therapies, 4) strengthen my writing and leadership/mentoring skills, and 5) solidify preliminary data for my first R01 application. In CF patients, pancreatic disease leads to diabetes mellitus in up to 50% of adult patients, resulting in reduced life expectancy. Progress in elucidating the pathophysiology of CFRD has been hindered by the lack of an animal model. Dr. Engelhardt's laboratory has recently developed a CF-knockout ferret - the first animal model that develops CFRD. However, CF ferrets have high perinatal mortality from abnormal intestinal and pancreatic function, altered glucose and lipid metabolism, and lung infections. Elevating GI pH appears to attenuate nutritional abnormalities, as the use of proton-pump inhibitors (PPIs) normalizes weight gain during the neonatal period. The CF ferret model appears to be a robust system for the development of CF pancreatic therapies and is the only animal model available for the study of CFRD. We hypothesize that effective treatment of CFRD will lead to reduced nutritional complications in CF ferrets, increased long-term survival, and reduced severity of lung disease. We will test this hypothesis by carrying out the following specific aims: 1) Identify the defects underlying the pancreatic and intestinal pathophysiologies associated with CFRD in the ferret CF model and 2) Establish a CFTR gene replacement therapy that will improve pancreatic function in CF ferrets. To address the goals of Aim 1, we will use standard pancreatic endocrine functional tests to confirm that a subset of CF ferrets develop CFRD and characterize the disease pathophysiology. We will determine if CFTR-mediated bicarbonate (HCO3-) secretion by the pancreatic duct and/or duodenal epithelial cells is reduced, and if so, how this impacts CFRD phenotypic severity. We will also evaluate the activity of compensatory, non-CFTR-based HCO3- secretory pathways using CFTR inhibitors to determine if these channels might influence the severity and/or disease onset of CFRD. To address the goals of Aim 2, we will adapt methods developed for the mouse (application of rAAV vectors encoding the WT ferret CFTR cDNA) for in vivo gene delivery to the pancreas in CFTR-/- ferrets. We will assess the effectiveness of pancreatic CFTR gene replacement in the CFTR-/- ferrets by evaluating serum glucose, insulin, glucagon, endocrine pancreatic function, and pancreatic duct cell physiology. We will also assess the impact of pancreatic CFTR gene therapy by evaluating ferret growth and nutritional parameters in the presence and absence of PPIs and pancreatic enzymes. Finally, we will evaluate the impact of pancreatic CFTR gene therapy on the progression of early pulmonary disease progression in CF ferrets following removal of antibiotics. These studies will assess lung inflammatory mediators (IL-1, IL-8, and TNFa), lung bacterial colonization, and lung histopathology. The major expected outcomes of this study are: 1) a more comprehensive characterization of endocrine pancreas abnormalities in the new CF ferret model, 2) an understanding of the basis of phenotypic variation in CFTR-associated pancreatic disease observed in both the CF ferrets and humans, and 3) the development of a pancreatic gene replacement approach that will aid in dissecting pathophysiology and treatment of CFRD. These results are expected to have a significant positive impact on the field of CF research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The combination of outstanding physical and chemical properties of polyurethanes (PUs) coupled with their biocompatibility have led to their use in a wide range of biomedical applications. In the last decade, concerns regarding the biostability of PUs were raised when some PU pacemaker leads were shown to undergo environmental stress-cracking in humans. Current studies in this laboratory directed toward a fundamental understanding of biocompatibility and biostability of PU elastomers have led to a hypothesis for the cell/polymer feedback control of in vivo biodegradation. In order to confirm and/or modify the hypothesis, experimental procedures will be carried out on PUs of known composition with and without specific additives that enhance biostability. The specific aims of the proposed research are: 1) To elucidate the chemical mechanisms by which cellular degrading agents (radicals, HOCl, acid)- cause hydrolytic and/or oxidative chain cleavage of polyetherurethanes, and other PUs with non-polyether soft segments where the mechanism(s) of biodegradation may be different. Correlative studies will be made with accelerated in Vitro conditions that mimic the in Vivo environment. 2) To characterize the surface degradation after in Vivo and in Vitro exposure, and correlate the results with the effect on performance properties. Methods of material characterization will include attenuated total reflectance infrared spectroscopy, photoacoustic infrared spectroscopy, ESCA, contact angle, gel permeation chromatography, scanning electron microscopy and scanning acoustic microscopy. 3) To ascertain the effect of stress and strain state on the rate of PU biodegradation. The effects of uniaxial and biaxial strain will be examined in vivo and in vitro, and the results will be correlated with creep and fatigue behavior as determined in vitro under oxidative and hydrolytic conditions. 4) To determine the effect of PU surface chemistry and the effect of additives on the adhesion of polymorphonuclear leukocytes and monocytes/macrophages under static and dynamic conditions, with and without the presence of complement. These studies will be correlated with in vivo experiments to examine the effects of antioxidants and macrophage inhibitors on biodegradation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objectives of this grant application are: 1. To develop new methods for the treatment of cholesterol cholelithiasis by investigating the efficacy of ursodeoxycholic acid (UDC) in the dissolution of gallstones in vivo. 2. To characterize the mechanism of the therapeutic action of UDC in comparison to that of chenodeoxycholic acid (CDC) by studying the effects of these two bile acids on bile acid metabolism, and the solubility of cholesterol in bile. 3. To investigate the source and site of the increases in the formation of UDC, which are found in some patients treated with CDC. 4. To study the role of bacterial alterations of bile acid metabolism in the pathogenesis of cholesterol cholelithiasis on the disease model of ileitis. 5. To determine the incidence and type of disturbances of bile acid metabolism in ileitis. Specifically this application includes the following proposals: 1) double blind controlled treatment trial of UDC 2) study of effects of UDC treatment on composition of biliary lipids (lighogenic index) and bile acid kinetics 3) study of the pathway and site of the interconversion between CDC and UDC using labeled CDC, UDC and 7-ketolithocholic acid 4) study of the role of intestinal bacteria in the inter-conversion between CDC and UDC using the cholyl-glycine-1-14C breath test and in vitro techniques 5) study of the relationship between bile acid deconjugation and changes in biliary lipid and bile acid composition in ileitis 6) study of the incidence and site of bile acid deconjugation in ileitis using the cholyl-glycine-1-14C breath test and the measurement of focal excretion of bile acid label.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Phosphatidylinositol-4,5,-bis-phosphate (PIP2) is the precursor of inositol-trisphosphate (IP3), diacylglycerol (DAG), and phosphatidylinositol-trisphosphate (PIP3). It also anchors cytoskeleton and numerous signaling molecules at the plasmalemma, and its metabolism is closely coupled to membrane trafficking. In addition, it modulates profoundly the function of several cardiac ion transporters and channels. This application addresses how PIP2 is regulated in heart and how PIP2 metabolism is related to membrane turnover at the cardiac sarcolemma. As suggested by Preliminary Data, we will test whether PI4-kinases are regulated by serine/threonine phosphorylation and whether lipid phosphatases are regulated by surface membrane insertion and oxygen-dependent proteolysis. As a new experimental model, we have generated transgenic mice with cardiac-specific over-expression of the type2alpha PI4-kinase (PI4K2alpha). This kinase localizes primarily to Golgi and internal membranes, and its over-expression is associated with high-grade cardiac hypertrophy and up-regulation of ECC. We will now test how membrane trafficking to and away from the cardiac sarcolemma is affected using (1) fluorescent membrane dyes, (2) a new amperometric method to monitor surface membrane fusion events, and (3) high resolution capacitance measurements in on-cell patch clamp configuration. Preliminary Data suggests that PKC's may activate PI4K2alpha on internal membranes, thereby initiating movement of vesicles containing lipid phosphatases to the sarcolemma. Insertion at the sarcolemma appears to be activated directly by DAG, whereby subsequent depletion of sarcolemmal PIP2 would prevent endocytosis and favor the expansion of the sarcolemma. Complementary to studies of PI4K2alpha, we will test whether the type 2beta PI4-kinase (PI4K2alpha) is the major sarcolemmal PI4-kinase and whether its regulation may be tied to the regulation of cardiac transporters and channels. Finally, the hypothesis will be tested that PIP2 metabolism is inherently sensitive to membrane tension and curvature, as well as to myocyte stretch (i.e. the cardiac preload).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objectives of this project are to understand the mechanisms and functions of DNA damage checkpoints in embryonic development and the chemotherapy response. Cells execute a protective response to chemotherapies which cause DNA damage or inhibit DNA replication. This involves a complex signaling network driven by the ATR (Ataxia-Telangiectasia and Rad3-related) kinase and its upstream regulator TopBP1 (DNA Topoisomerase ll-Binding Protein 1). Besides serving this protective function, ATR and TopBP1 also maintain genomic integrity during unperturbed DNA replication and embryonic development, and inherited mutations that disrupt the ATR signaling pathway cause human developmental disorders. Thus, pharmacologic inhibition of DNA damage checkpoints to sensitize cancer cells to chemotherapy is likely to cause side effects. TopBP1 has eight BRCT (BRCA1 C-terminal) protein interaction domains and is multifunctional, also acting in DNA replication, DNA repair, and transcription. TopBP1 likely participates in such diverse aspects of DNA metabolism by acting as a central component of multiple functionally distinct subcomplexes, but these various subcomplexes remain to be defined. The current proposal consists of three aims. The first Aim is focused on characterizing an interaction between TopBP1 and an entirely novel protein called TICRR (TopBP1-interacting checkpoint and replication regulator). TICRR interacts with the first two BRCT domains of TopBP1 and is required for DNA replication initiation and cell cycle arrest following DNA damage. The second Aim is focused on identifying and characterizing novel TopBP1 and TICRR protein interactions. The third Aim determines the functions of TICRR and TopBP1 during zebrafish embryonic development. Knowledge from these studies will lead to a better understanding of DNA damage responses to chemotherapy as well as illuminate the molecular underpinnings of human developmental disorders caused by defects in ATR signaling. Understanding differences between how checkpoint proteins function in normal cell proliferation versus in the chemotherapy response will facilitate the development of treatment strategies that inhibit checkpoint signaling in tumors, while leaving normal cells unperturbed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the previous period of grant support lithium (Li) was compared with carbamazepine (CBZ) for the acute and maintenance treatment of mania in patients who received no other adjuvant medications. The results of that study indicated that neither Li nor CBZ were very effective under these circumstances. Sixty-eight percent of patients wert dropped after 6 to 8 weeks of treatment for inadequate therapeutic response. Some differences were identified between the drugs, mainly in the greater degree of sedation and improved behavioral control with CBZ during the first five weeks of the study. Otherwise the two drugs yielded essentially the same therapeutic benefits. No significant toxicity was encountered. We now propose to compare Li-CBZ combined with Li-HAL in the acute and maintenance treatment of mania using essentially the same research design. Patients will again be treated for 8 weeks after which those who are unimproved or still meet study entrance criteria will be dropped from the study. The remainder will be followed for up to one year on double blind medications. At the end of that time patients who have been stable for at least 3 months will be tapered off CBZ or HAL and followed for up to a year on Li alone unless there is clinical worsening or recurrence. The aims of the study are to test hypotheses that Li-CBZ combined is a viable short and long term treatment for manic patients as compared with the standard Li-HAL. If this be so, then exposure to neuroleptics, which is a particular hazard in bipolar patients, can be significantly reduced. We will examine outcome of short and long term treatment in relation to sex, age at onset, other demographic variables, manic proneness, rapid cycling, EEG abnormalities and clinical characteristics. Safety data from the SAFTEE general inquiry, neurological ratings, clinical EEG, EKG and laboratory tests of hematologic, hepatic, renal and thyroid functions will be compared in the two treatment groups. Brain Surface Activity Mapping (BSAM) and neuropsychological assessments will be completed at baseline and after short and long term treatment to compare the two treatment groups and to relate to the degree of therapeutic response. Multivariate analysis will be employed to identify the characteristics of patients who respond favorably or unfavorably to the two treatments. Long term prophylaxis will also be evaluated in term of survival time on the study and continued status on Li alone after withdrawal of CBZ and FM.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Atherosclerosis is an inflammatory disease of artery walls in which innate immune cells play an important role. After infiltrating the endothelium, monocytes differentiate into macrophages, take up modified low- density lipoprotein (LDL), and progressively assume a lipid-loaded, foam cell phenotype. Foam cells secrete pro-inflammatory mediators and contribute to lesion progression. Foam cell formation and associated pro- inflammatory behavior involve transcriptional regulation of many genes, but the specific transcriptional regulatory mechanisms controlling these expression changes in response to various lesion-associated stimuli are not well understood. We will identify transcription factors that regulate foam cell formation using a systems biology approach. Murine macrophages will be stimulated with modified LDL in vitro and transcriptionally profiled. The gene expression patterns will be combined with findings from a variety of transcriptomic, proteomic, and genetic association studies to identify a core list of genes associated with macrophage foam cell formation and atherosclerosis. The promoters of these genes will be analyzed in light of genomic evidence for cis-regulatory function (including DNase I hypersensitive sites in macrophages that will be globally mapped by sequencing), to identify transcription factors that may regulate the core gene set. Four or more transcription factors will then be perturbed by knockout or knockdown, to determine their transcriptional regulatory function in the in vitro foam cell model. The long-term goal of this work is to identify the key transcription factors regulating foam cell formation and to characterize their function. This research project is a core element of a four-year career development plan for Dr. Stephen Ramsey to transition to a research career in which he will use the methods of molecular biology and computational biology to study cellular models relevant to cardiovascular disease. The project will be carried out at the Institute for Systems Biology under the mentorship of Dr. Alan Aderem and Dr. Ilya Shmulevich, and in collaboration with Dr. Elizabeth Gold. Drs. Aderem, Shmulevich and Gold have complementary domains of expertise spanning all aspects of the project, including macrophage biology/immunology, molecular cell biology, computational biology, and cardiovascular disease. They are ideally suited to guide Dr. Ramsey's research and career development as described in the training plan. This mentored career development plan will enable Dr. Ramsey to achieve his career goal of becoming an independent investigator in cardiovascular disease-related research, equipped to use experimental and quantitative methods in a synergistic manner. PUBLIC HEALTH RELEVANCE: Atherosclerosis (a leading cause of heart attacks and stroke) is a chronic inflammatory disease of the artery walls. Atherosclerosis involves accumulation of macrophage white blood cells in the artery walls and the transformation of these cells into lipid-loaded foam cells that promote inflammation. This project aims to identify molecules that regulate gene expression changes in foam cells, which may yield new macrophage-specific drug targets for atherosclerosis. (End of Abstract)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project is concerned with the biochemistry and genetics of heritable diseases of amino acid and organic acid metabolism. It is devoted to the detection and definition of new metabolic diseases and the study of metabolic interactions in these and other metabolic disorders. Emphasis is on identifying elevated or abnormal metabolites and relating them to the pathogenesis of the clinical disease. Characterization of defective or variant enzymes responsible for the altered metabolism is an integral part of the project. The knowledge gained from these studies is of importance in diagnosis and possible treatment of these inherited diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The cerebral cortex, the organ of human intellect, is the site of many mental and neurological dysfunctions, the causes of which can be traced to genetic and acquired developmental abnormalities in early prenatal life. A fundamental question in understanding this phase of corticogenesis is how, after their last cell division in the proliferative ventricular zone, postmitotic neurons achieve their laminar and radial positions in the cortical plate, differentiate into specific phenotypes, generate a repertoire of neurotransmitters and receptors, and establish functional synaptic circuits. Previous findings from research in this program Project include i) development of novel surface neuron-glia junctional proteins that are involved in neuronal migration, ii) demonstration of the role of voltage and ligand-gated Ca2+ channels in neuronal migration, iii) evidence of the role of the cytoskeleton in cell motility, iv) identification of specific proteoglycans that are involved in activity-dependent cortical differentiation, v) discovery of adrenergic receptors in the transient embryonic zones of the primate embryos, vi) demonstration of a cortical role for various neurotransmitter systems ina the regulation and generation of synaptic and neuronal activity in developing forebrain circuits. These findings have generated novel concepts regarding developmental mechanisms or opened new areas of research. The present proposal is based on the premise that these are causally interrelated and overlapping events that can be understood best in relationship to each other . With the support of this Program Project, a team of investigators with expertise in molecular biology, immunocytochemistry, laser microscopy, electron microscopy, neuroanatomy, receptor pharmacology, computer imaging, and electrophysiology have joined forces to explore five major themes: (1) Surface mediated mechanisms of neuronal migration; (2) Regulation of cell phenotype in the developing neocortex; (3) Transmitter regulation of cortical phenotype; (4) Neurotransmitter receptors as sites of morphogenetic activity in the transient embryonic zones of the developing primate cerebral cortex; (5) Development of network activity in thalamocortical circuits. Thr proposed experiments utilize some of the most advanced neurobiological approaches that can, at this time, be applied to the complex developmental problems involved in cortical ontogeny. Our experience is that a program project of this scope promotes communication between investigators with different backgrounds, fosters research collaboration, and enables comprehensive, multifaceted analyses that are either not possible or practical for individual investigators working alone.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It has long been a goal of neurobiologists to understand the mechanisms that regulate development of the mammalian forebrain, as the forebrain is often referred to as the structure whose complexity sets humans apart from other higher-order organisms. To date, numerous signaling pathways have been implicated in the proper patterning and development of the forebrain primordium, the prosencephalon. However, how these pathways are regulated at the molecular level is largely unclear. The retinoic acid (RA) signaling pathway is one such pathway. Though it is clear that retinoic acid signaling is required for proper development of the forebrain, it is unclear how cell type-specific responses to RA are modulated. Additionally, because of redundancy in RA-synthesizing enzymes and RA receptor molecules, it has been difficult to investigate the effects of regional loss of RA-dependent gene transcription. The goal of this study is to utilize a mouse line lacking enzymatic activity of the histone acetyltransferase Gcn5 (Gcn5hat) to better understand the role of RA signaling in border specification in the developing forebrain. Gcn5hat mutants display ectopic structures in the dorsal telencephalon that express thalamic markers, suggesting a massive rostral expansion of the dorsal diencephalon. Gcn5 has been previously implicated in regulating RA signaling and it has been hypothesized that rostral expansion of the developing thalamus is limited by localized dorsal production of RA. Preliminary data suggests that Gcn5 regulates RA signaling through non-epigenetic mechanisms, which calls into question the prevailing hypothesis that Gcn5-mediated histone acetylation is required to promote RA-dependent gene expression. This study aims to address two fundamental questions relating to Gcn5 and its role in RA-mediated forebrain development. First, I will investigate patterning and differentiation in the forebrains of Gcn5hat mutants to determine the exact role of Gcn5 in forebrain development. More specifically, it will determine the requirement of Gcn5 acetyltransferase activity for specifying the proper borders between forebrain structures and will also determine whether these structures go on to produce their appropriate neuronal subtypes. Second, this study will test the hypothesis that Gcn5 acetyltransferase activity is required for proper RA signaling in the developing forebrain and I will determine whether the developmental phenotypes can be modulated by changes in dietary RA levels. Lastly, this study will investigate the molecular mechanism by which Gcn5 regulates RA signaling. I will test the novel hypothesis that Gcn5 promotes RAR-mediated signaling via acetylation of TACC1. This investigation will broaden our understanding of how Gcn5 elicits tissue-specific signaling responses through novel, non-epigenetic mechanisms. Together, these experiments will shed light on new functions of a developmentally critical acetyltransferase, increase our understanding of RA signaling in forebrain development, and identify specific neurodevelopmental processes that require Gcn5, potentially implicating Gcn5 in human developmental, cognitive, and neuropsychiatric diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Aromatic amines are believed to induce tumors in humans and experimental animals because of their conversion to N-hydroxylated derivatives which are further metabolized toreactive metabolites that are capableof altering nucleic acid. The formation of N-acetoxyarylamines has been implicated as the latter metabolic activation step that is crucial to the carcinogenic and mutagenic activities of many of these agents. In the rat, mouse and rabbit, and perhaps other species such as the human and hamster, the terminal activation reaction, O-acetylation, involves the transfer of an acetyl group to the hydroxylamine from either acetyl coenzyme A or an arylhydroxamic acid. The same enzyme is also capable of N-aetylation to yield arylacetamides or arylhydroxamic acids from arylamines or arylhydroxylamines, respectively. The N-, O- and N,O-acetylation activities of the enzymes from different species vary in their rates and relative abilities to carry out these reactions. These differences are significant determinants of the susceptibilities of the target organs of the aromatic amines. The objective of this project is to explore the molecular features responsible for the differences in activation by O-acetylation and, consequently, how these factors mayinfluence the carcinogenic and mutagenic responses to these agents. The specific aims of the project are to: (1. Determine the primary sequence of the rat enzyme bymolecular cloning. Oligonucleotide probes and monoclonal antibodies will be used to screen cDNA libraries. (2. Identify the active site(s) of the enzyme by labelling with reagents specific for sulfhydryl groups and N-hydroxy-chloroacetylaminoarene substrates to identify amino acids that are crucial to the catalytic activity. the monoclonal antibodies will be used to help in establishing the areas of the molecule that are involved in catalysis. (3. Localize the acetyltransferase and mRNA in rat tissues by use of monoclonal antibodies and cDNA probes to identify those cells that may be at particular risk from these compounds. (4. Express acetyltransferase in bacterial and mammalian cells to verify the identity of the sequence and to provide a means by which cells can be provided with a metabolic activation system. (5. Compare the structures of acetyltransferases from other species that differ in their abilities to carry out N-,O- and N,O- acetyltransfer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The protocol subject to this grant application is Orphan Therapeutics Protocol OT-0401, entitled, \"A Double-Blind, Randomized, Placebo-Controlled, Multicenter Phase 3 Study of Intravenous Terlipressin in Patients with Hepatorenal Syndrome (HRS) Type 1.\" The primary objective of this trial is to demonstrate that intravenous terlipressin is safe and effective in the treatment of patients with HRS type 1 when compared to placebo. HRS is the development of renal dysfunction in patients with end-stage liver cirrhosis in the absence of any other cause of renal pathology. HRS type 1 is characterized by rapid deterioration of renal function, with marked increase in serum creatinine, azotemia, low urine output, dilutional hyponatremia and marked sodium retention, and a median survival time of less than 2 weeks. There is no available pharmacologic treatment for HRS type 1 in the U.S., therefore, there is a significant medical need for an effective and safe treatment. Terlipressin has shown activity in nine published clinical trials and retrospective studies (Hadengue 1998, Duhamel 2000, Uriz 2000, Mulkay 2001, Ortega 2002, Halimi 2002, Moreau 2002, Colle 2002, Solanki 2003). Protocol OT-0401 will enroll 120 patients of at least 18 years of age from both genders and all ethnicities and races. Patients' renal function, survival, and transplant status will be assessed to determine efficacy. Adverse events and laboratory assessments will be monitored and recorded to determine the safety of terlipressin in HRS type 1. The pharmacokinetics of terlipressin will also be assessed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "7. Project Summary- Resource Core 1 The goals of the Biostatistics, Informatics, and Translational Resource Core (BITR) are to (1) foster the design of interventions that promote the restoration of and prevent loss of function in older adults, (2) promote the search for mechanisms underlying successful interventions, (3) translate interventions from the laboratory to the clinic and community, (4) design studies so as to maximize treatment fidelity and promote the measure- ment of treatment fidelity in all OAIC studies, and, (5) educate and help junior faculty plan and execute studies. RC-1 will provide an informatics backbone, GERI, that will support all aspects of the University of Maryland Older Americans Independence Center's (UM-OAIC) work. RC-1 will provide biostatistical expertise to UM- OAIC investigators helping ensure that studies are optimally designed and adequately powered, will randomize subjects and help analyze study data. RC-1 will make sure that best statistical practices guide the design and execution of OAIC research, the analysis of data, and the interpretation of the results of the analysis. By pro- moting treatment fidelity we will help assure that the test of our interventions are as efficient as possible and will increase the success of our efforts to translate interventions to the clinic and community. The Core will par- ticipate in Research Working Groups (RWG), which will assist with the design, execution, and oversight of all OAIC research. Research Working Groups, composed of the investigator, the investigator?s mentor if the in- vestigator is a junior faculty member, the study coordinator, one or more members from each resource core, and ad hoc experts, will use the reports produced by GERI as they oversee studies and monitor study pro- gress. The reports will help the RWGs assure that studies are completed in a timely fashion, and will help the RWGs identify studies that have potential for translation from laboratory bench to the clinic and community. GERI will collect and track requests for OAIC services (Request for Services system), will assist with the re- cruiting of subjects (Registry), will track study progress (Tracking Database), collect information describing ad- verse events (Adverse Event Database), and will facilitate quality control (Quality Control and Outlier Detection Systems). We will educate faculty, OAIC scholars (junior faculty) and staff in the principals of statistics and ep- idemiology and education efforts in collaboration with the REC.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The University of Washington is well suited for effective and collaborative participation as a Clinical Center in the Idiopathic Pulmonary Fibrosis Clinical Research Network. The IPF research team at the University of Washington has the experience, access to patients, and collaborative approach necessary to exceed expectations as a clinical site. Specific aims of the IPF CRN at the University of Washington are to foster a national collaborative approach to IPF clinical research, establish an IPF clinical research center which efficiently advances clinical studies and knowledge of the disease, provide an environment for excellent state-of-the-art care for IPF patients including accurate diagnoses and patient follow-up capabilities, formalize the community network for patient recruitment and participation into clinical studies of IPF, improve outcome of IPF patients, and understand disease progression in new-onset and improve outcomes for all IPF patients. (End of Abstract)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mallory Bodies (MB) are cytoplasmic filamentous inclusions seen primarily in alcoholic liver disease, although they may also be found in association with other liver diseases. The primary structural components of MB are a group of epithelial specific intermediate filament cytoskeletal proteins termed keratin polypeptides 8 and 18 (K8/18). The biochemical basis for these abnormally aggregated keratins is presently unknown. A basic understanding of the nature of keratin modification in MB should allow designing rational experiments to determine if changes in keratins are essential for alcohol induced injury and if such changes can be reversed. Our hypothesis is that Mallory Body formation may result from alterations in glycosylation and/or phosphorylation of hepatocyte keratins. This hypothesis is based on finding discrete keratin cytoplasmic dots, that are morphologically similar to MB, when keratin phosphorylation and glycosylation increases during cell mitosis. In addition, desmin bodies which are found in familial myopathy and are derived from the intermediate filament protein desmin, were found to be hyperphosphorlated. In carrying out this proposal, we plan to take advantage of our studies involving the characterization of normal epithelial keratin glycosylation and phosphorylation. For example, we recently showed that keratins 8 and 18 are O-linked glycoproteins with glycosylation sites consisting of single N-acetylglucosamine (GlcNAc) residues. To test our hypothesis, we plan to study the phosphorylation and glycosylation of K8/18 in experimentally induced Mallory Bodies in mice, and in human liver specimen obtained from biopsies or surgical specimen. We also plan to determine at a detailed molecular level if other forms of posttranslational modification such as transglutamination account for MB formation. Finally, we plan to explore the possibility that primary amino acid structural changes occur in keratins of MB.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We hypothesize that a commercially competitive magnetic nanoparticle can be developed to eliminate debris in the preparation of cell cultures, such as stem cells. Current methods to remove unwanted material from healthy cultures rely on magnetic negative selection using Annexin V-based magnetic particles. Annexin V binds to the phospholipid phosphatidylserine (PS) present on the surface of dead cells and cell membrane fragments. A magnetic field is then applied to collect dead cells from the healthy culture. Although commercially successful, Annexin V-based technology has significant drawbacks: (i) it is restricted to removing only PS-containing membranes, (ii) stripped nuclei and oligonucleotides cannot be removed, (iii) Annexin V cannot be autoclaved, (iv) Annexin V activity can degrade over time, (v) Annexin V requires calcium-containing buffers, (vi) is difficul to use in serum, (vii) is inherently unstable due to the sloughing of Annexin V, (viii) is likely t have high endotoxin levels, (ix) is expensive to manufacture and maintain, therefore adding significant cost to the end- user. Our proposed product will eliminate dead cells and cell membrane fragments from culture as effectively as Annexin V-based magnetic particles. Our product will also (i) target and eliminate nucleic acids, (ii) provide greater flexibility in buffe selection, and (iii) tolerate autoclaving to assure sterility. Because the conditions under which a cell sample is prepared will become more stringent as technologies mature into clinical practice, successful completion of this application will have a profound and positive impact on many sectors of biomedicine, including reproductive medicine, diagnostic applications and cell therapy, such as autologus stem cell therapy. PUBLIC HEALTH RELEVANCE: The project will evaluate a novel technology to remove unwanted and harmful debris from healthy cell cultures. Because the conditions under which a cell sample is prepared will become more stringent as technologies mature into clinical practice, our product will have a profound and positive impact on many sectors of biomedicine, including reproductive medicine, diagnostic applications and cell therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY The molecular mechanisms that underpin the development of insulin resistance with age remain poorly defined, yet a better understanding of this process is critical, given the fact that obesity, metabolic disease and diabetes have reached epidemic status in the US. We discovered that alternative splicing of the insulin receptor in humans leads to truncated isoforms that have the potential to inhibit insulin signaling, by acting as decoy receptors or soluble binding proteins. Changes in the frequency of alternative splicing events could provide a novel mechanism by which insulin resistance and metabolic disorders develop. Determination of the physiological significance of these new isoforms is critical in establishing their role in the pathophysiology of insulin resistance with age. However, such studies in mammals are expensive, time consuming and risky. In contrast, the nematode C. elegans provides an economical system in which to rapidly determine their physiological relevance. To this end, we discovered a truncated isoform of the insulin receptor in the worm, termed DAF-2B, that is directly analogous to mammalian IR-C and IR-D. This discovery provides an opportunity to rapidly and economically define the physiological role and regulation of truncated insulin receptors in a genetically tractable organism, prior to initiating studies in mammals. Using a novel splicing reporter strain that permits visualization of differential splicing between the full length and truncated daf-2 isoforms in vivo, we find that daf-2b expression varies across tissues and through development, suggesting that these transcripts are subject to specific regulatory mechanisms. Importantly, over-expression of the daf-2b cDNA in worms produces phenotypes consistent with attenuated insulin signaling, including increased nuclear localization of the FOXO transcription factor DAF-16 and extended lifespan. Conversely, preliminary analysis of a daf-2b deletion strain, generated using CRISPR, indicates that loss of daf-2b confers insulin sensitivity. Finally, using our daf-2b reporter strain, we identified specific splicing factors that alter the expression of daf-2b, including one whose activity may be directly regulated by insulin signaling and Akt phosphorylation. Exploration of the mechanism by which splicing factor activity regulates the expression of truncated IR will identify novel points of regulation and intervention in this pathway. We hypothesize that aberrant or mis-regulated expression of truncated IR-C and IR-D isoforms in mammals could be causally involved in the pathogenesis of insulin resistance, diabetes and other forms of metabolic disease. Thus, determination of the physiological role and regulation of truncated insulin receptor isoforms in worms will provide a solid foundation from which to establish studies of equivalent isoforms in mammals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Proneural basic helix-loop-helix (bHLH) transcription factors function as transcriptional activators and are required for the development of specific retinal neuron subtypes. For example, the Ath5 gene is required for the differentiation of retinal ganglion cells (RGC), an early born cell type, while NeuroD contributes to the development of amacrine cells, a later born cell type. However, the mechanisms by which bHLH factors promote retinal neuron differentiation, and how they contribute to the ordered sequence of retinal histogenesis is less well defined. We have identified both transcriptional and post-translational mechanisms that regulate when during retinal development bHLH factors are active. If Ath5 or NeuroD are active during early retinal development they promote differentiation of RGCs, however if they are active later, they promote later born cell types, consistent with the idea that progenitor competence changes over developmental time. Furthermore we found that bHLH factors regulate retinal neurogenesis by activating the expression of downstream target genes that are involved in executing the neuronal differentiation program within retinal progenitor cells. [unreadable] [unreadable] Much remains to be learned about how the activity of bHLH factors is regulated and which genes are transcribed in response to bHLH activity. We propose to investigate both transcriptional and post-translational mechanisms for regulating bHLH activity during retinal neurogenesis. In addition, since our understanding of the genetic pathway downstream of proneural bHLH function is incomplete, we have performed a screen to identify additional genes that are regulated by Ath5 and NeuroD. We propose to analyze these putative targets and determine how they contribute to retinal neurogenesis. [unreadable] [unreadable] By defining the mechanisms controlling normal retinogenesis, we may gain a better understanding of how these problems can be disrupted in pathological situations. For example, a number of congenital disorders of vision, such as retinal dystrophies, may be caused by problems with the early genesis and differentiation of retinal neurons. Ultimately, this work may help us understand and manipulate the differentiation of retinal stem cells or progenitors for the treatment of degenerative diseases of the retina, such as glaucoma and retinitis pigmentosa. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Traumatic brain injury (TBI) is a major public health problem that raises the need to accurately measure its effects non-invasively. A biological target mechanistically linked to TBI and suitable for molecular imaging with diagnostic and prognostic implications could provide a quantitative non-invasive biomarker for TBI. Presently, such a biomarker does not exist. Multiple lines of evidence have demonstrated that TBI triggers a biochemical cascade that reduces the density of cerebral alpha7-nAChRs, one of the major subtypes of nicotinic acetylcholine receptors. Preliminary data from our laboratory has demonstrated that our novel positron-emission tomography (PET) radiotracer [18F]ASEM can accurately report the distribution of alpha7-nAChRs in control animals and that it exhibits strikin bilateral reduction of binding (32-73%) in rats with controlled cortical impact (CCI) TBI model. Currently, [18F]ASEM is the only alpha7-nAChR radioligand that has demonstrated excellent imaging properties with high specific binding in rodent and non-human primate PET studies. The main objective of this proposal is to validate the PET imaging properties of [18F]ASEM in two models of TBI in rats, focal - CCI and diffuse - impact acceleration (IA). 1) Cerebral alpha7-nAChR binding and distribution will be measured with [18F]ASEM - PET in CCI and IA models of TBI in rats. In agreement with previous in vitro alpha7-nAChR autoradiography data in animals with TBI reported by others and our preliminary PET results, our hypothesis is that the cerebral specific binding of [18F]ASEM in TBI will be significantly reduced. The reduction of the [18F]ASEM specific binding will correlate with the TBI severity and progression. 2) The PET results will be confirmed by immunofluorescent staining that will demonstrate significant alteration of the alpha7-nAChR protein in TBI rat brain tissue (in neurons and glia). The experimental focus of this proposal is to carry out proof-of-concept studies and obtain evidence of the suitability of [18F]ASEM for quantification of alpha7-nAChRs by PET in TBI models in rats. Our future goal is to evaluate alpha7-nAChRs as a potential biomarker of TBI in humans by studying the course of receptor changes in TBI and their sensitivity to the effects of various treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Malformations of cortical development are a major cause of epilepsy, mental retardation, autism, and related neurological disorders. Malformations arise from defects of cortical cell migration, axon tract formation, and differentiation. These fundamental processes are necessary to produce the characteristic laminar structure, precise neural circuitry, and specialized neuron types in the cortex, respectively. Tbr1 is a T-domain transcription factor that is necessary for fundamental developmental processes in the neocortex (the largest part of the cortex). Tbr1 deficient mice have a severe migration disorder (which resembles a form of human lissencephaly), as well as defects of axon pathfinding (affecting the callosal, corticospinal, corticothalamic, and thalamocortical projections) and neuronal differentiation. The broad goal of this project is to elucidate underlying mechanisms of cortical development by studying the role of Tbr1. Our hypotheses postulate that Tbr1 regulates specific aspects of glutamatergic neuron migration, axon pathfinding, and differentiation. This project has three specific aims. Aim 1 is to define the role of Tbr1 in cortical cell migration. This will be accomplished by transplantation studies. Aim 2 is to define the role of Tbr1 in formation of corticothalamic and thalamocortical axon connections. Using a novel in vitro assay, we will determine if Tbr1 regulates cortical and thalamic axon responses, guidance cues, or both. Also, by overexpressing Tbr1 in embryos, we will resolve whether Tbr1 specifies cortical axon connections cell autonomously. Aim 3 is to define the role of Tbr1 in glutamatergic differentiation and layer-specific fate choices. Gain-of-function assays will be used to determine if Tbr1 induces glutamatergic differentiation, suppresses GABAergic differentiation, and (at high levels) induces phenotypes of deep-layer neuron types. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research is designed to test the ability of a skeletal muscle-powered (rectus abdominis) assist ventricle to augment cardiac output in a chronic study in dogs. The research is based upon a series of studies performed in our laboratory and elsewhere that have demonstrated the feasibility of skeletal muscle powered circulatory assistance. The optimum conduit design in which selected skeletal muscles would most efficiently assist the circulation has not been determined, and the problems of muscle fatigue and lack of a suitable pulse generator to stimulate and coordinate skeletal muscle work are limitations to existing assist systems. The proposed research is divided into 3 separate studies which address these limitations. First we will compare 3 different regimens for converting untrained rectus abdominis muscle to a fatigue-resistant muscle without loss of strength. Feasibility data that show electrically stimulated tetanic contractions against a compressible pouch, at rates of 6-90/min, may prevent muscle injury and optimize muscle conditioning. Second, we will extend preliminary work on a custom designed pulse generator that can sense cardiac activity and be programmed to deliver a specific stimulus. Such a device would have multiple leads delivering a constant current pulse and would be powerful enough to chronically stimulate tetanic muscle contractions via the motor nerve in our rectus ventricle assist system. Third, we will combine our earlier findings, the findings of other investigators, and the results of proposed studies to determine the effectiveness of the rectus assist ventricle for chronically augmenting cardiac output in dogs. The results of this research will provide essential data for future clinical trials of this promising treatment modality for intractable chronic heart failure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hematopoietic stem cells (HSCs) give rise to each of the definitive blood lineages found in the adult vertebrate. Runxl is required for definitive HSC specification during embryogenesis and regulates adult HSC homeostasis. I recently performed a chemical genetic screen for modifiers of runx1+ HSC formation in zebrafish and identified the prostaglandin pathway as a conserved vertebrate regulator of HSCs. A stable prostaglandin E2 (PGE2) derivative was found to be a potent inducer of both zebrafish and murine HSCs and is currently being developed for use in human clinical trials for ex vivo expansion of HSCs prior to transplantation. To further characterize mechanisms controlling HSC induction in the vertebrate embryo, I plan to examine the interaction of the wnt and prostaglandin pathways in HSC formation in the AGM. I hypothesize that the prostaglandin pathway regulates wnt mediated induction of HSCs in the embryo and in adult HSC homeostasis. To characterize the role of the wnt pathway in HSC formation, I will use several heatshock inducible transgenic zebrafish lines that express activators or repressers of wnt signaling. Through chemical manipulation as well as morpholino oligonucleotide (MO) knock-down of PGE2production, I will test how prostaglandin signaling can enhance or suppress wnt mediated HSC formation and determine the conservation of this interaction in adult marrow and human cord blood HSC homeostasis. Taken together, these studies will enhance our understanding of the mechanisms regulating HSC formation and homeostasis. Stimulation of these pathways may prove beneficial for the treatment of patients with hematological disorders, bone marrow failure or in recovery after transplantation, while pathway inhibition might be useful in the treatment of leukemia. A K01 award will aid me in achieving my goal of becoming an independent scientist in an academic setting studying the regulation of HSC induction and function. PUBLIC HEALTH RELEVANCE: Hematopoietic stem cells form the foundation of our blood and immune system;the formation and function of these cells are carefully controlled in the body. The proposed research will help to identify mechanisms that regulate the birth and propagation of these stem cells. This work has great relevance for the development of novel agents to regulate leukemia, and for recovery from chemotherapy and bone marrow transplant.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "(3H)-labeled indicine N-oxide, indicine, retronecine, and retronecine N-oxide have been chemically synthesized for use in biological studies. The synthesis of (3H) indicine N-oxide proceeded from incidine N-oxide through a process involving reduction to indicine, ester cleavage, oxidation of the exocyclic carbon on the pyrrolizidine moiety, reduction with lithium aluminum tritide, re-esterification, and re-oxidation to the N-oxide. In the subsequent biological studies, the enzymatic, anaerobic reduction of indicine N-oxide to indicine has been demonstrated, and studies on the microsomal oxidation of (3H)indicine and (3H)retronecine to activated derivatives have been performed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Voltage-gated potassium (Kv) channels form a large and diverse family of ion channels that are involved in regulating the resting membrane potential, the action potential waveform, neurotransmitter release and rhythmic firing patterns of neurons. Their pivotal role is highlighted by several inherited human diseases caused by mutations in Kv channel genes. Among the many different types of Kv channels, Kv3-type channels display unique biophysical properties: very rapid activation and deactivation kinetics, high thresholds of activation and large unit conductances, properties that enable neurons to fire narrow actions potentials at extremely high frequencies. Among Kv3-type channels, subunits for Kv3.1 and Kv3.3 are highly expressed in the cerebellum, and some of the behaviorally observed phenotypes in Kv3-null mutant mice are characteristic of cerebellar dysfunction such as impaired motor performance and, of relevance to this proposal, very high alcohol sensitivity. We have previously shown that altered firing patterns of cerebellar Purkinje cells are responsible for impaired motor function yet not for heightened alcohol sensitivity. Here, we propose experiments to test the hypothesis that the extreme alcohol sensitivity of Kv3.1/Kv3.3-double mutants originates from changes in granule cell physiology, neurons that normally express high levels of Kv3.1 and Kv3.3 channel subunits. We will use a molecular biological approach to localize the neuronal origin of high alcohol sensitivity in the cerebellum of Kv3-mutant mice. In future work, this approach will enable us to study the altered neuronal physiology in brain-slice preparations and to correlate changes in neuronal firing patterns with the corresponding behavioral alterations, in particular with the intoxicating effects of alcohol. PUBLIC HEALTH RELEVANCE: We have recently developed potassium channel-mutant mice that are very sensitive to low concentrations of alcohol. Hence, these mice will serve as well-defined rodent models to study the electrophysiological changes, i.e., altered neuronal firing patterns, that cause extreme alcohol sensitivity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the last two years, there has been an explosion of novel technologies for the acquisition and interpretation of genomewide human molecular genetic data, including development of whole genome association (WGA) microarrays and statistical methods, as well as the publication of the HapMap to provide a context for accelerating the analysis and synthesis of such genomics data. Having published the first WGA study of schizophrenia (Lencz et al. 2007), we are cognizant of the technical and statistical complexities involved in applying these novel technologies. Therefore, the Special Scientific Procedures Core: Genomics plays a critical supportive role in the proposed CIDAR. The Genomics Core has three primary aims, necessary to the completion of CIDAR goals: (1) to provide basic laboratory services to CIDAR investigators such as blood draws, DMA extraction, creation of immortalized cell lines, and sample storage; (2) to provide methodological expertise in state-of-the-art genotyping and sequencing platforms, including high-precision QA/QC procedures; and (3) to provide advanced statistical support, including development of new techniques, relevant to this large scale, genomewide data collection. Laboratory services are well-equipped to store, track, and genotype large numbers (thousands) of patient samples rapidly and accurately using high-throughput technologies and robotics. Genotyping platforms include scanners for both Affymetrix microarray chip sets and Illumina Bead Arrays, and can support high density whole genome association studies and comprehensive SNP tagging strategies. In the last 18 months, more than 1 billion genotypes have been generated in our facility, and maintenance of high standards for genotyping QA/QC is a central focus of the Core, resulting in several top-tier publications. Statistical services provide expertise and published track records in critical issues for large genetic datasets, including: data reduction and complexity reduction methods, haplotype estimation and haplotype tagging strategies, gene-gene and genotype-phenotype interactions, and Bayesian and other multivariate modelling techniques. Development of novel methods (such as whole genome homozygosity analysis and analysis of copy number variation) is a priority of the Core. The Genomics Core will work together with both the Operations and Clinical Assessment Core, and the Research Methods Core: Cognitive Neuroscience, to apply genomics to prediction of both clinical treatment response phenotypes and neuroscientific endophenotypes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this research is to quantify the bone status, medical history, nutrient intake and caffeine intake of athe cohort of postmenopausal women who participated in the Penn State \"Women, Caffeineand Bone study\" two years ago. The initial study was cross- sectional in nature with a proposed recruitment of 150 healthy women ages 55-70 who had minimal or no exposure to hormone replacement therapy. By design, equal numbers of low (equivalent to 0-2 cups caffeinated coffee per day), moderate (equivalent to 3-4 cups of caffeinated coffee per day), and high (equivalent to 5 or more cups caffeinated coffee per day) caffeine users were recruited. Recruitment was performed such that each of the three subgroups was balanced with respect to habitual exercise patterns of the participants. A total of 146 women participated. Of these, eight were excluded for failing to meet one or more of the inclusion criteria. Studies of total body bone measurements and both hip bone measurements showed no relationship dietary caffeine intake and any bone measurement. The present study is a follow-up. We will make the same measurements on as many of the original cohort as possible, as as possible to the two year anniversary of each particpant's first clinic visit. The major anticipated finding from this follow-up study will be to establish whether older women with different caffeine intakes have the same or different rates of postmenopausal bone loss.In addition, we will construct the first longitudinal data base on the relationships among habitual caffeine intake, general health, and overall nutrient intake of postmenopausal women.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 60% of spontaneous abortions appear to result from chromosome aberrations associated with the zygote/fetus. One source of these abnormalities is inaccurate chromosome disjunction during the reduction division of meiosis. Genetic studies in a number of eukaryotes has clearly linked the heterodimeric meiosis-specific MutS homologs, MSH4-MSH5, to accurate chromosome segregation in meiosis I. In the last granting period we made significant progress in understanding the function of these genes/proteins. In this renewal application we propose to continue the biophysical analysis of the human hMSH4-hMSH5 heterodimeric proteins. We will determine the most relevant heterodimeric human MutL homolog (hMLH1-hPMS 1, hMLH1-hPMS2, or hMLH1-hMLH3) that is associated with hMSH4-hMSH5 function. In addition, we will expand our biochemical analysis of hMSH4-hMSH5 meiosis I functions to include biologically relevant recombination proteins (hRAD51; hDMC1; hRAD54; BLM1; hRPA) and synaptonemal complex proteins (SCP1; SCP2; SCP3). Finally, to confirm our in vitro studies, we will develop immunological and peptide competition reagent to examine the cellular localization and interaction of these proteins during meiosis. We propose four Specific Aims: I.) domain and mutational analysis of hMSH4-hMSH5; II.) determine the functional interaction (s) between hMSH4-hMSH5 and the heterodimeric human MutL homologs hMLH1-hPMS 1, hMLH1- hPMS2, and hMLH1-hMLH3; III.) examine the functional and biologically relevant interaction(s) between hMSH4-hMSH5 and meiosis-specific chromosome pairing and recombination proteins; and IV.) identification of meiosis-specific MSH/MLH pathway components. We will use innovative methods that to our knowledge are unique to this proposal, such as comparative real-time binding with Surface Plasmon Resonance and Total Internal Reflectance as well as interaction and surface mapping via free-radical footprinting and mass spectral analysis. These studies will provide a substantial foundation for understanding the meiosis-specific function(s) of the human MutS and MutL homologs in accurate chromosome segregation and the maintenance of mammalian fertility.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Bile acids may help to coordinately regulate cholesterol and phospholipid metabolism in the hepatocyte by modulating the expression of genes encoding bile acid biosynthetic enzymes and phospholipid transport proteins. The mechanisms by which bile acids coordinate the expression of these genes in order to maintain cholesterol homeostasis and the secretion of a specific ratio of cholesterol: bile acids: phospholipids into bile is unclear, but may have important implications in the pathogenesis of cholesterol gallstone formation, hypercholesterolemia, and cholestatic liver diseases. The objectives of the present grant application are: (1) To identify bile acid-activated signaling cascades within the hepatocyte and determine if the activation occurs via a protein kinase C-dependent or independent pathway; (2) To determine if bile acids activate receptor protein tyrosine kinases (RPTK) in primary hepatocytes and the bile acid structural requirements for activation; (3) To determine which bile acid-activated down-stream signaling cascade(s) regulate genes involved in maintaining cholesterol and phospholipid homeostasis in the hepatocyte, including: CYP7a1, CYP8b1, LDL receptor, neutral cholesterol ester hydrolase (CEH) and mdr2 phospholipid transporter; (4) To determine if the regulation of CYP7a1 by bile acids and cholesterol metabolism is altered in JNK-1 and JNK-2 knock-out mice and if over-expression of a dominant negative JNK-1 or c-Jun alters CYP7a1 regulation by bile acids in bile acid fed rats and in a chronic bile fistula rat model. Efforts will be made to determine if bile acids activated signaling pathways \"cross-talk\" with nuclear receptors i.e. farnesoid X receptor that are also activated by bile acids.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Using mouse model systems, studies have been carried out to determine how non-oncogene-containing retroviruses cause erythroleukemia and why certain mice are resistant to the pathogenic process. The Friend spleen focus-forming virus (SFFV) causes a rapid erythroleukemia due to the unique envelope gene that it carries. The SFFV envelope glycoprotein alters the growth of erythroid cells by rendering the cells independent of the erythroid hormone erythropoietin (Epo). In order to determine whether the SFFV envelope glycoprotein is activating the Epo signal transduction pathway, studies were carried out to compare erythroid cells stimulated with Epo or infected with SFFV for expression of tyrosine- phosphorylated proteins and DNA-binding proteins. Both Epo and SFFV were shown to induce the tyrosine phosphorylation of several common substrates, one of which appears to be a DNA-binding protein related to the class of transcription factors designated STAT proteins. No differences could be detected in the Epo signal transduction pathways between strains of mice that are resistant or susceptible to the effects of SFFV. Studies were also carried out on a mouse resistance gene, Rmcfr, that prevents the development of erythroleukemia induced by Friend murine leukemia virus (F-MuLV). It was previously shown that F-MuLV- induced erythroleukemia requires the generation and replication of polytropic MuLVs. Molecular cloning of the Rmcf resistance gene now indicates that it is the envelope gene of an endogenous modified polytropic virus. Expression of this gene apparently blocks infection of polytropic MuLVs by a mechanism analogous to viral interference.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We several projects. 1. A dogma of innate immunity is that neutrophils use chemoattractant GPCRs to chase bacteria through chemotaxis and then use phagocytic receptors coupled with tyrosine kinases to destroy opsonized bacteria via phagocytosis. Our current work has changed this dogma by showing that G-protein-coupled formyl peptide receptors (FPRs) directly mediate neutrophil phagocytosis. Mouse neutrophils lacking formyl peptide receptors (Fpr1/2-/-) are defective in the phagocytosis of E. coli and the chemoattractant fMLP-coated beads. fMLP immobilized on the surface of a bead interacts with FPRs triggers a Ca2+ response, and induces actin polymerization to form a phagocytic cup for engulfment of the bead. Chemoattractant GPCR/Gi signaling and phagocytic receptor/tyrosine kinase signaling work independently to promote phagocytosis of beads coated with either chemoattractants or IgG opsonins. Thus, in addition to phagocytic receptor-mediated phagocytosis, neutrophils also utilize the chemoattractant GPCR/Gi signaling to mediate phagocytosis to fight invading bacteria (Wen etal, 2019). 2. Neutrophils sense and migrate through a large range of chemoattractant gradient through an adaptation mechanism. Here, we reveal CPARI, a negative regulator of Ras, that controls GPCR-stimulated Ras signaling in human neutrophils. Cells lacking CAPRI (caprikd) exhibit significantly increased phosphorylation of AKT, GSK3, and cofilin, leading to excessive actin polymerization and subsequent defects in neutrophil chemotaxis. The caprikd cells display chemotaxis defects only in high concentration, but not in low-concentration gradient, remarkably, show better chemotaxis in sub-responsive concentration of chemoattractant gradient due to their higher sensitivity. Taken together, we reveal that CAPRI controls GPCR-mediated adaptation and downshifts the sensitivity of neutrophils for Chemotaxis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Brain death (BD) and ischemia reperfusion injury (IRI) are unavoidable consequences of heart transplantation. Brain death produces profound physiologic derangements and the systemic effects of this central injury, although little studied, are known to contribute to peripheral organ ischemia, the upregulation of adhesion molecules, cytokine expression and leukocyte accumulation within the donor heart. Brain death induced inflammation in the donor also renders the heart more susceptible to IRI in the recipient, and there is evidence to indicate that both of these injurious events have negative long-term consequences with regard to allograft survival. Our working hypothesis is that complement plays a central role in causing myocardial BD- induced inflammation and injury, and enhances IRI in the recipient. We propose that a complement inhibitory strategy applied to the donor (in addition to the recipient) will provide protection from inflammation and injury, and as a consequence, will improve long-term graft survival due to decreased graft immunogenicity and host alloresponsiveness. We propose to utilize relevant mouse models to determine the role of complement in myocardial brain death induced injury (BDI) and in IRI following heart transplantation. Our investigations will focus on complement effector mechanisms and in vivo interactions between complement and P-selectin, an adhesion molecule that is strongly implicated in myocardial IRI and that is expressed in the heart following BD. We further propose to develop novel therapeutic strategies based on targeted complement inhibition and P-selectin antagonism, and to characterize the inhibitors in our newly developed mouse model of heart transplantation incorporating donor BD. We specifically propose to: 1. Determine complement effector mechanism(s) involved in myocardial injury following brain death 2. Develop and characterize novel therapeutic strategies based on P-selectin targeted complement inhibition and P-selectin antagonism and, 3. Determine the effect of anti-complement therapy in the BD donor, the recipient, or both, on the severity of myocardial IRI after heart transplantation and on the development of an alloimmune response and acute rejection of the graft.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In addiction a set of perceptual representation (drugs and related paraphernalia) is tagged as salient and repeatedly selected for conscious processing. Understanding the neural systems by which perceptual representations are tagged as salient might allow better understanding of how these systems are disrupted in addiction. This proposal employs dense array ERPs and statistical, topographic, and dipole analyses to study the neural systems that attach salience to perceptual representations. Detecting task-relevant stimuli may require interaction between an orbito-frontal salience evaluation system and perceptual representations in the posterior brain. There is a potential ERP index of this orbito-frontal/posterior interaction: an inferior prefrontal P2a and a coincident posterior N2b. If the P2a indexes salience evaluation it should be independent of the stimulus and response modalities. If the N2b indexes perceptual processing it should be dependent upon stimulus feature. We test this model by manipulating target-defining feature, response type, and target salience. The task manipulations present relevant and irrelevant stimuli in different tasks: Passive, Silent Count, Keypress, and Withhold Keypress. In the Passive task there is no target detection so there should be no P2a. In the Count and Withhold tasks there is detection but no motor response; in the Keypress task there is both detection and a motor response. Since the perceptual and salience processing demands are equivalent we predict an equal P2a and N2b across active tasks. In the stimulus manipulation experiments the target-defining feature is changed. In one task subjects respond to visual stimuli occurring at a specific location while in another they respond to specific objects regardless of location. In a third task subjects respond to tones. We predict equivalent P2a's across target modalities but different topographic distributions of the N2b depending upon the salient feature. The final experiments manipulate the salience value of the stimulus. One study presents objects with no, small, or large reward value. The P2a should be larger to the more valuable stimuli with a constant N2b. The other study presents stimuli in different locations in salient and non-salient blocks. Constricted attention in the salient blocks should produce a larger P2a and N2b to foveal stimuli.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Autoimmune hepatitis (AIH) is a chronic disease characterized by recurrent hepatocellular injury, circulating autoantibodies, and autoreactiv T-cells. Despite established guidelines for the diagnosis of AIH, key triggers and immunologic pathways are poorly understood. Therapy remains dependent on chronic glucocorticoids with numerous associated adverse effects. Further, disease monitoring is sub-optimal and there is a need for predictive immune biomarkers to link immunopathogenesis to the clinical status. Macrophage Migration Inhibitory Factor (MIF) is a pro-inflammatory cytokine that mediates the host response to infection and stress by activating key innate and adaptive immune pathways. MIF's bioactivity and MIF polymorphisms have been implicated in many autoimmune disorders. Applying this to the field of hepatology, the applicant initiated the first investigation of MIF autoimmune liver diseases. This K08 proposal is directly based on that work, now published in Hepatology, under the hypothesis that MIF plays an active role in AIH by directly inducing and sustaining the inflammatory cascade through interactions with T-cells. Anti-MIF therapy and MIF modulation by a neutralizing MIF receptor (CD74) are hypothesized to be protective. Further, MIF polymorphisms are hypothesized to correlate with disease severity, and longitudinal serum MIF and CD74 levels may reflect AIH disease activity. Accordingly, the aims of this proposal are to: 1) Define the rol of MIF in signaling pathways of immune-mediated liver inflammation in a mouse model of T-cell hepatitis~ 2) Define the cellular mechanisms responsible for release of the MIF receptor CD74 and its modulating effect on MIF bioactivity~ and 3) Define the relationship of functional MIF genetic polymorphisms, and MIF and CD74 serum levels, to the disease course in AIH patients. Supporting data indicates that MIF absence in knockout mice protects against T-cell liver injury, and that a small-molecule MIF inhibitor is similarly protective. The circulating formof CD74 inhibits MIF bioactivity and is released from hepatic cells following stimulation i vitro. Finally, a genetic-clinical relationship between a high-risk MIF allele (-173*C) nd increase in both serum ALT and prednisone requirements was found in AIH patients from the US and from Japan. Based on these results, the K08 proposal will define a key mechanistic role for MIF in the immunoregulation of AIH, enabling new strategies for therapy and disease management. To accomplish this, a mentored and integrated five-year strategic plan for research and training will enable the applicant to experimentally test these hypotheses and to develop into an independent investigator devoted to translational hepatology. The application of MIF to AIH is novel and independent from the focus of both co-mentors, Drs. James Boyer and Richard Bucala. The collaborative environment at Yale University, combining training experience in hepatology and immunobiology, represents an ideal setting in which to conduct this mentored career development project in translational hepatology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During female meiosis, 3/4 of the chromosomes are eliminated and only 1/4 of the chromosomes are inherited by a single egg. In contrast, all chromosomes are distributed among 4 sperm during male meiosis. The elimination of 3/4 of the genome to allow inheritance of only 1/4 of the chromosomes is conserved in all animal phyla, suggesting some fundamental selective advantage. The long-term goals of this project are to elucidate the molecular mechanisms of chromosome elimination and elucidate the selective advantages of asymmetric meiotic division. Errors in meiosis lead to the absence of one chromosome (monosomy) or the presence of an extra chromosome (trisomy) in 10-30% of human conceptions with the majority of these aneuploidies leading to embryonic death. By elucidating the mechanisms of meiotic chromosome elimination in C. elegans, we will identify mechanisms likely to be defective during human meiosis. In this project we will pursue 3 specific aims: 1. Elucidate mechanisms that prevent the sperm from interacting with the oocyte meiotic spindle. 2. Determine how the meiotic spindle is attached at the cortex to facilitate extrusion of chromosomes into polar bodies. 3. Determine the mechanisms by which unpaired chromosomes are selectively extruded into polar bodies to correct trisomy. These aims will be addressed by time-lapse imaging of fluorescent protein fusions within meiotic embryos that have been depleted of key cytoskeletal regulators and cell-cycle regulators by RNA interference.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The main project during the period was a study of the influence of nearest neighbor interactions on enzyme kinetics. From the point of view of statistical physics, this work is an extension of the Ising problem (lattice of 2-state interacting units at equilibrium) to multi-state units at a steady-state arbitrarily far from equilibruim. Transients can also be studied, but we have spent little time on this aspect so far. The equilibrium problem itself is famous for its mathematical difficulty. Except for small systems and one-dimensional special cases, we have been forced to use the Monte Carlo method and the mean field (Bragg-Williams) approximation in the steady-state work. Several novel critical and phase transition properties have been found in these steady-state systems. The behavioral of the enzyme flux is of particular interest.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The topic of research is the neurobiology of affiliation, one of the domains of social behavior. The candidate's previous clinical research has provided empirical support for the use of diminished social affiliative behaviors as a marker for the primary, enduring negative symptoms, or deficit syndrome, of schizophrenia. The fundamental neurobiology of social attachment and asociality has potential relevance to the asociality of schizophrenia, and is of great importance to other mental illnesses. Understanding the basic neurobiology of affiliation and asociality could lead to the rational development of pharmacological treatments. Training will consist of formal course work, directed readings, and supervised research. The research will concern the neural basis of social affiliative behaviors in the highly social rodent, Microtus ochrogaster. Site-specific lesions of the nuclear groups of the amygdala will be used to begin investigation of this circuit.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "With increasing incidence of antibiotic resistance, development of new therapies against bacterial pathogens is essential for global public health. The bacterial type III secretion system (T3SS) represents an excellent drug target because it is externally accessible to small molecules and enables virulence of Pseudomonas, Salmonella, Chlamydia, and numerous other important pathogens. We have developed a high throughput screening pipeline to discover T3SS inhibitors and have shown the robustness of our approach through pilot screens identifying three classes of compounds active against the T3SS. We now propose to broaden our scope and screen three unique libraries comprising ~58,000 natural product fractions developed by members of our consortium as well as two commercial synthetic chemical libraries. According to the CDC, every year over 50,000 healthcare-associated Pseudomonas aeruginosa infections occur in the U.S., >6,000 of which are caused by multidrug resistant strains. To identify Pseudomonas T3SS inhibitors and potential future therapeutics, we will carry out the following aims. In Aim 1, we will implement our primary screen and counterscreens to identify natural product fractions and synthetic compounds with specific T3SS inhibitory activity. In Aim 2, we will validate hits identified in Aim 1, using three orthogonal distinct approaches. The identity and structure of bioactive natural products will be determined and initial structure activity analysis performed on identified synthetic scaffolds. Prioritized compounds will be purified or synthesized and evaluated for off target activity, if any, as well as breadth of activity against T3SSs in multiple relevant pathogens. In Aim 3, mode of action will be determined, using parallel genetic and biochemical approaches. This rigorous strategy will provide ~10 T3SS inhibitor chemical probes with identified molecular targets active against P. aeruginosa and potentially other pathogens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Novel Monte Carlo techniques are being developed to quantitatively determine the tissue volume sampled by non-invasive diffuse imaging modalities. Novel features of these methods include the use of expected value estimators and the explicit consideration of the number and location of photon interactions within the turbid medium. The computation of tissue sampling volumes and its variation with tissue optical properties and instrument configuration parameters greatly aid in the design of novel probes customized for a particular application.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of the proposed research is to continue a systematic investigation of the biotransformation of drugs and foreign chemicals (xenobiotics) in tissues of the human foeto-placental unit at various gestational stages. Special consideration will be given to the analysis of NADPH-dependent mixed-function oxidation reactions involving xenobiotics, steroids and lipids and studies of the interrelationships among these metabolic processes. An attempt will be made to define the characteristics of placental fetal hepatic and fetal adrenal cytochrome P-450 in terms of their spectral properties, catalytic functions, and pharmacologic-physiologic roles. Conjugation-hydrolysis reactions and their possible relationships to the transport of drugs between maternal and fetal circulations also will be studied in greater detail. The importance of such studies is indicated by the known role of environmental chemicals (including drugs and other xenobiotics) as principal interacting etiologic factors in the emanation of birth defects in humans. The drug-metabolizing capacity of the placenta and fetus would be expected to be an important determinant of the pharmacologic-toxicologic effects of drus on the developing conceptus as well as on the pregnant female. The long-term research objective is to provide a better understanding of the basic mechanisms of placental function such that a more rational approach to drug therapy in the pregnant woman, unborn, and newborn child might be developed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this core resource facility is to provide ongoing support for the clinical immunotherapy program in the Surgery Branch of the National Cancer Institute. The laboratory is managed by two co-investigators, Drs. Robert Somerville and John Wunderlich, and each investigator has submitted the same annual report. The major effort in the laboratory is producing, ex vivo, large numbers of human anticancer T lymphocytes that are used in adoptive cell therapy for patients enrolled in Surgery Branch clinical trials. All of the patients have metastatic cancer, primarily melanoma. More recent Surgery Branch clinical trials include cancers of the gastrointestinal tract, lung cancer, cancers induced by human papillomavirus, and cancers of multiple types that have particular immunologically targetable antigens in common. Frequently, ten to fifty billion cells are used for each treatment. The anticancer cells are generated in vitro from each patient's lymphocytes. The lymphocytes have natural anticancer activity, or anticancer activity induced or enhanced by genetic modification of the cells in vitro. Eighty patients with metastatic cancer have been treated with anticancer lymphocytes between August 1, 2013 and August 1, 2014. During this same period of time, twenty four different clinical trials devoted to these therapies have been active and supported by the core laboratory. The core laboratory has also carried out research activities to improve its services. Thus, efforts have continued 1) to simplify the cell production methodology and make the process easier and more cost effective, 2) to relate characteristics of the anticancer lymphocytes and their parent populations to clinical outcomes following their use for treating patients, and 3) to help translate preclinical adoptive immunotherapy models, developed in the Surgery Branch and elsewhere, into new clinical protocols. Finally, the core laboratory continues to process cells and sera collected from cancer patients for a variety of uses, in addition to generating the anticancer cells described above. The products are routinely analyzed by investigators in the Surgery Branch immunotherapy program to evaluate progress toward the goals of each immunotherapy clinical trial, as well as to address subsequent research questions that help identify changes needed in the clinical trials. The samples are also used by Surgery Branch investigators for specific laboratory research projects that may translate into new patient therapies. These research projects include 1) transducing patients T cells with new genes whose products will provide better tumor recognition or otherwise enhance the cells anticancer functions, 2) evaluating the ability of infused anticancer lymphocytes to survive and function in the patient, 3) identifying new cancer antigens that are recognized by patients anticancer cells, 4) identifying characteristics of infused anticancer T cells that relate to cancer regression as measured by standardized, objective criteria, 5) identifying common characteristics of patients with metastatic cancer who are more likely to respond to adoptive cell therapy, 6) evaluating selected biological response modifiers tested in Surgery Branch clinical trials, and 7) extending adoptive cell therapy to additional types of metastatic cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Recent work from our laboratory shows that in mice a subset of the olfactory sensory neurons (OSNs) that express the transient receptor potential channel M5 (TRPM5) respond to semiochemicals (chemicals involved in animal communication, from the Greek semeion for ``sign'') including putative pheromones, peptides from the major histocompatibility complex (MHC) and urine (2). Therefore, the response to putative pheromones and other semiochemicals is not mediated exclusively by the vomeronasal organ. In this grant we will test the hypothesis that this subset of OSNs expressing TRPM5 respond to semiochemicals using a specific subset of olfactory receptors. Importantly, despite the fact that humans do not appear to express a vomeronasal organ, data suggest that they respond to putative pheromones and other semiochemicals (4, 5). Therefore we will perform experiments searching for responses to semiochemicals in mice and humans. Using strong techniques involving recording from single OSNs, evaluation of olfactory receptor expression in these neurons using RNA-Seq, and recording responses of specific olfactory receptors to large numbers of semiochemicals and odorants we propose three specific aims: Aim 1. Test the hypothesis that olfactory receptors in mouse TRPM5+ OSNs are differentially expressed in the two genders. Aim 2. Are TRPM5+ OSNs involved in responses to semiochemicals at low concentrations in both human and mouse? Aim 3. Test the hypothesis that olfactory receptors expressed in TRPM5-expressing OSNs respond to semiochemicals at low concentrations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In salt-sensitive hypertension, an increase in NaCl intake raises vascular volume and therefore blood pressure. However, Cl- intake may be as much or more important than Na+ in the generation of salt-sensitive hypertension. Aldosterone increases renal NaCl absorption by increasing NaCl transporter insertion in the apical plasma membrane, which stimulates NaCl absorption, thereby raising blood pressure. This steroid hormone increases apical plasma membrane abundance of these renal Na+ and Cl- transporters, at least in part, through regulation of the ubiquitin ligase, NEDD4-2, expressed in the aldosterone-sensitive region of the nephron. Of the NEDD4-2-regulated NaCl transporters, the best studied are the thiazide-sensitive NaCl cotransporter, NCC, and the epithelial Na+ channel, ENaC. NEDD4-2 binds to ENaC, which ubiquitinates the transporter, resulting in its endocytosis and degradation. With aldosterone administration, Sgk1 is stimulated, thereby phosphorylating NEDD4-2, which prevents its association with ENaC. The hypertension observed in mice with NEDD4-2 gene ablation and in people with certain polymorphisms of NEDD4-L, the human homologue of rodent NEDD4-2, occurs, in part, from the increased renal Na+ and Cl- transporter abundance. In the cortical collecting duct (CCD) NEDD4-2 is expressed in both principal and intercalated cells. Within intercalated cell subtypes, NEDD4-2 protein expression is highest in type B cells, suggesting that Cl- transport in type B intercalated cells i modulated by NEDD4-2. While principal cell NEDD4-2 function has been well studied, little is known about NEDD4-2 in intercalated cells. As such, we generated intercalated cell NEDD4-2 null mice using Cre-lox technology. With intercalated cell NEDD4-2 gene ablation, we observed a substantial increase in both electroneutral Cl- absorption and HCO3- secretion in the CCD and increased mean arterial blood pressure. We hypothesize that NEDD4-2 associates with intercalated cell Cl- transporters such as pendrin and ClC-5, which leads to their ubiquitination, endocytosis and degradation. We hypothesize further that aldosterone increases Cl- absorption in the CCD, in part, by reducing the association of these transporters with NEDD4-2. This proposal will dissect the mechanism whereby intercalated cell NEDD4-2 alters Cl- transport by intercalated cells. Aims of the proposal are to determine the following: 1) If aldosterone increases CCD Cl- absorption by stimulating Cl- /HCO3- and Cl-/H+ exchange in tandem, 2) if aldosterone acts through NEDD4-2 to increase Cl- absorption by intercalated cells of the CCD and 3) what intercalated cell Cl- transporters are targeted by NEDD4-2 in the CCD. To accomplish these objectives, we will examine the effect of aldosterone and NEDD4-2 on intercalated cell transporter abundance and function in mice both in vivo and in vitro, using quantitative real time PCR, immunohistochemistry and immunofluorescence, immunogold cytochemistry, immunoblots, electrophysiology and transport studies in renal tubules perfused in vitro.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Vision begins when light is converted to an electrical signal in the photoreceptors. Increases in light decrease release of the neurotransmitter, glutamate, from cone and rod photoreceptor terminals, and decreases in light increase its release. These changes in synaptic glutamate concentration are detected by two classes of bipolar cells that then transmit the signal vertically through the retinal circuit to the ganglion cells. The neurotransmitter changes also are detected by horizontal cells that provide lateral transmission in the form of feedback and feedforward inhibition. There are two classes of bipolar cells, hyperpolarizing (HBCs) and depolarizing (DBCs). HBCs utilize ionotropic glutamate receptors and hyperpolarize in response to a light flash. DBCs utilize a metabotropic glutamate receptor, mGluR6, and depolarize in response to a light flash. Defects in transmission between photoreceptors and bipolar cells result in several forms of congenital stationary night blindness (CSNB). The incomplete form, CSNB2, results from mutations in genes critical to glutamate release in photoreceptors, including the 11F subunit of voltage dependent calcium channels. The complete form emerges from mutations in signaling in DBCs, which include mutations in mGluR6 and nyctalopin (a protein of unknown function). Signaling through DBCs is mediated via a metabotropic glutamate receptor, mGluR6, which modulates the activity of a non-specific cation channel of unknown identity. The details of this mGluR6 cascade are mostly unknown. The long term goal of this project is to characterize the molecular components required for synaptic transmission between photoreceptors and DBCs. This project has four specific aims: 1) determine the structure/function relationships of nyctalopin, 2) determine the state of the non-specific cation channel in several night blind mice, 3) determine the binding partners of nyctalopin, thereby elucidating new component of the mGluR6 cascade, and 4) create knockout mouse lines of the nyctalopin interacting proteins to determine if they result in nigh blindness. At the completion of this project, we will have identified new members critical to signal transmission in DBCs. Further, we will have identified new candidate genes for congenital stationary night blindness. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE: Vision requires a light signal to be converted to an electrical signal, which is then transmitted to the brain via a neuronal network. The group of diseases being studied is referred to as congenital stationary night blindness. They have defects in signal transmission between photoreceptors and the second neuron in the pathway. The studies in this proposal will characterize the nature of the defects and determine new proteins critical to function. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Curcumin, a yellow polyphenol compound found in the Indian spice turmeric (Curcuma longa) demonstrates potent anti-cancer effects against a wide variety of human tumor models in preclinical studies. A major pitfall forthe application of curcumin to human cancer therapy has been its limited systemic bioavailability, such that the few clinical trials conducted so far had administered mega doses (8-10 grams) of free curcumin daily to pafients in order to yield detectable levels. These high dosages significanfiy affect patient compliance and have stymied the application of this promising, and relafively non-toxic, agent to most visceral malignancies that would mandate high plasma levels of the active therapeutic. A novel nanoparticle encapsulated formulation of curcumin (nanocurcumin) has been engineered which bypasses the bioavailability pitfall of this compound, and enables strikingly higher circulafing and tissue levels upon parenteral administrafion. Nanocurcumin causes significant growth reduction of subcutaneous and orthotopic pancreatic cancer xenografts, and abrogates systemic metastases when co-administered with gemcitabine in a spontaneously metastatic animal model. The studies proposed herein have the overarching goal of performing the efficacy, pharmacokinetics, and toxicity data required for filing an invesfigafional new drug (IND) application with the USFDA. To this effect, nanocurcumin (as single agent or in combination with gemcitabine) will be tested in a panel of low-passage gemcitabine-resistant patientderived orthotopic xenografts (Aim 1) and in a uniformly lethal genetically engineered mouse model (Aim 2), and will undergo regulatory grade in vitro and in vivo toxicology testing by the federally-funded Nanotechnology Characterization Laboratory (NCL) (Aim 3), in preparation for a pre-IND application. Two hypothesis-driven aims that integrate unique CCNE resources will also be pursued, including the effects of nanocurcumin on vaccine efficacy and quantifiable measures of cellular immune response (Aim 4; in collaboration with Project 3) and engineering the next generafion of active-targeted anfibody-conjugated nanocurcumin (Aim 6; in collaboration with the Nanoparticle Engineering and the Validafion Cores).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal aims to characterize the effects of a variable genetic element in regulation of expression of the vole vasopressin 1a receptor (V1aR) gene. Vasopressin and the V1aR are involved in the regulation of species-specific social behaviors. Receptor distribution patterns are species-specific and correlate with social structure. Monogamous and non-monogamous voles differ in their behavioral response to vasopressin and expression pattern of V1aR in the brain. In addition, there is a tremendous amount of intra-specific variability in V1aR expression in monogamous prairie voles. A large, highly repetitive microsatellite region is found in the 5' flanking region of the VlaR gene of monogamous voles but not in non-monogamous vole species. Within prairie vole, the length of this microsatellite correlates with the level of VlaR binding in the brain. This research proposal will test the hypothesis that i) this polymorphic promoter sequence plays a role in the expression of the VlaR gene, and that ii) the individual variability in V1aR expression resulting from this variable promoter structure translates into individual variability in social behavior. Luciferase reporter assays in cell culture and a selective prairie vole breeding strategy with extensive behavioral characterization and subsequent molecular analysis will be used to test these hypotheses. This research will provide a possible molecular mechanism producing intra-specific variability in gene expression and behavior, which may be relevant to psychopathologies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The assembly of neural circuits in the developing brain is thought to require both genetically determined and activity-dependent mechanisms. Many of the long-term changes mediated by neural activity are dependent on calcium-activated transcription of new gene products. Thus, calcium-activated transcription factors provide a mechanism for linking extrinsic signals with genetic programs of neural development. NeuroD2 is a basic helix-loop-helix transcription factor previously isolated in our lab, using a screen for novel calcium-regulated transcription factors involved in cortical development. NeuroD2 expression is developmentally regulated in the hippocampus, peaking between P7 and P14, coincident with the primary period of synaptogenesis in this structure. Here, we demonstrate a role for NeuroD2 mediated transcription in the normal development of mossy fiber (MF) to CA3 connectivity. In NeuroD2 null mice, which have grossly normal brain structure, calbindin staining of the MF afferents revealed a dramatic and preferential reduction in the distal portion of the main MF bundle. Although the fiber tract extends the full length of the CA3 region, it fails to elaborate the characteristic enlargement known as the \"end bulb\" at its distal tip. TIMM staining for pre-synaptic terminals in the MF pathway revealed a similar, if not greater, reduction in synapse formation in this pathway. Analysis of post-synaptic morphology of CA3 pyramidal neurons by Lucifer Yellow dye injection revealed a similar reduction in the characteristic multi-headed spines of the proximal apical dendrites receiving MF input. Interestingly, the size and density of the more distally located classical spines was not affected in the KO. This raises the possibility that while NeuroD2 mediated transcription is required for the development of MF connectivity, synapses from other pathways onto the same neuron may form in a NeuroD2 independent manner. These studies begin to define a novel transcriptional pathway regulating the development of connectivity in a major excitatory component of hippocampal circuitry.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent studies have established the presence of an olfactory input to the mediodorsal nuclear complex of the thalamus, an area which has been known primarily as the thalamic dependency of orbitofrontal cortex. This new evidence for an olfactory thalamocortical relay brings olfaction into line with other sensory systems and provides fresh information on the largely unknown functions of MD and orbitofrontal cortex. The objective of the proposed research is a combined electrophysiological and autoradiographic delimitation of the thalamic and cortical olfactory projection fields and a determination of their associations with two other apparently related projection fields: amygdalar and gustatory. This work should serve as a necessary prelude to functional electrophysiological studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Obesity is a major cause of insulin resistance and rising rates of obesity are responsible for the increasing prevalence of type 2 diabetes mellitus. In addition to its function in the uptake and storage of energy as triglyceride, the adipocyte is a complex endocrine cell that regulates feeding behavior and insulin action. However, the relationship between adiposity and diabetes is complex; for example, thiazolidinediones increase fat mass, yet improve insulin sensitivity. It is vital to define factors that regulate adipocyte function to understand how the adipocyte modulates such diverse processes. We have previously shown that two nuclear receptor corepressors, the silencing mediator of retinoid and thyroid hormone receptors (SMRT) and the nuclear receptor corepressor (NCoR), repress adipocyte gene expression during adipogenesis. In the proposed studies, we will test novel hypotheses to explain how SMRT and NCoR regulate adipocyte differentiation, adipocyte function, and insulin sensitivity. In Aim 1, we will test the hypothesis that SMRT and NCoR dictate the function of the differentiated adipocyte. In Aim 2, we will develop mouse embryonic fibroblasts deficient in SMRT and/or NCoR to test the hypothesis that corepressors regulate adipogenesis and adipocyte survival in primary cells. In Aim 3, we will define the roles of SMRT and NCoR in adipocyte function and insulin sensitivity in vivo. By combining in vitro analysis of adipocyte cell lines, mouse embryonic fibroblasts, and isolated adipocytes, and by developing novel mouse models of corepressor deficiency to test our hypotheses in vivo, we will dissect the molecular mechanisms underlying SMRT and NCoR action in the adipocyte. Understanding the roles of corepressors in adipocyte biology is crucial if we are to design novel approaches to the treatment of obesity and diabetes. Relevance: Obesity is a risk factor for the development of type 2 diabetes mellitus, and diabetes is a major cause or blindness, kidney disease, amputation, and death. The goal of these studies is to test the hypothesis that two proteins, SMRT and NCoR, are important in fat cell (adipocyte) function and the ability of the body to respond to insulin. These results will be helpful in the design of new medications to treat patients with diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Admin Core Project Summary We propose to establish the Arizona Cancer and Evolution Center (ACE). Arizona State University will lead the Arizona Cancer and Evolution Center effort. Drs. Maley and Shibata, co-principal investigators, will direct the Center in collaboration with NCI program staff. ACE integrates a team of modelers, evolutionary biologists and cancer biologists with a successful track-record of collaborations. The objective of the Arizona Cancer and Evolution Center Administrative Core is to provide the necessary infrastructure for the overall organization and effective management of the Center to achieve the objectives of the CSBC and the NCI, as well as realize the scientific goals of the Center. The ACE Administrative Core will accomplish four Specific Aims during this funding period: Aim 1 Create and maintain a knowledgeable, skilled, and interdisciplinary leadership team that is connected to the research and the wider network; Aim 2 Design and implement a clear and effective communication plan across the Center; Aim 3 Identify and foster pilot projects; Aim 4 Solicit, coordinate, and respond to external advice for the scientific direction and opportunities for ACE. IMPACT: The Administrative Core will enable the Arizona Cancer and Evolution Center to transform current understanding and management of cancer by developing the theory, models, and measures of cancer that will be applicable and clinically relevant across cancers and species.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Air pollution is a significant cause of respiratory disease, and the most common gaseous air pollutants are ozone (O3), sulfur dioxide (SO2) and nitrogen dioxide (NO2). The primary goal of this study is to elucidate cellular mechanisms by which these gaseous air pollutants damage nasal and bronchial epithelial cells and promote inflammation in these tissues. Previous research suggests that these epithelia play an active role in the processes of health and disease through the synthesis of cytokines (protein mediators) and the expression of additional molecules which function as cell-cell, cell-substratum and cell-leukocyte adhesion receptors. The overall hypothesis of this proposal is that air pollutants act directly on the membranes of airway epithelial cells and, as a consequence, modulate adhesion receptor expression and cytokine production. An in vitro cell culture approach is proposed which will examine the responses of human and nonhuman primate nasal and bronchial epithelial cells. Both primary cell cultures and cell lines will be used. The first goal is to define the synthesis and expression of the cell-cell and cell- substratum receptors by these cells. These include proteoglycans, CD44, CD26, and beta1 integrins. Constitutive production of these receptors will be examined as well as changes caused by exposure to O3, SO2 and NO2. Exposures to individual gases and gases in combination will be done at ambient or near ambient concentrations (O3, 0.1-0.5 ppm; SO2: 0.5-5 ppm; NO2: 0.1-0.5 ppm). The hypotheses being tested are that respiratory epithelial cells produce adhesion molecules necessary to maintain normal epithelial function and that exposure to gaseous pollutants alter the synthesis of these products, adversely affecting tissue integrity. The second goal is to measure the expression of intercellular adhesion molecule-1 (ICAM-1) on nasal and bronchial cells and the effects of exposure to O3, SO2 and NO2 on the expression of this receptor. ICAM-1 is the primary leukocyte adhesion receptor on epithelial cells. The hypothesis being examined is that exposure to gaseous pollutants upregulates epithelial cell ICAM-1 expression thus enhancing the local inflammatory response by promoting leukocyte-epithelial adhesion. The third goal is to examine the types of cytokines produced by nasal and bronchial epithelial cells, and if the qualitative and quantitative synthesis of these cytokines is altered by exposure to O3, SO2, and NO2. The hypotheses being examined are that respiratory epithelial cells produce soluble factors which promote local inflammation and that exposure to pollutants stimulates these events. This study will advance our understanding of the functions of respiratory epithelial cells in maintaining normal tissue integrity and will determine whether gaseous pollutants produce respiratory inflammation and damage by modulating cytokine and adhesion receptor synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Analysis of 37-M-mode echocardiograms recorded during the period of active bacterial endocarditis involving the aortic valve in 17 patients disclosed or more echocardiographic abnormalities involving the aortic valve cusps or their immediate vicinity in 15 (88%), including \"shaggy\" echos indicative of vegetations in 12 (71%). Of the 12 patients with echocardiographic evidence of aortic valve vegetation, 11 developed overt congestive heart failure and either died or had aortic valve replacement and 7 had clinical events compatible with systemic emboli; of the 5 patients without echo-demonstrated vegetations, only 1 had congestive heart failure, none had aortic valve replacement, 2 died, and 1 had a systemic embolus. In comparison to our previously reported echocardiographic observations in patients with active infective endocarditis involving either the mitral (29 patients) or tricuspid valve (23 patients), infection involving the aortic valve was far more liable to produce overt congestive heart failure and systemic emboli, to necessitate valve replacement, and to cause death during the period of active infections.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The UCSF Rheumatoid Arthritis Panel is now ten year effort to trace the course, treatment, and outcomes of rheumatoid arthritis. 1,025 persons with RA from a random sample of Northern California rheumatologists have been followed through annual interviews, periodic up-dates on severity from their physicians, home examinations, and information from health care providers, insurers, and vital records. In addition, 194 persons matched to the persons with RA on age sex, and area of residence have been followed since 1989 as a control group. In the proposed period of funding, the number of persons with RA in the study will be expanded by approximately 250. The 250 will have disease of recent onset. The number of controls will be expanded by approximately 75. This data base has served six MAC projects, one supplement, and one D&F study over the ten year period. In addition, it has served thirteen projects funded by non-MAC sources. In the proposed period of funding, the data base will serve Project EEHSR 3, \"Fatigue and Rheumatoid Arthritis\", Project EEHSR 4, \"Stress, Coping, and Outcomes in Rheumatoid Arthritis\"; and Project EEHSR 5, \"Genetic Factors in Rheumatoid Arthritis Outcomes\". The Core Unit is responsible for the design and validation of all data collection instruments in the RA Panel Study; the training and monitoring of all data collection staff; the administration of telephone and face to face interviews to person wit RA, controls, and health care providers; the collection of sera and collection and analysis of radiographs; the entry of data via computerized telephone interviewing software and other methods; cleaning the data from each year; the management of data from all years of the study; performing data analysis; and statistical consulting. The foregoing activities are housed in a Core because several MAC projects and several non-MAC projects use RA Panel data, and because many of the functions of the Core are indivisible. These include: collective use of a fixed portion of all surveys; skills to design instruments; staff skills and effort to collect and process data; and software and hardware to collect and analyze RA Panel study data.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Statistics Core is responsible for the coordination of scientific activity among the various projects and cores, and for budgetary planning, data base management, and statistical services. In addition, will promote the development of new analytic procedures tailored to the special problems that danger of capitalizing on chance when many different measures are taken with relatively small populations. Our solutions to this problem include (a)constraints on the level of analysis to which a strong hypothesis applies, (b) methods for aggregating multiple variables into a single robust variable, (c) constraints on the order in which hypotheses are tested, and/or constraints on the way that individual variables are used within a single complex model, and (d) a clear and fruitful separation between hypothesis testing (confirmatory analysis) and hypothesis generation (exploratory analysis). We also outline plans for integrative analyses across the populations under study within the Multicenter, including the construction of models that can be used for systematic cross- population comparisons.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Type 1 diabetes (T1D) is one of the most prevalent chronic childhood diseases worldwide. In addition to its negative impact on quality of life for patients and their families, the disease also poses a significant financial burden on society. Thus, a curative solution, instead of the current maintenance therapy, is urgently needed. One such curative treatment shown to be very successful in mouse models is the infusion of autologous regulatory T cells (Tregs). Tregs are a small subset of CD4+ T lymphocytes that are primarily responsible for controlling pathogenic autoimmune responses in the periphery. Mounting evidence in animal models and patients demonstrates that T1D is associated with an imbalance between pathogenic T cells and Tregs. Treg therapy restores the balance and enables the immune system to regain self-control. With these encouraging results, a clinical trial of Treg-based therapy is being actively developed and is scheduled to start in 2009. At this juncture, it is important to understand the cellular and molecular basis of Treg function in controlling T1D. Our previous experiments demonstrate that a single infusion of islet-antigen-specific Tregs isolated from BDC2.5 T cell receptor transgenic mice (BDC Tregs) can prevent and reverse diabetes in the NOD mice. The BDC Tregs migrate to pancreatic lymph nodes and islets. In the pancreatic LN, they engage dendritic cells and effectively block further activation of pathogenic T cells by dendritic cells. How BDC Tregs halt 2 cell destruction in the inflamed islets has not been studied. In this grant application, we propose to systematically investigate the mechanism of T1D control in the NOD mice by BDC Tregs. We will determine the direct cellular target of BDC Tregs in vivo, and identify their impact on the ongoing inflammatory response in the islets at cellular and molecular levels. We will further determine the molecular profile of the therapeutic Tregs and identify molecule(s) responsible for their protective effect in vivo. Through the studies proposed in this grant application, we expect to gain better understanding of the pathogenic events critical for T1D progression and how therapeutic Tregs control these processes. The insight gained from these mechanistic studies will help to improve the design of Treg- based cellular therapy and to identify new targets for therapeutic intervention of T1D. PUBLIC HEALTH RELEVANCE: Type 1 diabetes is a chronic childhood disease that results from an immune-mediated destruction of the insulin-producing 2 cells. Regulatory T cells constitute a small population of immune cells that is mainly responsible for preventing unwanted immune response in healthy people. In animal models, regulatory T cell therapy can effectively prevent and reverse type 1 diabetes. Studies proposed herein are designed to understand the cellular and molecular basis for regulatory T cell control the disease. The insight gained from these mechanistic studies will help to improve the design of regulatory T cell-based therapy for patients and to identify new targets for therapeutic intervention of type 1 diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A synthetic version of a lipopeptide that is one of the first compounds to appear in HIV-infected cells was obtained from a commercial company and employed to generate a T cell line, using cells from an HIV patient donor. On further analysis it was found that the synthetically produced version of the targeted lipopeptide was not pure, but in fact a mixture, and further, the targeted lipopeptide was not antigenic, but instead two novel components were found to be antigenic. Mass spectrometric investigations indicate that the new antigenic components are very similar in structure to each other and have structures similar to the intended synthetic structure. Mass spectrometric studies at the BUSM MS Resource included the use of GC/MS, MALDI and ESI Q-o-TOF and MALDI and ESI-FTMS instrumentation, enzymatic digestion and chemical hydrolysis of the known and unknown compounds. The structure has now been solved and a synthetic analogs are being prepared. The manuscript has been accepted for publication in the Journal of Experimental Medicine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Embryonic stem (ES) cells provide an exciting opportunity in the field of regenerative biology and medicine, as they have the unique capacity to differentiate in culture into all somatic cells that make up an individual. Significant efforts have been made in generating functional hematopoietic stem cells (HSCs) from pluripotent stem cells. For instance, expression of HoxB4 (or Cdx4-HoxB4) during in vitro differentiation of ES cells has been shown to derive functional HSCs from ES cells (1-3). Despite these significant advancements, major challenges remain in utilizing in vitro derived HSCs and progenitors for regenerative medicine. For example, hematopoietic engraftment by ES derived hematopoietic progenitors remains consistently and disappointingly low. Even with HoxB4 expression, the frequency of HSCs and progenitors that can repopulate the recipient's hematopoietic system is estimated at best 1 in 2-5x106 among Cdx4-HoxB4 induced ES derived cells (3). Constitutive HoxB4 expression in transplanted cells could also be problematic. The overall objective is to robustly generate functional HSCs from ES and iPS cells. In achieving this goal, we posit that HSCs are generated from ES cells following the same molecular and cellular rules occurring in vivo. To this end, we demonstrated Flk-1 expressing (Flk-1+) hemangioblast, a common progenitor of hematopoietic and endothelial cells (or hemogenic endothelium), derived from in vitro differentiated ES cells can generate hematopoietic cells in culture (4, 5). In addition, we demonstrated that both primitive and definitive hematopoietic systems of the mouse are derived from the Flk-1+ mesoderm (6). Our preliminary studies indicate that when Flk-1+ hemangioblasts were sorted and further differentiated on OP9 cells, we were able to efficiently generate CD41+cKit+CD150+ cells, which have been previously identified as ES derived HSC/progenitors (3). Moreover, temporal co-expression of ER71, GATA2 and Scl positively induced CD41+cKit+CD150+ cells from ES cells: 1) by robustly inducing Flk-1+ hemangioblast formation from ES cells and 2) by independently inducing CD41+cKit+CD150+ cells from Flk-1+ hemangioblasts. Herein, we test a hypothesis that functional HSC generation from ES cells can be greatly improved by temporal ER71, GATA2 and Scl co-expression. We will determine if ES and iPS derived CD41+cKit+CD150+ cells could reconstitute hematopoietic system of the mouse and if temporal ER71, GATA2 and Scl co-expression would be superior to HoxB4 expression in generating functional HSCs. The ability to effectively direct ES and iPS cells to HSCs will provide a novel means of differentiation and will have a major potential impact in the field of regenerative biology and medicine. The outcome will also likely revolutionize the way we envision the cell lineage development and differentiation, thus will also have a major potential impact in the basic research field. Thus, the potential impact is high.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "von Willebrand Factor (vWF) is a large plasma glycoprotein which is required for platelets to recognize and bind to damaged endothelial surfaces. Bovine vWF binds directly to human platelets in a manner which is a model for the interaction of human vWF with platelets on subendothelial surfaces. The goals of the project are: (a) to identify the regions of the vWf molecule which are responsible for its binding to platelets, to collagen, and to glycosaminoglycans, (b) to identify the platelet receptor for vWF and determine the mechanism by which vWF binds to platelets, and (c) to describe the effects of vWF binding on the metabolic responses of the platelets, and, conversely, the effects of other agonists on platelet responsiveness to vWF. The functional domains on the vWF molecule will be identified and characterized by proteolytic fragmentation of the vWF, by chemical and enzymatic modification of vWF, and by use of monoclonal antibodies to vWF. The platelet receptor for vWF will be isolated by affinity chromatography and will also be identified in situ by photoaffinity labeling. Platelet responsiveness will be studied by measuring vWF binding, aggregation responses, shape change and the platelet release reaction. Alterations in platelet protein labeling will be monitored and the distribution of receptors on activated and non-activated platelets will be visualized by electron microscopy with gold-conjugated specific probes for the receptors. These studies should provide a better understanding of the basic mechanisms underlying platelet recognition of sites of damage in blood vessels. They may suggest new ways to deal with the thrombotic complications associated with diseased or damaged blood vessels or with artificial surfaces such as prosthetic heart valves, dialysis membranes, or vascular prostheses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Fibrosis of the hepatic subendothelial (Disse) space is a prominent component of the inflammation elicited by ethanol, and appears to correlate with clinically significant liver disease. The lesion consists of a collagen-rich new-matrix that replaces the low- density matrix normally present in this location. In recent and on-going studies from this laboratory, the lipocyte (fat-storing or Ito cell) has been examined in pure primary culture and found to be a major source of several matrix proteins including collagen, laminin and proteoglycans. In vivo, cells referred to as 'myofibroblasts' have been identified following chronic alcohol consumption and localized specifically to areas of fibrogenesis. Morphologic studies suggest that lipocytes and myofibriblasts represent the same cell in various stages of transition. The working hypothesis for the present studies is that ethanol 'activates' lipocytes, causing their conversion from resting, vitamin A-storing cells to matrix-secreting myofibroblasts. The pathway of lipocyte activation is the principal focus of the proposed research. Several possibilities will be explored, including stimulation by acetaldehyde or by soluble mediators of inflammation released as by-products of ethanol metabolism. The first series of experiments will establish wheter lipocyte activation occurs as a direct result of ethanol metabolism. This will be accomplished using a novel cell culture approach, in which hepatocytes and lipocytes are co-cultured as 'microorganoids' and incubated with varying concentrations of ethanol. Additional studies will examine whether lipocyte activation is indirect, involving compounds produced by neutrophils of Kupffer cells previously exposed to conditioned medium from ethanol-treated hepatocytes. Lipocyte activation will be judged with studies of ultrastructure as well as quantitative measurements of matrix protein secretion, matrix gene expression, and cell proliferation. Recently developed methods for evaluating matrix gene expression and for culture of individual parenchymal and non- parenchymal cell populations allow a direct approach to the pathogenesis of this common and important condition.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Core of this Program Project consists of four components that represent services that are provided to each of the three research projects. In addition, each of these components will continue to be engaged in development of the infrastructure required for the implementation of the methods in the research plan. The four components of the core described below are: 1. Administrative services 2. Neurological patient and subject recruitment support and development 3. Neuroscientific method support and development 4. Informatics and datacenter support and development.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of the proposed research is to better understand molecular mechanisms in the regulated production of ribosomal and messenger RNA. Huge advances have been made in the identification of proteins involved in transcription and RNA processing for RNA polymerase I and II, yet there are many important questions to be answered concerning basic mechanisms. Our unique approach offers a means to answer some of these questions by direct visualization of active genes using the Miller chromatin spreading method for electron microscopy. In brief, using Saccharomyces cerevisiae as a model system, we visualize specific genes in the presence and absence of key regulatory proteins (i.e., in appropriate mutants) to determine how the gene is changed in its absence. We are able to discern (and distinguish) effects on transcription initiation, elongation, termination, processing of nascent transcripts, chromatin structure, and template topology. Furthermore, for each of these topics, we are able to resolve patterns that are informative regarding the molecular mechanism affected. Our recent results have led us to focus on the following Aims: (Aim 1) Determine the role of histones and chromatin-associated elongation factors in elongation by RNA polymerase I;(Aim 2) Analyze topological abnormalities in rDNA that appear in the absence of topoisomerases and ask if and how these correlate with the increased recombination that occurs in these mutants;and (Aim 3) Develop and apply a system for the visualization of specific Pol II genes and analysis of co-transcriptional events. These studies are relevant to human health in the following ways: we will gain information on the regulation of ribosome production, which is important to cell growth control and is up- regulated in cancer;(b) we will gain information about the mechanisms of transcription elongation and splicing, mutations in which lead to many different human diseases, and (c) we may gain direct information on mechanisms of genomic instability by studying this process in topoisomerase-deficient cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A long term goal in the field of restriction-modification enzymes has been to generate restriction endonucleases with novel sequence-specificities by mutating or engineering existing enzymes. This will avoid the increasingly arduous task of brute-force screening of bacteria and other microorganisms for new enzymes. Our objective here is to generate rare cutters that would be valuable tools in genomic research and analysis. Over the past six years, we have studied FokI restriction endonuclease from Flavobacterium Okenokoites in great detail. FokI is a Type IIS endonuclease which recognizes the pentanucleotide duplex, 5'-GGATG-3': 5'-CATCC-3' and cleaves about 9/13 bp away from the recognition site. This implies the presence of two separate protein domains in this enzyme: one for sequence-specific recognition and the other for the endonuclease activity. Our studies o proteolytic fragments of FokI endonuclease have defined an N-terminal DNA- binding domain and C-terminal domain with non-specific cleavage activity (PNAS 89:4275-4279(1991)]. These results have been confirmed by the study of the C-terminal deletion mutants of FokI endonuclease [Gene 133:79-84 (1993)]. Furthermore, introduction of additional amino acid residues between the recognition and cleavage domains of FokI can alter athe cleavage distance from the recognition site within its DNA substrate [PNAS 90:2764-2768 (1993); J. Biol Chem.269:31978-31982 (1994)]. These results suggest that the two domains of FokI are connected by a liner region which appears to be amenable for repositioning of the DNA-binding domain with respect to the catalytic domain. Recently, we have successfully engineered th first chimeric restriction endonuclease by linking the Ubx homeodomain to the catalytic domain of (Fn) of FokI [PNAS 91:883-887 (1994)]. More recently, we reported the deliberate creation of novel site-specific endonucleases by linking two different zinc finger proteins to the cleavage domain of FokI endonuclease. Both fusions are active. This opens the way to generate many new enzymes with tailor-made sequence-specificities [Science, manuscript submitted for publication (1995)]. This work could lead to the development of an array of artificial nucleases with tailor- made sequence-specificities desirable for various applications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Atmospheric ultrafine particulates (UFPs, mass median aerodynamic diameter < 0.1 mu m) emitted from combustion processes and newly emerging nanotechnologies have recently been recognized as a significant global health hazard. Inhalation of these particles may lead to disruption of mitochondrial cristae and increased levels of reactive oxygen species (ROS) in lung epithelial cells. The respiratory and cardiovascular systems seem particularly vulnerable to the toxic effects of UFPs through a combination of the particles' characteristically large surface area and the presence of surface transition metals and polycyclic aromatic hydrocarbons (PAHs). Nevertheless, the surface chemistry of UFPs remains virtually unexplored and has not been correlated with possible toxicological mechanisms. The long-term objective of the present study is to identify the main contributors and processes that initiate the physiological detrimental effects of UFPs. The primary focus is on establishing that surface compounds that contribute to the production of ROS, principally iron and PAHs, can cause direct mitochondrial damage. UFPs will be collected in urban and rural areas of Washington State and analyzed for surface iron speciation and PAHs using x-ray photoelectron spectroscopy (XPS), time of flight secondary ion mass spectrometry (TOF-SIMS), and spectrophotometry. Simultaneously, their toxicological effects will be tested in vitro on beef heart mitochondria, with particular interest on whether the initial mechanisms of mitochondrial dysfunction are lipid peroxidation or ROS production and electron transport inhibition. Findings from this multidisciplinary project involving the collaborative expertise of atmospheric, surface, and biochemists will bridge critical gaps tying UFP characteristics to their toxicological mechanisms in cellular metabolism and contribute to current understanding of health effects of airborne particulate matter.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The somatostatins constitute a family of structurally related, small, peptides synthesized as large precursors within multiple organs including the islets, intestine, and central and peripheral nervous tissues. They serve both hormonal and paracrine functions in the regulation of secretions and probably as neuromodulators and/or neurotransmitters; their actions are essential for metabolic homeostasis. The production of the somatostatins is in turn regulated by hormonal and paracrine influences. We plan to investigate at a molecular level the organization and the expression of the somatostatin genes. We wish to determine the complexity (numbers and structures) of the genes encoding the somatostatin precursors and to determine how these findings are related to the post-translational processing of the precursors and to the expression of the biologic activities of the somatostatins. We will determine the nucleotide sequences of cloned cDNAs complementary to mRNAs encoding precursors of the somatostatins (pre-prosomatostatins) found in pancreatic islets, intestine, hypothalamus, andextrahypothalamic nervous tissues of the anglerfish and the rat. Knowledge of the nucleotide sequences will allow us to deduce and to compare the amino acid sequences of the somatostatin precursors synthesized in the different organs. From the primary structures we will be able to infer sites in the precursors that are cleaved during their co- and post-translational processing. The cDNAs will be used as hybridization probes to: (1) identify genomic sequences of RNA transcription, processing and turnover in islet and neural tissues in response to specific stimuli. The results of these studies may provide information relevant to the pathogenesis of diabetes mellitus, and other disorder of hormone deficiencies as well as insights into the functions of the somatostatins in the central and peripheral nervous systems.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the first year of the program the major elements of this have been implemented as set out in the application, notwithstanding the departure of the Principal Investigator, Dr. Bruce Ewald. Dr. Bedford was appointed in his place in October 1978 and two staff, Eileen Weiner, Veterinary Specialist and Ruth Manson, Veterinary Technician and Prosector have been brought into the program. Miss Weiner has taken responsibility primarily for the hematological monitoring of quarantined animals, of colonies, screening clinical cases and that involved in vendor evaluation. She also has overseen the procurement of samples for clinical chemistry of quarantine and colony subjects. Ruth Manson has been involved primarily in necropsy procedures, in preparation of tissues for histopathology processing, collection and monitoring of cultures obtained from animals sacrificed for vendor evaluation, sanitization procedures and intra-laboratory colony screening. The availability of the grant has fostered cooperation on a consortium basis between Cornell Medical College and Rockefeller University, begun this year and anticipated as continuing strongly in the future. This has involved not only discussion and exhange of opinion but formal meetings between all personnel involved in the diagnostic grant program, and exchange of facilities with Rockefeller assuming responsibility for culture work-up and Cornell the hematological aspects of the program. Histopathological evaluation of necropsy material in clinical and evaluation cases has been carried out by Dr. Frank Bloom.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Glial cells form a major portion of the central nervous system but remain relatively uncharacterized in terms of their roles in brain function. The long range goal of the proposed research is to determine the physiological properties of glial cells. We plan to develop procedures for isolating in culture the component cells of the rat brain and for studying their interaction in vitro. We will determine if homogeneous cultures of astrocytes or oligodendrocytes contain the biochemical means for communicating with neurons, and determine if glia cultured from specific sub-anatomical regions of the brain (cerebrum, cerebellum, hippocampus, caudate nucleus, etc.) have unique characteristics in regard to the potential for glia-neuron communication. Four specific projects will be carried out in order to determine: 1) the distribution of cell surface receptors for neurotransmitters on glia from different brain regions, 2) the relation between cell surface receptors and the regulation of cyclic nucleotide metabolism in glia, 3) the factors that regulate the membrane potential of glia and 4) the characteristics of ion transport systems in glia. We plan to develop a cell culture facility in support of these specific projects. The purpose of this facility will be two-fold; first, to provide each investigator with preparations of cultured cells that are standardized within the current limits of technology, and second, to evolve the technology of cell culture in terms of 1) application of current methodology to the culturing of cells from different brain regions, 2) characterization of the growth properties and growth potential in vitro of astrocytes and 3) characterization of the effects of various growth factors and differentiation factors on glial cultures. The systematic study of cultured glial cells should provide fundamental information on the biochemical basis of neuron-glia interactions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Clinical trials of a new treatment may proceed through three phases. Phase I trials are typically small studies that evaluate toxicity and determine a safe dosage range. The specific task is usually to determine the maximum tolerated dose. Once a safe dose of the treatment is chosen, its therapeutic efficacy will be tested in a phase II trial. Regimens shown promising in phase II trials will then be moved to large, multi- institutional phase III studies that compare their effectiveness to standard treatments. In situations when many regimens are possible candidates for a large phase III study, but too few resources are available to evaluate each relative to the standard, priority must be set to select only those with promise for successful evaluation in phase III trials. In this research, we propose novel statistical designs and strategies that utilize the observed clinical data in an efficient manner, which is hoped to translate into equally accurate clinical conclusions with fewer resources. Specifically, this application covers the following three clinical scenarios. First, we propose methods for phase I dose-finding trials based on multiple and repeated safety endpoints, and apply the methods to re-design a current stroke trial. Existing designs collapse the endpoints into a dichotomized indicator of toxicity or no-toxicity, and may do so at the expense of not utilizing all information available. Our proposed methods are expected to retrieve the information loss by using all endpoints in the conduct of a trial. Second, we propose methods for phase II dose-finding efficacy trials, in which patients will be enrolled in two stages. Having an interim analysis, we can shut down ineffective doses and reduce the number of patients treated at these doses. Third, we propose designs to select or screen treatments in phase II trials. Current designs are rigid and impose great practical difficulty during the planning and the implementation of a trial. Our methods provide, for instance, flexibility in the scheduling of interim analyses and the prior preference of treatments. These designs will be applied to design various clinical trials in patients with neurological disorders. Despite the efforts in the past decade, additional therapies for acute ischemic stroke and amyotrophic lateral sclerosis (ALS) are sorely needed. Upon successful completion of this research, we will extend our capacity to support early phase investigation of new treatments and enhance the statistical efficiency of selection and screening process in a variety of clinical trial settings. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Combinatorial Exploration of Cellular Regulatory Network Design Principles A major goal in systems biology is to understand how cells use molecular circuits to achieve complex physiological behaviors and how these behaviors break down in disease states like cancer. Although only a small fraction of existing networks have been well characterized, recurrent topologies have emerged for a variety of biological functions, suggesting that any particular behavior is encoded by a limited number of network structures. With the goal of rapidly identifying the structural 'solutions'for target behaviors, we propose to develop a synthetic biological (forward engineering) approach of screening combinatorial libraries of molecular circuits. My specific aims are to: 1) Develop methods to assemble combinatorial circuit libraries. As a testbed we will construct a library of synthetic positive feedback loops in the yeast mating pathway and screen these for memory behavior. 2) Develop a screen for a complex dynamic behavior-sensory adaptation. A recombination-based FACS assay will be developed to screen for cells that can transiently respond to a stimulus, but then automatically reset output despite sustained stimulus. Comparison of functional circuit architectures is expected to reveal global design principles-critical network topologies and parameters, and robust structural motifs-providing a quantitative blueprint for core circuit families capable of each target task. This analysis should provide fundamental insight into how complex circuits that achieve memory and adaptation can be built. In the long-term, the emerging map correlating network structure and function will guide the interpretation, therapeutic perturbation, and rational synthesis of biological systems. PUBLIC HEALTH RELEVANCE: Cells sense and respond to their environment using a molecular communication network composed of interacting proteins. Many diseases, including cancer, arise when alteration of a signaling protein changes the network's structure, resulting in a new cellular behavior. Our goal is to generate a map correlating network structure and function that will guide the understanding and therapeutic correction of complex biological systems.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research examines how English readers and listeners use their knowledge of the semantic structure of their language to comprehend sentences and discourse. It builds on earlier research examining how they use syntactic structure, extending it to topics that have been investigated in the field of semantics. In particular, the research investigates how people are able to comprehend sentences with variables, which are an essential source of the expressive power of human languages. It studies three distinct aspects of the processing of variables: a) true vs. 'fake' variable binding (\"Every man loves his wife\" vs. \"The men bought presents for their wives;\" b) the processing of `world variables,' essentially implicatures that some state of affairs is not true in the actual world; and c) the determination of the domain over which a variable is quantified (e.g. does \"The offices are mostly clean\" refer to most offices or to degrees of cleanliness?). Each of these topics is the subject of intensive analysis within the field of formal semantics and is ready for experimental psycholinguistic study. The experiments that are proposed study these topics with a variety of techniques, ranging from simple tasks of judging the meaning or the naturalness of sentences and discourses to measures of eye movements during normal reading. PUBLIC HEALTH RELEVANCE: The immediate goal of the research is to explore how normal adult readers and listeners compose the meaning of a sentence out of the meaning of its parts. This is a skill that is universal among intact humans, but that can be degraded or lost in cases of language disorder, most notably aphasia. A better understanding of aphasia must be based on a better understanding of the specific linguistic functions that are lost in aphasia, and it is hoped that such a better understanding will lead to better treatment - an important goal in an aging population with an increasing expected incidence of strokes leading to language impairments. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We will evaluate the effect of one week of daily, low dose prednisone therapy in CF patients by following airway responsiveness to mecholyl challenge and pulmonary function tests, pre- and post-bronchodilator therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "More people in the U.S. die from lung cancer than from prostate, breast, colon and rectum cancers combined. Of the estimated 187,000 people in the U.S. who were diagnosed with non-small cell lung cancer (NSCLC) in 2009, over 37,000 of them presented with localized or early stage disease. This fraction is expected to grow as methods of early detection improve and become more widely disseminated. Stereotactic body radiation therapy (SBRT) has been shown to provide excellent local control (85%-95%) for early-stage lung cancer patients. However, a recent phase II study found Grade 3 or greater toxicities in a significant fraction of the patients, particularly those with centrally located tumors. Studies have shown that decreases in SBRT margins can significantly reduce the probability of normal tissue complications. As SBRT is increasingly becoming the therapy of choice for early-stage, localized non-small cell lung cancer, reducing the harmful side effects becomes increasingly important. We are proposing a novel failsafe technology to reduce toxicity while retaining local control for this growing population of patients. Our hypothesis is that tracking lung tumors directly during SBRT, using beam's-eye-view (BEV) imaging coupled with a dynamic multileaf collimator (DMLC), will lead to clinically significant normal tissue sparing. The current proposal is the first to employ an advanced multi- template marker-less BEV tracking algorithm to derive the real-time tumor location for DMLC delivery. Clinical benefits of the multi-template marker-less innovation include 1) automatic selection of relevant landmarks, 2) continuous tracking during deformations, rotations and partial obscurations, 3) no additional imaging dose to the patient, 4) direct imaging of the entire tumor, and 5) no need for invasive fiducial implantations and the associated adverse effects. This represents a substantial improvement over previous techniques. The therapeutic advantage will be quantified experimentally in an anthropomorphic phantom system under clinical conditions. The end result will be an integrated real-time target tracking and dynamic delivery system for radiation therapy as well as quantification of the anticipated clinical benefits. Clinical integration with the DMLC tracking is a challenging engineering problem with a measurable positive impact on human health on a large scale. This project represents well the ideals of the NIBIB to support research and development in the physical sciences and engineering for the improvement of human health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The sexual behavior of American adolescents has become a national concern. One out of every ten females between the ages of 15 and 19 has a pregnancy each year, and, by age 21, approximately one out of every five young people has required treatment for a sexually transmitted disease (STD). Although education is but one among many interventions that should be employed to reduce teenage pregnancy and STD infection, new technologies, such as interactive multimedia, have rarely been investigated as delivery media for sexuality instruction. To exploit the potential of multimedia for effecting behavioral change among teenagers, American Research Corporation of Virginia proposes the ongoing development of \"Promotion of Responsible Sexuality with Multimedia\" (PRISM), a fourth generation sexuality education program presented by interactive multimedia and targeted for adolescents in grades 9 and 10. The Phase I program yielded a proof-of-concept presentation consisting of text and graphics that focused on pregnancy prevention. Phase II specific aims include development of additional instructional activities for pregnancy and STD prevention, integration of the instructional activities with full motion video and audio, and evaluation of the intervention with regard to changes in knowledge, attitudes, and behavior. PROPOSED COMMERCIAL APPLICATIONS With the phenomenal growth of home computer purchases, potential markets of the proposed software include families with adolescents as well as schools. In a school environment, the program could be integrated with existing school sexuality education programs or implemented as a stand-alone curriculum. With the increased popularity of juvenile electronic bulletin board services, the program could be advertised and distributed by state and private educational networks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Of the three TGF-beta isoforms, type 1 TGF-beta is both the most abundant in most tissues and the most acutely regulated in injury, repair, and the pathogenesis of various diseases. Although they lack any obvious developmental defects, mice in which the TGF-beta1 gene has been knocked out by targeted disruption die at about 3 weeks of age of multifocal inflammatory disease. This is accompanied by significantly elevated levels of nitric oxide in the serum of these mice. We have investigated the autoimmune phenotype in these mice and demonstrated, by backcrossing these mice onto an major histocompatibility antigen II null background, that generation of autoantibodies and tissue inflammation is dependent on expression of MHC class II antigen and the presence of CD4+ T-cells. This cross enhances effects of TGF-beta1 on hematopoiesis resulting in myeloproliferative disease with prominent extramedullary myeloplasia. In another approach, immunosuppressive treatments including rapamycin, dexamethasone, anti-CD4, and anti-CD8 are being used to prolong the life of the TGF-beta1 null mice. This has now made possible the study of wound healing in 4-5 week old mice in which maternally-transferred TGF-beta1 has been depleted. Finally, use of differential display showed that expression of ND2, a mitochondrially encoded component of the electron-transport chain, was suppressed in TGF-beta1 null mice. This has led to the demonstration that several mitochondrially-encoded components of this enzyme cascade are coordinately suppressed, a finding consistent with the observation of ultrastructural abnormalities in Golgi and mitochondria of cells of liver, heart, and lung of TGF-beta1 null mice which are strongly suggestive of an energy deficit and impaired vesicular transport. We have shown that treatment of either TGF-beta1 (-/-) and TGF-beta1 (+/+) cells with TGF-beta in vitro results in rapid upregulation of expression of mitochondrial genes. Present investigations are aimed at determining the mechanism whereby TGF-beta1 regulates expression of mitochondrial genes and its effects on cellular energetics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "HIV lacking Vif is susceptible to human APOBEC3 (A3)-mediated restriction, rendering the virus non-infectious. The archetypal family member, A3G, has a well-defined role in HIV restriction in T cells. Other A3s have been implicated but despite nearly a decade of study, no consensus has been reached regarding the restrictive repertoire. Following reverse transcription, nascent viral cDNA transcripts are deaminated by A3 proteins resulting in transition mutations that inhibit HIV replication. To counteract this, Vif targets A3s for proteolytic degradation. However, sequences derived from infected patients exhibit G-to-A hypermutation, a hallmark of A3 activity, suggesting Vif neutralization in vivo is incomplete. I hypothesize that these mutations promote HIV genetic variation and underlie beneficial phenotypes such as the acquisition of drug resistance. In the following proposal, I will define three vital characteristics of the A3 family: (i) which A3s are relevant to HIV restriction,(ii) the extent to which they contribute to the HIV mutation rate, and, (iii) whether drug-resistant virl isolates emerge following A3-mediated sub-lethal mutagenesis. I anticipate that when taken together these studies will supply definitive conclusions to the innate host-defense field while prioritizing efforts to develop future HIV/AIDS therapies especially for HIV infected substance abusers. PUBLIC HEALTH RELEVANCE: HIV has an intrinsically high mutation rate that can result in a virus that is resistant to currently available anti- retroviral therapies. A better understanding o the mechanisms of HIV mutation will lead to more effective treatment options for disproportionately affected populations, including those suffering from chemical dependency.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project 3 utilizes human and murine model systems to address the links between signature genetic events in melanoma progression (p16INK4a loss and N-RAS/B-RAF mutation) and the DMAdamage response in melanocytes. This project combines analyses of novel murine models of melanoma with a comprehensive molecular and immunohistochemical study of a large, clinically annotated human melanoma database. In specific aim 1, we combine two different RAS alleles, conditionally inactivatable p53 and p16INK4a alleles, and an inducible, melanocyte-specific CRE allele to produce new murine models of melanoma that are highly faithful to the human genetics of this tumor. In these models, all oncogenic events are restricted to the melanocytic compartment, and include somatic inactivation of p16/p53 or somatic activation of K-RAS, Two of the alleles (Tyr-CRE-ER-T and conditional p16INK4a) are newly characterized and unpublished. In specific aim 2, we combine these novel murine models of melanoma with neonatal UV-B exposure to facilitate in vivo melanomagenesis. Specific aim 2 also includes a detailed immunohistochemical analysis of the kinetics of the expression of markers of the DNA damage response and senescence with or without UV- B treatment in the setting of RAS activation, p16INK4a loss and/or p53 loss. This specific aim employs a similar approach to analyze human dermal reconstructs in immunodeficient mice with and without UV exposure using reagents supplied from projects 1 and 2. In specific aim 3, we extend our analysis of human primary formalin-fixed and paraffin-embedded melanocytic lesions to identify the relationship among RAS/RAF/p16INK4a mutation, ERK MAP kinase activation, senescence and the DNA damage response in the progression from nevus to metastatic tumor. This specific aim includes a comprehensive immunohistochemical analysis of several markers of the DNA damage response and senescence, as well as a mutational analysis of N-RAS, B-RAF and p16INK4a. This specific aim is powered (250 melanocytic lesions from nevus to metaststic melanoma) to account for molecular heterogeneity in primary tumors but still uncover biologically significant relationships among these signature genetic events, the DNA damage response and melanoma progression. We expect this work will further our basic understanding of human melanoma progression as well as identify new clinjcal predictors of disease progression and outcome.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is designed to provide the Principal Investigator, Anthony L. Guerrerio, with the necessary knowledge, skills, infrastructure and experience required to transition to a position as an independent researcher in the field of pediatric gastroenterology, focusing on mucosal immunology and its role in oral tolerance and the development of Eosinophilic Esophagitis (EoE). Dr. Guerrerio outlines a five-year plan to investigate the basic mechanism by which mutations in the TGF? receptor lead to immune dysregulation and the development of EoE. This work will be performed under the mentorship of Dr. Harry C Dietz, Victor McKusick Professor of Medicine and Genetics and Investigator in the Howard Hughes Medical Institute at Johns Hopkins University. Dr. Dietz has an impressive track record of mentoring young investigators for successful careers in academic medicine, including those funded under the K award mechanism. Dr. Guerrerio also has the support of a team of extraordinary physician scientists at Johns Hopkins including Drs. Cynthia Sears and David Huso. All of these individuals have committed their time, resources, and expertise to facilitate Dr. Guerrerio's career development and research goals. In addition, the candidate will acquire additional skills and training through didactic coursework at the highly regarded Johns Hopkins Bloomberg School of Public Health. The candidate's past academic experiences underscore his commitment to academic medicine and his desire to acquire rigorous and complete scientific training. He completed a MS in physics, then entered the NIH-sponsored Medical Scientist Training Program at Johns Hopkins University and did his graduate work under the mentorship of Dr. Jeremy Berg in the Department of Molecular Biophysics. There he performed basic research investigating the molecular events that allow intracellular zinc-sensing by a metal responsive transcription factor. He also developed a method of targeting proteins to a single-stranded region of DNA, which may have therapeutic applications. He completed both a residency in Pediatrics and a fellowship in Pediatric Gastroenterology at Johns Hopkins University. During fellowship, he became interested in the role of choline, a small quaternary amine now known to be an essential nutrient for humans. He completed a small clinical trial in children with intestinal failure that revealed a high prevalenc of choline deficiency in this population, and suggested that intravenous delivery of choline may be required to replete choline in this population. This study also defined normal plasma choline percentiles by age for the first time. He also demonstrated a role for choline deficiency in the development of fibrosis in post-menopausal women with nonalcoholic steatohepatitis (NASH). Returning to the lab, Dr. Guerrerio became interested in the mucosal immune system and how inappropriate immune activation can lead to EoE. This interest crystalized around the characterization of Loeys-Dietz Syndrome (LDS), a newly described autosomal dominant disorder caused by mutations in the genes encoding the receptor subunits for TGF?. LDS is associated with an increased risk of developing EoE and other more general food allergies. Remarkably, LDS mice also demonstrate increased Th2 inflammation, and the spontaneous development of EoE. Dr. Guerrerio joined the faculty at Johns Hopkins as an Assistant Professor in July 2009. His research experiences have afforded him with a unique background in molecular biology, biochemistry, immunology, and both human and murine models of disease. He is now poised to apply these skills, under the guidance of his mentors, to study the basic mechanisms underlying the development of EoE. The research in this proposal will focus on the role of the TGF? pathway and its interplay between the cell types of the esophagus using LDS mice as a model system. Aims include: 1) To define the role of lymphocytes and Tregs in the development of EoE in LDS and how signaling alterations in these cells lead to inappropriate production of Th2 cytokines, 2) Investigate the role of epithelia cells and other nonhematopoietic cells in the development of EoE in LDS, and 3) Define the signaling defect in lymphocytes carrying the LDS mutation and test whether pharmacologic inhibition of TGF? signaling mitigates the EoE phenotype. The scientific training obtained through this grant will lead to publications, data, and experience that will enable the candidate t secure independent NIH funding within the next 4-5 years and establish himself as an independent physician scientist.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cationic antimicrobial peptides (CAMP), 12-50 residues long eptides with high content of cationic amino acids, are characterized by strong activities directed against broad range of microorganisms including pathogenic bacteria, fungi and parasites. Despite intensive research and wealth of accumulated detailed information, their modes of action are not well understood. This proposal is a part of the ongoing research on elucidation of properties and mechanisms of action of MUC7 12-mer, an antimicrobial peptide derived from the low molecular-weight salivary mucin 7. This peptide has potent antimicrobial properties against opportunistic fungal pathogen Candida albicans and cariogenic oral bacterium Streptococcus mutans. We have recently employed a global approach enabling study of many different aspects of action of antimicrobial peptides at once. In this approach altered susceptibilities to the peptide of thousands deletion mutants of yeast Saccharomyces cerevisiae were tested simultaneously. Among identified deletions conferring hypersensitivity to the peptide, many involved genes associated with the RIM101 signaling pathway, best known as a response to changes in environmental pH. In the first Specific Aim we will confirm induction of this pathway by the MUC7 peptide and investigate its role in interaction of the peptide with target cells. In the second Aim we will test a hypothesis proposing existence of two distinct mechanisms of action of the MUC7 peptide, one fungicidal and the other fungiststic, and will investigate how the RIM101 signaling relates to both. We will also expand this study to the pathogenic yeast C. albicans and calcium signaling, which we have recently identified as playing a role in defense against the MUC7 peptide, analogous to that of the RIM101 response in S. cerevisiae. In the third Aim we will extend the original fitness profiling by direct selection for S. cerevisiae deletion mutants resistant to the peptide. Public Health Relevance: Due to the declining rate of discovery of new antibiotics and antifungal agents, and spread of resistant strains of pathogens, there is urgent need for development of new classes of antimicrobial therapies. Antimicrobial peptides are a potential source of novel drugs. Results of recently completed yeast deletion screen with MUC7 peptide provide new directions for studying its mechanisms of action.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The therapeutic effectiveness of 5-fluorouracil (5-FU) in the treatment of proliferative diseases such as basal cell carcinoma (BCC), actinic keratoses (AK) and psoriasis could be greatly improved by enhancing its dermal delivery. A prodrug approach utilizing a sequential combination of a labile, water solubilizing promoiety and a stable, lipid solubilizing promoiety is proposed to accomplish this objective. The hypothesis is that the combination prodrug, with the additional labile, water solubilizing promoiety attached, will partition from the vehicle into the skin more effectively than the prodrug with only the lipid solubilizing promoiety. In the skin the water solubilizing promoiety will be rapidly hydrolyzed to give the lipid soluble prodrug which will hydrolyze and partition through the skin relatively slowly to give more specific dermal delivery of 5-FU. In this proposal the labile, water solubilizing promoiety will be the 1-acetyl group and the stable, lipid solubilizing promoiety will be a 3-alkylcarbonyloxymethyl promoiety. A series of linear alkyl chain 3-alkylcarbonyloxymethyl derivatives (single promoiety) and combinations of the 3-alkylcarbonyloxymethyl derivatives with the 1-acetyl group will be synthesized, characterized (solubilities and stabilities), and the abilities of the single promoiety and combination prodrugs to deliver 5-FU and intact single promoiety prodrug into and through hairless mouse skin will be determined in diffusion cell experiments. The results from the diffusion cell experiments will be analyzed to determine if the combination prodrugs are more effective than the 3-alkylcarbonyloxymethyl prodrugs at enhancing the ratio of dermal (delivery into the skin) to transdermal delivery (delivery through the skin) of 5-FU. A prodrug approach that exhibits a better ratio of dermal to transdermal delivery than 5-FU while delivering more 5-FU dermally should be more effective than 5-FU in treating BCC, AK and psoriasis while reducing the risk of systemic side effects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this proposal is to understand how developmental mechanisms contribute to neural circuits controlling animal behaviors. The simple neuroanatomy and repertoire of stereotyped locomotor behaviors of larval zebrafish provide an ideal model to study the contribution of developmental genes to the formation and function of neural circuits underlying defined motor behaviors. In this proposal, I will use the zebrafish 'space cadet'mutant as a tool to investigate mechanisms of axon guidance in the context of forming neural circuits that modulate a particular behavior called the \"escape response\". Larval zebrafish perform this maneuver in response to tactile or acoustic stimuli. Mutations in the space cadet gene result in an errant execution of the escape response, caused by a guidance defect of a small subset of specialized commissural hindbrain neurons, called the spiral fiber neurons, which are part of a highly conserved \"brainstem escape network\" of neurons that control motor behaviors. In addition, retinal ganglion cell orientation and pathfinding defects within the retina suggest that space cadet plays a critical role in intraretinal pathfinding, a process that is poorly understood. In this proposal, I first will use recombination mapping, DMA sequence analysis, gene expression patterns, and gene misexpression techniques to determine the molecular identity of the space cadet gene to better understand its function in the context of molecular signaling pathways. Second, I will use in vivo timelapse imaging of retinal ganglion cell and spiral fiber axons in wild-type and space cadet mutant embryos to provide insight into the role(s) space cadet plays in mediating neuronal growth cone guidance in vivo. Finally, I will determine whether spiral fiber neurons and their downstream neuronal synaptic partners are required to mediate specific turning behaviors in response to environmental stimuli. Together, these experiments will provide insights into the functional implication of vertebrate axon guidance on animal behavior. My proposed experiments and analyses will result in novel information essential for understanding mechanisms of axon pathfinding and the control of motor behaviors by neural circuits. Moreover, the results from these studies will also provide a foundation on which to address the mechanisms underlying human congenital disorders causing visual and locomotor impairment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Suppressor of cytokine signaling (SOCS) family of proteins contain eight members (SOCS1 through SOCS7; and cytokine-induced SH2-containing protein or CIS), each of which contains a conserved central Src homology 2 (SH2) domain that is flanked by a variable length N-terminal domain and a 40-amino acid C-terminal domain called the SOCS box. SOCS proteins regulate signals transmitted by hemopoietin cytokine receptors with associated JAK kinases. Signals induced by Insulin and growth factors whose receptors possess intrinsic tyrosine kinase activity (IGF-1, FGFs, PDGF, EGF and erythropoietin) are also under feedback regulation by SOCS proteins. Significant interest in SOCS family proteins stems from the belief that SOCS proteins function to integrate multiple cytokine/growth factor signals and mediate cross-communication between antagonistic factors. Importance of SOCS proteins is underscored by the wide array of pathologic conditions that result from deletion of SOCS genes or dysregulation of SOCS genes expression. These include allergic and autoimmune diseases, insulin resistance, diabetes, liver degeneration, lymphoid deficiencies, polycystic kidney disease and cancer. Although the role of CIS, SOCS1, SOCS2 or SOCS3 in negative feedback regulation of cytokine and growth factor signaling is firmly established, functions of SOCS4, SOCS5, SOCS6 and SOCS7 are largely unknown. In this study, we have investigated the possibility that SOCS proteins are involved in mechanisms by which retinal cells are protected from oxidative stress-induced neuronal cell death, light-induced damage of photoreceptors and retinal degenerative effects of pro-inflammatory cytokines. We show for the first time that SOCS5, SOCS6, SOCS7 are constitutively expressed in the retina, while SOCS1, SOCS2, SOCS3 and CIS are undetectable but are induced by proinflammatory cytokines and growth factors. We further show that SOCS proteins are induced in the retina by high intensity light and following exposure of retinal cells to hypoxic conditions. Demonstration that retinal cells respond to cytotoxic cytokines, hypoxia or high intensity light by upregulating SOCS expression, suggest that SOCS proteins may mitigate injurious effects of environmental, chemical or oxidative stress in the eye and may therefore be a potentially important neuroprotective agents of the mammalian retina.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Understanding the immunological and genetic basis of control of AIDS virus replication should help inform vaccine design. As in HIV-infected humans, a limited number of macaques spontaneously control SIV replication to less than 1,000 copies/ml after infection (\"Elite Controllers\"; ECs) (18, 32). This is a greater than two log reduction from the typical plasma virus load after challenge with SIVmac239. Recently we identified sixteen ECs in a cohort of 196 Indian rhesus macaques (18). Genotyping for MHC class I alleles revealed that fourteen of these sixteen ECs expressed either Mamu-B*17 or Mamu-B*08, which appear to be the macaque functional equivalents of HLA-B57 and HLA-B27, respectively (2, 9, 12, 21, 26, 29). However, the monitoring and sampling of these animals, which were typically identified in the course of experiments run by different investigators, has not been consistent. Moreover, due to financial constraints, animals often must be euthanized at the end of specific studies. We plan to establish a sample bank, database, and to house existing and future EC macaques at the WNPRC. Making these unique resources available to the community of investigators working on SIV would be an extremely valuable service to the field. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A Respiratory Syncytial Virus (RSV) vaccine is not yet available. Avatar is developing a novel, di-tyrosine crosslinked, conformationally locked preF subunit immunogen (DT-preF) that, formulated with an adjuvant, will form the basis of a preF RSV vaccine. The risk of vaccine-enhanced disease (VED) came into sharp focus with the results of a 1960s formalin- inactivated RSV vaccine that resulted in numerous hospitalizations and the deaths of two children. A maternal- to infant vaccination strategy using a potent immunogen is therefore now considered the safest and most promising strategy for protecting infants. We have developed a maternal-to-infant vaccination mouse model that uses a highly virulent subtype of RSV line 19, and is uniquely able to elicit immune responses and pathology in mice that mirror many of the fundamental, age-dependent characteristics of enhanced RSV disease observed in humans. The model can distinguish between safe and unsafe vaccine immunogen formulations, and using this this model enables us to identify a DT:preF:adjuvant formulation that overcomes the risks of VED. In Phase I of this proposal, we will test four DT-preF:adjuvant formulations and identify the one that elicits the most protective responses that does not cause VED in vaccinated, pregnant dams or non-pregnant female mice, or lead to the development of enhance respiratory disease (ERD) in infant mice born to immunized mothers, following RSV challenge. In Aim I, we will rank and select the two most potent DT-preF:adjuvant formulations (based on neutralization titers) in immunized, adult female mice; in Aim II, we will identify a lead DT-preF:adjuvant formulation that most effectively protects postpartum dams, newborn pups and weanlings following maternal vaccination (based on lung viral titers); and in Aim III, we will demonstrate that our DT-preF adjuvant formulation elicits protective responses that do not lead to the development of ERD in dams or infant mice following RSV challenge. In Phase II, we will carry out additional preclinical studies in cotton rats as the next step toward human clinical studies Our lead formulation will be innovative in that it will (i) have sufficient stability for commercial development, (ii) provide infants safe, potent, long-lasting, and cost-effective protection against RSV, and (iii) enable maternal- to-infant vaccination.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Abstract The muscular dystrophies are the heterogeneous group of inherited muscular disorders characterized by muscle wasting and weakness, and are affecting both children and adults. The worldwide prevalence of inherited neuromuscular diseases estimated in a review study was of 28.6/100,000 and 33.3/100,000 if rarer disorders are included. As a group, these disorders, covers a considerable proportion of patients with chronic disease, however, recent studies of prevalence of the muscular dystrophies in the United States are lacking. For two funding cycles, the Muscular Dystrophy Surveillance, Tracking and Research Network (MD STARnet) has identified and tracked the health of patients with Duchennne and Becker muscular dystrophy (DBMD). This new funding cycle will seek to characterize the population of individuals who have a variety of muscular dystrophies through the expansion and modification of MD STARnet. The project will assess the feasibility of using the existing infrastructure of MD STARnet to expand to population-based surveillance for other types of muscular dystrophy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The computerized patient monitoring system developed in conjunction with Surgery Branch, NHLBI, provides realtime, beat-to-beat analysis of physiologic waveforms from patients within a four-bed intensive care unit. Based around a Xerox Sigma-3 Computer, the system monitors the electrocardiogram, arterial and venous blood pressurs, body temperature, urine output, and blood loss, as well as thermal and dye-dilution cardiac output. A Morphology Analsis of the electrocardiogram and arterial blood pressure waveforms is performed to detect fiducial markers. The application of cross-correlation techniques to the electrocardiogram allows the detection of premature ventricular contractions. Beat-to-beat data from the most recent thousand electrocardiographic complexes may be recalled at any time in the form of Joint Interval Histograms and Correlation Coefficient Scatter Diagrams. A five-minute electrocardiogram memory is maintained in realtime for use in the detection of transient ventricular arrhythmias. Vital signs are retrievable as eight-hour nursing shift summaries in tabular form, or as 12-, or 72-hour vital signs graphs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A critical driving force for optimal development of T helper type 1 (TH1) lymphocytes is signaling through the IL-12 receptor. The IL-12 receptor is composed of two subunits, with expression of the IL-12 receptor beta 2 chain (IL-12RB2) dictating a high affinity IL-12 receptor complex. Signaling through this high affinity IL-12 receptor controls the development of TH1 lymphocytes and the maintenance of this phenotype, while limiting lineage commitment to the TH2 phenotype. Since TH1 lymphocytes mediate cellular immunity, while TH2 lymphocytes enhance humoral responses, early expression of the high affinity IL-12 receptor is critical for a commitment to cell mediated immune responses. Salmonella is an intracellular pathogen of macrophages, epithelial cells and possibly dendritic cells, and requires cell-mediated immunity for clearance. Based on recently published work, we demonstrated that Salmonella-infected macrophages can significantly limit IL-12RB2 expression on T lymphocytes early in the response. This finding has profound implications for the early development and commitment of T lymphocytes to the TH1 lineage during Salmonella infection. The overall goal of this proposal is to define the mechanisms for Salmonella-induced reductions in IL-12RB2 expression in vitro and in vivo. At present, it is not clear whether induced reductions in IL-12RB2 expression are solely mediated by soluble factors or require macrophage-T cell contact. IL-12RB2 expression will be quantified at the level of mRNA using quantitative RT-PCR, and at the protein level using Western blot, FACS and radioreceptor analyses. Furthermore, reductions in T lymphocyte function associated with the loss of IL-12RB2 will be assessed, and a functional assessment of developing TH1 and TH2 lymphocytes will be determined by following STAT-4 activation, and T-bet, GATA-3 and c-maf mRNA expression, respectively. Whether infected dendritic cells can induce such alterations in CD4+ T cells will also be determined. Taken together these studies represent the first to define mechanisms whereby an intracellular bacterial pathogen can adversely affect the early development of TH1 lymphocytes upon infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The cytolytic T lymphocyte (CTL) specifically recognize and kill by direct lysis target cells which are diseased or foreign. The clinical benefits of manipulating the CTL response are clear, however, safe manipulation will almost certainly require an understanding of CTL-target cell recognition at the molecular level. Other than the qualitative requirement of foreign antigen and MHC proteins, target cell components necessary for optimal CTL recognition have not been defined. The overall aim of this project is to more precisely define the receptor ligand interactions between membrane proteins on the CTL and target cell. We will do this by constructing and using a new type of lipid vesicle (an artificial target cell) which is cell size and has a sealed outer membrane. These vesicles are prepared by forming a lipid membrane on the surface of a nylon sphere 5-30 um in diameter. In initial experiments, we have used anti-CD3 monoclonal antibody covalently attached to these vesicles to trigger CTL and demonstrated CTL mediated permeability changes in the vesicle membrane. We have also developed an approach for examining receptor ligand interactions by incorporating cell surface proteins into the artificial target cell (ATC) membrane and examining cell-ATC adhesion. Using this approach we have shown directly that LFA-3 is a ligand for the T cell surface protein CD2. We hope to use this methodology to biochemically define receptor-ligand interactions. These studies should result in a better understanding of the molecular basis of CTL-target interaction and serve as a basis for designing a vaccine able to trigger an in vivo response.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of the proposed work is to investigate the role of bacterial colonization/infection in the etiology of gastric and urinary bladder cancer. Studies in human will explore the relationship of bacterial colonization of the stomach and severity of precancerous lesions with the concentration of carcinogens and their precursors in gastric juice as well as with resultant DNA damage in stomach tissue. Total N-nitroso compounds in human gastric juice will be determined by thermal energy analysis and genotoxins, assayed using the SOS chromotest, will be isolated and identified. Studies in vitro and in vivo in rodents with experimentally induced bacterial colonization/infection of the stomach and bladder will explore the mechanism by which bacteria and inflammatory changes produce alkylation and oxidation of Dna. Alkylated DNA bases in tissues will be determined by immunostaining and immunoassays and levels of 8-hydroxydeoxyguanosine, as a measure of oxidative DNA damage, will be detected by HPLC with electrochemical detection. Unknown carcinogen spectrometry. Bacterial enzymes responsible for nitrosation of amine/amides will be characterized. This approach should lead to a better understand of molecular mechanisms by which bacterial colonization/infection is involved in human carcinogenesis. Once markers for DNA damage that are associate with microbial colonization/infection in the stomach and urinary bladder have been identified, they could be exploited in subsequent epidemiological and intervention studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will track the course of three mosquito invasions. One of these, the spread of Aedes albopictus in North America, began with the shipment of used tires to the US from Asia in the early 1980's. We are interested in its interaction with resident mosquitoes, especially competition with the treehole mosquito Aedes triseriatus. Two other invasions, by Aedes albopictus and Aedes aegypti, are early in the colonization phase in Bermuda. Tests for interspecies agonistic effects will be conducted by tracking population change in 100 US localities concurrently, with the help of an extensive network of school children and their teachers, who will send substrates from egg traps to us for species identification. Substrates from egg trap sites in Bermuda, part of a vector surveillance program, will be sent to us for species identification, using PCR methodology. Rates of change for each population will be calculated, and tests for density effects by each species will be conducted. This study, which will be among the largest concurrent census-based studies of natural populations, will attempt to link experimental work with actual changes at the population level. An independent test for competition between species will be done by sampling female adults in eight locations in the US and eight in Bermuda, selected for their relatively high and low densities of each species. Females will be measured, with the expectation that competition will result in smaller females. Tests for the influence of larvae on female habitat choice will also be conducted under field conditions in the US and Bermuda. Habitat choice can be either a mitigating or an intensifying factor for other species interactions, including competition. All species involved in this study are involved in the transmission of important human diseases, including La Crosse virus, Dengue virus, and yellow fever. All have potential to transmit West Nile virus.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Genetics and Regulation training program at The University of Chicago is a broad interdisciplinary program to train Ph.D. scholars in advanced rationales and methods of genetic analysis for careers as independent scientists in basic and applied biomedical research and education. The training faculty members are comprised of tenure-track members of six basic science departments: Biochemistry and Molecular Biology, Ecology and Evolution, Human Genetics, Microbiology, Molecular Genetics and Cell Biology, and Organismal Biology and Anatomy, plus selected faculty in the Departments of Medicine, Psychiatry, Radiation Oncology and Surgery. Most of the trainers also have secondary appointments in the Committee on Genetics. The training grant is administered by the Director, and a Steering Committee, constituted of trainers from the major participating departments and representing various sub disciplines of genetics. Trainees are selected from students admitted to various Departments and degree-granting Committees, on the basis of previous academic record and specific interest in genetics. They take courses in molecular, transmission and population/evolutionary genetics and meet uniform program standards. Progress is monitored on a regular basis by individual Departmental/Committee advisory committees, and the Genetics and Regulation Training Grant Steering Committee. Common features of the program are course work, laboratory rotations, teaching and assisting in courses, seminar programs, journal clubs, joint research meetings, student seminar programs and Mini-Symposia focused on areas of current interest in genetics. A total of 18 increasing to 24 trainees is proposed for the next five year period. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Marital distress and divorce is of great concern in the U.S., as the divorce rate for first marriages is around 40 to 45%, and marital problems are a risk factor for a variety of child and adult problems. To combat disturbing trends of marital dissolution and distress, as well as the associated problems and costs, there is increasing government support for the wide spread implementation of marriage education. Although this movement is supported by existing empirical research, we still need to know much about marital processes over time and the effects of marriage education, particularly across diverse samples and outcomes. We propose to implement a community level, controlled, longitudinal, randomized investigation of an ongoing marriage education program for Army couples delivered by Army chaplains. The intervention is an Army adaptation of the Prevention and Relationship Enhancement Program (PREP) for young married Army couples. This adaptation is entitled Building Strong and Ready Families (BSRF). BSRF is designed to target empirically identified risk and protective factors for marital conflict and distress, which are also strongly associated with a range of individual and family functioning problems. We seek to test the impact of BSRF on a range of relationship risk and protective factors, and the effects of intervening with these factors on a wide range of outcomes which have not been studied in the marriage education literature, including depression, substance abuse, parenting, and child behavior outcomes. We will also evaluate a uniquely wide range of important, yet understudied, relationship constructs. We also have access to a uniquely diverse population, one not typically reached by traditional mental health services, due to the demographic make up of the Army and the use of Army chaplains as service providers (hence, a less stigmatizing resource for prevention and counseling for many people who will not seek services from a mental health professional). Our longitudinal design will allow tests of bi-directional effects of variables such as mental health and relationship health over time, enabling us to test and refine our theoretical model of relationship risk and protective factors. Thus, the proposed research investigates underlying factors of healthy marriage, processes of marital distress and dissolution, bidirectional effects of marital conflict and individual and family functioning, and the immediate and long term effects of marital education. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In Phase I of this study, Anatomix developed an interactive 3-dimensional computer graphic model of embryonic chick development that illustrated the early phases of the developing head-fold, intraembryonic coelom and heart tube fusion. This model, which is the first of its kind to show the temporal aspects of development in 3-dimensions, was tested via presentation to first year medical students. In Phase II of this study, we propose to use the techniques pioneered during Phase I, along with student feedback on the chick embryo models, to develop similar, yet more sophisticated models that show the full range of human cardiac development. Human models will be more directly relevant to medical education and the necessary histological material is available. Increased sophistication will be achieved through (l) the use of high end computer graphic hardware and software (2) use of a professional science writer to develop textual materials and (3) a scientific advisory board including teachers, clinicians and textbook authors. The materials developed in phase II will constitute a core of anatomically and temporally accurate models of embryonic heart development that will be used to produce instructional materials targeted to a wide variety of teaching levels. PROPOSED COMMERCIAL APPLICATIONS: The core graphics technology produced by this research will be used to produce 3D embryology references for basic science research, interactive multimedia learning tools for students, and virtual embryos for clinicians to study the pathogenesis of cardiac malformations. The products of this research will he delivered to the educational markets as CD-ROM titles and VHS or DVD video.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Precise phenotyping depends significantly on the quality of animal subjects. Stable housing and husbandry minimize the influence of exogenous factors that could distort phenotyping results. Additionally, mice submitted to the UMass MMPC share space with cohorts from microbiologically diverse sources. They, as well as mice housed in resident colonies, must be protected against potential cross-infection by adventitious agents. The Animal Care Core operates under the supervision of Dr. Jerald Silverman as the Core Director, who oversees the animal care facility of UMass Medical School. The Animal Care Core provides facilities and services relevant to these priorities under the following specific aims: (1) to provide stable, biocontainment housing, husbandry and health care for mice (2) to provide a dedicated room for housing (Podl-124 of LRB) and on-site phenotyping procedures, (3) to assess the microbiological profile of each cohort and provides the Center Director with interpretation and advice regarding the results, and (4) to provide to the users ofthe phenotyping service, at the discretion ofthe Center Director, results and advice on the health status of submitted mice. The goal of the Animal Care Core is to support the operation of UMass MMPC with issues pertinent to animal care and quality of phenotyping mice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This grant proposal requests a fourth cycle of funding to explore and expand our understanding of the genetic basis of Hirschsprung disease (HSCR). From the genesis of these studies until today, we have established HSCR as a model complex disorder by: (1) demonstrating that HSCR shows oligogenic inheritance; (2) discovering at least 6 (RET, GDNF, EDNRB, EDN3, SOX10, SEMA3) of the 12 genes known for this disorder; (3) demonstrating genetic interactions between RET and EDNRB signaling (in humans and mouse models) and their downstream transcriptional consequences, as an explanation of the reduced penetrance; (4) discovering the role of common polymorphisms at RET and SEMA3 on HSCR and the molecular basis of this association by in vitro and in vivo analyses; and, (5) assembling and maintaining a database/sample repository (International Hirschsprung Disease Consortium) and a public information resource for ascertaining new cases and families. This grant describes four sets of specific aims that have the goal of identifying a large number of susceptibility genes for HSCR by virtue of them harboring many more dosage and sequence mutations than would be expected by chance. First, we will confirm our recent discovery of SEMA3 as a HSCR locus; second, we will perform focused genetic screens of dosage mutations at all human genes and a genome-wide association study for common susceptibility variants; third, we will target DNA sequencing on a set of 2,000 genes and target loci (~10% of the coding genome) that have emerged from these prior genetic screens. A fourth aim will be to continue sampling new cases and families, over and above those being collected by members of the International Hirschsprung Disease Consortium, and expand our information resource. We have focused this research on gene discovery and on primary functional analysis leading to specific identification of genes, rather than a deeper understanding of mechanisms of pathogenesis. This choice, a minor departure from our tactic in the last cycle and not possible before, is occasioned by the realization that we have identified very few of the HSCR genes and that many more can now be discovered using new technologies. A better map of this 'HSCR gene universe' will allow a better choice of which mechanisms to investigate in a deeper fashion.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The CA125 antigen is a well-characterized serum marker for epithelial ovarian carcinoma and is utilized clinically to monitor response to therapy and recurrence of this malignancy. Recent identification and cloning of a cDNA molecule encoding the core peptide of the CA125 antigen revealed considerable homology to a family of membrane-associated mucins, and the CA125-encoding gene was designated MUC16. The cloning and characterization of MUC16 provides multiple new opportunities for a molecular genetic-based, mechanistic evaluation of the role of this molecule in ovarian neoplasia. The long-term goal of this project is to test the hypothesis that MUC16 plays a critical role in the development and/or maintenance of the malignant phenotype in ovarian carcinoma. This goal will be accomplished through a broad range of experimental approaches designed to gain insight into the function of MUC16 in normal development and cell biology, as well as in the neoplastic transformation of ovarian epithelial cells. The specific aims of this project are to: 1) Annotate the genomic structures and analyze promoters of the MUC16 and Muc16 genes. Exon-intron boundaries, coding regions, untranslated regions and transcription factor binding sites will be characterized physically and functionally. 2) Characterize the function of Muc16 in vivo. Protein expression during embryonic development and in the neonatal and adult mouse and in murine ovarian cancers will be determined, as will the effects of transgenic knockdown of Muc16 in a temporal- and tissue-specific manner when crossed with transgenic mice prone to invasive ovarian carcinoma. 3) Using previously generate data on the regulation of MUC16 expression, correlate implied molecular pathways with promoter structure, and test the hypothesis that CA125 secretion results from post-translational processing of MUC16, exploring the mechanism of this processing. 4) Determine whether a reduction in MUC16 expression in human ovarian carcinoma cells affects the neoplastic phenotype. An RNA interference approach will be used to determine whether reduction of MUC16 mRNA levels in human ovarian carcinoma cells affects cell proliferation, differentiation, anchorage-independent growth or tumorigenicity in vivo. It is anticipated that studies of the function of MUC16 in multiple biological contexts will prove most effective in ultimately elucidating the role of this molecule in ovarian tumorigenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION Despite advances in critical care, patients with major burns remain at significant risk for septic complications. Previous work from our laboratory has shown that in the murine model of thermal injury and superimposed sepsis there is a bone marrow myeloid shift resulting in increased monocytopoiesis and arrest in granulocytopoiesis. Although sympathetic stimulation is known to accompany both clinical and experimental thermal injury and adrenergic stimulation modulates immune cell function, cytokine release and myelopoiesis under non-stressed conditions there is a lack of information on bone marrow noepinephrine (NE) release during trauma and its impact on myeloid commitment. Therefore, we hypothesize that burn injury and sepsis lead to increased bone marrow NE release which influences myeloid commitment and thus contributes to the subsequent cytokine dysregulation. We will test this premise in Specific Aim #1 by establishing that bone marrow sympathetic nerves are activated with thermal injury and sepsis leading to increased release of norepinephrine. Using pharmacologic, surgical and genetic models of sympathetic denervation we will establish that monocytopoiesis and granulocytopoiesis consequent to bum with infection are mediated by adrenergic stimulation in bone marrow. Specific Aim #2 will establish that bone marrow sympathetic stimulation with burn plus sepsis changes monocyte as well as monocyte progenitors to release enhanced amounts of cytokine in response to bacterial endotoxin. Using both tissue macrophage and those derived from bone marrow progenitors, cytokine responses to endotoxin as well as phagocytosis and chemotaxis will be determined in mice with denervated compared to neural intact bone marrow. Having established the role of sympathetic stimulation in mediating changes bone marrow myeloid commitment and function Specific Aim #3 will focus on the elucidation of cellular mechanisms by which NE mediates the observed changes. This aim will use paradigms involving bone marrow monocyte progenitor cells subjected to adrenergic agomsts and antagonists to examine the proliferation and differentiation. Such findings will be extended using murine and human monocytic cell lines to investigate specific cell-signaling mechanisms. Results of the proposed research will provide important new information on the pathophysiology of thermal injury and sepsis and will lead to therapeutic approaches to enhance the survival of the critically injured.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This protocol is intended to cover screening of individuals for treatment of their psoriasis when those individuals do not enter a study, for one reason or another. These are not \"off-protocol\" visits, but neither can the visit(s) be attributed to a particular protocol. With new patients a medical history is obtained, the skin is examined visually, the nature of participation in research studies is discussed. Current protocols are described. With old patients the skin is examined, interim medical history discussed and current protocols are described. No biopsies are taken or blood drawn; other protocols cover these activities. If individuals enter a particular study the screening visit is attributed to that protocol. Some people may be potential candidates for a later but not current study, and some individuals are deemed unsuitable for any study; in both these cases we would attribute the visit to this screening protocol.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The precise control of neurotransmitter release is mediated by proteins specific to synapses. One family of essential regulators is SV2, a family of transporter-like proteins that have emerged as a target of new therapies to treat nervous system disorders. Synaptic vesicles also contain a related protein, SVOP (SV2-like protein), about which very little is known. To study the role of SVOP in neurotransmission, we propose to establish mouse lines in which SVOP expression can be universally or selectively disrupted. These mice will be a crucial reagent in determining both how SVOP contributes to neuronal functioning and whether it holds promise in the development of new therapeutics. PUBLIC HEALTH RELEVANCE: This pilot project will generate a genetically modified mouse line that will be used to identify the function of SVOP, a transporter-like protein present on synaptic vesicles about which very little is known. SVOP has significant similarity to SV2, a small family of synaptic vesicle proteins that are essential regulators of neurotransmitter release. SV2A is the binding site of a new class of antiepileptic drugs that are also finding use in the treatment of other nervous system disorders including neuropathic pain and dyskinesias. The proposed mouse line will play a crucial role in vetting SVOP's potential as an alternate drug target.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Develop and evaluate an evidence-based, user friendly, and culturally sensitive training module, \"Dementia Care: Sleep Behavior Disorders\" targeting formal caregivers in nursing homes and assisted living facilities as well as informal (family) caregivers providing care in the home, using DVD/Web DVD technology. AIMS: Translate current evidence based research on sleep disturbances among dementia patients into a comprehensive training module of all-inclusive strategies aimed at improving sleep for nursing home and assisted living caregivers and home based caregivers in Spanish and English versions. Evaluate the usability, cultural acceptability and workplace benefit of both English and Spanish versions of the module. Pilot test the revised module as to the ability to increase knowledge in and impact on both formal and informal caregivers. Evaluate the final product with a large and diverse population. INNOVATION: Translation research of existing research findings (not anecdotal care information) related to sleep behavior disorders of demented individuals into information that can be readily used by a caregiver. An Interactive Training Module: Sleep Behavior Disorders in a DVD/Web DVD format that provides enhanced interactivity and an ability to update training materials. This product re-creates the training program in the Spanish language and culture. OUTCOMES: Effective but inexpensive method of translating the best of dementia care research in the management of sleep behavior disorders in individuals with dementia for the formal or informal bedside caregiver. This quality training will benefit the long term care industry by relieving the burdens of existing workers, facilitating recruitment of new workers, and reducing worker attrition. Better training for coping with sleep behavior disorders extends the ability of families to care for their loved ones in the home, significantly reducing excess patient disability and caregiver stress. COMMERCIAL POTENTIAL: A Market Feasibility Study identifies 6,329,754 total institutional and/or individual buyers including, among others, the following market segments: institutional caregivers, nurses, educational institutions and the lay public. This study projects a first year market penetration rate of 1.5 percent of those markets. PUBLIC HEALTH RELAVENCE: The US government spends over $100 billion a year on AD and, as of 1996, the average lifetime cost of care was $175,000. Alzheimer's disease costs corporate America $33.16 billion a year. The business community is often unaware of its annual expenditures impacted by Alzheimer's disease, primarily because of the popular belief that the disease affects only individuals who have retired from the workforce. That misconception overlooks the millions of currently employed people caring for a family member with the disease. Alzheimer's Cost to U.S. Businesses: Caregiver absenteeism: $ 7.89 billion, Productivity loss: $13.22 billion Replacement of caregivers who leave: $3.59 billion, Continuing insurance for workers on leave and fees to temp agencies: $1.32 billion, Cost of Employee Assistance Programs: $0.0055 billion and Caregiving total: $26.0255 billion. The LightBridge Sleep Behavior Disorders Care Program: (i) improves quality of life for the Alzheimer patient and their care providers, (ii) provides less costly care for longer periods of care for families, local, state and federal agencies, and (iii) is pro-active in its outreach to minority groups such as the Hispanic population. OUTCOMES: Better training for coping with sleep behavior disorders extends the ability of families to care for their loved ones in the home, significantly reducing excess disability and caregiver stress. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The decline in adaptive immunity, nave T-cell output and a contraction in the peripheral T cell receptor (TCR) repertoire with age are largely attributable to thymic involution and the loss of critical cytokines and hormones within the thymic microenvironment. To further understand the various processes at work in the involution process, we have focused our efforts on characterizing several of the cellular and molecular changes that occur within the thymus and thymocytes during aging. We have approached these efforts in several different ways: (1) Molecular profiling: We have examined the transcriptomes of progressively aging mouse thymi and thymocytes of different sexes and on caloric-restricted (CR) vs. ad libitum (AL) diets. Genes involved in various biological and molecular processes including transcriptional regulators, stress response, inflammation and immune function significantly changed during thymic aging. These differences depended on variables such as sex and diet. Interestingly, many changes associated with thymic aging are either muted or almost completely reversed in mice on caloric-restricted diets. These studies provide valuable insight into the molecular mechanisms associated with thymic aging and emphasize the need to account for biological variables such as sex and diet when examining the molecular pathways responsible for thymic involution. Moreover, in the thymocyte profiling studies, genes associated with oxidative phosphorylation, T- and B- cell receptor signaling and antigen presentation were observed to significantly change with thymocyte age. Interestingly, several immunoglobulin chains were found to have increased expression in thymocytes with age. While the increased expression of immunoglobulin genes in aged thymocytes may have been attributed to the thymic B cells (which were found to be actively producing IgG and IgM antibodies), further analysis of highly-purified thymic T cells derived from aged but not young thymi demonstrated high levels of IgM on their cell surface suggesting the possible presence of auto-antibodies on the surface thymocytes with advancing age. Whether there are autoantibodies targeting proteins on thymic subsets with advancing age remains to be determined. We are currently examining several gene pathways that were elucidated by these studies and are also utilizing pro-thymic hormone infusions and various transgenic and knockout mouse models to more fully understand the role of certain genes in maintaining thymic integrity or facilitating thymic loss. We believe array analysis of the thymi and thymocytes of hormone treated mice may yield valuable data on the common molecular processes involved in thymic regeneration. (2) Proteomic profiling: We have also examined the proteomic changes that occur within the thymus during the involution process. Over 25,000 peptides were identified using mass spectroscopy in these studies to examine specimens derived from young and aged thymi and thymocytes. While the majority of the proteins identified were not significantly different between the age groups, there were several hundred proteins that were differentially expressed between the young and old thymic specimens. Many of the identified proteins were associated with oxidative stress, T cell differentiation, signal transduction and adipogenesis. These profiles are currently being further analyzed and processed and several of the identified proteins are being verified. (3) Lipomic profiling: Here, we analyzed possible age-related lipid metabolism alterations in total thymus and thymocytes from mice with 4 to 18 month-old as well as in thymus of aged mice treated or not with growth hormone (GH), known to partially reverse thymic involution. Using electrospray tandem mass spectrometry, we were able to increased amounts of tryglicerides (TG), free-cholesterol and C24:1 sphingomyelin in total thymus of old mice as compared to young ones. Moreover, we verified a decrease in sulfatide C24:1 and in unknown long-chain ceramides, comparing aged with young thymi. In addition, a significant increase in 4-HNE and 8-epi-prostaglandin F2a (PGF) in thymi of older animals. By immunohistochemistry, we verified an increase in the expression of 4-HNE as well as of tumor necrosis factor-alpha (TNF-a) and CD204 (a marker for oxidized LDL receptor) in aging thymus. In our analysis of isolated thymocytes of various ages, we found a decrease in TG, free cholesterol and in sphingomyelin species with age. Interestingly, we detected a significant amount of ceramide C24:1 in thymocytes from old mice when compared to their younger counterparts. Following GH treatment, we found a significant decrease in the amount of TG, free-cholesterol and PGF in the thymus of old mice. In addition, GH treatment increased significantly the amount of dihydroceramides in aged mice and promoted a trend to decrease 4-HNE, as compared to control. These data suggested to us that there is increased oxidative stress in the thymus as an animal ages and that such changes may be attenuated upon growth hormone treatment. The overall goal of this project is to produce comprehensive molecular and biochemical profiles of the thymus during the aging process to identify unique and common changes in gene, protein and lipid expression and functionally relate such changes to the physiological and functional alterations that occur within the thymus with age. While such profiles may also be valuable for other lymphoid organs and cellular subsets, we have focused our efforts on the thymus, as its involution is believed to be one of the most significant obstacles to overcome in addressing the immunological deficits associated with aging.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In response to infection, CD8+ T cells are selected by interaction with APC to undergo significant expansion and differentiation into effector cells. These effector cells participate in clearance of the infection and then undergo a rapid death phase where the majority (80-95 percent) of the effector population is lost. The remaining cells represent the memory pool and are maintained at relatively constant levels for life. Additionally, CD8+ T-cell responses to different epitopes from the same pathogen exhibit reproducible immunodominance hierarchies. Together, the expansion, death, memory and immunodominance relationships define homeostasis of Ag-specific CD8+ T cell in response to infection. Surprisingly, little is known regarding the regulation of these facets of Ag-specific CD8+ T cell homeostasis. In particular, essentially no information has been obtained regarding the regulation of the death phase of Ag-specific CD8+ T cells. Homeostasis of Ag-specific CD8+ T cells in response to infection of mice with Listeria monocytogenes (LM) or Lymphocytic Choriomeningitis Virus (LCMV) has been well documented. Our preliminary results demonstrate that all facets of Ag-specific CD8+ I cell homeostasis-expansion, death, memory and immunodominance are aberrant in perforin/IFN-y double deficient mice after LM infection. Examination of Ag-specific CD8+ T cell homeostasis in the parental perforin- and IFN-y-deficient mice reveals that perforin regulates the expansion phase after LM infection while IFN-y regulates immunodominance hierarchies after LM infection and the death phase and after LM or LCMV infection. The overall goal of this application is to further our understanding the role of IFN-y in regulation of immunodominance and the death phase of CD8+ T cell homeostasis and to determine if IFN-y also regulates the death phase of CD4+ T cells. The following Specific Aims address the overall hypothesis that IFN-y regulates immunodominance and the death phase of Ag-specific T cells independently of its role as an effector molecule in resistance to infection. SA-1. Determine the role of IFN-y in regulation of immunodominance. SA-2. Analyze the role of IFN-y regulation of the death phase of CD8+ T cells. SA-3. Determine the impact of IFN-y on regulation of Ag-specific CD4 T cell homeostasis. Since memory levels of Ag-specific T cells result from the magnitude of expansion minus the death phase understanding the regulation of the death phase has important implications, not only to increase our fundamental understanding of the immune system, but also to enhance the process of rational vaccine design.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of this work is to understand the relationship the structure of external and middle ears and their function in auditory signal transmission. The approach is to measure acoustic performance is selected species that vary in aural structure and to construct analytical models in which structural components are explicitly represented in networks of mechanical elements. From these measurements and models general rules for structure- function relations are derived. During the current grant period attention was focused on cat and lizard. In cat measurements of the properties of the external and middle ear have determined the performance of these parts of the ear as collectors and transmitters acoustic power to the inner ear. The lizard studies have shown that the usual presumption that sound enters the ear primarily though the ear canal must be modified; accessory sound pathways through large areas of the lizard's skin also couple sound to the middle ear, most effectively at low frequencies. Comparison of the measurements on cat and lizard shows that the tympanic membranes have distinctly different acoustic properties; at high-frequencies the cat TM is resistive whereas the lizard TM is masslike. These results suggest that substantial interspecies differences exist in the processes that couple acoustic stimuli into tympanic- membrane motion. The projects proposed here are focused on these processes and on the structural basis of variations in these processes among different vertebrate classes. Three areas will be examined: 1) The performance of the mammalian external ear, both as a collector of acoustic power and as a source of information about the location of sound sources, will be studied through measurements in species whose external ears have interesting structural similarities and and differences. 2) Accessory pathways to the middle ear will be studied in lizard, chicken, and also in some mammals (pig and sheep) in which the tympanic-membrane structure differs from cat, guinea pig, and human. In pigs and sheep a large structurally-distinct section of the tympanic membrane is flaccid rather than stretched. We will explore the hypothesis that this \"pars flaccida\" functions as an accessory pathway to the middle ear. 3) The relationship between structure and function of the tympanic membranes of cat, guinea, pig, chinchilla, alligator lizard, and chicken will be studied through measurements of their \"mechanical transformer\" properties. Since the tympanic membrane plays a key role in producing the transformer-like properties of the middle ear and its structure varies dramatically among vertebrates, specification of the TM's structure-function relations is key. The understanding of sound reception that results from this work will be of practical significant to audiologists in the diagnosis of conductive hearing disorders and to otologists in the surgical correction of abnormalities of the ear.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have instituted routine dynamic Intensity Modulated Radiation Therapy (dIMRT). This is the most complex of IMRT treatments and requires considerable pre-treatment verification to ensure accurate execution. These treatments are delivered using our twin Clinac-21EX accelerators. A Hi-ART system (Tomotherapy Inc.) for delivering treatments via helical tomotherapy has been installed, accepted and is currently treating patients. This system provides ROB with cutting edge treatment technology to support future clinical trials. We have started a series of initiatives with our Nucletron High Dose-Rate Afterloader (HDR) using a small, high intensity 192Ir source for interstitial and intercavitary brachytherapy. The oldest Clinac-21EX accelerator has been replaced with a new machine is a state-of-the-art Varian TrueBeam STX. This accelerator will be equipped with the new high definition multi-leaf collimator with leaf widths as thin as 2.5 mm. This will render it capable of performing frameless stereotactic radiosurgery (SRS), although a set of circular collimators with diameters ranging from 4mm to 25mm and an SRS planning module are included in the acquisition. The patient couch also provides 6 Degrees-of-Freedom (6DOF) to greatly improve positioning accuracy by permitting corrections for pitch, roll and yaw, in addition to the traditional longitudinal, lateral and vertical position adjustments. This machine includes the Edge Extracranial SABR Package that will permit the unit to deliver frameless stereotactic body radiation therapy (SBRT) using advanced motion management based on radiofrequency local positioning using implanted fiducial transponders and infrared cameras. The revised Edge package includes a Calypso radiofrequency positioning system, utilizing implantable fiducial transducers and real-time tracking capability. ROB has acquired a second TrueBeam STX unit to replace its RapidArc Clinac-21EX. This machine provides treatment redundancy with the older unit, as well as having additional photon energies of 8 MV and 18 MV. Work is now progressing on upgrading the OSMS system on the older unit and the clinical implementation of a prostate radiotherapy protocol using Calypso RF transponders tomonitor prostate position and correspondingly gate the treatment delivery. ROB has acquired a new Elekta Flexitron 40 channel High-Dose-Rate (HDR) remote brachytherapy afterloader to replace its previous unit. Additionally, ROB has acquired a new B-K ultrasound unit, which has vastly improved imaging capabilities relative to the previous unit. We are currently acquiring a couch-mounted stepper-stabilizer to improve needle insertion methodology. As a next step, ROB is planning to remove its Tomotherapy unit from B2-3643 and to modify the room into a brachytherapy suite which is optimized for minor surgical procedures such as this. .", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The five-year relative survival rate of cancer patients has improved from 50 to 68% in the last thirty years. The survival rate of pediatric cancers has reached 80%, and the NCI predicts that 1 out of every 250 adults will be a survivor of childhood cancer by 2015. As the number of cancer survivors increases, so does the concern for their quality of life post-treatment. One of the most serious problems associated with radiation and chemotherapy is the off-target effect on germ cells. For young women, this is a particularly serious concern because the immature oocytes that comprise the ovarian reserve are highly sensitive to chemo/radiation therapies. Since the ovarian reserve represents ALL the follicles available for future fertility as well as endocrine function, the loss of immature oocytes and subsequent sex steroid deficiency have significant physical and psychological consequences. The goal of this project is to better understand the mechanisms associated with survival and death of immature oocytes caused by cancer treatments, and in doing so, identify neo-adjuvant medical approaches for preserving fully functional ovaries in young women undergoing treatment. A recent breakthrough discovery by Gonfloni et al. demonstrated that the ovarian reserve could be protected against the chemotherapeutic drug, cisplatin, by co-administration of kinase inhibitor imafinib mesylate. Although its precise mechanism was not demonstrated, their study suggested that cisplatin-induced oocyte death was mediated by p63, a member of the p53 transcription factor family. Additionally, it was proposed that imatinib mesylate rescued oocytes from apoptosis by blocking activation of p63 by c-Abl. Our preliminary studies of p63-null oocytes confirmed the essential role of p63 in cisplatin induced death of oocytes. In addition, our studies suggested that the gene dosage of p53 and p63 determines the death/survival of oocytes. Moreover, p73, another member of the p53 family, was also highly expressed in oocytes undergoing apoptosis in response to cisplatin. Therefore, the overall hypothesis of this proposal is that the balance between survival and death of oocytes in primordial and primary follicles is controlled by the interaction between p53 family members, and thus the p53 family members are targets for fertoprotective agents. To develop safe and effective fertoprotective therapies, it is essential to elucidate the mechanism through which p53 family members are activated by chemotherapy. To advance our understanding, we have developed an assay system that consists of in vitro culture and subrenal grafting of mouse ovaries. We will dissect the molecular pathways involved in the death of immature oocytes by applying this assay system to genetically engineered mouse models. The results of these proposed studies will be critically important for understanding the mechanisms that underlie physiologic and iatrogenic follicle death and may reveal new strategies for protecting the fertility of young women diagnosed with cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite reasonable clinical experience with mixtures of L-amino acids in the management of Phenylketonuria (PKU), L-amino acids which are the source of approximately 75% of dietary protein equivalent in this condition have not been systematically studied to determine their efficacy in supporting longterm growth and in maintaining normal amino acid profiles. This study is an attempt to evaluate the ability of L-amino acid mixtures such as that of the Ross Metabolic Formula System (RMFS) to support normal nutritional status of infants with PKU.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this research is to develop advanced magnetic resonance spectroscopy (MRS) and imaging techniques and to apply them and other complementary methods to studying brain metabolism, neurotransmission and enzyme activity. MRS allows measurement of the neurotransmission of glutamate and GABA in vivo, which plays important roles in many major psychiatric diseases, including depression and schizophrenia. During 2016-2017, we made significant progress in the development and application of novel spectroscopic and imaging techniques for studying metabolism and neurotransmission in vivo in the brain. We invented and validated a computational method for quantifying spatially localized MRS data involving multiple nuclear spin systems (Zhang et al, Med. Phys., 2017, doi: 10.1002/mp.12375). This method can reduce the time for calculating chemical basis function, using a typical desktop personal computer, from days and weeks to just a few minutes. As such, design and optimization of proton MRS by computers have become practical and highly useful. By optimizing localized proton MRS using extensive computation to simultaneously maximize spectral resolution and sensitivity to nuclear spin relaxation we have developed a novel multiparametric technique that can simultaneously measure concentration, longitudinal and transverse relaxation times of many brain chemicals, including glutamate, in a single scan session within ten minutes of MRS data acquisition. This multiparametric approach allows us to measure both concentrations and the microenvironments of brain chemicals (e.g., glutamatergic neurons) in a typical clinical setting (An et al, Magn Reson Med. 2017. doi: 10.1002/mrm.26612.). In addition, a novel radiofrequency pulse-driven steady state method has been developed that can measure transverse relaxation of chemicals without the need to change echo times. Significant progress has also been made in studying glutamate and GABA neurotransmission and in vivo enzyme activities in both human and animal experiments. In particular, we demonstrated, for the first time, the feasibility of measuring carbonic anhydrase activity in the human subjects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dental cementum covering the tooth root is critical for tooth attachment, long-term stability, and function of the dentition, yet regulation of root development and cementum formation remains poorly understood, hampering efforts towards periodontal regeneration. This application is a Pathway to Independence Award proposal that aims to advance fundamental knowledge on cementum biology by addressing gaps in our current understanding of cementum formation, focusing on areas of: extracellular matrix (ECM) proteins and local phosphate/pyrophosphate regulation implicated to be central to the process of cementogenesis. We propose that a coordination of local matrix proteins and phosphate regulating factors is crucial for proper development and mineralization of cementum. This hypothesis will be tested by three specific aims: 1) To define the role of ECM protein bone sialoprotein (BSP) in cementum formation, using null and conditionally null mice and in vitro approaches; 2) To determine the function of ECM protein osteopontin (OPN) in cementum development, by evaluating OPN-pyrophosphate interactions in tooth development; 3) To establish the role of sodium- phosphate co-transporter PiT1 (SLC20A1) during cementoblast differentiation and function, by mapping its expression during periodontal development, and analyzing the phenotype in PiT1 conditional null mice, including effects on cementum regulatory factors. The applicant is a postdoctoral research fellow highly qualified to lead this research program based on his training studying cementoblasts in vitro and cementum formation in vivo, reporting on phosphate/pyrophosphate metabolism in tooth formation, driving the paradigm- shifting findings regarding pyrophosphate control of cementum formation, and discovering the necessity of ECM protein BSP for cementum mineralization. The applicant's long-term career goal is to lead a productive research program that provides significant insights into the molecular mechanisms driving tooth root formation and mineralization, and translates those insights into novel approaches for regenerating periodontal tissues and restoring function. This project is an ideal starting point for the applicant to transition to his independent research carer because it provides a framework for development of further skills necessary to accomplish the long-term goal. The proposed career development plan incorporates didactic coursework, laboratory training, and a structured mentorship plan to facilitate accomplishment of short-term goals, including development of a skill set for mineralized tissue research, establishing an independent research project, providing a pathway for a faculty position, and affording a future opportunity to build on this proposed work. The applicant's institutional environment provides strong support in areas of postdoctoral development and transition.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Serine/threonine kinases Akt (also known as PKB), are among the most important and versatile protein kinases at the core of human physiology and disease. Akt regulate numerous cellular processes, including cell growth, glucose and lipid metabolism, differentiation, proliferation, and apoptosis. Three highly homologous Akt isoforms, encoded by three separate genes, are expressed in mammals. Combined deletions of the three Akt isoforms in the mouse suggest compensatory and complementary roles of isoforms. However, the three isoforms also have distinct functions, even though the mechanisms of specificity are only starting to emerge. Studies of the role of Akt, and specifically of Akt isofors, in atherogenesis are very limited. It has been recently shown that genetic ablation of Akt1 promotes coronary atherosclerosis via the enhanced expression of proinflammatory genes in the artery wall in a macrophage independent manner. We have recently explored the role of Akt3 in atherosclerosis using mice with a genetic ablation of the Akt3 gene. Our studies have demonstrated a specific, macrophage dependent, atheroprotective role for Akt3 in hyperlipidemic ApoE-/- mice. In a step-by-step fashion, we tested various potential mechanisms and demonstrated that the absence of Akt3 transforms macrophage into cell which aggressively accumulates cholesterol esters via two mechanisms: increased lipoprotein uptake via pinocytosis, and parallel increased protein stability of cholesterol esterifying enzyme ACAT1 (also known as SOAT1). Multiple lines of evidence suggested that Akt3 suppresses atherosclerosis by restricting cholesteryl ester accumulation and foam cell formation in macrophages, an early and critical step in atherogenesis. The mechanism is Akt isoform specific, and our preliminary studies suggest that one mechanism for specificity is via differentia subcellular localization of Akt1 (a major isoform in macrophages) and Akt3 in macrophages. The long-term goal of this proposal is to investigate the molecular mechanism of Akt3 regulation of macrophage function. As a specific hypothesis, we propose that Akt3 specifically suppresses transformation of macrophage into lipid accumulating phenotype by inhibiting macrophage pinocytosis and by regulating a particular branch of cholesterol metabolism - cholesterol esterification via direct regulation of ACAT1 protein stability. We will investigate the role of AK3 in atherogenesis in vivo and in vitro with a particular focus on the molecular mechanisms of regulation of Akt3 activity, ACAT1 expression and pinocytosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Voltage-gated Na+ channel (VGSC) SCN1B is a multi-functional gene involved in modulation of VGSC transcription, cell surface expression and subcellular localization, regulation of Na+ current (INa), and cell adhesion. SCN1B is expressed as two splice variants in brain: 21 and 21B (initially called 21A). 21B is predominantly expressed in developing brain while 21 expression prevails after the first postnatal week. 21 and 21B share the N-terminal immunoglobulin domain in SCN1B that is required for cell adhesion. Mutations in SCN1B in humans are linked to Genetic Epilepsy with Febrile Seizures Plus (GEFS+)-spectrum disorders. A mutation causing complete functional loss of SCN1B results in Dravet Syndrome, a pediatric epileptic encephalopathy that is the most severe disorders of the GEFS+ spectrum. Scn1b null mice have severe, spontaneous epilepsy and early lethality, a phenotype similar to Dravet Syndrome. In addition, loss of Scn1b in mice results in neuronal migration and pathfinding abnormalities, disrupted axonal fasciculation, INa abnormalities, altered VGSC expression in the hippocampus, and mistargeting of Nav1.6 to the axon initial segment (AIS) in cerebellum. Thus, disruptions in SCN1B expression during development may lead to a spectrum of neurological diseases. The long-term goal of this work is to understand the physiological role of SCN1B in brain. The objective here is to determine the roles of 21 and 21B in neuronal excitability. The central hypothesis to be tested is that SCN1B is crucial for establishment of excitability in brain. It is proposed that 21B, which is expressed during late prenatal/early postnatal brain development, is essential to SCN1B function. Further, it is proposed that disruptions in SCN1B expression during this critical developmental period result in defects in brain circuitry, as well as aberrant VGSC subcellular localization in neurons, that lead to secondary changes in excitability. The rationale for this work is that understanding the role of SCN1B in brain development may lead to novel treatments targeting SCN1B for epilepsy. Three specific aims will be pursued: 1: To test whether Scn1b expression during brain development is critical for establishment of excitability in vivo. 2: To determine whether Scn1b-mediated epilepsy is an interneuronopathy. 3: To use human induced pluripotent stem cell (iPSC) neurons to understand the role of Scn1b in human epilepsy. This work is expected to reveal the physiological roles of Scn1b (21 and 21B) in brain development and how defects in Scn1b expression contribute to epilepsy in mice and humans. These results will have positive impact because these identified roles will lead to greater understanding of the mechanisms of GEFS+-spectrum diseases and may lead to novel therapeutic agents for epilepsy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studying an exceptionally rare well-aged population represents a logical progression for a new phase of the BLSA. By including a new cohort of participants who have aged exceptionally well, the BLSA will be able to utilize fully the extensive set of measures recently introduced in the study. In addition, the study of IDEAL aging is consistent with the original mission of the BLSA concerning the study of healthy aging, independent of the effects of disease. Despite excellent health at the time of enrollment only a small proportion of the nearly 500 current BLSA participants aged 80 years or older meet IDEAL criteria. By enrolling the IDEAL cohort in the BLSA their biologic, physiologic, behavioral and functional characteristics will be evaluated with using the same methods used with the current cohort who will serve as a type of control group. As is customary in the BLSA, we plan to follow this cohort for life with yearly visits. STUDY AIMS The first aim is to identify factors and characteristics that distinguish IDEAL from non IDEAL individuals. In the current BLSA, an extraordinarily wide range of information is collected on biomarkers, physiological measures, behavioral and environmental risk factors and functional and disease-related outcomes. We intend to compare the two groups to identify factors that discriminate IDEAL aging from non IDEAL aging individuals. The second aim is to identify physiological, environmental and behavioral characteristics that are risk factors for losing the IDEAL condition over several years or longer. We postulate that the mechanisms of extreme longevity probably differ from those associated with delay or escape from disease and disability. For example, we hypothesize that in older persons who escape diseases and disability, homeostatic mechanisms (integrity of anti-oxidant mechanisms, low level of oxidative damage to macromolecules, low inflammation, unimpaired autonomic reflexes, normal hormone levels etc.) are relatively intact, with little accumulated damage. DESIGN The operational definition of IDEAL aging is living until age 80 and meeting the current BLSA enrollment criteria which in brief requires the absence of chronic disease and conditions and mobility and cognitive impairment evaluated using a rigorous screening process. It can be conservatively estimated that approximately 1% of those 80 years or older are IDEAL aged individuals. IDEAL participants will be recruited from the Baltiore-Washington area, which has over 600,000 persons aged 80 years or older of whom an estimated 6,000 should be IDEAL candidates. Enrollment of IDEAL participants in the BLSA will be conducted under and R&D contract, which has been awarded to WESTAT. Recruitment has been initiated and the first few participants have been enrolled.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (provided by candidate): My long-term career goal is to become an independent researcher and obtain an academic faculty position at a top-tier academic research university, where I may teach and mentor students in the fields of environmental and molecular toxicology. My primary research interests focus on determining chemical origins for human health problems, particularly how environmentally relevant endocrine disrupting chemicals perturb vertebrate development. Thus far my background training has been primarily in environmental health sciences and toxicology, with additional training in biochemistry and molecular biology. With this grant application, I will adhere to a rigorous mentored training curriculum to acquire expertise in emergent techniques, including next- generation RNA sequencing, genomics, and computation biology. Using these techniques and new molecular methods, systems biology and bioinformatics will be a cornerstone of my independent research career. Environment: The research and career development plans described in the grant application will be carried out at Oregon State University (OSU). There, I will engage with highly successful faculty and research staff, and take advantage of unique state-of-the-art facilities and outstanding institutional training environment. OSU has a rich training environment, and is therefore an ideal location to achieve my career goals. My selected mentors are top scientists in the fields of developmental and systems toxicology and computational biology, which will provide me with the best possible training and opportunities to succeed as an independent scientist. My primary mentor, Dr. Robert Tanguay is widely recognized toxicologist that has championed zebrafish as a relevant and modern model for systems toxicology and human health related diseases. Dr. Tanguay has strong collaborative relations with my co-mentors, Dr. Susan Tilton and Dr. Katrina Waters, both of whom are highly regarded computational biologists. This unique opportunity will afford me with training in one of the best aquatic laboratories in the world and top-notch computational biology facilities at the OSU-CGRB/PNNL. Research: The goal of this research project is to determine how endocrine disrupting chemicals adversely affect development of the reproductive system. Concerns regarding human exposures to historical and emerging AHR environmental ligands are compounded by evidence that AHR activity modulates endocrine pathways. Specifically, a systems biology approach will be used to study how aryl hydrocarbon receptor (AHR) ligands perturb neuroendocrine- and sex-steroid pathways early in development, leading to reproductive health effects later in life. Zebrafish are an ideal developmental vertebrate model for use with cutting edge deep sequencing technologies to dissect molecular events that lead to reproductive toxicity, particularly the involvement of non-coding regulatory RNAs in modulating the ER signaling. These studies will not only enhance our understanding of model AHR ligands, but also for a variety of ligands which lack toxicity data.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Inflammatory lung injury is a significant cause of morbidity or mortality. Recent evidence suggests 1) that accumulation of neutrophils (PMN) in the lungs is associated with acute as well as chronic inflammatory lung injury and 2) that alveolar macrophages (AM) may release mediators that recruit and activate PMN in the lungs. Recognizing the potential detrimental consequences associated with the accumulation and activation of PMN in the lung as well as the need for controlling and delimiting inflammatory responses; one might speculate that mechanisms exist which under normal circumstances decrease or suppress PMN recruitment and activation in the lung. This premise is supported by the observation that PMN are rarely present in the extravascular space of the lungs. To address this possibility we hypothesized that under normal conditions, AM release factors that suppress recruitment and activation of PMN in the lung. Our preliminary findings have supported this premise by showing that supernatants from unstimulated AM contain factors that decrease PMN chemotaxis, adherence and oxygen radical production. Our immediate specific aims are: 1. To determine and partially characterize factors released by AM in vitro then suppress PMN function in vitro. 2. To determine if these factors exist in vivo. 3. To determine if in vivo PMN suppressor factors are biochemically similar to in vitro AM derived PMN suppressor factors. Since PMN can injure the lung, suppression of needless PMN recruitment to and activation in the lung may be essential to maintaining healthy lungs. Furthermore, defects in AM synthesis and/or release of PMN suppressor factors may well be the basis of certain inflammatory disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "People living with HIV infection are now recognized as an important target population for HIV prevention efforts. Unfortunately, the sexual behavior of HIV-positive persons has only recently become a frequent subject for scientific study. One important aspect that remains largely unstudied is the pattern of sexual relationships among people living with HIV. In this project, investigators will develop a typology of the sexual relationships of HIV-positive adults and examine how relationship patterns are related to sexual risk and risk-reduction behaviors. Data will come from the Risk & Prevention Study, collected in late 1998 from a national probability sample of 1,421 adults drawn from the HIV Cost and Services Utilization Study (HCSUS). The specific aims are to: (1) Develop and apply a classification scheme for describing the sexual relationships of HIV-positive adults in terms of number of partners and type and concurrency of relationships; (2) examine how sexual relationship patterns vary by age, gender, sexual orientation, race/ethnicity, and fertility/parenthood status; (3) examine how relationship duration and exclusivity are related to other relationship characteristics, such as emotional closeness, drug use, and violence; (4) test for differences between relationship types in sexual risk and risk reduction behaviors, including condom use and avoidance of anal or vaginal intercourse, and in related behaviors, such as disclosure of HIV status to sexual partners; (5) estimate the prevalence among HIV-positive adults of HIV risk-reduction strategies that involve partner selection, and examine the correlates of these strategies; and (6) examine the bivariate and multivariate associations between relationship patterns (number of partners and concurrency of relationships) and self-reports of having been recently diagnosed with a sexually transmitted disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "B lymphocytes are the central mediators of humoral immunity, with aberrant B cell function contributing to multiple autoimmune diseases including rheumatoid arthritis and lupus. B cell function is regulated by cell-surface molecules that generate transmembrane signals, regulate intercellular communication, and direct lymphocyte localization within tissues. The aim of these Studies is to examine the in vivo functions of CD20 and CD22, two B cell-specific celt-surface proteins. CD20 is a membrane-embedded component of an oligomeric complex that regulates transmembrane 2+ Ca transport and cell cycle progression. Anti-CD20 immunotherapy has become a standard treatment for non-Hodgkin's lymphoma, and shows great promise for the treatment of autoimmunity. Despite this, relatively little is known about the function of CD20 in vivo and why it is such an effective target for immunotherapy. CD22 is a lectin-like member of the immunoglobulin superfamity that functions as an adhesion molecule for diverse sialylated cell-surface and soluble ligands. Ligand binding by CD22 regulates transmembrane signals and the survival of peripheral B cells. Genetic alterations in CD22 ligand binding may contribute to autoimmunity. Based on these findings, we propose that CD20- and CD22-directed monoclonal antibodies (mAbs) will be effective therapeutics for autoimmunity with individual unique properties. Since mechanistic and outcome studies are difficult and expensive in humans, we have developed mouse models for anti-CD20 and -CD22 mAb therapy to test this hypothesis. There are four specific aims designed to further determine how CD20- and CD22-directed therapies regulate B cell function in vivo and in autoimmune mice. In Specific Aim 1, the functional significance and functional consequences of CD20 and CD22 mAb therapy will be assessed in normal mice. In Specific Aim 2, the efficacy of mAb therapy will be assessed in mouse models of autoimmunity. We will determine whether anti-CD20 and anti-CD22 therapy influences the onset or severity of autoantibody production and disease. Specific Aim 3 will assess whether and how mAb therapy affects the development of an autoimmune repertoire. Specific Aim 4 will determine whether combined anti-CD20 and anti-CD22 therapies synergize to regulate the development of autoimmune disease. Since CD20 and CD22 provide important regulatory checkpoints for ablating or adjusting humoral immune responses, modulating their in vivo functions may provide mechanisms for modulating humoral immunity and effectively treating autoimmunity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research involves the histological study of human temporal bones with special emphasis on Meniere's disease and other types of endolymphatic hydrops (such as cochleo-saccular degeneration). These abnormal cases will be studied with particular attention to the vascular supply and pattern of the endolymphatic duct and sac, as well as to the para-vestibular canaliculus and its contained large vein.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This grant requests support for travel and expenses of fellows to attend the 2007 Clinical Immunology Society (CIS) School on Systemic Autoimmune Diseases. This grant is requested to help defray travel costs for the U. S fellow travel and expenses to attend the School. Additional funding will be solicited from pharmaceutical and biotechnology companies. [unreadable] [unreadable] This School will be an intensive four-day residential course to be held in March 14-18, 2007 in Santa Fe, New Mexico. The program will be advertised through academic program directors and members of CIS, The American College of Rheumatology and American Association of Allergy and Applied Immunology as well as through other major societies with interests in clinical immunology. [unreadable] Advertising emphasis will be given to attract applicants representing minorities. [unreadable] [unreadable] The topic of the School will be Systemic Autoimmune Diseases, geared toward fellows in training holding either an MD an/or a PHD, within their last years of fellowship training. The primary goal of the school is education on the diagnosis, pathogenesis and treatment of systemic autoimmune diseases such as SLE, Scleroderma, Arthritis, and Vasculitis as well as gene therapy. These presentations will include topic overviews, state of the art establishes treatments, laboratory approaches, pathogenesis and future biologics. Emphasis will be given to new treatments that are under development and future possibilities. Sufficient time will be allocated for fellows to present either interesting cases or their work. [unreadable] [unreadable] By the end of the School, participants should be able to better diagnose and treat systemic autoimmune diseases, have an enhanced awareness of clinical immunology processes as they relate to systemic autoimmune diseases and its importance in scientific discoveries and clinical application; and become ambitious in pursuing academic careers to further our understanding, diagnosis and treatment of human systemic autoimmune diseases. Lastly, the School should stimulate future collaborations between young investigators in different medical centers and countries, and between young investigators and experienced physician/scientists in the field. [unreadable] [unreadable] The school will be residential in character and held in an informal setting and will attract applicants from the North America, Europe and Asia. Ten faculty and 25 fellows will be selected to attend. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Programmed cell death occurs in many tissues of our bodies throughout life. The quick and efficient removal of dying cells is a critical step that protects neighboring cells, and is also an important part of tissue remodeling and wound healing. The failure to remove apoptotic bodies has been implicated as a cause for certain types of chronic inflammation and autoimmune disorders. In mammals, macrophages and dendritic cells, as well as many other cells types can carry out the engulfment of apoptotic corpses. The cloning of the engulfment genes in the model organism C. elegans, and the identification of their homologues in mammals, has provided an exciting opportunity to delineate this process in better detail. Our work during the previous funding period defined a role for a novel engulfment protein ELMO in regulating the cytoskeletal reorganization to mediate engulfment. Specifically, ELMO associates with another protein Dock180 and together they function as a novel type of Rac-GEF to activate Rac, and thereby modify the cytoskeleton. Using a combination of biochemical analyses and phagocytosis assays in mammalian cells, we have shown that the ELMO/Dock180/Rac pathway represents an evolutionary conserved mode of signaling that promotes engulfment. We also identified two new players (RhoG and TRIO) that function together with ELMO during engulfment. However, a number of key questions remain. The upstream receptor(s) on phagocytes that links to the ELMO/Dock180/Rac module during engulfment is not known. In Aim 1 we will characterize a novel type of receptor that we have cloned through a yeast two hybrid screen with ELMO that promotes phagocytosis of apoptotic targets. Aim2 will address how phagocytes are attracted to apoptotic cells in vitro and in vivo, and the role of phagocyte proteins in this process, including ELMO and Dock180. Thus far, the relative importance of the ELMO/Dock180/Rac signaling module in a mammalian model system, such as in mouse models, have not been addressed. Aim3 will address the importance of ELMO in engulfment in vivo, using conditional knockout of ELMO1 and conditional dominant negative ELMO1 transgenic mice we have generated. Taken together, through these studies, we expect to gain a better molecular understanding of the fundamentally important process of clearing apoptotic cells and define the role of specific players during engulfment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will expand upon the applicants' previously developed computer simulation model of advanced HIV disease and OI's to derive one which encompasses the full natural history and spectrum of HIV disease, and incorporate HIV-RNA as a predictor of disease progression. They will incorporate into the model data from randomized trials of combination antiretroviral therapy as well as from recent OI prophylaxis trials. Using this expanded model and up-to-date clinical, cost, and quality of life data, they will estimate the cost and cost-effectiveness of different HIV management strategies for individuals and groups of patients throughout the course of the disease. Building upon the intellectual foundation and working relationships they have previously established, they propose to examine specific hypotheses related to: 1) the comparative efficacy of early, aggressive intervention with antiretroviral therapy; 2) the relative performance of combination antiretroviral therapy compared to OI prophylaxis, and 3) the impact of new developments in OI research in refining the role of prophylaxis. Finally, they will use the results of these analyses to derive implications for clinical practice. The ultimate objective is to provide timely information which will be useful to clinicians and policy makers in developing rational approaches for the management of HIV.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The marmoset, a small South American primate, is a unique animal in which to study the early embryology of multiple births, and the differentiation of germ cells from early fetal life through adulthood. The marmoset normally produces biovular twins which show complete chorionic fusion prior to somite formation. The embryological processes which lead to blastocyst fusion and early blastocyst form, i.e., prior to fusion, are unknown. It is the aim of this research to describe the pre-fusion blastocyst and determine how and when blastocyst fusion occurs. As a first step in carrying out this study, it will be necessary to determine when, during the marmoset reproductive cycle, ovulation occurs, and to develop a simple and reliable test for early pregnancy. During pregnancy in the marmoset, inter-twin vascular anastomoses develop and allow the exchange of cells which migrate or are motile during embryonic development. This gives rise to individuals which are chimeric for hematopoietic, lymphoid, and, most especially, germinal tissue. Thus, we have available a primate model in which to examine sex-determining factors and in which to evaluate the roles of somatic and germinal elements in gamete differentiation. Data indicate that approximately one-half of all males possess germ cells with an XX sex chromosome constitution. Preliminary evidence indicates that germ cell chimerism also occurs in female marmosets.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study is collecting \"hard to get\" data about the actual economic behavior of street-level opiate addicts. The virtual absence of quantifiable data about the street behavior of addicts is noted in almost all literature reviews. This lack of data is a major obstacle for testing theories about and developing drug policies toward opiate addiction. This research will begin to provide necessary data and analysis of addict income, expenditures, crime, and drug use behavior. This grant is a pilot study investigating the economic behavior of street-level opiate addicts in the East Harlem area of New York City. The study will obtain data about the total income and expenditures of addicts. The porportion of total income derived from crimes of different kinds and the proportion of total expenditures due to opiate consumptin will be estimated. Changes in addict lifestyle due to arrest, increase in drug use, etc. will be related to economic behavior. During the first year, the project began testing different ethnographic and interviewing techniques in order to develop the best and most cost-effective techniques for obtaining quantifiable data on the addict's daily income, expenditures, crime and drug use. This methodological experimentation will continue in the second year. A sample of street-level addicts selected to represent differing economic lifestyles, ethnicities and sexes are being followed longitudinally for two years. A census of addicts living in or routinely active on the streets in a given geopraphical area will be completed and basic demographic data on these addicts will be obtained.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application requests support through an NIAAA Mentored Research Scientist Development Award (K01) to promote the career development of Dr. Cathy Lau-Barraco in the area of alcohol secondary prevention research for emerging adults. As part of the Candidate's transition to become an independent investigator pursuing this line of research, she has developed an intensive training plan that aims to build upon her existing skill set and fill in important gaps in her training. The Candidate's proposed training goals are to: (1) broaden and strengthen her alcohol knowledge base, particularly as it relates to the emerging adulthood developmental period, (2) develop skills in qualitative and advanced quantitative research methods, (3) develop expertise in the development and evaluation of secondary prevention programming for emerging adult drinkers, (4) increase proficiency in the principles and skills of motivational interviewing, (5) enhance scholarly productivity via secondary data analyses and manuscript preparation, and (6) develop a competitive R01 grant application testing the personalized feedback secondary prevention program in a larger efficacy trial examining both short-term and long-term effect on alcohol-related outcomes. The primary aims of the proposed research plan are to: (1) adapt a feedback-based secondary prevention program for nonstudent emerging adults, and (2) conduct a pilot study comparing alcohol-related outcomes between emerging adults randomly assigned to the tailored personalized feedback (PF) condition or assessment-only (AO) control condition. The proposed research plan is divided into two phases. The goal of Phase I is program development and refinement. The goal of Phase II is to evaluate the newly adapted program in a controlled pilot study. The new program will be evaluated against an AO control over a 9-month follow-up period. A total of 220 men and women between ages 18 to 25 will be recruited from the community through newspaper advertisements, flyers, and internet postings. In the pilot trial, PF participants will receive personalized feedback regarding their alcohol use and related risk factors delivered within the context of a 50-minute session using motivational interviewing strategies. The AO participants will complete the assessment measures only. It is hypothesized that participants in the PF condition will reduce their alcohol use and alcohol-related negative consequences more than participants in the AO condition over the 9-month follow-up period. PUBLIC HEALTH RELEVANCE: The aim of the proposed research is to develop prevention programming specifically for nonstudent emerging adult drinkers, as this group may be particularly vulnerable to alcohol-related problems in later adulthood. Implications of the findings may be profound given the disparity in the existing scientific literature with regard to the understanding and prevention of alcohol-related risk among nonstudents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The work proposed represents a continuation of our exploitation of heritably different inbred mouse strains to uncover and understand genetic factors involved in the regulation of the immune response. We have determined that spleens from different mouse strains differ in the number of plaque forming cells responding to certain well characterized antigens. Moreover, mouse strains differ in their in vivo vs in vitro (cell suspension) response in characteristic ways. The genetic bases of these responses have been partially characterized with a series of hybrid crosses and backcrosses. We propose to utilize this information to experimentally attack the following questions: 1) Do genes which control the number of antibody- producing cells in vivo act exclusively within immunocompetent cells or do they also act in other cells which then decisively determine the extracellular environment of immunocompetent cells? 2) Are such genes expressed equivalently or differentially within thymus derived vs bone marrow derived cells? 3) Are the genetically controlled effects on the in vivo response related to other described regulatory mechanisms: the suppression of antibody synthesis by thymus cells or by cells possessing histamine receptors on their surface?", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our long-term goal is to further our understanding of the molecular mechanisms by which FSH initiates ovarian follicular development. Following the elevation of intracellular cAMP levels by FSH, the adenylyl cyclase enzyme exhibits a transient attenuation response. In specific aim one, we will investigate possible mechanisms responsible for the transient attenuation of the granulosa cell adenylate cyclase enzyme. cAMP Induces all of the differentiation-associated effects in immature granulosa cells attributed to FSH; thus cAMP Is believed to mediate these actions of FSH. Yet, identification and characterization of the enzymes which mediate the effects of cAMP, the cAMP dependent protein kinases, is not available. In the next three specific aims, we will identify the subunit composition of the cAMP-dependent protein kinases present in immature porcine granulosa cells, we will elucidate the acute regulation of this enzyme by cAMP in response to FSH, and we will identify proteins which are phosphorylated in this cascade. Although relatively low levels of cAMP are needed to promote granulosa cell maturation, pharmacologically high levels of cAMP are not deleterious. In specific aim five we will investigate mechanisms which protect the granulosa cells from excessively high intracellular cAMP levels. It is hoped that these studies will extend our understanding of the molecular mechanisms by which FSH via cAMP regulates granulosa cell function.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Rat fetuses exposed to bis-diamine develop various cardiovascular anomalies. The relation between anomalies of the heart and the aortic arch arteries were quantitatively analyzed in 65 treated rats at a late fetal stage under a dissecting microscope. There were 26 rats (40%) with a ventricular septal defect in the perimembranous portion, of which 14 rats (22%) had tetralogy of Fallot, four (6%) had truncus arteriosus, and eight had relatively small septal defect with no other major anomalies. Of the four rats with truncus, two had a vestigial vasculature on the truncus root and three had a rudimentary infundibulum. In 44 rats (68%) the middle latitudinal muscle bundle of the ventricular septum was not continuous with the left ventricular free wall but rather with the right ventricular free wall. There were, isolated or in association, a double or right aortic arch in six rats (9%), aberrant subclavian arteries in nine (14%), right ductus arteriosus in 12 (18%), and agenetic ductus in four (6%). The cross-sectional area of the ductus, as corrected by that of the aortic isthmus, was abnormally small in 47 rats (72%). The rats with a septal defect and/or an abnormal myocardial architecture usually had a small ductus; it was very small or absent in the rats with tetralogy. It is speculated that the defect may be the anomalous and reduced development of the 6th arch arteries, which, imposing pressure load on the fetal right ventricle, may have led to the persistence of ventricular septal defect as a vent and/or to the formation of myocardial architecture favorable for the generation of pressure in the right ventricle.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Alcohol consumption is a major etiology of chronic liver disease worldwide. The morphological spectrum of human alcoholic liver disease includes fatty liver, alcoholic hepatitis, and cirrhosis. In rodents, intragastric infusion of ethanol for 4-5 weeks leads to steatosis, inflammation, and to a less extent fibrosis in the liver, whereas feeding Lieber-DeCarli liquid diet containing ethanol does not cause significant liver injury except steatosis. In humans, interestingly, only a small percentage of heavy drinkers (10-15%) developed alcoholic liver injury, strongly suggesting that alcohol is a cofactor for developing chronic liver disease. Accumulating evidence suggests that many genetic and acquired factors are implicated in the susceptibility of the individual to alcohol-induced liver injury. These factors include chronic viral infection, nutritional factors, the dose and duration of alcohol consumption, pattern drinking, age of onset of drinking, genetic polymorphisms of cytokines and alcohol-metabolizing enzymes, gender, histocompatibility antigens, immunological factors, genetic predisposition to alcohol addiction, and hepatic iron overload. It has been well documented that alcohol consumption accelerates the development and progression of liver disease induced by hepatitis virus infection. Our lab is to study how chronic ethanol consumption potentiates liver injury induced by other toxins or viruses and to study the molecular mechanisms underlying alcohol-induced liver injury. We have demonstrated that alcohol consumption accelerates T cell- and MCMV virus-mediated hepatitis and that treatment with IL-6 ameliorates alcoholic fatty liver disease in mice. Currently, we are studying (1) the hepatoprotective effect of IL-6/STAT3 in alcoholic liver injury and transplantation, (2) the effect of alcohol drinking on liver fibrosis via modulation of the innate immunity, (3) the effect of alcohol drinking on liver progenitor cells. We are also collaborating with Dr. George Kunos to investigate the role of canabinoid in alcoholic liver disease, and with Dr. Jake Liang from NIDDK to study the interaction of alcohol and hepatitis viral proteins in liver injury.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: None of the established therapies for patients with Pulmonary Arterial Hypertension (PAH) reduce mortality or reverse established pulmonary vasculopathy. However, patients with Hereditary PAH (HPAH) inherit heterozygous, autosomal dominant mutations in the BMP type 2 receptor gene, BMPR2, suggesting that strategies to correct BMP signaling defects could present a new therapeutic approach for this disease. Over 300 HPAH BMPR2 mutations have been identified, each with potentially different effects on BMPR2 function. However, these can be classified into 3 functional groups: i) Class I: mRNA is degraded by non-sense mediated RNA decay, (NMD+); ii) Class II: receptors are expressed (NMD-), but mis-folded and are not expressed on the cell surface; and iii) Class III: receptors are expressed (NMD-) and correctly localized but non functional. This classification is important since NMD inhibitors may restore BMP signaling with Class I mutations, while compounds that correct protein folding may restore signaling with Class II but not Class III BMPR2 mutations. Importantly, both NMD inhibitors and protein folding agents, are effective, safe and being evaluated in patients with other heritable diseases, such as cystic fibrosis. These studies will determine whether mutation-specific therapies also correct BMPR2 expression and function in HPAH patients. As an initial step to establish the applicability of this paradigm in HPAH, we will use HPAH patient-derived endothelial cells (ECs). Ideally we would use pulmonary endothelial cells (PECs) from HPAH patients for all of these studies. However these cells are in limited supply and have short lifespan in culture. For this reason we will supplement initial studies using HPAH PECs obtained from the PHBI Cell Core, with studies using late outgrowth endothelial progenitor cells (LEPCs) obtained from peripheral blood of HPAH patients with known BMPR2 mutations. Some of these cells have been generated and will be provided by a collaborator. However, we are in a unique position at Vanderbilt to supplement these studies using samples derived from the largest cohort of HPAH patients with defined BMPR2 mutations in the USA. In this way we will be able to study a cohort of HPAH patient-derived ECs that include a representative distribution of BMPR2 mutations from all 3 functional classes of mutation. Using these cells therefore, we will determine the nature and functional classification of BMPR2 mutations, and evaluate the effects of NMD pathway inhibitors and protein folding agents on BMPR2 expression and function in these HPAH patients. These studies therefore will identify BMPR2 mutant sub-types and responsiveness to NMD pathway or protein folding therapy in a relevant cell type, and will form the foundation to classify therapeutically targetable BMPR2 mutations and more extensive evaluation of NMD inhibitors and/or protein folding agents in HPAH. PUBLIC HEALTH RELEVANCE: Pulmonary Arterial Hypertension is an invariably fatal disease for which there is no curative treatment. A major challenge for the scientific community therefore is to develop therapeutic approaches to prevent the progression of pulmonary vascular disease that develops in these patients. Our studies will provide important insight into a common pathway that regulates this process and identify approaches to interfere with this pathway and prevent the development of irreversible pulmonary vascular disease in these patients. (End of Abstract)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A comparative neurobiology of aging resource is being developed to amplify understanding of normal aging processes and to promote prevention and treatment of age- related neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. The resource provides scientific access to the brains of great apes following natural death in zoological gardens, research centers, and retirement sanctuaries as a means of studying aging in non-human primates most genetically, neurobiologically, behaviorally, and cognitively similar to humans. Noninvasive prospective assessments of behavior, cognition, and locomotor patterns, along with detailed clinical histories enhance the value of brains obtained opportunistically at autopsy. No animals are killed to obtain their brains. The repository allows direct comparisons of ape and human brains and uses methods similar to those employed in human brain banks maintained for the study of neurodegenerative diseases. An expert Advisory Review Board has been established to assure wise and optimal use of the resource by reviewing applications. A detailed chimpanzee brain atlas will be produced during Phase II. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Applicant's Description-edited) The Dallas County Familial Breast Cancer Registry at UT Southwestern Medical Center will comprise a population-based repository of clinical data, demographic and pedigree information as well as biological specimens obtained from breast cancer families and appropriate controls. The biological specimens to be stored in this repository will include serum, extracted DNA, lymphoblastoid cell lines, cryopreserved mononuclear cells, paraffin block specimens and, in some instances, fresh tumor and normal tissue materials. The registry is designed to facilitate collaboration involving the identification and characterization of new genes which predispose to breast cancer. Funding of this application will facilitate correlation of genetic predisposition factors with demographic and reproductive risk factors. The registry extends the already existing UT Southwestern Familial Cancer Registry, a joint effort of UT Southwestern Medical Center; Texas Oncology, PA, a large private practice network of oncology specialist; Baylor University Medical Canter, a large private hospital; and the Susan G. Komen Breast Cancer Foundation. This unique partnership, known as \"The Komen Alliance\" has to date identified and analyzed over 300 families with a probable predisposition to breast cancer. The population base to be studied will correspond to Dallas County, Texas, a population of 2 million which includes approximately 1080 new cases of breast cancer annually. We will attempt to screen every newly diagnosed case of breast cancer in Dallas County over a two-year period and will strongly encourage participation by women thought to have a familial form of breast cancer in the Familial Breast Cancer Registry.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Through the use of (1) binding studies with purified hepatocyte membranes (2) liver perfusion studies with the isolated organ and (3) whole animal studies, the proposed investigation seeks to examine the interaction of the glucoregulatory hormones: insulin and glucagon, and the lipoproteins VLDL, IDL and HDL. The binding studies will concentrate on a potential interaction at the membrane bound receptor level. The perfusion studies will examine uptake and catabolism of lipoproteins by the liver under the influence or lack of insulin and glucagon and conversely, check potential lipoprotein influence on insulin and glucagon extraction. Glucose removal and production by the perfused liver will also be examined to see if lipoprotein-hormonal interaction can influence these processes. Finally, animals will be treated with 4 amino pyrazolopyrimidine to reduce lipoprotein levels, after which labelled glucose will be introduced via duodenal catheter. Glucose extraction will be measured and the influence of supplemental lipoproteins assessed. This work addresses potential links between diabetes and heart disease by proposing an interactive role for glucoregulatory hormones and lipoproteins. It may also serve to elucidate glucose handling by the liver in Maturity Onset Diabetes and possibly reveal some relationships fundamental to insulin and glucose homeostasis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Electrophysiologic analyses during chronic application of opioids to organotypic explants of fetal mouse spinal cord with attached dorsal root ganglia (DRGs) have provided the first demonstration that isolated mammalian CNS tissues can, in fact, develop a marked tolerance after several days' exposure in vitro.* Attempts will be made to clarify cellular mechanisms regulating development of tolerance and dependence in these unique CNS tissue cultures by introduction of specific metabolic and pharmacologic blocking agents and by other selective manipulations of the rigorously controlled physicochemical environment of these cultures. Extracellular recordings of the changes in DRG-evoked dorsal-horn network responses during chronic exposure to opioids will be correlated with intracellular recordings from the DRG neurons to detect possible functional alterations in presynaptic DRG terminals during development of tolerance, e.g. decreased opioid sensitivity, increased calcium conductance. Factors leading to the marked increase in opiate binding that occurs in our cord-DRG explants after chronic exposure to naloxone will be analyzed and similar methods will be used to detect possible changes in opiate receptor levels in tolerant cultures. Further studies of cross-tolerance between morphine and opioid peptides will be made to clarify the degree to which tolerance selective for receptor subtypes may develop in the dorsal-horn networks of cord-DRG explants. Development of cross-tolerance to the acute depressant effects of serotonin, and possibly to other monoamines, in opioid-tolerant cord-DRG explants will be analyzed with specific transmitter antagonists. Development of physiologic properties of opioid dorsal-horn networks and their DRG inputs will be studied in normal culture media and following chronic exposure to opioid agonists and antagonists introduced at early stages of maturation of fetal cord-DRG, and isolated DRG, explants in vitro. Correlative studies will be made with cord-DRG tissues explanted from pregnant rodents after chronic exposure to opioids in utero. The proposed studies should provide valuable insights into mechanisms of tolerance and plasticity in opioid systems of the CNS and to problems in maternal narcotic addiction. Crain et al., Life Sciences 25:1797-1802, 1979: 31:241-247, 1982.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Arthropods serve as vectors for several emerging and reemerging infectious disease causing agents including Rickettsia felis. Since first being described in colonized cat fleas, R. felis has also emerged as a human pathogen. To understand the potential risk of this pathogen, we must first understand the biology of the organism. Our long-term objective is to delineate the role of the flea vector in the maintenance and transmission of R. felis in nature. Defining the mechanisms of transmission within populations of fleas is critical for our understanding of the epidemiology of flea-borne spotted fever and widely impacts our approach to control measures. The proposed studies will assess the role of invertebrate and vertebrate hosts in transmission of R. felis via the specific aims: (1) delineate the mechanisms by which R. felis spreads through individual cat fleas, Ctenocephalides felis, and throughout flea populations. Parameters of R. felis infection and transmission in fleas; specifically, a) horizontal transmission (contribution of vertebrate hosts and concurrent feeding); b) vertical transmission (dynamics of replication and dissemination through adult tissues and immature stages of life cycle); c) flea fitness (fecundity and eclosion rates); and d) sex ratio distortion (comparative analysis of male:female ratios) will be measured in a controlled laboratory R. felis/C. felis model; and (2) examine the ecology of flea-borne rickettsioses by assessing the influence of the vertebrate host bloodmeal and flea species on vector competence. Studies proposed will assess the vector competence of cat fleas for R. felis in the presence of alternate bloodmeal sources, as well as the competence of flea species other than cat fleas to acquire and transmit R. felis. Rickettsioses are diseases caused by members of the typhus group and spotted fever group of Rickettsia. Pathogenic Rickettsia are typically transmitted to humans by arthropods (insects and ticks). This project seeks to examine the interplay between Rickettsia felis and the vector flea, to better understand the ecology of flea-borne spotted fever. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "XPS is used to characterize dextran-coated glass coverslips. The dextran is immobilized by activating its hydroxyl groups and then covalently attaching it via the activated groups to the substrate. Once immobilized, the remaining activated functional groups on dextran can be used for attachment of molecules such as peptide sequences for specific cell-type recognition.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We couple \"On the fly\" Quantum Dynamics with the Semiclassical Approximation Initial Value Representation (SC-IVR) to calculate molecular power spectra. Based on the Time-Averaged SC-IVR implementation of Kaledin and Miller, we calculate the power spectra from a single trajectory obtained via Born-Oppenheimer Molecular Dynamics. My job is to perform BOMD calculations with Q-Chem. Other than positions, energy and momenta, SC-IVR also requires Hessians for each time step along the trajectory, which is the rate determining step. For small molecules such as CO2 and H2O, we need approximately 2000 steps. For bigger systems such as benzene we are currently working on, it is crucial for us to have access to a cluster for the frequency calculations at each step. We use Q-Chem 3.1.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The broad objective of the proposed research is to obtain a basic understanding of endocrine changes during sexual maturation of the rhesus monkey. The specific objectives are a) to continue studies of circulating LH, FSH, testosterone, and 17 Beta-estradiol from birth through early adulthood; b) to determine whether gonadotropin secretion if under hypothalamic control in the infant; c) to determine when negative feedback of gonadal steroids on LH and FSH secretion becomes established; d) to determine whether chronic androgen treatment results in precocious maturation of hypothalamo-hypophyseal-ovarian function. Experiments conducted thus far by the Principal Investigator indicate that circulating LH, determined by an O-O radioimmunoassay for monkey LH, is extremely high in both sexes during infancy. These concentrations gradually decrease and by 6 months of age, are comparable to those of the adult. These increased levels of LH in the infant are not observed when another radioimmunoassay (HCG-Rh) for monkey LH is used. Initial studies with exclusion chromatography indicate that the material is similar in molecular size to adult LH when elution patterns were determined by the O-O system. However, no LH activity was detected in the same column fractions by the HCG-Rh system. Because of the marked discrepancy between these two systems, the initial phase of the proposed research will be conducted to examine some of the biological, immunological, and physicochemical properties of the material detected by the O-O system, and to validate a radioimmunoassay(s) for measurement of biologically active LH (and \"infant LH\" if such a form exists in the sexually immature monkey.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This K23 application proposes a carefully planned program of mentored patient-oriented research and concurrent multidisciplinary didactic training in the methods of clinical and laboratory investigation. The candidate plans a career as an independent clinical investigator focusing on patient-oriented research related to molecular determinants of chemoresistance and prognostic outcome. The Tulane/LSU General Clinical Research Center, including the GCRC Core Laboratory, will serve as the primary performance site and will provide research support. The Department of Pathology and the Tulane/LSU GCRC will provide structured mentoring. Dr. David's research plan entitled \"Prognostic Significance of Activated Akt/PKB in Non-Small Cell Lung Cancer: A Case-Control Prospective Study\" will incorporate the basic principles of Epidemiology, Biostatistics, Community Health, Human Genetics and Molecular Medicine which have formed the basis of the Master of Public Health in Clinical Research degree that she is concurrently pursuing from the TU School of Public Health and Tropical Medicine under the auspices of the TU NIH-sponsored Clinical Research Curriculum Award Program (K30 Award). The proposal is predicated on the hypothesis that overexpression of activated, i.e. phosphorylated, Akt/PKB (phosphoAkt) is a pro-survival signal transduction mechanism in human non-small cell lung cancer (NSCLC) which may antagonize therapy-induced apoptosis. It is further postulated based on results of preliminary studies that loss or inactivation of the tumor suppressor protein PTEN permits unregulated Akt phosphorylation, reduced apoptosis and increased survival in NSCLC tumor cells. The specific aims are: 1)To demonstrate the predictive value of phosphoAkt and PTEN status at diagnosis with respect to therapeutic response and survival in newly diagnosed NSCLC patients and 2)To directly test the mechanistic roles of phosphoAkt and PTEN in chemoresponsiveness by differentially manipulating their expression with siRNAs in human NSCLC cell lines shown to be chemoresistant or chemoresponsive to Iressa (gefitinib).The objective of the study is to drive the development of targeted tumor-specific therapies for NSCLC. Dr. David will recruit and follow case and control subjects from a population of patients undergoing diagnostic bronchoscopy for suspected primary lung cancer. She will follow all study patients for up to 36 months. She will assess tumor response after two cycles of chemotherapy in all case patients who receive chemotherapy. The performance of this study will enhance and expand the activities of the GCRC Core Laboratory and the Lung Biology Group. This research program is central to Dr. David's career goals, which are to perform independent and original research that is relevant to identifying signal transduction proteins which may antagonize therapy-induced apoptosis in non-small cell lung cancer. To this end she will be provided the protected time and institutional resources necessary to achieve these goals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The object of this project is to study diseases of the exocrine and endocrine portions of the pancreas with emphasis on diabetes mellitus. At the present time, an animal model for viral induced diabetes is being investigated. Special attention is being given to: 1) genetic differences in susceptibility to viral induced diabetes; 2) purification, cultivation and characterization of beta cells from both animals and man; and 3) effect of various factors, including viruses, on the functional capacity of beta cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Estrogens interact in a variety of ways with the liver. Not only is the liver an estrogen-responsive tissue, but it is also responsible for the interconversion, metabolism and excretion of the estrogens. The effect of liver disease on these processes has not been studied well, except in the specific case of alcohol-induced liver disease. Therefore, using five well-characterized animal models of liver diseases, we will assess the effect of a variety of liver diseases on several aspects of estrogen metabolism in the male rat. First, the potential responsiveness of the liver to estrogens will be evaluated by quantitation of cytosolic receptors in both the diseased and normal livers as well as by determination of the affinity of the receptors for estrogen. Second, we will assess changes in the levels and/or affinity of male-specific estrogen binding protein whose proposed role is that of an estrogen scavenger. Third, the synthesis and tissue levels of the catechol estrogens (2-hydroxy estrogens) will be examined in rats with diseased and normal livers. These compounds are of interest because they are major metabolites of the estrogens in the liver, and, additionally, because they have been shown to interfere with P-450-related drug metabolism in the liver. Therefore, since alterations in any one of these various aspects of hepatic estrogen metabolism could adversely effect individuals with liver disease, an evaluation of various types of liver disease on these individual processes should be meaningful. Moreover, this study should provide a basis for the understanding of the effect of specific types of liver disease upon various aspects of estrogen metabolism and thus the effect of such alterations upon the individual.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "An accurate medication history is an essential part of patient assessment and can have vital impact on the person's care. However, manually-acquired histories are prone to inaccuracies. The problem is especially serious in emergency care and in disaster situations due to the lack of time, overloaded staff and special patient conditions (e.g. comatose or confused patients, unaccompanied minors or elderly patients). This study is partially funded by the Bethesda Hospitals Emergency Preparedness Partnership. It focuses on patients attended by the Emergency Department (ED) of a regional hospital (Suburban Hospital, Bethesda, MD) and evaluates the potential added value of electronic prescription history information from Surescripts. We established a secure electronic connection between the hospital and Surescripts so that prescription-filling reports could be retrieved from Surescripts in real-time, based on four pieces of patient identifying information (name, date of birth, gender and zip code) obtained from the registration process. For three months we collected the Surescripts information in parallel with the medication history manually acquired by the ED nurse. We also retrieved demographic, administrative (e.g. class of insurance, mode of arrival) and clinical (e.g. vital signs, Glasgow coma score) information from the hospitals database as predictors. All the information was de-identified before being sent to NLM for analysis. This research only involved de-identified data collected for quality assurance purposes. We conducted the data analysis retrospectively; there was no intervention or patient contact. The ED-collected records of prescriptions were manually typed in by triage nurses, so the drug names were subject to variation and typos. Surescripts data provided standardized names. To make the two sources comparable, we mapped all drugs to their standard names in RxNorm, the U.S. standard reference drug terminology. Mapping was done largely by automatic text matching algorithms followed by manual review of the unmapped items. About two-thirds of all ED patients were registered in the Surescripts database, and for about half of all patients, Surescripts returned some medication history information. We have completed the analysis of this data, and in short, Surescripts provides more complete medication history than the manual history when the Surescripts database has any information about the patient. However, it has no information for about a third of the patients who have medications. The Surescripts information contains drugs that the patient is currently taking but also the full history of prescriptions being filled in the past year. ED providers found the summaries very useful in spotting problematic behaviors (e.g. narcotic drugs abuse, poor drug compliance). We have submitted a paper to the journal Annals of Emergency Medicine, which has been accepted and is pending publication. With the Surescripts prescription data we obtained in the first phase, we performed preliminary analyses of the number of different prescribers per patient as a measure of care continuity, and it seemed quite high. We have also examined the number of interruptions that physicians would experience due to drug interaction messages under different levels of interaction importance. While doing this analysis, we have acquired, a much larger de-identified database of prescriptions covering the entire Washington, D.C. metropolitan area. With this database we will seek to answer many questions about prescribing patterns, including, but not limited to: the degree to which drug interacting pairs written by different providers from different offices would not be seen as an interaction by the provider in either office; the drugs most frequently responsible for drug interactions; the frequency of provider reminder interruptions according to different commercial drug interaction knowledge bases at various levels of threshold settings, and; identification of events (e.g. ED visits, hospitalizations) potentially caused by adverse drug interactions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Evidence from our studies on the effects of systemic injections of pharmacological agents in behaving monkeys has led to the proposal that the formation of stimulus memories depends on interaction between the cholinergic and glutamatergic systems. More specifically, our evidence suggests that the critical event for storage of the trace or representation of a stimulus is the potentiation exerted by activation of the cholinergic muscarinic receptor on activity mediated by the glutamatergic NMDA receptor. To test this hypothesis, we have been examining the effects of infusing pharmacological agents directly into the perirhinal cortex, which is known from lesion studies to be the most critical area in the temporal lobe for stimulus recognition. Our results thus far have shown that, like systemic injections of a cholinergic muscarinic receptor antagonist (scopolamine), microinjecting this drug into perirhinal cortex impairs recognition memory. By contrast, recognition memory is unaffected by either systemic or perirhinal injections of dopaminergic receptor antagonists (e.g. haloperidol). We have also demonstrated that, like systemic injections of an NMDA receptor antagonist (MK-801), perirhinal infusions of such an antagonist (D-AP5) impairs recognition memory. Again by contrast, recognition memory was unaffected by perirhinal injections of a kainate/AMPA receptor antagonist (CNQX). These results provide preliminary support not only for the hypothesis that stimulus memory depends on the interaction between muscarinic and NMDA receptor activation, but also for the notion that such interaction occurs within the neurons of the perirhinal cortex. The cholinergic/glutamatergic hypothesis of cognitive memory formation has been challenged on the basis of the finding that pretraining rats in the Morris water maze in one environment eliminates the impairing effects of muscarinic and NMDA receptor antagonists on learning the maze in another environment (for review, see Cain, Neuroscience and Biobehavioral Reviews 22: 181-193, 1998). In many of these studies, pretraining is used to familiarize animals with the procedural aspects of the task, and so it is commonly assumed that subsequent training in a novel environment should therefore be particularly sensitive to cognitive spatial processing. However, our recent behavioral findings in rats suggest that the pretraining promotes subsequent use in the novel environment of stimulus-response habits rather than cognitive memory. Therefore, corticostriatal habit circuits relying on intact nigrostriatal dopamine function may contribute to water maze performance following pretraining. Lesion and drug experiments will be conducted to test this hypothesis. Earlier findings in monkeys suggested that systemic injection of haloperidol, but not of scopolamine, retards the learning of a set of concurrent visual discriminations in which the stimulus pairs within the set are each presented just once every 24 hours. In a new study, using a version of this task in which the stimulus pairs of the set are each repeated a few times within each session, systemic injections of both drugs was found to retard learning. If confirmed, the differential results on the two versions of the task would support the notion that discrimination learning with pair-repetition just once every 24 hours can be mediated only by a dopaminergic-dependent corticostriatal habit system (and, hence, is susceptible to disruption only by haloperidol), whereas learning with pair-repetition within a session is mediated by both the latter system and a cholinergic-dependent cortico-limbic memory system (and, consequently, is susceptible to disruption by both pharmacological agents). The circuitry underlying the formation of stimulus memories is thought to involve a series of projections from the high-order sensory processing areas through structures in the medial temporal lobe, from there to the anterior group of thalamic nuclei and the magnocellular division of the medial dorsal nucleus (MDmc), and then to the ventral prefrontal and cingulate cortices. The parallel circuit underlying habit formation is thought to involve a series of projections from the neocortex through the basal ganglia, from there to thalamic nuclei within the ventral and intralaminar groups, and then to the premotor and supplementary motor areas. However, in the course of investigating medial thalamic efferents in macaques, we uncovered other thalamo-cortical routes that could contribute to stimulus memory and habit formation. The medial thalamic injection sites for anterograde tracers covered the midline nuclei, as well as MDmc, medial portions of the magnocellular ventral anterior nucleus (VAmc) and the intralaminar paracentral nucleus (Pc). These injections yielded terminal labeling in the outer half of layer I across an extremely large cortical expanse, sparing only the premotor and supplementary motor areas, precentral and postcentral gyri, and primary auditory cortex (the primary visual area in the occipital pole was not examined). In complementary studies, in which retrograde tracers were injected into various cortical areas, we searched for groups of neurons within the above medial thalamic region that were consistently labeled by the different injections and were therefore a potential source of the widespread projection to cortical layer I. Numerous retrogradely labeled neurons were seen in the midline group of thalamic nuclei after prefrontal, cingulate, and rhinal injections, suggesting that this particular thalamo-cortical projection could participate in the acquisition of stimulus memories. In addition, Pc and the medial portion of VAmc contained labeled cells from all the injected fields except rhinal cortex, suggesting that the widespread thalamo-cortical projections from these two nuclei, which belong to the ventral and intralaminar groups, might participate in habit formation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT The prevalence of heart failure (HF) is increasing, and it is estimated that there will be 9.6 million U.S. adults who have HF by 2030. More than 50% of HF patients have a type of heart failure with preserved ejection fraction (HFpEF), also termed diastolic HF. The hallmark symptoms of HFpEF are lack of energy, fatigue, shortness of breath, exercise intolerance, and decreased quality of life. These patients have impaired cardiac muscle relaxation and diastolic filling, which investigators have associated with cellular energetic impairment. However, current HF guidelines offer no effective treatment to address these underlying pathophysiologic mechanisms. Thus, patients with HFpEF suffer with severe lack of energy and other debilitating symptoms that affect their overall health-related quality of life. We have pilot data on patients with HFpEF showing that two supplements (ubiquinol and D-ribose) added to their usual HF care enhanced their myocardial energetics and diastolic function resulting in decreased symptom burden. Both ubiquinol (reduced form of coenzyme Q10) and D-ribose play a vital role in mitochondrial adenosine triphosphate (ATP) production. Thus, we propose a biobehavioral symptom science study using these two supplements (therapeutic interventions) to target mitochondrial bioenergetics to reduce the complex symptoms experienced by patients with HFpEF. Using a randomized, double-blind, placebo-controlled study, the overall objective is to determine if administering ubiquinol and/or D-ribose to HFpEF patients for 12 weeks would decrease the severity of their complex symptoms and improve their cardiac function. There are two aims. AIM 1: To determine the effects of oral ubiquinol, D-ribose, or a combination of the two administered over 12 weeks on symptoms accompanying low bioenergetics in patients with HFpEF. The measures used to assess patients' perceptions of their health status and level of vigor (energy) will be the Kansas City Cardiomyopathy Questionnaire (KCCQ) and Vigor subscale of the Profile of Mood States. The KCCQ includes five domains such as physical limitations, symptoms, self- efficacy, quality of life, and social interactions. AIM 2: To determine the effects of oral ubiquinol, D-ribose, or a combination of the two over 12 weeks on biological measures in patients with HFpEF. The 6-minute walk test will be used to test exercise tolerance. Left ventricular diastolic function will be assessed using innovative advanced echocardiography software called speckle tracking. We will measure B-type natriuretic peptides (secreted from ventricles in HF) and lactate/ATP ratio (measure of cellular energetics). These measures will determine if patients with HFpEF experience improved functional and clinical outcomes. Measures will be obtained at baseline, 6 weeks, and 12 weeks. The projected patient outcomes are improved health status, enhanced energy, increased 6-minute walking distance, improved cardiac performance, lower B-type natriuretic peptide, and increased ATP production. Improved clinical outcomes will enhance the patients' self- care activities of daily living, leading to better quality of life.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Hypericin is a naturally occurring photosensitizer that exhibits potent anti-viral and anti-tumor activity when activated by light. The antiviral activity of hypericin is effective against a number of enveloped viruses, including HIV-1, the causative agent of AIDS. However, in vivo application of hypericin for antiviral or cancer therapy is hampered by its photodependency. The long range goal of these studies is to generate chemiluminescence as an in vivo light source for activation of the virucidal and tumoricidal activity of hypericin and other photosensitizers. The objectiv of this proposal is to determine the feasibility of chemiluminescent activatio of hypericin in cells. The chemiluminescent reaction of firefly luciferase and luciferin can activate the antiviral activity of hypericin in a cell free system; however, the efficiency of energy transfer was found to depend on the localized concentrations of the energy donor and acceptor. In the first specific aim, the proximity of the energy donor and acceptor, i.e. luciferin and hypericin, will be increased by means of a chemical tether. The first type of tethered molecular will be a \"caged\" luciferin, wherein the carboxylic acid group present in luciferin is capped as an activated ester. The second type of tethered molecules will be those which maintain their integrity in the presenc of esterases. This design will take advantage of a very efficient intramolecular energy transfer between hypericin and the chemiluminescent intermediate of the reaction of luciferin with luciferase. All tethered compounds will be tested initially for photochemical properties, and promising molecules will be further characterized biologically. In the second specific aim, stable cell lines expressing the luciferase gene under the control of a lentivirus promoter will be isolated, infected with virus, and treated with tethered molecules. Confocal and fluorescence microscopy will be used to localize hypericin tethers and luciferase within virus-infected cells. Reactions between luciferase and tethered luciferin will be monitored as a function of hypericin activation using fluorescence spectroscopy. Finally, cells will be assayed for production of infectious virus. The results of these studies will determine if chemiluminescent activation of hypericin or other photosensitizers can be applied in vivo. The proposed studies represent a multidisciplinary approach to the development of novel antiviral and anti-tumo therapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This Program Project asks: Can dietary restriction (DR), long known to retard aging and diseases in rodents, do so in nonhuman primates? Our hypothesis is that DR will similarly retard aging in a primate species, as reflected by attenuated rates of change of most biological indicators of aging and increased health span and life span. Two overall Specific Aims continue to be addressed. Sp. Aim 1 is to contribute to the development of the rhesus monkey as a model for the study of aging. Our longitudinal study should lead to an improved understanding of biological aging in this model. Ongoing study of conventionally-fed animals from our large aging colony will be continued as will investigation of three Groups of DR and Control rhesus monkeys, all of which were young adults (-10 yrs. old) when entering the study. Group 1 (n=15/group, males) began in 1989 with R01 funding. In 1994, with PPG support, we added 46 animals to increase statistical power for the key outcome measures on health span and life span. These were Group 2 (n = 15/group, females) and Group 3 (n = 8/group, males). These three Groups allow us to address Sp. Aim 2: to determine the influence of DR on the rate of aging in a primate species. This PPG's three Projects depend on the two Cores (Core A: \"Administrative &Biostatistical Support\";Core B: \"Animal Health and Sample Procurement\"). Our highest priority is to determine health span, functionality and life span in these monkeys. Each of our projects has been designed to address this goal and characterize multiple features in the progression of age and effect of DR. Project 1, (\"Role of DR-induced Metabolic Alterations in Sarcopenia\") expands our studies to explore a mechanistic hypothesis that an adjustment of energy metabolism with DR is a major mechanism in aging retardation. Project 2 (\"Energy Balance and Metabolism\") will continue to probe relationships among age, DR, body composition (with an emphasis on adipose tissue), and energy expenditure and metabolism. Project 4 (\"Brain Structure and Function\") combines MRI and behavioral studies to explore the relationship between cerebral and cognitive changes with age and DR. The proposed work which utilizes the outstanding Primate Center and this precious animal resource will extend our 16-year-long study of aging and DR, an issue at the heart of gerontology and of widespread interest to the general populace.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A Comparative Cancer Center is being established at the College of Veterinary Medicine at Cornell with the aim of bridging basic research efforts in cell biology, signal transduction and virology with translational research and the clinical treatment of animal patients with cancer. The intention is for the Cancer Center to integrate the strengths of the Cornell community in structural biology, as well as new initiatives in Chemical Biology and Genomics, with basic and translational cancer research. Through the combined efforts of ten laboratories working in different areas of cell biology, signal transduction and virology, and five structural biology groups (three using X-ray crystallography and two using NMR), the three dimensional structures will be determined for a variety of proteins relevant to cell growth control and cancer. The ultimate goal is to use this structural information, together with small molecule chemistry to develop novel modulators/inhibitors of the protein activities, and examine the usefulness of these molecules as therapeutics in the veterinary clinic. An important step toward achieving this goal will be to establish a Recombinant Protein Expression Laboratory that will serve as a direct link between the cell/molecular biology groups affiliated with the Comparative Cancer Center and the structural laboratories. The facility will be managed by a Research Associate Professor and a full-time Research Support Specialist and will contain all of the necessary equipment and reagents for performing high level recombinant protein expression for structural determinations. The plan is to develop the Recombinant Protein Expression Laboratory in stages, with the ultimate goal being to establish a facility with full capability for the generation of recombinant proteins in E. coli, insect cells and mammalian cell systems for structural analyses. Each of the participating laboratories will have full access to the facility and its overall direction will be advised by an executive committee consisting of members from each of the major participating research areas and departments. The expectation is that the development of the Recombinant Protein Expression Laboratory will represent a key first step in coordinating campus wide efforts in basic research that will ultimately prove beneficial to animal cancers that are highly relevant and/or approximate the human disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Tumors shed their cells into the peripheral blood, generating metastases that confer their lethality. Circulating tumor cells (CTCs) have been detected in patients of many types of cancers, even those with no signs of clinically overt metastases. Because CTCs are accessible using minimally invasive procedures, the detection and characterization of CTCs are pivotal for early diagnosis, prognosis, and treatment monitoring, as well as for understanding of the molecular and cellular events that lead to metastases. Our objective is to establish a sensitive and reliable method for detecting and capturing CTCs of all tumor types, regardless of their tissue origins, in research and clinical settings. We hypothesize that the telomerase-selective adenoviruses specifically label cancer cells and provide a simple method for efficient CTC detection and isolation. Our specific aims are (1) to generate replication-competent adenoviral vectors with optimized hTERT promoters for selective labeling of viable cancer cells; (2) to generate and characterize a tet-regulatable telomerase-selective adenovirus for the detection and isolation of viable CTCs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We wish to improve treatment for patients with deep-seated brain tumors. With the help of the MRI-guided and nanoparticle-enabled system we are constructing, therapeutic access to these tumors will be provided through the delicate nooks, crannies, and membranes of the skull. In preliminary studies, we have demonstrated the ability to bypass the blood-brain barrier traumatically and to manipulate magnetic nanoparticles (MNPs) in mouse brains. By the end of Phase I, we will have built and tested an MRI-compatible gradient insert system capable of delivering magnetic nanoparticles (MNPs) to selected regions in normal mouse brains. Using the data collected from the Phase I mouse study, we will have run simulations of a large-scale magnetic insert system, in order to demonstrate the feasibility of targeting deep- seated brain and skull-base targets in humans. Discussions are well underway with investors to match the Phase II NCI contribution (see Letter of Support). Strategic partner Chemicell is conducting toxicity and other studies in support of an eventual investigational new drug application (IND) of the drug to be used in Phase II of this study. The potential market size for this product can be estimated to be in the range of several hundred million dollars, based on either the number of potential surgical users and practice sizes (i.e., 4,000 neurosurgeons, averaging 8 physicians/practice, at an expected price per system of $1 million) or the purchase price and valuations for companies with similar products (Sentinelle Medical, acquired for $85 million by Hologic, and $2 billion valuation for Intuitive Surgical).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Diabetic retinopathy (DR) is traditionally regarded as a microvascular complication in diabetic retinas. However, it is increasingly recognized that the loss of retinal neuronal function and viability occurs before the onset of retinal microvascular abnormalities in diabetic humans and animals. Diabetes induces photoreceptor degeneration. Cone degeneration is particularly devastating, as cone photoreceptors are responsible for bright and color vision. At present, the mechanisms governing diabetes-induced photoreceptor degeneration are largely unexplored. Recent studies suggest that vascular endothelial growth factor (VEGF), a major angiogenic factor that is up-regulated under hypoxic and diabetic conditions, may play a role as a survival factor for retinal neurons, including photoreceptors. However, VEGF is a therapeutic target of diabetes-induced retinal vascular complications and anti-VEGF agents have been used to develop therapeutics for DR. As VEGF receptor-2 (VEGFR2) and neuropilin-1 (NRP1) are the most abundantly expressed VEGF receptor-like proteins in neuronal cells, identifying the roles and mechanisms of VEGF signaling through VEGFR2 and NRP1 in retinal neurons is paramount to the safety of long-term anti-VEGF treatments for DR. To test our central hypothesis that VEGF signaling through VEGFR2 and NRP1 plays an essential role in preserving photoreceptor function and viability in DR, we will use both in vitro and in vivo models to determine the function and mechanisms of VEGFR2 and NRP1 signaling in photoreceptors. In Aim 1 we will examine VEGFA/EGFR2 signaling activated pathways, regulation of VEGFR2 activity, and the involvement of NRP1 in VEGF signaling cascade. In Aim 2 we will determine if the loss of VEGFR2 exacerbates ischemia- or diabetes-induced photoreceptor dysfunction by examining cone density, retinal morphology, and photoreceptor function in rod- or cone-specific VEGFR2 knockout mice. In Aim 3 we will generate rod- or cone-specific NRP1 knockout mice and characterize photoreceptor function, morphology, and density in these conditional NRP1 knockout mice under ischemic or diabetic conditions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Administrative Core (AC) provides oversight of all scientific and fiscal aspects of the PPG as well as management of all collaborative activities and coordinating. It will coordinate and organize the meetings of the projects and cores leaders and staff. The core will also organize and facilitate meetings of the Internal and External Advisory Committees. The AC will be responsible for the implementation of decisions, corrective actions, suggestions, and initiatives recommended by the Advisory Committees as well as the project leaders. The AC will provide tools and platforms that will foster the integration, collaboration, and communication within the program. The core will be responsible for all interactions with University of Colorado central resources as well as other institutions involved in this project including the University of Colorado Hospital, National Jewish Hospital, and Stanford University.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Among the cancer screening tests known to significantly reduce mortality, screening for colorectal cancer (CRC) is utilized least. Rates of CRC screening are lowest among the underserved, those who are non-white or non-English speaking and/or of low socioeconomic status. Late stage diagnosis for CRC is more common among African Americans, Asians, and Hispanics than non-Hispanic whites. More than other cancer screening tests, CRC screening is dependent upon effective communication between provider and patient, an interaction that can be problematic under any circumstance but is further complicated when provider and patient are from different cultures. The proposed research represents the formative phase of an intervention to adapt electronic tailoring to produce guidelines and messages that will aid providers in effective communication across cultures on CRC screening. Our specific aims are, for African American, Latino and Vietnamese patients, to: 1.) Identify the cultural, social, and psychological factors that influence the effectiveness of provider-patient communication on CRC screening; 2.) Develop and test communication strategies that enable providers to bridge the most important cultural, social and psychological differences for a CRC screening referral; 3.) Develop a research plan for a controlled trial of tailored provider-patient communication across cultures regarding CRC screening. The conceptual framework for the study draws from subjective culture theory, communication theory and behavior science theory. This three-year qualitative study consists of two phases of research. Phase I, Concept Identification, will include 20-40 video-taped observations of actual provider-patient visits in which the physician recommends CRC screening, followed by unstructured interviews with the patients. Phase I also includes 4 provider focus groups; key informant interviews with experts on the three targeted cultures; and 8 patient focus groups, 2 in each of the 3 targeted culture and two with Anglos. Phase II, Refinement and Testing of provider guidelines and messages will include another set of 8 patient focus groups and a round of 10-20 observations to pilot test guidelines and messages. Results will inform an intervention to be tested in a subsequent study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Compact Disc Incorporated proposes research and development for reducing dietary cancer risk in children and families using a successfully developed Phase I prototype Compact Disc-Interactive (CD-I) video disc incorporating NCI and HHS/USDA food pyramid dietary guidelines. The goal is a commercially viable game-oriented CD-I disc that greatly enhances educational and behavior change results versus passive videos or books. Titled Pyramid Adventures it includes games specially designed to increase child and family cancer preventive behaviors relating to dietary choices. The prototype Triminator game turns players into a time-traveling robot that must identify and eliminate high fat foods in the refrigerator. Phase II aims: 1) use Phase I findings to inform final product refinement, 2) produce game segments for ages 6-12, 3) test acceptability of designs with 2nd and 5th grade children, 4) test-efficacy in changing children's knowledge, attitudes and intended behaviors toward including a greater variety of fruits/vegetables and low-fat foods in their diet, Aim 5) secure and implement distribution commitments from Philips for Phase III commercialization of the final CD-I. After interacting with Pyramid Adventures, children will select increased proportions of fruits, vegetables and whole grains for snacks and regular meals. Phase III objectives call for national distribution of Pyramid Adventures CD-I, produced and tested during Phase II.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The abiding concern of this project is to characterize several naturally occurring slow viral diseases of domestic animals that resemble important, etiologically obscure diseases of man. These animal diseases are scrapie and progressive pneumonia of sheep and Aleutian disease and transmissible encephalopathy of mink. Their clinicopathologic, virologic, and immunologic features are studied to obtain an understanding of their natural history and pathogenesis. Such studies should provide not only information on mechanisms by which viruses cause protracted disease but also investigational techniques that may help to determine whether certan subacute and chronic diseases of man are caused by viruses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The new NIH roadmap lays out a streamlined bench-to-bedside philosophy that should be applied to future biomedical research. Within the field of Neural Engineering (NE), Neurotech Reports projects that the overall worldwide market for neurotechnology products will be $3.1 billion in 2006. They predict this will reach $7.6 billion in 2010. For the typical academic researcher engaged in more traditional \"basic\" research, these trends represents both an opportunity and a challenge: an opportunity to more directly impact clinically relevant problems via commercialization, but a challenge to adapt from more traditional ways of practicing academic science and engineering. Although the need to shift towards a translational emphasis affects all areas of biomedical research, the field of Neural Engineering (NE) is uniquely impacted by virtue of the biologically protected status of the brain and nervous system. Not only do interventions here face some of the greatest scientific challenges of all biological systems, they can also raise serious ethical concerns. The goal of the proposed program is to provide support for a half-day special-topic symposium on Translational Neural Engineering at the annual meeting of the Biomedical Engineering Society. The symposium is titled \"Translational Neural Engineering: Multiple Perspectives On Bringing Benchtop Research Into The Clinical Domain\". The main objective of the special symposium is to consider the question: What constitutes a viable model for translational research in the field of Neural Engineering? Implicit in the formulation of an answer to this question is the need to: define translational NE research, identify successful and unsuccessful strategies within NE, and highlight areas of need and opportunity within NE. To achieve this, a diverse and broad- ranging list of speakers has been assembled. Successful academic science and engineering researchers who have had translational research success, the clinicians who have been involved in the process, a bioethicist and a representative from a government funding agency have already agreed to participate. The symposium is particularly well-suited for educating students (who constitute 65% of the >2100 attendees) and junior faculty. The long-term objective of the symposia is to equip traditional researchers with the skills, methodologies and mindset for success in translational neural engineering. A white paper on translational neural engineering will be submitted as a manuscript to Journal of Neural Engineering after the meeting. The proposed symposia becomes relevant to public health when the long- term objectives are met and traditional research engineers are adequately trained to navigate the waters of successful translational research in neural engineering. This will effectively provide a smoother/faster path to market for the clinical application of exciting neural engineering technological breakthroughs aimed at improving diagnostic and therapeutic success for diseases of the nervous system. The proposed symposia titled \"Translational Neural Engineering: Multiple Perspectives On Bringing Benchtop Research Into The Clinical Domain\" becomes relevant to public health when the long-term objectives are met and traditional research engineers are adequately trained to navigate the waters of successful translational research. This will effectively provide a smoother/faster path to market for the clinical application of exciting technological breakthroughs aimed at improving diagnostic and therapeutic success for diseases of the nervous system. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In this study we are performing phenotype/genotype studies in women with congenital deficiency in the secretion or responsiveness to gonadotropin releasing hormone (GnRH). Isolated GnRH deficiency is a rare disorder, with an estimated incidence of 1/50,000. Patients present with failure of normal pubertal development and hypogonadotropic hypogonadism (low LH, FSH and estradiol or testosterone), indicative of a hypothalamic or, less frequently, pituitary abnormality. The disorder is referred to as isolated hypogonadotropic hypogonadism (IHH). Approximately 50% of these patients lack a sense of smell (Kallmann Syndrome, KS) while the other half are normosmic (nIHH). Studies in KS/nIHH patients have been particularly informative in helping us understand the GnRH requirements for normal reproductive function and have led to development of a physiologic regimen of GnRH replacement in men and women. While pulsatile GnRH serves as treatment for patients interested in conceiving or in their potential to conceive, these patients have provided a clinical model that allows us to address key questions in reproductive physiology . When synthetic GnRH first became available for clinical study, there was not yet an adequate understanding of the physiology of GnRH secretion in the human to support its potential therapeutic application. As a result, early attempts at ovulation induction were unsuccessful. It was soon appreciated that an episodic mode of delivery was essential for normal pituitary stimulation by GnRH. Studies which defined the frequency of pulsatile GnRH secretion in normal women at different stages of the menstrual cycle were key to designing a physiologic program of pulsatile GnRH administration that resulted in successful ovulation induction in patients with GnRH deficiency. Additional studies demonstrated that while replacement of GnRH using the subcutaneous route was adequate to reproduce normal physiology in GnRH-deficient men, the intravenous route is superior in women. We have now determined that the dose of GnRH that induces ovulation of a single dominant follicle, followed by normal luteal phase dynamics for the majority of women is 75 ng/kg administrated intravenously. Moreover, normal gonadotropin and gonadal steroid dynamics are faithfully re-created using a frequency that mimics the frequency in normal women and changes across the cycle from once per hour to once every four hours. Replacement of GnRH induces ovulation in approximately 95% of GnRH-deficient women. However, a subset of women with KS/nIHH do not ovulate in response to the physiologic 75 ng/kg dose. Thirty-seven women with KS/nIHH who received a physiologic replacement regimen of pulsatile GnRH (75 ng/kg at a physiologic frequency) were studied during their first cycle of GnRH replacement. In those who ovulated in the first 20 days of treatment, LH, FSH and estradiol levels rose quickly to the range of normal women in the early follicular phase. Patients with LH and/or FSH below the 25th centile of ovulatory patients were defined as pituitary resistant; 90% of these patients ovulated with a longer treatment duration or a higher dose of GnRH. Surprisingly, mutations in GNRHR were found in search for genes conferring pituitary resistance in patients with KS/nIHH. With GnRH treatment, three patients demonstrated FSH levels above the 75th centile of the ovulatory women, providing evidence of ovarian resistance once GnRH was replaced. With GnRH treatment, three patients demonstrated FSH levels above the 75th centile of the ovulatory women, providing evidence of ovarian resistance once GnRH was replaced. None of the three had evidence of mutations in genes known to be associated with GnRH deficiency although some are known to be present in the ovary, again providing fertile ground for future gene discovery. This study is being conducted in patients identified by the PI with the support of investigators at Massachusetts General Hospital as is part of the PIs long-term collaborative efforts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "E. coli MutY and its mammalian homologues (MYH) play an important role in the prevention of mutations associated with 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG) by removal of misincorporated adenine residues from OG:A mismatches. Recently, a direct correlation between mutations in the gene encoding human MutY (hMYH) and colorectal cancer has been uncovered. This highlights the importance of base-excision repair, and the repair of OG:A mismatches in the prevention of carcinogenesis. My laboratory has developed a rigorous and multifaceted research program aimed at providing a detailed understanding of the functional properties of MutY and hMYH. Importantly, our functional analysis of hMYH variants has been important for establishing the connection to colorectal cancer. In order to further the understanding of the relationship between hMYH and colorectal cancer, as well as continue our understanding of the complex features of mismatch recognition by this unique BER glycosylase, we propose the following in this research grant application: (1) We will determine the functional properties of variants of hMYH that are correlated with colorectal cancer. Specifically, this will involve analyzing the repair of OG:A mismatches by a representative set of hMYH variants in bacterial and mammalian cells. (2) We will evaluate the adenine glycosylase activity of this same representative set of hMYH variants to determine if reduced OG:A repair is due to defects in the intrinsic adenine removal activity of the enzyme. Aims 1 and 2 will provide important information on the relationship of these variants to colorectal cancer. (3 ) We will provide insight into specific steps that are involved in OG:A mismatch recognition and adenine removal by MutY using fluorescence spectroscopy with substrates and substrate analogues. This will allow us to reveal the important features of OG and A that are involved in initial base-pair recognition, and how recognition of OG is coupled to adenine extrusion and removal. This aspect will utilize substrate analogues that we have characterized previously and build upon the kinetic framework we have developed for analyzing MutY. (4) We will provide insight into the mechanism of adenine excision by examining the structural properties of Bacillus stearotheromophilus MutY with transition state and substrate analogue-containing duplexes using X- ray crystallography. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies from this laboratory have demonstrated that calcitonin (CT) exhibits hypocitricemic and hypercitraturic effects. However, the physiological relationship of these effects to the functional aspects of CT action needs to be elucidated. This proposal is concerned with an investigation into the possible contribution of the citraturic effect of CT to its hypocitricemic action. Animal studies involving CT induced changes in plasma citrate in relation to the renal effects will be performed. Simultaneously, the mechanism of the citraturic effect will be studied by ascertaining changes in citrate clearance, renal citrate utilization, and renal A-V differences. Also, the hypocitricemic effect of CT in nephrectomized animals will be ascertained to determine the renal involvement in this effect. With the use of in vitro (renal cortex slice) systems the possible effects of CT on renal citrate oxidation, as well as tubular citrate levels will be studied. Consequently, these studies should establish the possible physiological action of CT on kidney regulation of citrate, the contribution of the renal effect to the hypocitricemic action, and the mechanism of the renal effect.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The work proposed here is intended to elucidate the mechanisms by which cells that fail to adopt appropriate fate induce programmed cell death. Normal development and homeostasis requires proper specification and cellular differentiation. However, interference with the establishment of cellular fates and cellular functions can lead to the induction of cell death. This strategy might guard the organism against developmental errors, because if these cells were not removed, they might cause malignancies such as cancer. However, under pathological conditions the same strategy may cause the inappropriate death of cells giving rise to congenital defects during development or other conditions such as neurodegenerative disorders. Thus, a detailed investigation of the underlying mechanisms will provide new insights into human diseases in which deregulation of apoptosis is known to occur and may lead to new strategies for therapeutic intervention. It is unknown why a cell dies that receives no or an incorrect developmental signal. In the genetic model organism Drosophila melanogaster, a number of mutants exist that block normal cellular specification and differentiation. Subsequently, these cells undergo cell death. Thus, these mutants provide an excellent genetic model to study the regulation and onset of this form of cell death. We have determined that cellular mis- specification is the underlying cause of cell death in these mutants. The cell death-inducing gene hid is specifically up-regulated in mis-specified cells, suggesting that mis-specification-induced cell death is the result of an active gene-directed process. We postulate that a mechanism monitors the cell's ability to develop correctly. If the cell fails to do so, the monitoring mechanism triggers the transcriptional induction of hid and induces cell death. To genetically and molecularly characterize the postulated monitoring mechanism we will (1) analyze the promoter of the hid gene and identify the factor(s) binding to it in response to mis-specification, (2) analyze the genetic requirement of a number of genes identified in a microarray analysis for mis- specification-induced cell death, and (3) perform genetic screens to identify genes which are required for this process. It is the goal of this proposal to gain a comprehensive understanding of the mechanisms underlying this interesting biological phenomenon, and to exploit this knowledge for therapeutic purposes. [unreadable] [unreadable] This project investigates the mechanisms by which cells die if they do not receive the correct developmental information. These incorrectly informed cells resemble cancer cells, and the observation that they die may protect the organism from several common diseases. Understanding the mechanisms of this process may lead to new therapeutic interventions for cancer, and may also be relevant for treatment of neurodegenerative diseases. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this contract proposal is to label monoclonal antibodies with astatine-211 for use in radioimmunotherapy. The goal of the proposed research is to develop a method to incorporate astatine into antibodies such that the astatine is not labile in vitro or in vivo. In the investigation, two different approaches to labeling will be compared. One approach will be direct labeling with astatine and the other approach will involve the use of astatinated small molecules. The later approach will include development of methods to attach the astatine to the small molecules such that the astatine is stably attached to them.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Maintenance and plasticity of synaptic connections are fundamental to all higher cognitive processes. The proper trafficking of postsynaptic glutamate receptors (GluRs) is critical for this process. It is suggested that their misregulation can impair memory formation, as in Alzheimer patients, and can potentially contribute to other neurological disorders, such as epilepsy and schizophrenia. Our goal is to understand the mechanistic steps of receptor trafficking. This is critical for both understanding basic neuronal function and potentially for treatment of neurological disorders. By using a combination of EM and two-photon imaging techniques, along with electrophysiology, we will determine at high resolution the site of membrane and synapse insertion of specific receptor subunits in tissue. Post-embedding immunogold will be used to localize proteins under EM, and a pH-sensitive eGFP, super ecliptic pHluorin, will monitor surface delivery of receptors. Working in young rat hippocampal slices we will first study the trafficking of GluR4, a process critical for early synaptic strengthening. Next, we will track the coordinated syanptic insertion of GluR1 and replacement by GluR2 following the induction of long-term potentiation (LTP). These phenomena will be studied in preexisting and nascent synapses with the goal of understnading the role each plays in maintaining synaptic circuits.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mitotic entry is driven by the explosive activation of the kinase MPF (M phase promoting factor; Cdk1/cyclin B), which phosphorylates hundreds of target proteins at thousands of phosphosites. When cells subsequently exit mitosis, many of these phosphorylations are removed by the phosphatase PP2A associated with the regulatory subunit B55 (PP2A/B55). In order to protect the MPF-mediated phosphorylations from premature dephosphorylation, PP2A/B55 is shut off during M phase by a pathway involving a kinase called Greatwall (Gwl) and its effector Endosulfine (Endos). MPF phosphorylates and thus activates Gwl; Gwl in turn phosphorylates and thus activates Endos, and phosphorylated Endos (pEndos) binds to and thus inactivates PP2A/B55. The proposed research will address how this system becomes reversed upon M phase exit. The first two Specific Aims will expand upon preliminary results indicating that: (1) the major phosphatase that removes the Gwl-driven phosphorylation on pEndos is surprisingly PP2A/B55 itself; and (2) a simple mechanism we call inhibition by unfair competition can explain how pEndos can simultaneously act as an inhibitor and a substrate of PP2A/B55. The experiments described in the first Specific Aim will dissect the molecular basis for the unfair competition mechanism, investigate the means by which pEndos action may be influenced by events other than Greatwall phosphorylation, and model this aspect of M phase exit both in vitro with purified components and mathematically in the more complex environment of the cell. In the second Specific Aim, we will explore the possibility that inhibition by unfair competition reflects an ancient regulatory module through which kinases belonging to the AGC family (like Greatwall) regulate phosphatases belonging to the PPP family (like PP2A/B55) via the phosphorylation of small intermediary molecules (such as Endos). We will test whether known phosphoprotein regulators of the phosphatase PP1 operate through the unfair competition mechanism, and we will search for novel regulators of other PPP phosphatases using a strategy based on our insights from pEndos and PP2A/B55. In the third Specific Aim, we will identify the phosphatase(s) that inactive Gwl kinase when cells exit M phase. Preliminary results indicate that at least one of these Gwl- inactivating phosphatases is insensitive to the drug okadaic acid, and thus cannot be PP2A/B55. We will ask if this okadaic acid-insensitive phosphatase corresponds to an enzyme already known to be involved in M phase exit, or instead whether the Gwl-inactivating enzyme defines a novel phosphatase activity. The work described in this proposal has the potential to deepen our understanding of key cell cycle transitions and the biology of phosphatases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is a fundamental gap in knowledge regarding how mutations in the genes encoding cyclic nucleotide- gated (CNG) ion channels can produce achromatopsia, cone dystrophy and macular degeneration in humans. Our long-term objective is to understand the mechanisms controlling the activity of these channels and the pathophysiology of retinal diseases associated with CNG channel mutations. The core objectives of this application are to determine the cellular mechanisms responsible for the effect of cone CNG channel gating or trafficking mutations on cell viability, and the structural features critical for control of channels by phosphoinositides. Recently, we have functionally characterized several disease-associated mutations in the CNGA3 and CNGB3 subunits of cone CNG channels and discovered dramatic effects on channel gating, regulation and/or trafficking, but the cellular consequences of these defects have not been determined. The central hypothesis is that gain-of-function mutations in cone CNG channels lead to photoreceptor death via enhanced or uncontrolled channel activity, disturbance of intracellular calcium (Ca2+) homeostasis and subsequent Ca2+-dependent apoptosis. Conversely, trafficking defects are expected to impair cell viability via endoplasmic reticulum (ER) stress. The rationale for the proposed research is that developing an understanding of photoreceptor dysfunction and loss associated with abnormal CNG channel activity will provide insight into possible treatments for several related cone dystrophies. Guided by strong preliminary data, we will address these issues by pursuing two specific aims: (1) identify the connection between disease associated functional changes in cone CNG channels and the cellular mechanisms leading to photoreceptor dysfunction and death;and (2) determine the mechanisms and interactions underlying the ability of CNGB3 subunits to confer sensitivity to channel control by phosphoinositides. These studies will utilize molecular and cellular manipulations, biochemical approaches and/or electrophysiological studies of human CNG channels expressed in cone photoreceptor derived 661W cells or Xenopus oocytes, and as transgenes in zebrafish cone photoreceptors. The proposed research is innovative in that informative in vitro studies will be extended to transgenic expression of mutant CNG channels in vivo. Overall, the proposed work is significant because it is expected to enhance our understanding of the mechanisms that lead to retinal degeneration and blindness, and to provide insight into potential approaches for prevention of photoreceptor loss. PUBLIC HEALTH RELEVANCE: The proposed research has relevance to public health, because completion of these studies will provide important insight for preventing and treating vision loss. Our work is focused on understanding vision at the molecular and cellular levels and how mutations in genes coding for proteins critical for vision can lead to dysfunction and retinal degeneration. We study ion channel proteins that are responsible for generating electrical signals ultimately interpreted by the brain as visual information. The major goal for this new project is to elucidate specific mechanisms linking pathogenic changes in channel function or control to cell death.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Imaging mRNA turnover in P-bodies and its application to brain ischemia Injured cells adapt to an insult by removing their mRNAs for storage or destruction. Both these processes occur in the same cytoplasmic foci referred to as P-bodies. The storage and destruction of transcripts cause the translation block and their relative intensity determines the fate of the cell. The ability to separately visualize these processes directly in P-bodies on a cell-by-cell basis is essential for the assessment and future control of cellular response to injury. However, in spite of the enabling nature of such an in situ technology, at present no method can selectively label both the decay and the storage of mRNA transcripts in P-bodies using standard fixed tissue sections. In this project we will resolve this limitation and will introduce the first histotechnology which separately visualizes mRNA decay and storage in P-bodies. We will then apply the new assay to study mRNA turnover in focal brain ischemia in rat. Specific Aims of the proposal are: 1. To develop the first histotechnology for in situ labeling of mRNA turnover in P-bodies. The assay will be applicable to fixed tissue sections. The approach will encompass visualization of the activity of mRNA decay in P-bodies by using the new random DNA nanomer probes. It will permit the co-labeling of P-body core proteins for the assessment of mRNA storage occurring in P-bodies. 2. To test and validate this newly developed in situ labeling technique in sections of experimental stroke. To optimize its specificity, sensitivity and reliability of detection. By making use of fixed tissue sections, the assay will enable the analysis of the archived pathology samples. The assay will be broadly applicable in biomedicine because it will allow visualization of previously undetectable molecular pathways in a convenient histotechnology format. Its application to brain ischemia will provide useful information about the dynamics of ischemic cell injury.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "BRCA 1 is a putative tumor suppressor gene located on chromosome 17q21, conferring an 85% lifetime risk of developing breast and/or ovarian cancer on women carrying BRCA1 germline mutations. The isolation of BRCA1 will likely provide insight into the biology of breast cancer as well as lead to predictive testing for genetic susceptibility to breast cancer. Immortalized lymphocytes from families with apparent autosomal dominant transmission of breast cancer are essential reagents in our efforts to identify BRCA1.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Post-replication DNA mismatch repair is an essential and highly conserved mechanism to ensure genetic fidelity. The first step in DNA mismatch repair is recognition of DNA base mismatches and insertion or deletions by MutS in bacteria and MSH complexes in eukaryots. The structure for the MSH2-MSH6 complex which recognizes base mismatches and short base insertions has recently become available from crystallography and will be used as the starting point for extensive computational studies to elucidate atomistic details of the mechanism by which mismatch repair is initiated. One aim is to carry out molecular dynamics simulations of MSH2-MSH6 over 100 ns time scales to study allosteric coupling between mismatch recognition and ATPase activity that would lead to conformational changes in MSH2-MSH6 to signal repair initiation. A second aim is to examine the detailed interaction between MSH2-MSH6 and DNA to understand the mechanism by which defective DNA is distinguished from regular DNA by employing biased simulations with a new biasing potential that facilitates translocation of MSH2-MSH6 along DNA. The immediate goal of the development account is to determine the best choice of simulation software and TeraGrid platform in preparation for an MRAC proposal.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Eyelid dynamics during blinking will be investigated in detail in normal subjects, particularly relating to the interaction between lid motion, tear spreading, and corneal wetting. A specialized high-speed camera system will be employed, using framing rates up to 500 pictures/second, and capable of working at subject-to-camera distances of 10 feet or more. In this manner, the camera system can be remote from the subject, allowing for natural blinking and tearing. Associated studies will be performed to quantify the surface-chemical events associated with tear spreading, and the effectiveness of commercially available artificial tear preparations in duplicating the wetting and spreading properties of natural tears will be investigated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The mission of the Analytical Laboratory for Pharmacogenetics, Pharmacodynamics and Pharmacokinetics (3P) is to advance clinical cancer research by providing accurate and efficient state of the art data and analyses of pharmacogenetics, pharmacodynamics and pharmacokinetics parameters. The 3P Laboratory has three services available to Cancer Center members, instrumentation, analytical service and the sample acquisition service. 1. Instrumentation The 3P Laboratory maintains and instructs Cancer Center members on the use of all laboratory equipment housed in the 3P Laboratory. The 3P Laboratory has 1,200 square feet of laboratory space and 240 square feet of office space which house over 25 laboratory instruments. The space was specially designed for efficient use and maintenance of analytical instruments. The laboratory design also facilitates timely and efficient processing of clinical samples. 2. Analytical Services The 3P Laboratory is designed as a facility for developing and executing a wide variety of bioanalytical methods, including processing and analyzing samples for PK/PD/PG studies in support of Phase I, II, III and chemoprevention clinical trials. This service includes development and performance of original assays for new agents, assays to monitor endogenous biochemicals, genotyping and mutation assays and adaptation of existing methodology for analyses where applicable. 3. Sample Acquisition Services (SAS) The 3P Laboratory also offers a Sample Acquisition Service for all UWCCC clinical trials. This service is offered to all investigators participating in clinical trials requiring acquisition, processing, storage and shipping of samples for PK/PD/PG studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our previous studies have demonstrated the importance of human sis/PDGF-2 gene deregulation in its activation as an oncogene in cells responsive to PDGF stimulation. Current studies have focused on the structure, regulation, and function of this gene. We have shown that human tumor cells arising from PDGF- responsive cell types express sis/PDGF-2 mRNA and mitogenically active sis/PDGF-2 homodimers. Utilizing cDNA cloning, S1 nuclease mapping, and primer extension techniques, the normal human sis/PDGF-2 transcriptional unit has been defined. These studies also suggested the presence of transcriptional and translational regulatory signals within the sis/PDGF-2 locus, and have provided an approach for elucidating mechanisms by which this gene is controlled. Knowledge that the v-sis oncogene encodes a PDGF-related product whose transforming activity requires functional interaction with the PDGF receptor has suggested the importance of identifying the active site of the v-sis translational product. Site-directed mutagenesis of v-sis has localized an 89 codon stretch as its minimum transforming region and has shown a requirement for each of 8 cysteine codons within the region for proper folding of the v-sis gene project. These studies have also predicted three testable models for the active conformation of this protein are represent an important step in identifying the receptor binding domain of this oncogenic growth factor.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The BLAST family of protein and DNA database search programs constitute one of the key services offered by the NCBI. These programs are currently run on NCBI servers about 70,000 times during a typical weekday. This project represents an ongoing effort to improve and extend the functionality of these programs. Efforts this year have focussed primarily on the use of compositionally adjusted amino acid substitution matrices with \"Blastp\", the program for comparing protein queries to protein databases. Preliminary results indicate that such score matrices are useful primarily for the comparison of sequences with similar lengths (less than a factor of two difference), and for sequences whose compositions are relatively similar, as measured by relative entropy. Algorithmically, we have needed to rewrite the heuristic for producing gapped alignments when the substitution matrix used is modified. This is due to the fact that the rescaling procedure employed for compositionally adjusted statistics essentially never produced longer alignments, whereas the new compositionally adjusted matrices frequently yield extended alignments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our goal is to determine the optimal media for the growth of various human tumors in primary culture. This entails determining the hormone and factor requirement for growth in serum-free medium. We wish to determine the similarity or dissimilarity of the requirements of individual tumors of the same class, such as colon tumors. Optimal growth media would be useful in attempts to develop in vitro tests for drug sensitivity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Nature uses an amazing array of enzymes to make small molecule natural products. Among the most interesting but least understood enzymes making these compounds are the iterative polyketide synthases (IPKSs) found in filamentous fungi. In contrast to the well-studied bacterial type I PKSs that operate in an assembly-line fashion, IPKSs are megasynthases that function iteratively by using a single set of catalytic domains repeatedly in different combinations to produce structurally diverse fungal metabolites. Bioinformatics analysis of the genomes of recently sequenced fungal species revealed that each genome contains a large number of genes encoding IPKSs. The total numbers of IPKSs significantly outnumber the known polyketides and polyketide-nonribosomal peptides isolated from these species, suggesting that a majority of biosynthetic genes are silent in these fungi under cultivating conditions. This in turn suggests that the fungal species may have untapped potential to synthesize a much large number of natural products. Furthermore, analysis and engineering of IPKSs have been hampered by inability to obtain sufficient amounts of the functional purified megasynthase from either the native fungal host or heterologous Aspergillus hosts. As a result, the programming that governs metabolite assembly by IPKSs is not understood. Key aspects that remain to be elucidated include: 1) the catalytic and structural roles of each domain in the megasynthase; 2) substrate specificities of the catalytic domains and their tolerance to perturbation in megasynthase functions; and 3) factors governing the choice of different combinations of catalytic domains during each iteration of catalysis. The objective of this proposal is to develop the genetically superior Saccharomyces cerevisiae as a heterologous host for reconstitution, analysis and engineering of IPKSs, especially the enigmatic highly-reducing IPKS, such as LovB associated with Lovastatin biosynthesis. We have accumulated a significant body of preliminary data to demonstrate that S. cerevisiae is a highly robust host for expressing these megasynthases in functional forms, and can facilitate the production of polyketide products both in vivo and in vitro with purified enzymes. The following specific aims will be pursued: 1) Engineer and optimize S. cerevisiae towards producing fungal metabolites and megasynthases; 2) Reconstitution of fungal megasynthases in S. cerevisiae; 3) Biochemical analysis of fungal PKS using S. cerevisiae; and 4) Genome mining of filamentous fungi using S. cerevisiae as a host. PUBLIC HEALTH RELEVANCE: Filamentous fungi are a rich source of natural products. Among them, polyketides represent an important family of structurally diverse natural products. Polyketides are produced by polyketide synthase (PKSs). Genome sequencing of many fungal species has revealed each organism contains a large number of PKSs, yet many of these PKSs have unknown functions or produce unknown metabolites. Therefore, filamentous fungi can be considered underachievers of natural product producers. We propose here to use Saccharomyces cerevisiae as an expression host to heterologously produce fungal PKSs. We will use this genetically superior host to reconstitute fungal PKSs of both known and unknown functions, perform genome mining of sequenced fungal species and engineered biosynthesis of new fungal-derived natural products.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. While liver disease is currently ranked as the seventh leading cause of adult death in the United States, the gold standard to diagnose and monitor the progress of patients afflicted with hepatic pathologies remains limited to liver biopsy. Liver biopsies can be associated with sampling variability, poor reproducibility and major complications, rendering it essential to develop non-invasive techniques able to diagnose different liver conditions. With the advent of high performance magnetic resonance (MR) systems and advanced sequences, MRI shows potential for improved and non-invasive assessment of liver disease. Fast spin echo or single shot techniques, often combined with fat suppression, are the most common T2 weighted sequences used in liver MRI procedures. Liver MRI is highly dependent on the administration of contrast agents, especially when detection and characterization of focal lesions are the issues. Currently, there is no single MRI technique that is optimal for detecting the different liver pathologies. The objective of this project is to develop CEST and MT MRI techniques for monitoring the progression of chronic liver diseases in human and animal models.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research seeks to illuminate the mechanisms of human learning and memory and, in particular, the role of brain oscillations in this vital human capacity. While it has recently been shown that certain brain oscillations increase during learning and memory tasks (an oscillatory subsequent memory effect, or SME), the exact nature of their involvement in memory function remains unknown. In the proposed studies brain waves will be recorded from patients undergoing surgical/invasive monitoring as part of the clinical treatment of medically refractory epilepsy. By performing cognitive tasks that pose no health risks whatsoever, these patients can provide exquisite data on the involvement of brain waves in memory formation. The proposed research aims to determine: (1) the relative roles of oscillations in hippocampus and neocortex in supporting memory formation, (2) the relation of the phase of oscillatory activity and the cognitive processes underlying memory formation, (3) the relation between the oscillatory SME and the traditionally studied event related potential SMEs, and (4), the roles of temporal and semantic information in producing SMEs in free recall. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The cadherin-based adherens junction (AJ) is a critical cell-cell adhesion apparatus, misregulation of which leads to development failures and cancers. The formation of AJs is known to be dependent on the assembly of a local actin network, but the regulation of this remains unclear. Our preliminary data suggest that talin, a protein known for its role in the regulation of the actin network associated with integrin-based cell-matrix adhesion, is also involved in the AJ through a novel talin proteolytic fragment containing the vinculin- and actin- binding domains and a dimerizing region (referred as the VAD fragment). This fragment also undergoes posttranslational arginylation in vivo. We hypothesize that the proteolysis alters the domain architecture of talin, allowing the resulting VAD fragment to localize to the cadherin- based AJ. There the fragment enhances local actin assembly with its functional domains and stabilizes the AJ. Furthermore, arginylation changes the affinities of the VAD fragment to its binding partners and enhances its physiological function in AJs. In this proposed study we will test this hypothesis in these three specific aims: (1) How is the talin VAD fragment recruited to the cadherin-mediated AJ, and how does it enhance actin assembly? (2) Is the formation of cadherin-based AJs dependent on the VAD fragment? (3) How does arginylation enhance the function of the VAD fragment? Through this study, we will elucidate a novel regulatory mechanism for cadherin-based cell-cell adhesions mediated by the novel processing of talin, which was traditionally considered only associated with integrin-based adhesions. Also this project will provide new insight into how the conserved but poorly understood posttranslational arginylation modification affects protein function in cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "1 There is a fundamental gap in our understanding of the circuit mechanisms underlying even simple naturalistic 2 behaviors, such as making a cup of coffee, which proceed through a sequential execution of sub-behaviors. 3 Continued existence of this gap represents an important problem because obtaining a circuit-level understand- 4 ing of complex multi-step behaviors is a necessary step toward unlocking the mysteries of healthy brain func- 5 tion and of disorders. The overarching goal is to obtain a circuit-level understanding of such naturalistic behav- 6 ior. The research objective here is to unravel the logic of sensorimotor transformation in the context of odor- 7 modulation of locomotion in Drosophila. The central hypothesis is that, like many of our own everyday actions, 8 control of odor-modulation of locomotion is hierarchical. A fly?s locomotion is built from simpler elements called 9 locomotor primitives, each of which lasts between 1-3 seconds (or 10-30 steps). Odors, instead of acting on 10 instantaneous locomotor parameters such as speed and angular speed, act on these locomotor primitives and 11 change the probability that the fly spends performing a given locomotor primitive. This hypothesis was formu- 12 lated on the basis of our previous work and preliminary data. The rationale for the proposed research is that 13 understanding odor-guided locomotion?a complex, flexible behavior?in the context of a genetically tractable 14 system will allow a precise delineation of the steps that underlie sensorimotor transformation in the context of a 15 naturalistic behavior. The hypothesis above will be tested by characterizing the circuit basis of modulation of 16 locomotion by food odors using a combination of techniques including imaging, electrophysiology, quantitative 17 behavior and computation. The proposed research has three specific aims. 1) To extract the locomotor primi- 18 tives and test the hypothesis that odors modulate locomotion by changing the time a fly spends performing dif- 19 ferent locomotor primitives. 2) To test the hypothesis that different ORN classes modulate the time spent in 20 distinct locomotor primitives. 3) To elucidate the role of lateral horn in odor modulation of locomotion. The re- 21 search is innovative because it employs sophisticated statistical tool (Hierarchical Hidden Markov model, 22 HHMM) and cutting-edge experimental tools in the context of a genetically tractable model organism to obtain 23 insights into naturalistic behaviors. The proposed research is significant because it will vertical advance our 24 understanding of sensorimotor processes involved in naturalistic behaviors. Insights from multiple fields have 25 all come to the conclusion that behavior is organized into discrete packets or behavioral primitives. Actions un- 26 fold by a sequential recruitment of these discrete packets. A critical barrier to the study of natural behavior is 27 that in most cases there is enough variability in these discrete packets to make them unrecognizable without 28 the help of sophisticated statistical tool. By deploying HHMM, we overcome this critical barrier. Besides repre- 29 senting a vertical advance in our understanding of naturalistic behavior, another possible positive outcome of 30 this study is better diagnosis of neurological conditioning that occur through improper sequencing of actions. 31 32 33", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": ": The long-term goal of this laboratory is to understand the sensory input and brainstem neural circuits involved in processing taste information. Based on results obtained in the prior grant period, there is a much greater understanding of the mechanisms involved in neural processing in the rostral nucleus of the solitary tract (rNST). In the proposed grant, properties of afferent gustatory neurons, neurons in rNST, and neurons in the pontine taste relay, the parabrachial nucleus (PBN), will be examined. In addition effects of neuromodulators, known to influence feeding behavior, on the synaptic activity of rNST neurons will be studied. Specifically, whole cell recordings of neurons in in vitro brain slice preparations and acutely dissociated neurons will be used to study: 1. The biophysical characteristics of primary afferent neurons involved in conveying taste information to the brainstem. 2. The mechanisms of inhibition in rNST by investigating the biophysical characteristics of rNST interneurons. 3. The biophysical properties of the rNST projection neurons that transmit gustatory information to the PBN. 4. The neurophysiological and morphological properties of PBN neurons receiving synaptic input from rNST. 5. The effects of neuromodulators known to influence feeding behavior on synaptic activity of rNST neurons. Since the sense of taste has a primary role in feeding, in regulating food and fluid intake, and in the initiation of oral reflex activity such as salivary secretion, it is important to learn how sensory information originating in taste receptors is processed and distributed by taste relay nuclei. It is also important to understand the mechanisms of action of neurochemicals that influence feeding at the taste relay nuclei. The experiments detailed in this proposal will provide important new information on the neural circuits, neurotransmitters and neuromodulators responsible for integrating, modifying and distributing the information that originates in taste receptors, and will thereby provide greater understanding of the role of the gustatory system in feeding and feeding-related disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Successful treatment of metastasis continues to be the main challenge in cancer. Local chest wall recurrences of breast cancer often occur in the context of metastatic disease, presenting the challenge of treating both. In this setting, salvage chemotherapy/hormone therapy result in overall response rates of only 20-30%, warranting novel treatment strategies. Our group has demonstrated that local ionizing radiation therapy (RT) can be successfully combined with immunotherapy (IT) strategies to generate an anti-tumor immune response, enabling not only rejection of the treated tumor, but also of metastases outside the irradiated field (abscopal effects). Imiquimod (IMQ) is a small molecule immune response modifier which triggers immune activation via toll-like receptor (TLR)-7 and is FDA approved, based on its clinical activity against a variety of skin cancers. In preclinical models of breast cancer, the syngeneic murine tumors TSA and 4T1, we have shown that the combination of local RT with topical IMQ successfully delayed both primary tumor growth and metastases. We propose a clinical trial that translates the preclinical experience of combining topical imiquimod and local radiotherapy to the setting of metastatic breast cancer patients who also have chest wall recurrences. The strategy is that of vaccinating patients against their own tumor to generate an immune response that, if successful, can be detected outside the field of local therapies. The specific aims are to: 1. Determine the safety and therapeutic efficacy of topical IMQ and concurrent local RT in breast cancer patients with cutaneous metastases. 2. Determine the effects of TLR7 agonist IMQ and RT on tumor-specific T cell immunity and tumor immune signature. The selection of breast cancer patients with chest wall recurrences is based on the following rationale: 1) the accessibility of the lesions to topical agent (imiquimod) and to radiotherapy;2) the ease to visually monitor local response and to perform Fine Needle Aspiration biopsies for mechanistic studies. A Phase I/II trial will be conducted to evaluate the safety of the combination of RT and IMQ, topically applied to skin metastases of breast cancer. Efficacy is measured with respect to local tumor regression as well as systemic responses (outside the field of combined therapy), as a demonstration of an effective anti-tumor immune response. Immunomonitoring will consist of an analysis of tumor-specific T cell immunity and an assessment of immune signatures in the tumor, to learn whether the proposed treatment induces tumor- specific T cell responses and generates a tumor immune signature consistent with immunological rejection. The proposed studies will create the basis for a potential paradigm shift with direct clinical implications: harnessing a novel combination to effectively immunize the patient to her own tumor, and thereby achieving a response of the systemic disease. PUBLIC HEALTH RELEVANCE: Breast cancer that has recurred to the chest wall is often concomitant with visceral metastases and does not respond readily to standard therapy. In a clinical trial that tests the novel combination of two local therapies, radiotherapy (RT) and topical drug imiquimod (IMQ), we propose to generate (and monitor) an anti-tumor immune response, capable to reject and control the disease locally as well as at distant metastatic sites. This combination has shown efficacy in a mouse model of breast cancer by harnessing the immunostimulatory effects of RT and those of IMQ, a FDA approved biologic agent for topical treatment of skin tumors. In this model immunized animals also rejected lung metastases. If successful, this strategy could be translated to other metastatic tumors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cardiovascular diseases are the leading cause of death world wide. Although traditionally thought of as diseases of \"industrialization\", there is growing recognition of the importance of cardiovascular diseases and cardiovascular risk factors such as obesity, high blood pressure, and smoking as major contributors to morbidity and mortality in developing countries, including Argentina. Nevertheless, epidemiologic studies of factors associated with cardiovascular risk in developing countries remain extremely rare. Identifying factors associated with cardiovascular risk may help guide policy or may suggest vulnerable groups which should be targeted for intervention. In industrialized countries there has been growing awareness of the role that features of living environments, including characteristics of residential environments, may play in the development of cardiovascular risk factors. However, information on the social and geographic patterning of the cardiovascular disease epidemic in countries like Argentina is virtually non-existent. Using death certificate and survey data from Buenos Aires, one of the largest urban areas in Latin America, we propose to investigate the geographic patterning of cardiovascular risk and the socioeconomic factors associated with this patterning. The specific aims of this proposal are: (1) To describe the spatial patterning of small-area variations in cardiovascular disease mortality in the city of Buenos Aires.(2) To investigate associations of area socioeconomic characteristics with cardiovascular disease mortality in the city of Buenos Aires. (3) To investigate spatial variations in the prevalence of obesity, smoking, self-reported hypertension, and physical activity in the city of Buenos Aires; and (4) To investigate associations of both area characteristics and personal socioeconomic indicators with the prevalence of cardiovascular risk factors. This research will be carried out in collaboration with Hugo Spinelli of the Universidad de Lanus in Buenos Aires, Argentina as an extension of NIH grant R01 HL071759-01 (Diez Roux, PI, June 2,2003-June 1 2007).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Left ventricular (LV) enlargement (remodeling) after myocardial infarction (MI) is a consequence of infarct expansion, extension of the infarct borderzone (BZ), and hypertrophy of the remote myocardium in addition to the effects of neuro-humoral compensatory pathways. Post-infarct LV remodeling is a strong predictor of mortality and a high incidence of heart failure (HF). Acellular hydrogels that limit LV remodeling when injected into an MI or BZ are receiving increasing attention. We term this therapy matrix-assisted infarct stabilization (MAIS) and suggest that MAIS works by decreasing myofiber stress in the infarct, BZ and remote myocardium thereby decreasing infarct expansion, improving BZ contractility and decreasing hypertrophy of the remote myocardium. Optimal design parameters of MAIS need to be determined and although initial work using finite element (FE) simulations and animal experiments to determine the effect of injection volume and hydrogel stiffness have been performed, a systematic investigation of MAIS design parameters is clearly needed. Therefore, Aim 1 seeks to determine optimal MAIS design parameters using FE element models validated with diastolic and systolic regional LV strain and LV volumes measured with cardiac magnetic resonance imaging (MRI) before and after MAIS in sheep. Sheep for validation studies will have MAIS performed with a tunable non-degradable hydrogel (semi- interpenetrating polymer networks; sIPN) injected 1) directly into the MI and BZ and 2) by intracoronary injection. In Aim 2 the effect of MAIS on regional LV stress, infarct expansion and contractility will be calculated in sheep with MI + BZ sIPN injection, intra-coronary sIPN injection and in the optimal MAIS design determined in Aim 1. The proposed work will continue our focus on BZ contractile dysfunction that was begun during the previous funding period where recent studies in sheep after antero-apical MI revealed a critical role for intracellular (IC) matrix metalloproteinase-2 (MMP-2) as a mediator of contractile protein and mitochondrial dysfunction. Further, recent studies have characterized two IC MMP-2 isoforms. First, full length IC MMP-2 cleaves contractile proteins troponin I and myosin light chain 1 (MCL-1) in the setting of oxidative stress (reactive oxygen species: ROS). Second, an N-terminal truncated constituitively active isoform (NTT-MMP-2) is induced by ROS and predominately located in the mitochondria where it is associated with abnormalities in mitochondrial ultrastructure and respiration. Further, transgenic mice with NTT-MMP-2 develop progressive LV hypertrophy and eventual HF. Based on preliminary studies, we believe that MAIS will prevent BZ injury by interrupting a process mediated by mechanical stress, ROS, and activation of the IC MMP-2 that causes damage to BZ contractile proteins, mitochondrial dysfunction and impaired ATP production. Aim 3 therefore seeks to determine the effect of optimized MAIS on ROS, IC MMP-2 expression, mitochondrial function and ATP production.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The epidermis and stratum corneum will be studied by means of the freeze-fracture technique. The nature of the thickened horny cell membrane complex and the lamellar structure of the membrane coating granule and its product will be given particular attention. Scanning electron microscope studies of human pathological disorders such as eczema and lichen planus will be correlated with light and transmission electron microscopic observations of these tissues. The mechanism of cell stacking and its maintenance will be investigated in embryonic skin and during the development and remission of essential fatty acid deficiency. Studies will be made of the permeability of normal and essential fatty acid deficient mouse skin to ultrastructural tracer molecules.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have shown that targeted therapy to the type 1 insulin-like growth factor receptor (IGF-IR) is very effective in decreasing tumor size in preclinical models of prostate cancer (CaP). It is also clear from phase 1 trials that human monoclonal antibodies (hmab) to the IGF-IR are well tolerated. We also show that the efficacy of IGF-IR hmabs are dependent on the clinical stage of tumor e.g. AD vs. Al and alterations in components of the IGF-IR signaling pathway. Therefore, as we proceed with.our clinical trials, there is an urgent need for basic and preclinical data to guide these trials forward. The hypothesis of this proposal is: the effectiveness of targeting the IGF-IR in treating CaP depends on the status of the androgen responsiveness of the tumor and the integrity of the IGF-IR signaling pathway in the cancer. This hypothesis is addressed by the following aims: Aim 1. Determine mechanisms of resistance to IGF-IR targeted therapy by the human monoclonal antibody A12. This aim will use the 27 unique human prostate cancer xenografts available in the Biospecimen Core as well as our preclinical finding of the significant synergy between castration and IGF-IR inhibition. Aim 2. Analysis of the functional interactions of the AR on IGF-IR gene expression as a determinant of response to A12 in CaP. This aim will define the mechanism of androgen regulation of the IGF-IR . Aim 3. Determine the potential of tumors that remain dormant following treatment with castration plus A12 to re-initiate tumor growth. This aim will define the mechanisms and characteristics of those CaP that remain dormant after combination therapy with castration and IGF-IR inhibition. Aim 4. Determine how the protease activity of TMPRSS2 influences IGF signaling and the response to A12. As an interaction with Project 2, this further develops the effects of TMPRSS2 cleavage of IGFBPs -3 and -5. Aim 5. Role of androgens and IGF in establishing the survival and growth of dormant CaP cells in an osseous site. This interaction with Project 1 will determine the role of IGF-IR inhibition on DTCs. The long term goals are to define the characteristics for our clinical trials targettting IGF-IR to succeed. RELEVANCE (See instructions); IGF-IR inhibition will become part of the clinical armementarium for tretament of prostate cancer. Project 3 will define the appropriate setting for the most efficacious clinical use of this targeted therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Administrative Core is the Center's planning and decision-making body and provides overall scientific direction and leadership. It will serve as CIRA's central administrator, fiscal executor and coordinating body;ensure that the Center is a primary source of information on HIV/AIDS related topics;monitor Center activities and develop, and implement and evaluate annual strategic plans. A Director, two Deputy Directors, an Executive Director, and two Assistant Directors, will comprise the Center's Leadership Group (LG), which will meet monthly to guide its administrative and scientific direction and activities. The LG will meet semi-annually with all Core Directors to discuss Core activities and plans and financial resources for implementing them, monitor Core progress in following its strategic plan, and facilitate coordination and collaboration among Core Directors. The Core will convene four advisory bodies to provide guidance in its activities. The Executive Committee will advise on all aspects of the Center's operations, including Center policies, priorities, and future scientific directions. The External Advisory Board will bring together individuals of national prominence to review the Center's progress and plans on an annual basis and provide advice on its future development. The Deans'Committee will advise on how CIRA can best involve scientists from different Yale graduate and professional schools and establish linkages throughout the University. The Community Advisory Board, comprised of representatives from community agencies and individuals infected and affected by HIV/AIDS, will help to ensure both the relevance of the Center's research to the needs of the community and the input of the community in CIRA's research. The Admin Core will oversee the affiliation process and help facilitate CIRA science by providing a range of services, including: grant preparation and, for CIRA scientists with primary appointments in the School of Public Health, grant management;assistance with personnel planning;development and maintenance of centralized databases for information sharing, program monitoring, and strategic planning;publication of the weekly email bulletin;and development and maintenance of CIRA's internet and intranet websites. The Administrative Core will also organize CIRA's AIDS Science Day.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract Cardiovascular disease (CVD) is the leading cause of death in the US, however African American residents of rural areas in the south and southeast regions have the highest prevalence of CVD of any group. African Americans carry a significant burden of CVD risk factors that often co-occur; this burden is amplified in rural communities. CVD disparities at the intersection of race and geography are driven by individual risk behaviors and complicated by factors such as limited access to quality healthcare, socio-economic inequities, limited healthcare infrastructure and environmental barriers to behavior change. Interventions to ameliorate CVD burden in rural African American communities will require placing the individual in the context of the larger community and taking advantage of new technologies to support behavior change. However, how best to integrate mobile technology into existing evidenced based interventions (EBIs) is still an emerging field and social and physical environmental factors important in rural communities are rarely considered in existing EBIs. The proposed study will address this gap in the literature by determining the feasibility and efficacy of adapting EBIs to consider the social and physical environment in important in rural African American communities and determining the acceptability of mobile technology in these communities to support behavior change. The proposed study is built on the strong foundation of Project GRACE's 8-year history of designing and testing interventions using a community-based participatory research (CBPR) approach, and individual and collaborative expertise in community-based CVD outreach, service and research. We have developed a phased CBPR study with a long-term goal to reduce rates of CVD in Eastern NC. The overall objective of this proposal is to assess feasibility of implementing an EBI, adapted to the needs and interests of a rural community in order to plan a large scale study. To that end our specific aims are to 1) expand and sustain a Project GRACE CVD coalition of community and academic stakeholders to develop successful CVD risk prevention strategies in rural communities; 2) conduct a mixed-method community needs and assets assessment based on: a) assemble, review and assess existing sources of CVD data; b) identification of community strengths and resources using a web-based survey of community, faith based, social service and healthcare organizations; c) determine the acceptability of components of CVD risk reduction EBIs and community members' perceptions of possible targets for intervention using focus group interviews; d) determine specific family influences (barriers and facilitators) on acceptability of EBI acceptability; 3) adapt PREMIER, a multi-component EBI using intervention mapping; and 4) conduct a small-scale randomized control trial to assess a) efficacy; and, b) feasibility and adaption of implementing adapted PREMIER in rural settings.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "It is requested that funds be provided to cover the expenses of approximately 8 out of 20 symposia that will make up a major part of the program for the First International Congress on Cell Biology scheduled for Sept. 5 - 10, 1976, in Boston, Massachusetts. Approximately 3500 persons are expected to register and benefit from the unique communication that large congresses alone can provide. The symposia, planned for the morning hours, will cover a wide range of health related topics, including the following: Molecular Cytogenetics of Eukaryotes, Heterogeneous Nuclear RNA, Functional Organization of Chromosomes, Chromatin Structure, Cytoplasmic Control of Nuclear Expression, Cells and Hormone Action, Eukaryotic Cell Cycle, Plasma Membrane Organization, Transducing Membranes -Structure and Function, Cell Surface Immuno-Receptors, Cell-Cell Interactions, Cell Surface in Neoplasia, Molecular Basis of Motility, Gamete Morphogenesis, Somatic Cell Hybridization. They will be followed in the afternoons and evenings by sessions of short contributed papers assembled and scheduled to extend the discussion of topics reviewed in the morning symposia. Workshops and poster sessions will further enrich the scientific program. As is customary, abstracts of the contributed papers will be published in advance of the Congress. Additionally, arrangements will be made to publish, either in the Journal of Cell Biology or in book form, the papers presented at the symposia. Thus, the proceedings of this First International Congress on Cell Biology to be held in the U.S. will be made available to the scientific community at large. The Congress will combine the annual meeting of the American Society for Cell Biology with the first meeting of the new International Federation for Cell Biology. The latter organization comprises three major societies of cell biologists - the American Society for Cell Biology, the European Cell Biology Organization, and the Japan Society for Cell Biology. The American Society is hosting the Congress and making all of the arrangements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this project is to cover basic and applied aspects of various pathogenic fungi including their morphology, taxonomy, pathology, epidemiology, biochemistry, and genetics. The topics of present interest include: 1) biochemical differences among four serotypes of cryptococcus neoformans; and 2) structure of the capsular polysaccharide of serotype C.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To develop cutting-edge research that helps improve the ability of medical institutions, public health agencies and government institutions to plan and respond to pandemics. The center will focus on broad influenza surveillance and characterization of virus strains and development of pathways and approaches to rapidly link basic research findings with clinical and public health practices to better prepare for seasonal and pandemic influenza outbreaks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Idiopathic anterior uveitis in man is characterized by a breakdown of the blood-aqueous barrier which is acute and often recurrent. Left untreated, it can frequently lead to secondary glaucoma. Currently available objective measures of ocular inflammation necessitate undesirable invasive procedures such as paracentesis. Thus, clinical assessment of the severity of anterior uveitis and its response to therapy, has relied heavily upon subjective grading of such parameters as flare and cells. A major aim of the proposed studies is adaptation of the technique of aqueous fluorophotometry to the objective, but non-invasive measure of anterior segment inflammation. Eventual direct, clinical application of the procedure is our ultimate goal. Aqueous fluorophotometry will be used to measure the changes in severity of a bacterial endotoxin-induced model of anterior uveitis in rabbits and later in monkeys. By taking advantage of a new method developed in our laboratory, we can compare fluorophotometric estimates of severity with results of ultrastructural tracer studies on the same eyes. The ability to distinguish between sites which do and do not contribute to aqueous fluorescence at each stage of uveitis severity provides a valuable tool with which to refine pharmacokinetic models necessary to interpret fluorophotometric data in a clinical setting. Using the same procedures, in conjunction with the freeze-fracture technique, we will examine the process of re-establishment of the blood-aqueous barrier during resolution of anterior uveitis, both with and without treatment using steroidal and non-steroidal anti-inflammatory agents. Particular emphasis will be given to the mechanisms involved in the reassembly of tight junctions, which constitute the anatomic equivalent of the blood-aqueous barrier. Incomplete or faulty barrier reformation, after uveitis, could contribute to recurrence. Thus knowledge of the process of barrier reformation may be critical to an understanding of recurrent anterior uveitis in man. Additional animal models of increased blood-aqueous barrier permeability without appreciable inflammatory cell responses will be exploited to better understand the relationship between flare and fluorophotometrically measured aqueous fluorescence. Finally, studies in rabbits will determine if changes in intraocular pressure caused by inflammation are paralleled by changes in the size or number of gap junctions in the ciliary epithelium of these eyes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite the centrality of motor learning to basic and clinical neuroscience, we know very little about the quantitative role neural systems play in the transformation of senses into adapted control. The experiments presented here will challenge normal human subjects with perturbations of varying strengths, durations, frequency of application, biases, and spatial complexities both within and across movements. The variability of these perturbations will generate a template with which we will identify subtleties in trial-by-trial adaptation. These interrogations, coupled with novel state space analyses, will enable a thorough understanding of the transformation of specific sensory experiences into immediate, incremental adaptations in predictive control, greatly enhancing our quantitative understanding of human motor adaptation. Experience enables us to build internal dynamic models of our movement environment. Investigators of this internal dynamic adaptation have hypothesized two components of learning: the abstraction of an error signal from previous movements and the application of this error to either specify or generalize learning across movement space. Human trial-by-trial adaptation has, to date, suggested that adaptation constantly scales with sensed error and generalizes broadly across movement space. However, preliminary results from the PI have discovered surprising flexibility in both components of learning: sensory feedback can induce adaptation strikingly disproportional to movement error, and environments can induce narrowing of generalization across movement space. Here we propose to identify the necessary sensory experiences to induce these newly established changes in the fundamental computations people execute to transform single movement sense into incremental adaptation. These results will illuminate how the nervous system performs real-timed signal processing to improve motor performance. The resultant models will help the neuroscience and biological modeling community to better connect behavior to its underlying physiological basis. These insights will also be of use to investigate the full repertoire of normal motor control and how control fails in disease states. We aim to formulate the scientific basis of how rehabilitation can optimally help patients generalize beyond clinical training to improve motor function in their daily lives.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mixed-function oxidases containing different forms of cytochrome P-450 are the key enzymes that metabolize a wide variety of drugs, chemicals and carcinogens. The focus of this project is the utilization of specific inhibitors and inducers of aryl hydrocarbon hydroxylase (AHH) to probe the multiplicity, diversity, and different catalytic properties of the cytochromes P-450. Seven,8-benzoflavone and L-maackiain acetate are inhibitors that are effective against different forms of AHH. We are using these inhibitors to probe the active catalytic sites of the multiple forms of cytochrome P-450. Also, other compounds, i.e., medicarpin and maackiain, are being studied and their structural and inhibitory effects on cytochrome P-450 are being determined using microsomes and purified cytochromes P-450.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "New, powerful interventions such as early treatment and pre-exposure prophylaxis are emerging as options for HIV prevention globally. However, their inclusion in HIV prevention strategies at the national level is not warranted, given health systems limitations; and stakeholders' concerns. Moreover, the ways in which such strategies could be incorporated not fixed, and strategic combination prevention programs must be designed. This proposal intends to promote, and study, a process through which (1) HIV prevention stakeholders in Peru can express their views and concerns about existing and emerging (e.g. Pre-Exposure Prophylaxis, Early Treatment for Prevention) HIV prevention interventions focused on MSM/transwomen; (2) the capacity of the health system and changes needed to incorporate emerging interventions, and associated costs, will be assessed; (3) combination prevention scenarios associated with effectiveness and cost-effectiveness estimations will be developed; and (4) stakeholders will seek to agree on one or more feasible and acceptable prevention scenarios, for potential implementation. Methods to be used include stakeholder analysis, health systems and cost assessments, mathematical modeling based on global evidence of effective interventions and on local evidence of stakeholders' views and of health systems conditions, and scenario analysis to discuss proposed scenarios and seek consensus on the most viable one(s) for future implementation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] [unreadable] The Twenty-Fifth International Neurotoxicology Conference (NEUROTOX 25) addressing the theme of \"Environmental Etiologies of Neurological Disorders: Scientific, Translational and Policy Implications\" will be held October 12-16, 2008 in Rochester, New York. The meeting locations are the Hyatt Regency Conference Center and the University of Rochester Medical Center located nearby. The International Neurotoxicology Conference is an annual event that focuses on a timely theme while providing an opportunity for presenting new data related to the general interdisciplinary field of neurotoxicology. The International Neurotoxicology Conference Series will reach its silver anniversary in 2008, with its 25th meeting. The Specific Aims identified in the proposal will be accomplished by organizing and conducting a 4 1/2 day conference according to a varied format similar to previous conferences in this series which has proven to be highly successful. Each conference has its own \"special touch\" and consists of Tutorials, Symposia, Workshops, Panel Discussions, Roundtables, Debates, Poster Session, Pre- and Postdoctoral Student Competition and Awards and social events as well as novel approaches to facilitate networking and mentoring. NEUROTOX 25 will not be simply an occasion to present data. It will also coalesce human, experimental animal, methodological, and epidemiological research, as well as risk assessment, social and policy implications, by offering a well-recognized venue where previously unconnected investigators can come together and where commonalties can emerge. Traditionally the Neurotoxicology Conference places special emphasis on nurturing, promoting and rewarding pre- and post-doctoral students and young investigators. Extra effort is extended to mentor students and help them network with the leaders in the field. The health relatedness of this application is that the results of this conference will contribute to a fuller understanding of the true risk posed by environmental toxicants for human neurological diseases or disorders. It has become evident that many human neurological diseases arise from complex interactions of multiple risk factors, of which environmental chemical exposures may serve as one contributing risk. Other environmental and host factors, such as genetic background, stage of development, dietary status, immune status, obesity, stress, socioeconomic status, gender, aging, behavior, and intercurrent disease state, as well as simultaneous chemical exposures, can also contribute. Science, policy, and translation can no longer be treated as independent entities. NEUROTOXICOLOGY 25 is poised to view and address them as they truly are: a multidimensional interdisciplinary problem. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application requests funding for the study coordination, data management, and biostatistical support for a Phase III clinical trial of Coenzyme Q10 (CoQ) in Huntington's disease (HD). This is a companion application to that of Merit E. Cudkowicz, M.D. from Massachusetts General Hospital entitled \"Coenzyme Q10 in Huntington's Disease (2CARE).\" The primary aim of the project is to conduct a multicenter randomized, double-blind, placebo-controlled trial of CoQ at a dosage of 2400 mg/day involving 1100 ambulatory subjects with HD followed prospectively for 5 years. CoQ is a cofactor involved in mitochondrial electron transfer and an antioxidant that has demonstrated increased survival in a transgenic mouse model of HD and promising results in human trials in HD and Parkinson's disease. It is hypothesized that chronic treatment with high-dosage CoQ will slow the progressive functional decline in HD. The specific aims of our application are (1) To provide project coordination for the trial including: development and implementation of web-based randomization and enrollment procedures, provision of study documents (protocol, model consent and assent forms, operations manual, case report forms), training of site personnel and standardization of study procedures, facilitation of recruitment and retention of a diverse sample of research subjects, and packaging and distribution of study drug; (2) To provide data management for the trial including: receipt and entry of all data in a relational database, generation and resolution of data queries, monitoring of important study events, and maintenance of a compliant and secure data environment; and (3) To provide biostatistical support for the trial including: refinement and implementation of the statistical analysis plan to address the specific aims of the trial; design and implementation of an interim monitoring plan that will allow modification or termination of the study on the basis of early evidence of either (i) safety concerns, (ii) overwhelming evidence of efficacy, or (iii) futility, in conjunction with the NINDS-appointed Data and Safety Monitoring Board (DSMB); and preparation of open- and closed-session reports for the DSMB.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (From the applicant's abstract): Beta-amyloid precursor protein (Beta-APP) is an integral transmembrane glycoprotein whose proteolytic processing results in the generation and secretion of soluble Beta-APP amino-terminal peptides (sBeta-APP) as well as amyloid-beta protein (A-Beta), the major constituent of the amyloid deposited in the brain of Alzheimer's disease patients. The protein interaction domain of two adaptor proteins, X11 and Fe65, bind to the YENPTY motif within the cytoplasmic domains of Beta-APP. Since this motif has a role in the internalization of Beta-APP, and A-Beta is generated in part following Beta-APP internalization, the effect of X11 and Fe65 binding on Beta-APP trafficking and processing have been studied. It was hypothesized that A-Beta and sBeta-APP are physiological ligands with reciprocal effects on neuronal integrity, and imbalance in the relative amounts of A-Beta and sBeta-APP may have beneficial or deleterious effects on neuronal survival and growth. The goal of this application is to test the hypothesis that complexes among the adaptor proteins and Beta-APP reflect a signaling pathway that regulates the processing of Beta-APP. Specifically, Aim 1 is to examine the effect of Fe65 binding to Beta-APP on Beta-APP processing and secretion and to compare this effect to that of X11; Aim 2 is to examine the effect of X11 and Fe65 binding to Beta-APP on cell associated Beta-APP trafficking; Aim 3 is to establish a transgenic mouse model overexpressing human X11 in the formation and deposition in transgenic mice overexpressing human X11 in the brain; and aim 4 is to examine the effect of X11 association with Beta-APP on A-Beta secretion and on amyloid formation and deposition in transgenic mice overexpressing both X11 and Beta-APP. Study of the molecular function of Beta-APP within a signal transduction pathway may lead to insights on pathological processes activated or inhibited in Alzheimer's disease patients. Manipulating the interaction of Beta-APP with an effector protein, such as X11 or Fe65, may provide a novel approach for the pharmacological modulation of Beta-APP processing in Alzheimer's disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The GRIP project has provided partial support for UNC's advanced technology tracker work and is a customer for trackers designed by this team. As noted last year, the UNC optical tracker is attractive in the PIT because it is immune to the electrical noise created by the motors in the PHANToM. This electrical noise causes significant error in our current commercial magnetic trackers. The new lightweight optical tracker was deployed early in the reporting period and has undergone testing, calibration and algorithmic refinement during this reporting period with RMS noise of 0.1 mm in position and 0.03 degrees in orientation. The system measures position and orientation in a 26 by 30 foot working area and makes 1500 to 2000 measurements per second with less than 1 millisecond of latency. Software has been developed and tested for multiple simultaneous trackers operating under the same ceiling (n = 2 currently) to support deployment of this tracker in the PIT. Experiments were conducted to determine an optimal pattern of tracker ceiling tiles (the tracker's LED beacon system is located in the tiles) for the PIT two-user configuration. We expect to deploy the tracker over the PIT before the end of March 1998.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "FIU is a public, research institution (RU/H) located in Miami. It has an enrollment of ~50,000 students (>73% UR), emphasizes research in its mission and includes health and diversity as strategic themes. FIU provides an excellent opportunity to develop a significant number of well-rounded, underrepresented (UR) scientists for biomedical research careers. In 2007, FIU launched a rigorous pilot biological sciences curriculum for freshmen called Quantifying Biology in the classroom (QBIC). Having impressively graduated three freshman-through-senior cohorts of graduates, QBIC successfully passed all academic and administrative evaluation and review steps, and is now institutionalized, beginning Fall 2013, as a Quantitative Biology Track in FIU's Biological Sciences Department. QBIC offers a pool of talented & exceptionally prepared scholars for FIU's MARC U*STAR Program. For each year of our proposed 5-year grant period, we request funding to support 8 juniors, through graduation, for two years (totaling 40), who will be selected from a pool of talented and well-prepared honors students in QBIC and traditional curricula, since not all of our talented students are in QBIC. QBIC is a 4-year program developed by FIU's biological sciences, statistics, mathematics, chemistry, physics, computer science and biomedical engineering faculty. It is an integrated lock-step cohort block program where students take classes together, and courses within blocks have identified & coordinated areas of overlapping synergy. Scholars get to see how biological data is analyzed, how mathematics can model biological processes, and teaching units show them how sound biological theories are generated. Part 1 (freshmen & sophomores) integrates calculus & statistics with biology, genetics, ecology & chemistry using a Teaching Pentagon; Part 2 (juniors & seniors), offers tailored-upper level courses. MARC U*STAR trainees will actively participate in directed scientific research & enrichment activities that include modeling workshops, journal club activities, professional development workshops, science cafes, scientific presentations & participation in extramural research. Thus, the three-prong short and long term goals and measurable objectives for FIU's MARC U*STAR Program include: (1) provide motivated, honors students with the tools & skills that allow them to be academically competitive & successful; (2) immerse them in supportive & stimulating research environments with activities that enhance & nurture their interest in science, (3) increase the number of UR who continue on to, excel in and complete biomedical or behavioral sciences Ph.D. or MD/PhD programs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sickle cell anemia is an autosomal recessive disorder and the most common genetic disease affecting African-Americans. Approximately 0.15% of African-Americans are homozygous for sickle cell disease, and 8% have sickle cell trait. Acute pain crisis, acute chest syndrome (ACS), and secondary pulmonary hypertension are common complications of sickle cell anemia. Pulmonary hypertension has now been identified as a major cause of death in adults with sickle cell disease. Similarly, pulmonary hypertension has been identified as a chronic complication of hemolytic disorders such as thalassemia, hereditary spherocytosis and paroxysmal nocturnal hemoglobinuria. Sildenafil has been proposed as a possible therapy for both primary and secondary pulmonary hypertension and recent phase I/II studies from the intramural NIH suggest it is well tolerated and efficacious in this population. Furthermore, a number of recent studies have suggested that NO based therapies may have a favorable impact on sickle red cells at the molecular level and could improve the abnormal microvascular perfusion that is characteristic of sickle cell anemia. This clinical trial was designed with three major objectives: 1) to assess cardiopulmonary function in patients with sickle cell disease and thalassemia with and without pulmonary hypertension, to better characterize the effects of chronic hydroxyurea therapy on cardiopulmonary function, 2) to determine the relative acute vasodilatory effects of sildenafil, and inhaled NO in patients with hemolysis-associated pulmonary hypertension and 3) to determine the chronic effects of the addition of inhaled NO on pulmonary hemodynamics and functional capacity in patients with hemolysis-associated pulmonary hypertension chronically treated with sildenafil. Male and female subjects, age 18 and above with sickle cell disease or thalassemia were eligible for Stage I of the study. Stage I evaluated 25 subjects and included pulmonary function testing, chest x-rays, VQ scan, 6-minute walk, sleep study, cardiac MRI, MRI assessment of liver iron burden, high resolution CT scan, and blood tests. After the Stage I testing subjects who were found to have mild to severe pulmonary hypertension were asked to breathe nitric oxide for 20 minutes through a facemask while undergoing the right heart catheterization. Then they received one dose of sildenafil and their pressures were monitored for 4 hours followed by another 20 minutes of breathing nitric oxide. Eighteen subjects completed Stage II of the study. Stage III was open to those individuals who had completed Stages I and II and had been on chronic sildenafil therapy for at least 3 months. Stage III consisted of breathing nitric oxide through the INO Pulse device for 6 weeks while continuing to take the oral sildenafil therapy. Three subjects completed this stage. The NHLBI IRB approved this study on 6/6/06. Our first patient was enrolled on 9/11/06. Thirty-one subjects were enrolled in this study. Three subjects completed all three stages;18 additional subjects completed stages I and II only;7 subjects completed stage I only;1 subject completed only part of Stage I, 1 subject did not return for the study after giving informed consent;and 1 subject withdrew for medical reasons. Stages I and II of this protocol provided the necessary work-up needed to determine if a subject had pulmonary hypertension. It also provided the research subjects for Stage III, which evaluated an intervention for this disease. This study is now closed to accrual and is open for data analysis and sample sharing only.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neisseria gonorrhoeae causes about 800,000 new infections each year in the United States, with health-care costs approaching 2 billion dollars/year. Various surface components, including lipooligosaccharide (LOS) and opacity protein(s) (Opa(s)), are important in mediating disease. The objectives of this proposal are to understand how Opa and LOS function in disease, and determine if their interaction enhance virulence. The central hypothesis of the proposed research is that expression of specific Opa/LOS combinations promotes GC- GC interactions to produce biofilms with different disease potentiating properties and antibiotic resistance profiles. We intend to test our hypotheses by pursuing three specific aims: We will determine how Opa and LOS function cooperatively to promote bacterial-bacterial interactions, how bacterial-bacterial interactions effect the antibiotic resistance properties of GC biofilms and how bacterial aggregation influences adherence, invasion and/or transmigration. The results from our study will allow us to define how LOS and Opa variation contribute to disease pathogenesis. The impact on human health is expected to be significant, because with the new knowledge gained, we will be better positioned to understand what is needed to make a successful vaccine and what challenges we will face in developing new approaches to the treatment of gonorrhea.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Membranes have been subject to increased investigation because of their role in cellular growth, division, and differentiation. The cell surface is also implicated in the expression of the neoplastic state, perhaps as the major phenotypic factor. Lectin-mediated agglutination, disappearance of a large surface protein (LETSP), increased fibrinolysis, increased transport of 2-deoxyglucose, and changes in morphology are expressions of the neoplastic state useful in understanding the membrane alterations involved in the transformation of normal cell behavior. We propose to study the effect of enucleation and metabolic inhibitors on the expression of these characteristics. The use of cells infected with temperature-sensitive viruses provides the necessary system to study. Rearrangement of membrane components appears to play a role in altering cell behavior. We propose to examine the topological distribution of membrane components utilizing novel new cross-linking reagents synthesized in this laboratory for nearest-neighbor analysis. In addition, lectin-binding to cell surfaces of normal, transformed, and trypsinized cells will be re-examined by means of equilibrium dialysis. Previous binding studies have not met this requirement. The absolute number of receptor sites (Scatchard plot), the heterogeneity (Sips distribution), and thermodynamic parameters describing lectin:receptor interactions can be obtained and compared in terms of experimental cell manipulations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Acute Myeloid Leukemia (AML) is a heterogeneous malignancy that arises from the bone marrow, resulting in less than 50% of patients with AML being cured. The molecular mechanisms of AML are not well understood. The specific aims of my previous R01 grant addressed the overall hypothesis that the cAMP Response Element Binding Protein (CREB) contributes to leukemogenesis. In the initial funding period, we described the overexpression of CREB in AML. We successfully completed the three specific aims and demonstrated that 1) CREB is expressed in less differentiated normal hematopoietic stem and progenitor cells; 2) CREB overexpression is associated with an increased risk of relapse and decreased event-free survival; 3) overexpression of CREB in myeloid cells results in increased proliferation and survival in vitro; 4) hMRP8-CREB transgenic mice develop myeloproliferative disease; and 5) CREB knockdown in vitro and in vivo inhibits leukemic progression but not long-term hematopoietic stem cell engraftment. In this revised competitive renewal application, we propose to examine the molecular pathways to understand the mechanism of CREB function in normal and aberrant hematopoiesis. We propose four specific aims. In Aim 1, we will study the role of CREB on hematopoietic dynamics by transplanting bone marrow from CREB transgenic mice into lethally irradiated mice and assessing molecular changes in the myeloid compartment. Our collaborators and I found that the microRNA, miR-34b, recognizes a sequence in the 3' UTR of the CREB gene. In Aim 2, we will characterize the regulation of CREB by the microRNA, miR-34b. Our preliminary data showed that Meis1 and Pbx1 are upregulated 33- and 28-fold by expression profiling. Meis1 and Pbx1 are direct targets of CREB. Thus, Aim 3 describes experiments to study the requirement of Meis1 and Pbx1 downstream of CREB. Finally, we identified sox4 and gfi1 in a retroviral insertional mutagenesis screen in which CREB transgenic mice were infected with the MOL4070LTR retrovirus. CREB transgenic mice infected with this retrovirus developed AML with a shortened latency compared to control mice. In Aim 4, we will examine the role of sox4 and gfi1 as genes that cooperate with CREB and contribute to the development of AML. These studies will provide new insights into how CREB regulates normal hematopoiesis and contributes to leukemogenesis. PUBLIC HEALTH RELEVANCE: The goal of this R01 application is to study the role of CREB in the development of acute myeloid leukemia. This project is relevant to public health because it focuses on identifying new pathways and targets for development of drugs to treat AML. Therefore, results from this work will improve the quality of life of AML patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This program project builds on and expands the research conducted in the RAND Corporation's previous NIA program project by renewing it for five years with a focus on the intersection of health and economic status. The research conducted within the program project will contribute to the following long-range goals: understanding how health and socio-economic status are related in older populations, how those relations are influenced by the institutional structures put in place by different nations, how wealth accumulation and savings behavior change in response to events, and how housing ownership is influenced by risk, as well as contributing to the improvement of public policies intended to enhance the health and wealth of the elderly population. As did its predecessor projects at RAND, this program project will have an international scope;researchers will examine societies that have some socio-economic similarities to that of the United States but with differing institutional contexts. The program project will include seven component projects on these topics: consumption dynamics near and after retirement, socio-economic status and health, comparing work disability across countries, institutional effects on health and economic status, causes and consequences of obesity among the elderly, expectations and realizations of retirement benefits, and housing price risk, home ownership, and wealth. The program project also includes an administrative core and a data management and computing core. These two cores and the program project approach that unites all components will permit efficiencies in research support and facilitate interaction with researchers elsewhere while attracting young researchers to the study of aging issues. Project staff conserves the expertise accumulated through the predecessor program projects and includes researchers who assisted in collecting some of the datasets to be used----experience that enhances understanding of the data's potential and limitations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We plan to develop novel tools for assessing GPCR ligands modulating G-protein-dependent and ?-arrestin- dependent ERK signaling pathways. Many GPCRs display a phenomenon in which different ligands can differentially activate different signaling pathways via the same receptor. It has become clear that single signaling pathway approaches for identifying drug candidates are not adequately suited to detect the full repertoire of compounds that may have other signaling pathway activities beyond the target receptor. One has to examine multiple signaling pathways to link pathway activities to physiologic functions. ERK signaling cascades are important components of GPCR signaling pathways. Both G-protein and ? -arrestin mediated signaling pathways can lead to ERK activation. G-protein activation of ERKs results in the translocation of active ERK to the nucleus, where it can phosphorylate and activate various transcription factors. In contrast, when ERK is activated via an arrestin-dependent mechanism, active ERKs remain largely in the cytoplasm, where they can phosphorylate non-nuclear substrates. Subcellular locations of phosphorylated ERKs determine downstream signal transduction cascades. Current ERK activation assays are all antibody-based methods. The antibody-based assays measure the phosphorylation status of ERKs using cell lysates, but lack the ability to distinguish G-protein-dependent or arrestin-dependent ERK activation. Critical information on subcellular location and distribution of activated ERKs, which determine specific signal transduction cascades, is missing. In addition, some of antibody based assay methods are not HTS friendly. Currently, pathway- specific ERK activation assays with the HTS capability do not exist. We propose to develop these novel tools by applying newly developed cell-based protein-protein interaction LinkLight assay technology. The assays can measure specific subcellular ERK activation signaling pathways. ERK signaling pathways are implicated in LTP, memory, learning, mood stabilization etc. Therefore, developing these tools for identifying compounds modulating pathway-specific ERK activation should facilitate new GPCR drug discovery efforts for treating mental disorders. We propose four aims for the project. Aim 1. Develop G-protein-dependent ERK2 activation pathway assay. We plan to utilize phosphorylated ERK2 interaction with imp7 as signal readout for assay development. We plan to validate the assay by using the m-opioid receptor (MOR) and its ligand morphine for nuclear location of the ERK2 activation pathway, since morphine is known to activate G-protein-dependent ERK pathway. Aim 2. Develop arrestin-dependent ERK2 activation pathway assay. We plan to utilize phosphorylated ERK2 interaction with receptor-associated ? -arrestins as signal readout for the assay development. We plan to validate the assay by using the MOR and its ligand etorphine for arrestin-dependent cytoplasmic location of the ERK2 activation pathway, since etophine is known to activate arrestin-dependent ERK pathway. Aim 3. Generate and characterize D2R ERK pathway-selective activation assay cells. Dopamine D2 receptors (D2R) include a long (D2L) and a short (D2S) isoforms. We plan to generate stable D2L G-protein- dependent and D2S arrestin-dependent ERK activation assay cells. It is known that D2S activates ERK signaling by an arrestin-dependent pathway and D2L activates ERK signaling by an arrestin-independent pathway. The cells will be used in aim 4 for assessing D2R compounds on ERK pathway activities and for assessing HTS feasibility. Aim 4. Assessing D2R compounds on specific ERK pathway activities and assessing HTS feasibility by conducting a small pilot screening. Brain-derived dopamine D2 receptor-regulated behaviors have been associated to ERK activation. However, it is largely unknown D2R compounds on ERK activation especially specific ERK activation cascades. We plan to profile D2R compounds (listed in ref. 44) to look for their ERK pathway-selectivity preferences. Dr. Sam Kongsamut, our consultant for the project, having over 20 years experience in antipsychotic drug discovery programs in pharmaceutical industry will correlate the specific ERK pathway assay potencies with potencies of in vivo studies (animal models and clinic data). Pathway-selective compounds could have functional selectivity. We also plan to demonstrate assay feasibility for HTS. Dr. Wei Zheng's group in NCATS will conduct the pilot screenings using the LOPAC library. Based on the results of the aim 4, we will plan next phase research proposal (phase II) and collaborate with partners for developing new leads for treatment of mental health disorders. Once developed, these cells would be the unique products on the market. We plan to market specific GPCR ERK LinkLight assay cells, provide custom-services for compound profiling and screening, and license ERK pLuc reporter host cells with an annual fee. The end users can use these host cells to develop their interested GPCR ERK LinkLight assays.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Mental Health Clinical Research Center at NYU has provided a new organizational structure in the medical school that has enabled investigators to work together to improve our understanding and treatment of three major psychiatric illnesses: organic mental syndrome, affective disorders and schizophrenia. Specifically, the Center has been organized in order to: (1) provide string and effective leadership in the development of collaborative studies involving clinical investigators and basic scientists, (2) develop control procedures to insure that the best possible research is carried out, (3) provide Core resources to investigators in the center in order to facilitate research design, diagnostic and behavioral assessment, statistical analysis and access to a computer facility, (4) provide Core laboratories to investigators in the Center so that specialized tests and procedures can be carried out in conjunction with clinical and basic science studies, (5) provide seed money in order to encourage quality pilot proposals, particularly by young investigators, (6) facilitate recruitment of normal volunteers, symptomatic volunteers and patient volunteers, (7) provide a setting in which both didactic and preceptor type training can be provided to students, postgraduates and other relevant groups, (8) provide an institutional framework for the training and development of young investigators, (9) establish a resource, in the designated areas of interest of the Center, for the school, the community and the broader scientific community and (10) carry out periodic rigorous external review and criticism in order to maintain the most effective leadership and quality of scientific research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Tissue and Peptide Resource Core serves four primary functions encompassing human tissue, animal tissue, Abeta peptides and data storage/distribution. The Core provides postmortem human tissue to Program investigators to facilitate individual research. The Core staff will process human autopsy tissue to meet the special needs of the investigators (e.g. fixation suitable for electron microscopy) and will select and characterize a subset of cases for distribution to all investigators so that they are working on a common set of staged cases. Each of the five projects will require such human tissue. The Core also provides postmortem animal tissues (canine and transgenic) that serve as model systems of brain aging. Tissues and embryos from investigators and the Core will facilitate crossing Hsiao APPsw and LaFerla APP and Abeta mice with other transgenic mice (e.g. Fas receptor mutant-lpr) as needed by Program investigators. All of the five projects will require canine or transgenic mouse lines. The Core will continue to produce high quality synthetic Abeta peptides and derivatives for us in vitro experiments, and in the production, characterization, and purification of specialized Abeta antibodies. All of the projects make use of peptides from the Core. Finally, the Core will quantify pathology within tissues distributed to Program investigators and maintain records of the distribution of all tissues and peptides, ensure the usage of common tissues and adjacent sections where appropriate and provide quantitative data in a format usable to all investigators. Statistics and power analyses for all Program Project Investigators will be guided by the Data Management and Biostatistics Core for the Institute for Brain Aging. To facilitate in case selection and quantification, the Tissue and Peptide Resource Core will also optimize staining parameters using Core generated antibodies and assist Program investigators with immunocytochemistry protocols.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The BIENESTAR study is a longitudinal cohort study of the effect of drug abuse on nutritional status and outcomes among Hispanics with HIV infection. The cohort is comprised of 3 groups: HIV+ drug abusers, HIV- drug abusers, and HIV+ non-drug abusers. Efforts will focus on expanding and maintaining this unique cohort. New hypotheses addressed in this competitive renewal reflect current trends, as HIV becomes a chronic disease. Liver disease is an increasingly common complication in patients with HIV. While it is generally accepted that HCV is largely responsible for liver disease in HIV infection, studies suggest that drug abuse itself is a risk factor for liver disease among persons with HCV. Animal studies have shown that cocaine can cause liver toxicity by the induction of oxidative stress with tissue damage. Hispanics of the NE USA preferentially inject cocaine more than other races, and therefore may be at increased risk of oxidative stress and liver dysfunction. Low serum antioxidant micronutrient levels are common in drug abusers, further increasing the risk of oxidative tissue damage. This study proposes to examine antioxidant status, oxidative stress, and liver dysfunction in Hispanic drug abusers with HIV infection. There are 4 specific aims: 1) To examine the association between the type (cocaine versus heroin) and frequency of drug abuse with plasma antioxidant capacity and oxidative stress. The specific interest is to know if cocaine differs from heroin in its ability to alter antioxidant capacity and cause oxidative stress; 2) To examine the association between the type and frequency of drug abuse and liver dysfunction. If cocaine or heroin cause oxidative stress to hepatic tissues this may result in detectable alterations in liver function; 3) To examine the association between oxidative damage and liver dysfunction. An association between oxidative stress and liver dysfunction will imply that drug abuse alters liver function through a mechanism involving oxidative stress; and 4) To examine the influence of infection with HIV, infection with HCV, and alcohol consumption on the associations described in specific aims 1 to 3. This will determine if drug abuse is an independent risk factor for liver dysfunction or whether, like alcohol, it is an accelerator of liver dysfunction in persons with HIV and/or HCV infection. The proposed study will build on the infrastructure of the BIENESTAR study (NIDA DA 11598) and the BIENESTAR-micronutrient study (NIDA DA 14501).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "NISC currently operates the following suite of production sequencing machines: 1 PacBio RS II, 4 HiSeq2500s, and 3 MiSeqs. Using these platforms, we have generated over 620 billion reads in the past year alone. Though we remain consistently at a level of a mid-scale genome sequencing center, we have maintained advantageous economies of scale while remaining relatively agile. In keeping with the Comparative Sequencing interests, several years ago NISC implemented an amplicon-based Sanger sequencing pipeline designed to focus on intra-species variation. Numerous clinically relevant projects were designed to amplify and sequence specific genes and regions of interest in small groups of human subjects, yielding great insights into disease related genotype/phenotype combinations. As an early model for application of genome sequence data to medical research our flagship ClinSeq Project greatly advanced the study of atherosclerosis by providing sequence data for 250 genes in over 500 volunteers (http://www.genome.gov/20519355). While this approach was extremely productive, we evaluated and then adopted the NextGen sequencing platform to more efficiently and rapidly collect whole exome data for the ClinSeq Project, followed by many other medically relevant projects. As a consequence of these advances, NISC no longer offers Sanger-based amplicon targeted sequencing in production mode. One publication related to our earlier Sanger-based efforts is listed in the publications section of this report (8). The adoption of many new sequencing protocols in production created the commensurate need for dramatic changes to sample tracking, flow control and primary analysis pipelines, as well as, project management and cost accounting. Rapid design, development and implementation of new Laboratory Information Management System (LIMS) by a dedicated NISC team has met the initial challenges and continues to evolve quickly to adapt to a continuous flow of changes in sequencing technologies. A combination of talented IT staff and bioinformaticians have met the challenges of extremely large and complex data sets by implementing and continuously adapting pipeline programs to support rapidly evolving software associated with each of the sequencing platforms. Beyond primary analysis that results in DNA basecalls and quality scores, NISC has worked closely with members of other NHGRI research groups to implement and support high-throughput production of biologically relevant secondary analysis. One shining example of these efforts is the production scale processing of Whole Exome Sequencing (WES) data to all of our clients, the end product of which is distilled sets of variants of interest that are accessible in user-friendly fashion by the use of the in-house developed VarSifter program. The success of these programs has led to an increasing number of projects from a growing number of investigators. Last year we added a CLIA compliant pipeline for WES of samples originating from the NIH Clinical Center through the Clinical Center Genomics Opportunity program (https://www.genome.gov/27558725) and have completed sequencing of over 100 samples. Just recently, the Exome with Secondary Findings Analysis Test was CLIA certified. The implementation of improved project management tools is helping to address the challenges associated with such growth, which is now yielding results as publications for WES (n = 6) (4, 5, 7, 10, 11, 14), Whole Genome Sequencing, Assembly and/or Annotation (n = 1) ( 1), RNAseq (n = 1) (13), microbiome study (n = 1) (6), and HIV antibody studies (n=4) (3, 9, 12, 15). Finally, of all the publications over the last year, the one that pushed the envelope on use of cutting-edge sequencing technologies was the study of plasmid diversity from a hospital associated outbreak of carbapenemase producing enterobacteriaceae (2). For this study we used the PacBio RSII single-molecule sequencer to generate complete and highly accurate assemblies of these bacterial genomes along with their associated plasmids. A key aspect of this project was the exhaustive validation of the PacBio assembled genomes conducted at NISC which involved expert manual review, genome-wide comparison of sequence data from an orthogonal sequencing technology, and optical mapping to confirm the long-range structure of the assemblies. This analysis estimated the assemblies generated by PacBio were at least 99.9999% accurate, thus providing key evidence that the PacBio system is a powerful method to generate complete and accurate genomes of drug-resistant bacteria. In the foreseeable future, NISC is well positioned to provide next-gen sequence data for several large, multi-year projects, for example, the Skin Microbiome Project, and Mouse Methylome Project, a collaboration with NIEHS, as well as expanding the access to sequencing by Intramural NHGRI investigators through continued sequencing support of their most promising projects. Our focus is to increase operational efficiencies of the next-gen pipeline, refine existing protocols, implement additional protocols as new sample/experimental types are requested from researchers and continue to expand the value added data analysis packages available. The PacBio sequencing instrument is now in production for a variety of library types, and we are testing specific applications for the BioNano Genomics/Irys physical restriction mapping platform. Furthermore, we will continue to monitor developments in the rapidly evolving sequencing and informatics technologies, implementing those we deem most appropriate for the sequence data we produce for collaborating investigators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This work proposes a fully integrated approach to the study of mechano-electrical transduction in the sense of touch and focuses on the development of a novel microsystems-based tool[unreadable]namely, a force clamp that uses appropriately-scaled piezeoresistive cantilevers as direct force sensors (PR force clamp). The proposal includes work to fully characterize the capabilities of a first-generation device, to design and build a second- generation device capable of operating at higher bandwidth, and to use these devices to analyze and model the biomechanics offeree transfer and mechano-electrical transduction by a model mechahoreceptor cell, the touch receptor neurons that innervate the body wall of the nematode Caenorhabditis elegans. C. elegans is an excellent biological platform for developing the proposed tools, which are likely to find application in the study of mechano-electrical transduction in other mechanosensory cells including vertebrate hair cells and dorsal root ganglion. A set of only six touch receptor neurons is responsible for behavioral responses to touch in C. elegans (compared to tens of thousands in vertebrates) and such responses are initiated by activation of the MEC-4 channel complex by forces applied to the body wall. Prototype force clamp systems use PR force clamps and piezoelectric actuators with programmable controllers to apply calibrated nano- to micro-Newton point load profiles with > 1kHz bandwidth. A critical and innovative aspect of this work is the integration of the PR force clamp with patch-clamp electrophysiology for synchronous force-displacement- physiological recordings. This technology will enable the first direct measurement of dynamic changes in tissue stiffness that may occur during mechano-electrical transduction and give rise to adaptation. Relevance to public health. The sense of touch can be degraded by both inherited and acquired disease, including AIDS and diabetes, as well as by chemotherapeutic drugs. Complications from such peripheral sensory neuropathies are estimated to cost more than $4 billion annual in health care costs. Despite this, the sense of touch and its degradation in disease remain poorly understood. The research tools developed in the course of this work have the potential to significantly advance our understanding.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Description: The Clinical Protocol Review and Monitoring Committee (CPRMC) consists of 22 members, appointed for three year terms, representing the major research and clinical disciplines of the VCC. The membership includes both M.D.s and non-M.D.s (e.g., investigators in the areas of biostatistics, preventive medicine, chemotherapy pharmacy, basic science, and nursing). Meetings are held twice a month as dictated by the number of submissions for review and a quorum must be present. The CPRMC reviews all protocols executed under the auspices of the Cancer Center and which use VCC resources.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of the proposed studies are to investigate the relationship between immune responsiveness and leukemogenesis in congenic strains of mice. One portion of study will be directed at evaluating the immune responsiveness of HRS/J hr/hr, hr/plus, and plus mice. These lines, which are congenic at the hairless locus, have significantly different leukemia incidences which are paralleled by altered general immune responsiveness. This study will evaluate their immune responsiveness to tumor- and murine leukemia virus-antigens and relate these specific responses to rates of leukemogenesis. Also, three strains in which spontaneous mutations is hairlessness were recently found will be evaluated to determine the effect of the presence of the hairless gene on different genetic backgrounds. Experiments are described which will evaluate the leukemia incidence and general immune responsiveness of the new mutants and their respective \"wild\" types. Similar studies are planned uing the congenic resistant strains AKR.L-H-2b/l and L.AKR-H-2k. The AKR.L-H-2b/1 line has an AKR/J background and is congenic for the H-2b genotype derived from strain C57L/J. These mice have been shown in our laboratory to have delayed and reduced leukemia incidence with respect to AKR/J and preliminary evidence incicates that they also have an altered immune responsiveness. This study is designed to confirm and expand upon these findings. Also, the L.AKR-H-2k line, which has C57L/J background and is congenic for the H-2k genotype derived from AKR/J will be evaluated to determine its leukemia incidence and overall immune responsiveness. Thus, I plan to examine the relationship between immune responsiveness and leukemogenesis using several congenic mouse strains which are uniquely suited to this type of study. I hope information gained opens the door to future studies in which the immune system can be manipulated in a manner beneficial to the tumor-bearig host.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this project, and related progress in the past year, are reviewed below. (1) Development of a reverse genetics system that can be used to modify the antigenicity and virulence of rotaviruses (RVs) and to develop new vaccine candidates. The RV genome consists of 11 double-stranded RNA segments. The positive-sense RNA of each segment typically contains short 5' and 3' untranslated regions (UTRs) that flank a single open reading frame (ORF). Occasionally, individual RV genome segments undergo rearrangement due to partial head-to-tail duplication of a segment sequence. In most cases, rearrangements initiate after the stop codon, leaving the ORF and the encoded protein unaffected. During virus replication, naturally occurring segments with sequence rearrangement are preferentially packaged over wild-type segments. The mechanism for the improved-replication phenotype is unclear, but may be connected to duplication of RNA sequences involved in packaging of the RV genome. As an extension of these observations, we used reverse genetics to engineer duplications into the segment encoding NSP2. The duplications were 25, 50, 100 and 200 bases in length and were inserted into the 3' UTR of the NSP2 segment such that ORF was unaffected and retained the capacity to encode NSP2. Recombinant RVs containing the duplicated sequences grew to high titer and were genetically stable. They were, however, unable to compete with wild-type virus in mixed infections, suggesting that the location and /or length of the engineered sequence duplication did not provide for preferential packaging of the segment. These results indicate that sequence duplication alone (or increased segment length) is not the only factor responsible for enhancing the packaging activity of an RNA segment. Indeed, our work raises the possibility that many rearrangements that arise in vitro and in vivo go undetected and are eliminated from the virus population. (2) Elucidation of mechanisms that RV proteins use to antagonize the interferon signaling pathway. The RV nonstructural protein NSP1 inhibits the expression of type I interferon (IFN), thereby promoting viral spread. NSP1 is predicted to function as an E3 ubiquitin ligase by inducing polyubiquitination and subsequent proteasome-mediated degradation of the IFN-regulatory factors IRF3, IRF5, and IRF7. All IRF proteins share an N-terminal DNA-binding domain, while IRF3, IRF5, and IRF7 contain a C-proximal IRF-association domain (IAD), which mediates dimerization. IRFs are maintained in an inactive state in the cytoplasm through an autoinhibitory domain that interacts with the IAD, burying key residues required for IRF dimerization and preventing IRF nuclear accumulation. C-terminal phosphorylation induces charge repulsions, causing the autoinhibitory domain to unmask the IAD, allowing formation of transcriptionally active IRF dimers. To identify the region of IRF proteins targeted for degradation by NSP1, we generated a series of IRF3 and IRF7 truncation mutants and co-expressed each with NSP1. We found that the IAD was necessary and sufficient for degradation by NSP1. Disruption of conserved residues required for dimerization of IRF3 did not interfere with NSP1-mediated degradation of the protein. Similarly, constitutively-expressed dimeric forms of IRF3 were targeted for degradation by NSP1. These results indicate that NSP1 recognizes the dimerization domain of both the monomeric and dimeric form of IRF proteins for degradation. The ability to target IRF proteins in both states suggests multiple steps of the IFN signaling pathway are targeted by NSP1, allowing for more efficient inhibition of IFN expression. (3) Analysis of the diversity and evolution of the RV genome: (a) Characterization of asymptomatic and symptomatic G10P11 RV Infections in Vellore, India, suggest a possible role of Aichi virus in diarrheal disease. Human G10P11 RVs causing symptomatic and asymptomatic infections have been described in neonates living in Vellore. Towards the goal of identifying possible genetic determinants responsible for the different phenotypes, the genomes of G10P11 viruses in stool samples from neonates/infants with asymptomatic infections (n=20) and with symptomatic infections (n=19) were recovered and analyzed using a combination of Sanger, 454, and Illumina sequencing methods. The results of RV sequence alignments and phylogenetic analysis showed no differences in the nucleotide or amino acid sequences of G10P11 RVs associated with asymptomatic and symptomatic infections. Surprisingly, deep sequencing revealed the presence of Aichi viral RNA in 2 of the 20 asymptomatic infections and 6 of the 19 symptomatic infections. These results indicate that genetic determinants are not responsible for differences in symptomatic and asymptomatic G10P11 RV infections. Our results raise the possibility that other infectious agents, such as Aichi virus, may be responsible for some diarrheal disease previously associated with symptomatic strains of G10P11 RV. (b) Increased incidence of G2P4 RV infections as Vanderbilt University Medical Center (VUMC) during 2010-2011 reveal distinct genotype 2 alleles. As part of the CDCs New Vaccine Surveillance Network, stool specimens were collected from infants and young children presenting with acute gastroenteritis at VUMC during the season preceding the introduction of rotavirus vaccines in the US and prospectively since. Most laboratory-confirmed rotavirus infections at VUMC were caused by G1P8 rotaviruses during the 2005-06, 2006-07, and 2007-08 seasons and G3P8 strains during the 2008-09 season. Relative to the preceding years, an increase in rotavirus infections were noted for the 2010-11 season, for which G2P4 viruses were largely responsible (12 G2P4, 13 G3P8, 1 G9P8, 1 G12P8). To better understand the diversity and relationships of the G2P4 viruses, their genomes were sequenced. All the G2P4 viruses contained complete genotype 2 (DS1-like) constellations (I2-R2-C2-M2-A2-N2-T2-E2-H2), indicating a lack of reassortment with more commonly circulating genotype 1 (Wa-like) viruses (I1-R1-C1-M1-A1-N1-T1-E1-H1). Phylogenetic analysis showed the genes of the G2P4 viruses could be resolved into at least two subgenotype alleles and most of the viruses could be resolved into either of two clades based on the conserved nature of their allele constellations. Our findings suggest, like G1P8 and G3P8 viruses, a limited amount of genetic exchange occurs between the G2P4 virus clades co-circulating in the same geographical location during the same year. (c) Vaccine-derived NSP2 segment in RV from vaccinated children with gastroenteritis in Nicaragua. The efficacy of RV vaccines can be low in developing countries. For example, a vaccine efficacy against severe diarrhea of only 58% was noted in a 2007-2009 Nicaraguan study with RotaTeq. To understand the basis of vaccine failure, the genomes of RVs isolated from vaccinated Nicaraguan children with gastroenteritis were sequenced. The results revealed that all the viruses had typical genotypes (11 G1P8, 1 G3P8) and that nine of the G1P8 viruses and the single G3P8 virus had genome constellations common to human RVs. However, two of the G1P8 viruses had atypical constellations, G1P8I1R1C1M1A1N2T1E1H1, due to the presence of a genotype-2 NSP2 (N2) gene. The sequence of the N2 NSP2 gene was identical to the RotaTeq N2 NSP2 gene, indicating that the two atypical viruses originated by reassortment of human G1P8 RVs with RotaTeq viruses. These data suggest that the high level of vaccine failure in Nicaragua is neither due to antigenic drift nor to the emergence of new antigenically-distinct virus strains. Our data also suggest that the widespread use of RotaTeq has led to the introduction of vaccine genes into circulating human RVs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT ABSTRACT Substance abuse is associated with a more severe course of HIV disease as shown by the poor immune reconstitution in cART treated subjects and an increased rate of co-morbidities. The mechanisms that are involved in these poor clinical outcomes are not well understood most likely a consequence of poorly- controlled confounding factors in study cohorts, lack of quantitative assessment of drug levels,, incomplete measurement of immune function and poor understanding of the mechanisms that lead to HIV persistence. We have used systems biology a n d shown that t h e balance between pro and anti- inflammatory pathways a s major drivers of HIV persistence. Disruption of this balance leads to poor immune reconstitution in HIV infected cART treated subjects and to increased frequencies of latently infected cells. Cocaine use as shown by our preliminary results accounts for further perturbations of the balance between pro and anti inflammatory effector molecules and cells which results in lower levels of immune reconstitution in these subjects and perturbations in T cell homeostasis which have led to heightened HIV persistence in other cohorts. We have assembled a multidisciplinary group that will allow us to overcome these aforementioned hurdles and will use unbiased OMICs approaches to address our major objective which is to delineate the molecular mechanisms triggered by cocaine that lead to HIV persistence. In specific aim 1 we will use three independent state of the art assays to quantity the size and cellular localization of the HIV reservoir; we will use transcriptomics on specific cell subsets and other high density readouts including flow cytometry and system serology to identify the effector cells and molecules that are associated to increased HIV reservoir size. In specific aim 2 we will apply a global and targeted proteomics approach to validate molecular pathways triggered by cocaine that lead increased HIV persistence. Metabolomics will be used to identify the role of metabolites and particularly short chain fatty acids and cholesterol on HIV persistence. In Specific Aim 3 we will validate the mechanisms identified by gene expression and proteomics which we have shown in aims 1 and 2 to be associated to the impact of cocaine on HIV persistence; we will use a novel primary cell assay of HIV latency which allows for the first time to measure latency reactivation in multiple T cells subsets. LARA will allow us to confirm in vitro that cocaine and its metabolites can mobilize the above identified pathways and monitor their impact on the establishment of HIV latency in different memory T cell subsets and on the response to Latency Reactivating Agents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "SPID#: 23 Despite almost 13 years of research on HIV and its role as the etiologic agent of AIDS, the mechanisms of pathogenesis remain mostly unknown. Specific in vitro effects have been identified that may contribute to the overall course of disease development. These include direct cell killing by HIV (syncytia formation); the induction of apoptosis; and the development of autoimmune responses; among others. While all of these effects have been observed in vitro and in vivo, the only mechanism consistently identified in vivo has been apoptosis. Because the exact time of infection can only be estimated in HIV-infected patients, a majority of the studies involving pathogenesis have been conducted during middle or late stage disease, at the least, following seroconversion. While much research has shown that a number of changes, including viral tropism changes, occur late, the early stages of infection may be as (or more) important in pathogenesis. The initial stages of infection involve viral amplification and seeding of lymphoid tissues, and can possibly dictate later effects, i.e. rapid or slow progression to disease. The SIV/macaque model represents the best system currently available to study AIDS pathogenesis, due to the striking genetic and biologic similarities. This model provides an excellent mechanism to investigate early pathogenesis of lentiviral infection. A recently characterized SIV isolate from a sooty mangabey (SIVsmmFGb) displays varied pathogenesis in pig-tailed and rhesus macaques. In pig-tailed macaques, this virus rapidly induces AIDS, whereas in rhesus macaques, disease progresses at a typically slower rate. The progression in pig-tailed macaques is associated with a high plasma viral load (as measured by p27 assay) an insufficient immune response (no anti-SIV antibody detected in most cases). In contrast, the rhesus macaques show a lower viral load, and make high levels of anti-SIV antibody. The pig-tailed macaques all succumbed to AIDS by 9 months post infection. In all cases, SIV- encephalitis was detected, illustrating the neurotropism of this virus. The differences in pathogenesis of this virus between rhesus and pig- tailed macaques should provide an excellent system for investigating determinants of AIDS pathogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Environmental hazards and stress, carcinogens, and anti-cancer therapeutics cause apoptotic cell death through the intrinsic or mitochondrial death pathway. Abnormal apoptotic response often contributes to tumor development and defects in apoptosis are intimately associated with tumor cell resistance to anti- neoplastic agents. Caspase activation lies in the core of apoptotic cell death. In the mitochondrial death pathway, cytochrome c (CC)-initiated Apaf-1 apoptosome formation represents a key initiating event in caspase-9 (an initiator caspase) activation, which ultimately activates effector caspases such as caspase-3 to execute cell demise. How Apaf-1 apoptosome is formed and regulated in vivo (i.e., in the stimulated cells) remains poorly understood. Recently, we provided evidence that physiological levels of nucleotides inhibit the CC-induced, apoptosome-mediated caspase-9 activation by binding directly to CC and preventing CC from interacting with Apaf-1 (Chandra et al., Cell 125, 1333-1346, 2006; Appendix I). Consequently, the CC- mediated apoptosome assembly and activation are blocked. Co-microinjection of nucleotides and CC renders cells resistant to the CC-induced apoptosis in vivo whereas experimentally reducing nucleotides enhances both CC and apoptotic stimuli-induced cell death. These observations lead us to hypothesize that physiological levels of nucleotides, in addition to their well-established roles in nucleic acid synthesis, intermediate metabolism, and maintenance of bioenergetics, also function as critical prosurvival factors by directly inhibiting the CC-mediated apoptosome formation and caspase activation. There are several critical unanswered questions related to Apaf-1 apoptosome regulation by nucleotides. We propose the following three Specific Aims to address some of these questions: 1) To further study nucleotide interaction with CC in vitro and to elucidate nucleotide interaction with CC in vivo; 2) To investigate nucleotide interaction with Apaf-1 and its impact on apoptosome activation; and 3) To study the effects of Apaf-1 isoform expression and nucleotide interaction on Apaf-1 apoptosome activation. These goals will be achieved by a combination of cell biological, biochemical, and molecular approaches. The accomplishment of the proposed goals will greatly advance our understanding of apoptosome regulation in vivo and help us understand cellular response to environmental stress and cancer cell response to anti- tumor therapeutics. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Secretory granules are the subcellular organelles in which the salivary parenchymal cells store their secretory products until such time that they are released into the oral cavity. Secretory granule maturation is believed to be an important step in the formation of these granules and is thought to be comprised of the condensation of macromolecular and other secretory products into these discrete package. Little is known of this step in the secretory cycle of salivary glands, and yet it may be an essential and highly significant aspect of normal salivary gland function, and hence, oral health. The overall objective of this study is to determine the nature of the changes that secretory granules undergo during maturation, and to elucidate the cellular mechanisms by which this maturation process occurs. This will be accomplished in part, but not totally, by utilizing a model system consisting of rat parotid secretory granules of differing buoyant densities, the less dense of which appears to be a precursor of the precursor of the denser population of granules based on pulse labelling studies. The specific aims are to characterize these two types of secretory granules by direct measurement of their volumes and concentrations of macromolecular and inorganic components. Qualitative comparisons of the proteins, glycoproteins, and sulfated macromolecules such as glycosaminoglycans, in addition to quantitative determinations, will be carried out. Two possible mechanisms for secretory granule maturation via the lowering of the colligative properties of the contents will be tested for. These are: 1) the complexing of macromolecules with or without the intermediation of ions, and 2) the pumping of ions out of the granules so that water will follow, or into the granules in order to promote intermolecular complexing. In situ granule studies will also be carried out in order to verify the findings using isolated secretory granules.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal seeks funding to create and distribute a nationally representative 1 in 100 Public Use Microdata Sample of the 1920 United States Population Census. Public Use Samples already exist or are in preparation for the census years 1850, 1880, 1900, 1910, 1940, 1950, 1960, 1970, 1980 and 1990. Although the historical samples for the period 1880- 1950 have only been available for a few years, they have already led to an outpouring of new research on the nature of long-term social change. As each new sample is created, the value of the previous census files has been enhanced, because they become increasingly useful for cohort analysis and studies of social change. Microdata files allow researchers to make tabulations tailored to their specific research Questions and to avoid many of the incompatibilities in the published data for different census years. In addition, the public use samples have allowed researchers to move beyond simple tabular analysis and apply increasingly sophisticated multivariate techniques. These data haven dramatically increased the power of quantitative social science research. A new public use sample for the 1920 census will bridge the existing gap between the 1910 and the 1940 public use samples. When the 1920 sample is complete, we will have a continuous series of microdata for every census year in the twentieth century, with the sole exception of the 1930 census, which is still protected by the Census confidentiality rules. The case for a sample from 1920 is especially compelling because the enumerator's manuscripts include a great deal of information on demography and social structure that can only be taken advantage of through the creation of a new microdata set. The period from 1910 to 1940 is critical for the study of topics such as fertility decline, urbanization, immigration, household composition, and occupational structure. Besides converting a sample of the 1920 population into machine-readable form, the project will evaluate the sample quality through consistency checks, random blind verification, and comparison with aggregate statistics in the published census volumes; edit and allocate missing, illegible, and inconsistent data through logical rules and imputation procedures; construct new variables on household composition and relationships within families; create alternative coding schemes to ensure that the 1920 sample is compatible with all other public use samples currently available; and prepare documentation for the user file, including detailed descriptions of the sampling and data processing procedures, and a guide to the use of the sample.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the Center for Reproductive Science and Medicine is to develop understanding of the mechanisms that govern normal and disordered function of the hypothalamic-pituitary-ovarian axis. This application represents our renewal for year 25 and our productivity has been outstanding over the most recent granting period with 86 papers published. We will continue to produce novel and important contributions to the reproductive sciences, integrating multidisciplinary clinical and basic research to facilitate and accelerate the translation of promising new discoveries into clinical medicine. We have chosen three innovative and complementary Research Projects, all with experienced, internationally renowned leaders. We will address the hormonal control of the gonadotropin beta-subunit genes in the pituitary, focusing on regulation by activin, GnRH, and steroid hormones. The emphasis will be on understanding the synergy and interdependence between these hormones in controlling transcription in model immortalized gonadotrope cells and genetically modified mice. We will chart new territory in the role of metabolic homeostasis in infertility. A broad based, in vivo and in vitro approach is proposed to elucidate the mechanisms of insulin, adiponectin and PPAR gamma action in impaired reproductive function, using novel animal models and immortalized hypothalamic GnRH-secreting immortalized neurons and gonadotrope cells. We will focus on the influence of gonadal steroids and insulin in the endocrine control of the ovarian granulosa cells in vivo in normal women and patients with polycystic ovary syndrome [PCOS], addressing fundamental mechanisms underlying PCOS. All three projects have teams of highly experienced investigators and all three Project Leaders serve as Co-investigators on at least one other component of the Center. These three projects are highly interactive and synergistic and create a coherent mechanistic and translational program for the Center. The only core will be the Administrative Core. The Administrative Core facilitates and supports all of the activities of the Center and provides the Enrichment Program. It also houses the Human Ovary Tissue Bank, which provides well-characterized human ovary tissue to all NIH-funded investigators in North America.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The association of recipient blood transfusion and improved allograft survival is almost universally accepted. However, the mechanism of this phenomenon is unclear. If immunological enhancement is the mechanism, the potential recipient should receive blood from several donors which would expose him to a wide range of foreign antigens, some of which might be shared with the organ donor. However, the price of intentional transfusion is sensitization of the recipient against HLA specificities causing a delay in transplantation in some cases and, in others, precluding it altogether. The specific aim of this proposal is to determine whether or not sharing of a major or minor histoincompatibility between blood and organ donor is necessary to achieve the effect. Using the primarily vascularized heart transplant model in inbred congenic lines of mice, donor-recipient combinations will be selected in which blood donors and heart donors will share no antigens which are foreign to the recipient, another combination will present only foreign H-2 differences to the recipient, another multiple H-2 and non-H-2 antigens, and another a single K-region difference only. Preliminary work in this laboratory has shown that \"non-specific\" blood administered six weeks prior to transplantation delays rejection significantly, whereas donor blood (K-region difference) does not delay rejection but is associated with a recovery of heart graft rejection. Rejection of these beating heart grafts can be precisely determined at the time of a marked decline of palpable impulse (hearts are palpated daily through the thin abdominal walls of the mice). Timing of administration of whole blood, red cells, platelets, lymphocytes, and serum will be tested and survival curves will be constructed and compared to controls in all groups of experiments. The health-relatedness of this proposal is timely in that clinical transfusion protocols are being developed on a widespread basis for dialysis patients awaiting transplantation. A clearer understanding of the mechanism of recipient blood transfusion should lead to a more logical approach to the most appropriate clinical transfusion protocol, principally an understanding of whether donor blood or non-specific is more effective.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The underlying hypothesis of this study is that the complexity of cancer dictates that compared to single modalities, combination therapies that are mechanistically independent and which are directed at nonoverlapping molecular targets will additively or synergistically enhance treatment outcome. Toward achieving this goal, the current Program Project Grant (PPG) application builds on recent advances in the understanding of cancer biology, in mechanisms of current and emerging therapies as well as the enormous progress made in imaging technologies, to propose new photodynamic therapy (PDT)-based combination treatments for pancreato-biliary (PCBC) and non-melanoma skin cancers (NMSC) in an approach we term Combination Photodynamic Biologic Therapy (CPBT). PDT is a photochemistry-based modality that is approved for the treatment of a number of cancer and non-cancer pathologies. Strategically, these treatments response-enhancing CPBTs will be achieved in two ways: (i) by preconditioning tumors with agents which initiate molecular events that augment PDT, and (ii) by administering a second treatment specifically tailored to a particular molecular response elicited by the PDT. Optical imaging techniques will be invaluable for the online monitoring of light and photosensitizer dosimetry, treatment planning and molecular response. Project 1 will investigate molecular pathways affected by compounds that enhance ALA-PDT and can be exploited by a combination regimen to increase treatment response in skin carcinomas in models and in humans. Project 2 proposes to improve the survival and quality of life of patients with pancreatic or biliary tract carcinoma using PDT in clinical studies. Project 3 will build upon results from the previous funding cycle that PDT instigates molecular responses that can be targeted for much improved treatment outcomes. Preclinical models of pancreatic cancer (PanCa) will be used. Project 4 takes advantage of the advances in optical imaging to develop quantitative tools to monitor molecular features of tumors and their response to treatment, and to integrate them for the development of molecular-based combination therapies for standard clinical procedures. The proposed program will feature three cores that will provide support in three areas: Core A, through administration, scientific integration, education and career development, intellectual property development, and administration; Core B, through biological models, molecular pathology, and biostatistics; and Core C, through in vivo imaging, dosimetry, and the transfer of developed technologies to industry. It is anticipated that such rational, mechanism-based treatments will significantly benefit patient survival, quality of life and/or disease palliation and, combined with real-time imaging to monitor tumor progression and treatment response, provide patient-specific treatments that can be realistically envisioned. Potential benefits to public health: this program impacts on the treatment of three cancers: two of these PanCa and BTC which have few treatment options and are usually fatal. NMSC on the other hand has many options but the high incidence puts a heavy burden on society in terms of cost and suffering.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronic inflammation is a common ground to cancer development - accounting for ~20% of the world-wide cancer load. Hepatocellular carcinoma, the third leading cause of cancer mortality, is a prototype for inflammation-associated cancer, as most of the cases develop on a background of chronic hepatitis. While eliminating the underlying cause (e.g., hepatitis) is many times hard to attain, recent elucidation of inflammation-activated molecular pathways suggests that therapies directed at disrupting these pathways could be useful for cancer prophylaxis in chronic inflammatory states. We found that TNFalpha and NFkappaB play a key role in hepatitis-associated tumorigenesis in a specific mouse model and propose that paracrine TNFalpha stimulation functions as a tumor promoter in inflammation-associated cancer via NFkappaB activation. Short-term anti-TNFalpha treatment induced premalignant hepatocyte apoptosis, and NFkappaB inhibition via a dominant NF-kappaB represser suppressed tumor progression in a mouse model. We will assess the value of anti-TNFalpha prophylaxis in HCC development in a mouse model and determine if it could stabilize the premalignant phase and delay tumor progression. During the course of the proposed study we aim to achieve the following specific goals: 1. Assess the effect of in vivo TNFalpha inhibition on tumor development in Mdr2 knockout mice - a model for hepatitis-associated liver cancer. 2. Study the source of TNFalpha in the Mdr2KO microenvironment in relation to hepatocyte NF-kappaB activation. 3. Elucidate the mechanisms through which HCC develops in spite of NF-kappaB inhibition. Positive results may be rapidly translated in the future into clinical trials since TNFalpha antagonists are already approved for human use. Understanding the mechanisms underlying the response to this treatment and the mechanisms of escape may help to plan effective treatment regimens for clinical studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "More than 100 million Americans are obese and 17 million are affected by type 2 diabetes. While diet and lack of physical exercise are the main risk factors for these diseases, several epidemiological and animal studies have shown that the intrauterine nutritional conditions can also influence body weight and glucose homeostasis in adulthood. We hypothesize that maternal nutrition influences the intrauterine environment and can induce genome-wide DNA methylation changes in multiple organs of the fetus. We hypothesize that this epigenetic programming is responsible for translating intrauterine stress into molecular responses that can durably affect the health of the offspring. We propose to test this hypothesis by studying a mouse model of diet-induced maternal obesity using a unique combination of high-throughput genomic tools: the characterization of the genome-wide DNA methylation patterns, using a novel sequencing developed in our lab, and extensive gene expression profiling in multiple tissues. We will study embryos and newborns from C57BL6 dams fed on high- fat or low-fat diet prior and during pregnancy to characterize the molecular basis of intrauterine programming. We will also analyze older animals to understand the lasting molecular and phenotypic consequences of maternal obesity. Our findings will provide a better understanding of the mechanisms responsible for the life-long metabolic consequences of maternal obesity, which currently affects 20-40% of pregnant women in the US.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-range goal is to understand how aryl hydrocarbon receptor (AHR) signaling pathway activation affects prostate development and prostate disease. The developing prostate is highly sensitive to 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD), the prototypical AHR agonist. In mice, TCDD inhibits prostatic bud formation by the fetal urogenital sinus (UGS), prevents the ventral prostate lobe from forming, and permanently alters the structure and function of the other prostate lobes. Preliminary results show that ?-catenin is essential for normal prostate development and implicate aberrant ?-catenin signaling as the key mechanism by which TCDD causes prostate abnormalities. Hypotheses to be tested in wild-type and transgenic C57BL/6J mice are as follows: Aim 1: TCDD inhibits prostatic bud formation by decreasing ?-catenin signaling in basal urogenital sinus epithelium (UGE). Aim 2: TCDD down-regulates ?-catenin signaling by decreasing expression of canonical Wnts and Rspos, and/or by increasing expression of non-canonical Wnts, in urogenital sinus mesenchyme (UGM). Aim 3a-c: TCDD epigenetically alters gene expression in the fetal UGE, and does so via ?-catenin down-regulation; Aim 3d: In utero and lactational TCDD exposure causes the senescent prostate to retain the epigenetic signature seen in TCDD-exposed fetal UGE and to inappropriately retain androgen responsiveness characteristic of young control adults. Aim 4: In utero and lactational TCDD exposure accelerates prostate adenocarcinoma progression in Nkx3.1; Pten mice and TCDD treatment in adulthood does likewise. Results from Aims 1 and 2 will significantly advance our understanding of the mechanisms by which AHR agonists affect early prostate development, and may help explain how TCDD and related chemicals cause abnormal development of other organs that also develop via mesenchyme-induced epithelial budding and branching (e.g., breast and lung). As for long-term effects, it has already been established that in utero and lactational TCDD exposure causes the dorsolateral prostate to retain abnormally high androgen dependence into senescence, and increase susceptibility to a type of prostate tumor (neuroendocrine) that is uncommon in men. Aims 3 and 4 will in part determine if low-level maternal TCDD treatment also increases (a) androgen responsiveness in the senescent dorsolateral prostate, and (b) susceptibility to prostate adenocarcinomas, the most common prostate tumor type in men. Aim 4 will also determine if Nkx3.1; Pten mice are an appropriate model for studying the increase in prostate cancer seen in Vietnam veterans exposed to the TCDD-contaminated herbicide Agent Orange. The proposed research has a high likelihood of establishing that ?-catenin signaling abnormalities are key contributors to TCDD toxicity in mammals, and of establishing that fetal TCDD exposure creates an epigenetic signature in the prostate, one associated in adulthood with altered responses to androgens and with cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Gene expression is regulated by auxiliary factors in both a positive and negative manner to increase or decrease expression of specific genes. This regulation often involves direct interaction of factors with the transcription complex to modulate different stages in the transcription process. This proposal addresses the mechanism of transcriptional antitermination of the ribosomal RNA (rRNA) operons of E. coli responsible for the prevention of transcriptional polarity of these genes. Several E. coli proteins (Nus factors) and specific sequences in the DNA have been identified that are involved in modification of transcription complexes initiated from the rRNA promoters. It has been proposed that the antitermination transcription complex is assembled via direct and specific interactions between one or more of these Nus factors, and the nascent RNA complementary to these antitermination sequences in the DNA (boxA and boxB sequences). Photochemical crosslinking will be used to test this model and identity the nature of specific protein-RNA interactions. A new series of nucleotide analogs that contain photoreactive crosslinking groups on the base in positions that do not interfere with normal basepairing will be incorporated in vitro into the nascent rRNA. Active transcription complexes will then be irradiated with long wavelength ultraviolet light to cause covalent attachment of the modified RNA to adjacent proteins, and specific interactions with the transcription factors and RNA polymerase will be examined. Protein-RNA interactions will be identified at the level of specific nucleotides in the RNA and domains and/or amino acids in the proteins. Specifically, the following questions will be addressed: 1. Do Nus factors interact directly with the RNA? 2. Do Nus factors alter the interaction between the polymerase itself and the RNA? 3. Does the termination resistent transcription complex retain the RNA encoded by the antitermination sequences? 4. Does the putative antitermination protein in E. coli also required for antitermination of the rRNA operons interact with the RNA, and can it be identified by crosslinking?", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Homologous recombination is a critically important process responsible for repairing double-strand DNA breaks and exchanging genetic material between homologous chromosomes in cells undergoing meiosis. Our goal is to understand the mechanisms of homologous recombination in humans. The mechanism of homologous recombination involves forming a cross-stranded structure between two homologous chromosomes called a Holliday junction. The Holliday junction possesses a remarkable property, in which this structure can freely translocate along the DNA axis in a process termed branch migration. The central role of branch migration in homologous recombination and DNA repair has only recently been fully appreciated. Branch migration drives 1) the resolution of homologous recombination intermediates, 2) the retrieval of information lost at sites of DNA damage, and 3) the restart of stalled DNA replication forks. In human cells, branch migration is catalyzed by RecQ helicases, including BLM and WRN. Mutations in the BLM and WRN genes cause a high degree of genome instability, which lead to premature aging disorders called Bloom's and Werner's Syndromes. Recent genetic analysis has demonstrated that Rad54 also plays an important role in homologous recombination of eukaryotes. Double RAD54 BLM knockouts show a dramatic increase in chromosomal instability over the level observed in either single knockout, indicating a functional redundancy between these two proteins. Indeed, our lab recently discovered that Rad54 can also promote branch migration of Holliday junctions. We will use purified human proteins in an array of biochemical assays specifically designed to investigate the branch migration activity of Rad54 and BLM. The goals of this proposal are to understand how Rad51 affects the function of Rad54 and BLM in homologous recombination and investigate the non-canonical helicase activity of Rad54 that functions during branch migration. These assays will be combined with a mutational analysis of Rad54 to determine which motifs are crucial to its biological activities. Our goal is to better understand how the branch migration activities of these two similar, but distinct enzymes contribute to the maintenance of genome integrity in human cells. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Two strains of mice were used in this project. Female C3H and CBA mice were exposed to a total body dose of radiation of 3 Gy with or without Tempol supplementation in the animal's food. Immediately following the radiation exposure, animals were be placed on either control or TP-containing food. The groups include: a) no radiation, control food, b) 3 Gy, control food, c) no radiation, TP food, and d) 3 Gy, TP food. For C3H mice, one additional group was added receiving 3 Gy total body irradiation where the administration of the TP containing food was delayed one month post-irradiation. Preliminary data show TP food supplementation after radiation: a) did not alter food consumption compared to animals on a control food diet, b) compared to animals on control food diet, the TP diet resulted in decreased weights in both mouse strains (40% for C3H and 20% for CBA), and c) TP food supplementation post-radiation significantly enhanced the survival of both mouse stains. Median survival values for 0 Gy, 3 Gy, 0 Gy TP, and 3 Gy TP for C3H mice was 706, 434, 764, and 670 days, respectively. For CBA mice median survival values for 0 Gy, 3 Gy, 0 Gy TP, and 3 Gy TP for C3H mice was 901, 660, 939, and 782 days, respectively. The incidence of hematopoietic neoplasms (predominantly lymphomas) was significantly reduced in both mouse strains by TP treatment and both the onset and incidence of solid neoplasms was significantly reduced in CBA mice treated with TP. These preliminary data would encourage further research and development of TP as a chemopreventive agent. The second hypothesis is also being tested that mice protected from lethal total body irradiation by administration of a radioprotector immediately before radiation exposure will experience an elevated risk of cancer induction. Mice were exposed to a total body radiation dose of 10.8 Gy, a radiation dose that results in 100% lethality. Ten minutes prior to the 10.8 Gy exposure the animals will be injected with a radioprotector. The control for this group, another set of animals was exposed to 5.4 Gy total body irradiation. This radiation dose was derived from the radiation dose modification factor (2) when the radioprotector is administered 10 min before total body irradiation. These animals will also be followed for their entire lifespan for tumor induction as outlined above. Preliminary data show that the median survival for mice receiving 0, 5.4 or 10.8 Gy were 706, 460, and 491 days, respectively. There was no difference between the 5.4 and 10.8 Gy groups (p = 0.42);however, the median survival of both irradiated groups was significantly shorter compared to unirradiated mice (p &lt;0.0001). Cancer incidence (hematopoietic plus solid tumors) was similar between the 5.4 and 10.8 Gy groups and was significantly greater than for unirradiated controls. However, the ratio of hematopoietic to solid tumors differed between the two groups, with the 5.4 Gy group having a higher incidence of hematopoietic neoplasms compared to the 10.8 Gy group (1.8 fold). A greater incidence of solid tumors was observed in the 10.8 Gy group. These preliminary results suggest that mice protected from lethal whole body radiation have a shortened lifespan, due in large part, to cancer induction post-radiation compared to unirradiated controls. Lastly studies have been initiated to determine if metabolites in the urine of mice receiving whole body radiation can predict for radiation-induced cancer induction prior to the observation of tumor mass.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Vascular complications in diabetes mellitus are the result of metabolic and hormonal abnormalities caused by insulin deficiency. The identification of specific factors and the mechanisms by which they can cause vascular cell dysfunction and subsequent pathologies is very important in the treatment of diabetic patients. Several mechanisms by which hyperglycemia can adversely affect vascular cell function have been postulated. We have been investigating the particular effect of hyperglycemia on increasing levels of diacylglycerol (DAG) and protein Kinase C (PKC) in tissue cultured vascular cells, in retinal vascular tissue from the diabetic rat, and the effect of PKC elevations on retinal blood flow. Retinal blood flow was measured using our video fluorescein angiography (VFA) system. We have demonstrated decreased retinal blood flow in the early stages of diabetes in both humans and in different diabetic rat models. More importantly, we have shown a direct relationship between decreased retinal blood flow in the rat and the activation of PKC. These results provide evidence linking biochemical alterations caused by hyperglycemia with functional changes in retinal circulation in early diabetes. The studies proposed here will focus on characterizing the relationship between DAG and PKC levels, assayed from blood samples, and retinal blood flow changes in Type I insulin dependent diabetic patients with different levels of diabetic retinopathy. Diabetic patients will be studied cross- sectionally from onset of diabetes through the development and progression of diabetic retinopathy. The results from these studies will aid in our understanding of the mode of action of PKC, at a cellular and physiological level, in affecting retinal blood flow changes and the development of retinal vascular complications in diabetic patients. The elucidation of the natural history of these changes in early diabetes will aid in possible future inhibition therapies for treating diabetic patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project uses electrophysiological and biochemical methods to study a new model of status epilepticus. This model uses pretreatment with lithium followed by administration of subthreshold doses of convulsants. The basic model entails treatment with LiCl (3 mEq/kg; ip) and pilocarpine (30 mg/kg; sc) which invariably produces generalized convulsive status epilepticus which lasts for several hours and is fatal. There are three major goals of this proposal. (1) We will test the hypothesis that lithium lowers the seizure threshold to other convulsive stimuli, including ECS, bicuculline, picrotoxin, pentylenetetrazole, kainic acid and chemical (carbachol) and electrical kindling. (2) We will test the hypothesis that function of the GABAergic system is impaired during status epilepticus by measuring the state of the GABA/benzodiazepine/picrotoxin chloride channel complex during seizures. (3) We will test the hypothesis that alteration of calcium homeostatis plays a role in status epilepticus by testing the anticonvulsant and proconvulsant actions of calcium antagonists and a calcium agonist, respectively, and the effects of seizures and drug treatments on calcium homeostasis in synaptosomes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Because of their elevated frequency, ability to self-renew and rapid acquisition of effector function following re-activation, memory T cells have an enhanced ability to protect from secondary challenge. The generation of memory T cells is the focal point of numerous vaccine and immunotherapeutic strategies. Most (90-95%) effector T cells die after pathogen clearance, but those fated to become memory cells can be identified during the primary effector response, showing that activated T cells receive differentiation cues during the primary response to infection that influence memory fate differentiation. We have recently shown that T cell receptor (TCR) signals play a key role in driving CD4+ memory T cell differentiation. TCRs that are able to engage MHC Class II-bound antigen (pMHCII) in sustained interactions are biased towards the formation of long-lived memory, while TCRs that engage in short-lived interactions with antigen are biased towards terminal effector cell differentiation. We will build on those studies by using diverse infectious models to define aspects of the TCR-dependent activation and transcriptional program that leads to the formation of lymphoid-resident, circulating and tissue-resident CD4+ memory T cells. We pose three key questions. First, what is the role of TCR signal strength in the formation of memory T cells? We will explore the hypothesis that increasing TCR signal strength in vivo drives terminal effector T cell differentiation, while weaker TCR signal strength allows memory T cell formation. Second, what are the TCR binding parameters associated with memory T cell development? We will measure 2D affinity and bond lifetime with the application of force for TCRs at that are effector-biased or memory-biased. We will test the hypothesis that bond lifetimes will predict TCR-dependent memory differentiation. Third, what are the transcriptional programs that control memory formation? We will test the mechanistic role of molecules that are differentially expressed in memory T cell precursors during the primary effector response, including TCF-1. We anticipate that resolution of the questions posed in this study will provide a framework for determining in greater mechanistic detail how memory T cells form and identify therapeutic approaches for directly modulating CD4+ effector and memory T cell differentiation in vivo.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "CD4, the T-cell receptor for the human immunodeficiency virus, depends upon glycosylation for proper surface expression. Initial studies employing acute lymphoblastic leukemic cells were extended by the successful transfection of a plasmid containing the CDNA for CD4 into mutant and wild type Chinese hamster ovary cells. Characterization of these clones suggests that CD4 contains biantennary unsialylated complex-type oligosaccharides. The data also suggest that the inability of CD4 to reach the cell surface when it lacks carbohydrate is due to the selective degradation of unglycosylated CD4. The REV protein of HIV has been recently shown to lead to the premature transport of the partially spliced MRNA encoding the envelope glycoprotein from the nucleus to the cytoplasm. To develop a.system to examine the effects of REV, a means has been devised for generating nuclei in vitro around exogously added DNA. This method involves using extracts from Xenopus laevis where it has been demonstrated that nuclear assembly can be reconstituted. The availability of such preparations should allow examination of the mechanism of REV function. In other studies, examination has begun of the structure of the nuclear pore complex, across which REV, TAT, and other viral proteins cross following viral infection. The structure of the pore complex is explored using recombinant DNA techniques. The major nuclear pore glycoprotein np62 has been cloned, sequenced and expressed in cultured cells. The gene encoding this protein has been isolated and found to be 2.95 kb in length and devoid of introns. The availability of the primary sequence of this protein has allowed the preparation of anti-peptide antibodies which react with p62. These antibodies should prove useful for probing the function of the pore complex in the HIV life cycle.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although high density lipoprotein cholesterol is a strong risk factor for accelerated atherosclerosis, little is known about its function and metabolism. One function of HDL may be to deliver cholesterol to the steroid hormone producing tissues, adrenal, ovary and testes. Our previous studies indicate that the adrenal cortex accumulates cholesterol from lipoprotein particles in the high density range by a receptor-mediated process not associated with lipoprotein endocytosis and lysosomal degradation. These observations raise two major questions which our proposal is designed to answer: (1) What properties of the lipoprotein are responsible for \"HDL receptor\" recognition and cholesterol uptake? (2) What is the mechanism by which adrenocortical cells take up HDL cholesterol? To answer the first question we will examine the ability of well characterized homologous and heterologous lipoprotein particles from several density classes and lipoprotein families as well as HDL subfractions to compete with human 125I-HDL3 for receptor binding and to deliver cholesterol to rat and swine adrenal cells. Further studies using isolated apo A-I and apo A-I lipid recombinants will be performed to better define the role of this apoprotein. We view adrenal accumlation of HDL cholesterol as the result of three sequential events: i) lipoprotein binding, ii) transmembrane movement of cholesterol, iii) intracellular cholesterol movement to sites of storage or utilization. Properites of each of these events will be further examined. The effects of pH, ionic strength, lipoprotein charge and ACTH stimulation on binding will be characterized. The effects of energy inhibition on cholesterol uptake will be determined. We will determine if both esterified and free cholesterol as well as phospholipids may enter the cell. Finally, further studies to better define control of the uptake process will be performed. these studies will provide important information in two areas: (1) HDL structure, function and metabolism and (2) cholesterol metabolism in steroidogenic tissues. Better understanding of HDL metabolism may aid our understanding of the atherosclerotic process and how to prevent or reverse it.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this proposal is to demonstrate the capabilities of the University of Pittsburgh to participate as a clinical site in the NICHD-sponsored Pelvic Floor Disorders Network (PFDN). Our site has a longstanding track record of successful contribution to multicenter studies of urinary and fecal incontinence, and pelvic organ prolapse. We are particularly well suited to be a clinical site in the PFDN because of our volume, research infrastructure and track record, basic and translational experience and expertise. Access to large numbers of nulliparous women enables us to contribute uniquely to studies of the role of pregnancy and parturition in the etiology and prevention of pelvic floor disorders (PFDs). Magee-Womens Hospital (MWH) is the central resource for gynecologic specialty care for the 19 hospital University of Pittsburgh Health System serving a very large aging population. Our site brings expertise in urogynecology, physical therapy, geriatrics, urology, gastroenterology and mental health. We offer unique technical expertise in genomics, proteomics, tissue regenerative techniques, biochemical and biomechanical impact of meshes on the vagina and central neuronal control of bladder function. We propose to establish a comprehensive, scientifically rigorous clinical and translational research program within the PFDN for prospective comparative studies of mesh materials used in prolapse and incontinence procedures. The program will generate data of immediate clinical relevance as it will present scientifically sound, vendor independent evidence to guide surgeons'selection of specific graft materials and evidence-based practice guidelines for management of mesh complications. The 3 major components of the proposal are: 1) mesh specific infrastructure for implementation in PFDN clinical trials employing mesh inclusive of the development of a Mesh Morbidity Index and establishment of a Biospecimen Repository 2) the first RCT of meshes selected through rigorous analyses of biomechanical and biochemical properties and 3) translational studies on the cellular response to mesh materials and pathophysiology of mesh complications. The RCT will serve to pilot the database, compare clinical outcomes of meshes whilst providing specimens for translational studies. PUBLIC HEALTH RELEVANCE: Though prosthetic mesh enhances durability of prolapse and incontinence repairs, escalating complications have prompted public health warnings. Understanding the pathophysiology of mesh related morbidity and identifying modifiable risk factors through comparative and translational studies will inform on complications, support guidelines on mesh selection and route of delivery, and direct innovations in mesh technologies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Uganda faces problems of rapid population growth and low contraceptive prevalence, as well as reproductive health problems including sexually transmitted diseases, genital tract infections, and a generalized HIV epidemic. The Institute of Public Health (IPH) and Department of Population Studies (DPS) at Makerere University have collaborated with the Department of Population and Family Health Sciences and the NIH- funded Population Center at Johns Hopkins and Columbia Universities, collaborated on the Rakai Project, and conducted population-based epidemiologic, demographic, behavioral and biomedical studies in rural Rakai District, southwestern Uganda. The Rakai Project currently has several NIH grants (RO1s and UO1s), and has benefitted from Fogarty Foundation funding. This proposal builds on our prior Fogarty support to continue builds Population and Health research and training at Makerere University. Three previous Fogarty Masters graduates will complete doctoral training, and 3 new candidates will complete masters training at JHU. These graduates will develop Advanced Research Training proposals on their return to Uganda. We also propose 7 short-term trainees (5 in population/reproductive health, and 2 in laboratory methods). On their return, trainees will rotate through the Rakai Project field site and other field projects, to obtain practical experience. There will be 8 postdoctoral Fellows (3 research Fellows, 3 teaching Fellows to work on training and curriculum development for Makerere, and 2 clinical Fellows in reproductive health). We propose 5 in-country workshops to develop research training and curriculum development for Makerere, and a Conference in Year 5 to disseminate research findings. The program will be coordinated with other JHU Fogarty programs, the JHU Gates Institute (which will provide cost-sharing), and the JHU Mellon Distance Learning Program to facilitate internet access and transfer of training materials. The US and Uganda-based training, research and curriculum development will enhance development of and coordination between DPS and IPH, to provide an integrated long-term, self-replicating program of Population and Reproductive Health in Uganda. The goal is to establish a critical mass of researchers and trainers in Uganda who will sustain training programs submit high quality NIH research proposals, and conduct independent research. Ultimately, we see Uganda and Makerere University providing a resource for regional research training in Africa.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overarching goal of this proposal is to provide the candidate with the means, mentorship and structure to transition to an independent investigator focused on examining and improving communication between female patients and their health care providers. She is Assistant Professor in Ob-Gyn with fellowship training in health services research. Many conditions treated by obstetricians-gynecologists deal with sensitive social and behavioral issues that necessitate patient disclosure and the provision of information and advice. The candidate's interest in patient-provider communication represents a shift in research focus and a natural extension of her previous work on health care screening and interventions for intimate partner violence (IPV). Using IPV as an example of a challenging topic faced by women's health care providers, she intends to understand what components of patient-provider communication in IPV are associated with increased patient disclosure, comfort and motivation to seek help. She intends to design a communication intervention to improve provider IPV discussions and health and safety outcomes for affected women. Aims of the research project are: 1) to describe the content and style of patient-provider communication regarding intimate partner violence during the initial obstetric visit; 2) to describe association between communication and IPV disclosure; 3) to describe the patients' perspectives on the discussion including how it affected their intentions to attempt to change their situations to increase their safety; and 4) to develop a provider communication intervention on how to ask and counsel about IPV. The candidate's career development plan incorporates coursework, tutorials with methodological and topic experts, conferences, and mentored research to meet her educational objectives: 1) to acquire knowledge and skills to assess patient-provider communication; 2) to learn how to develop and implement interventions to improve patient-provider communication; and 3) to enhance understanding in measuring outcomes of communication intervention. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Organisms capable of dramatic regeneration maintain adult cells that have the flexibility, or plasticity, to change their fate. This feature enables thm build replacement parts, like organs or limbs, to recover from damage or disease. Previous work on the project and other studies have revealed a broad propensity for specialized cells to display some properties of stem cells; they give rise to new cell types in response to injury. The dynamic changes that cells undergo during the transition from one fate to another could reveal the mechanisms that cells employ to enable them to switch their fates. Such insights will reveal basic features of cellular plasticity that could improve regeneration therapy in human cells. However, due to limitations in analyzing the global status of regenerating cells in vivo, little is known about how cells make the traversal from one state to another. The experimental plan proposes to provide a comprehensive view of cells as they transition to new fates during regeneration. The study utilizes plants because the model system provides a rare opportunity to model cellular plasticity. In addition, the mechanisms that regulate plasticity are well conserved across plants and animals and the plant's adept ability to regenerate will illustrate the full potential of cells to change their identity. The proposal takes advantage of a powerful model system that permits continual imaging of regenerating tissue and the analysis of the transcriptional contents of single cells as they traverse fates. This system includes an inventory of active genes for each cell fate, permitting quantitative analyses to track the complex identity of cells as they regenerate. Thus, the complete analysis will generate a model of the trajectory of regenerating cells in order to address basic questions about regeneration: What is the origin of the highly plastic cells that participate in regeneration? Do regenerating cells show proliferative behavior that resembles stem cells? Do fate transitions require losing all identity and reaching a ground state or younger developmental stage (dedifferentiation)? Or, do cells traverse directly to new fates? The results will point to processes that can be targeted to make regeneration more efficient. The innovation of profiling the complete transcriptional dynamics of cells and projecting them onto images of regenerating tissue is widely applicable to many developmental model systems. Thus, overall, the proposed work has broad impact on the field of regeneration. PUBLIC HEALTH RELEVANCE: One important goal in therapeutic regeneration is to enhance the ability of existing cells to replace damaged cells. Despite their differences, plants and animals share mechanisms that control a cell's ability to switch identities. The goal of the projec is to use a powerful system in the highly regenerative plant to understand how a cell can change its identity with the aim to provide insights for new approaches in regenerative medicine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Core B: Genetics Core The Genetics Core continues to build on the highly-successful service functions provided to PPG researchers in the previous grant. With highly-trained personnel and specialized instrumentation, this Core provides our researchers with transgenic mouse production, knock-out mouse production, high quality DMA sequencing and microsatellite genotyping. These facilities were well-known among national Core labs for their outstanding technical competence and cost-effectiveness. The transgenic animal lab, under the management of Dr. Thomas Saunders, has accumulated 18 years of experience producing transgenic mice, and 15 years experience in genetic knock-out methodology. The DMA sequencing lab has been in operation for 14 years, the last 12 of which were under the direction of Dr. Lyons. That laboratory has an outstanding reputation for reliability and cost-effectiveness. They recently acquired an Illumina BeadStation 500GX, and have begun providing high-throughput SNP typing services. All four of the PPG projects depend heavily on the Genetics Core. The services described here will be provided on a recharge basis, with members of this Program Project receiving preferential discounts.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are major gaps in our understanding of how DNA damage is sensed and repaired and how DNA repair is coordinated with other dynamic chromatin functions (e.g. transcription, DNA replication) to maintain genome stability. DNA repair prevents environmentally-induced cancer in normal cells and can confer resistance to chemotherapy and radiotherapy in cancer cells. Therefore, knowledge of genome maintenance mechanisms is crucial for us to understand, prevent, and successfully treat cancer. The immediate objective of the proposed exploratory project is to define a new role for the DNA repair protein Rad18 in chromatin modification and regulation of gene expression. Our long-term goal is to determine how Rad18-mediated regulation of gene expression contributes to genome maintenance and tumor suppression (in normal cells) and chemo/radio-resistance (in cancer cells). Previously, Rad18 has been implicated only in regulating DNA repair proteins. Based on strong preliminary data, the central hypothesis is that direct mono-ubiquitination of Histone H2A at K119 by Rad18 is a novel mechanism for transcriptional repression that contributes to genome maintenance. The rationale is that defining mechanisms of Rad18-mediated genome maintenance will provide a better understanding of environmental carcinogenesis and new opportunities for improving cancer therapy. The Specific Aims (SAs) of this exploratory R21 proposal are: (1) Define novel Rad18 functions in epigenetic control of gene expression. (2) Identify Rad18-responsive histone marks and transcriptional events in vivo. For SA1 we will perform genome-wide analyses to identify Rad18-dependent histone marks associated with altered gene expression following exposure to environmental and therapeutic genotoxic agents (Polycyclic Aryl Hydrocarbons, or PAH, and Ionizing Radiation respectively) in primary human cells. SA2 will determine the extent to which Rad18 influences gene expression and histone modifications in bone marrow (BM) progenitor cells, known targets of genotoxicity from environmental PAH and radiotherapy. To determine the contribution of H2A-ubiquitination to Rad18-mediated genome maintenance we will use 'separation-of-function' Rad18 mutant specifically lacking H2A-directed E3 ligase activity. The proposed ideas and research are innovative because they seek to provide new paradigms where none exist: chromatin modification by Rad18 is novel and indicates that Rad18 may have new genome maintenance functions involving transcriptional repression. The proposed mouse studies are innovative because physiological Rad18 functions in genome maintenance have not previously been studied in BM progenitor cells (or other somatic cells) in vivo. The work is significant because it will define new Rad18 functions that protect normal cells against environmental exposures, and confer resistance to chemotherapy and radiotherapy in cancer cells. This work will identify new biomarkers of environmental and therapeutic genotoxicity. We seek to validate Rad18 and its effectors as druggable targets whose inhibition sensitizes cancer cells to chemotherapy and radiation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Type 2 diabetes mellitus is predominantly a disease of Western societies. Although a number of factors may be working in synergy to account for the difference in prevalence between Eastern and Western cultures, the role of dietary composition cannot be overlooked. In Western populations, the daily consumption of phyto estrogens of the isoflavone class is less than 1 mg per day, compared to 50-100 mg per day in average Asian diets. At physiologically relevant concentrations, isoflavones have been shown to enhance beta cell insulin secretion, and to increase their sensitivity to glucose in vitro. In addition, an isoflavone-rich diet increases liver and adipocyte mRNA insulin receptor expression. Thus, isoflavones may enhance beta cell insulin secretion and peripheral tissue insulin responsivness in vivo; the two major defects associated with type 2 diabetes. The specific aims of this proposal are to: 1) Systematically evaluate the concentration-dependent effect of three dietary isoflavones- genistein, didzein and glycitein- on beta cell function in vitro. Response will be measured by insulin gene expression, intracellular insulin accumulation and insulin levels in the medium; 2) Determine insulin dependent responsivness of human adipocytes in the presence of these three isoflavones. This will be quantitated by glycogen synthase activity and glucose uptake 3) Quantitate the effect of dietary isoflavones on parameters of glycemic control when given in the acute setting to subjects with Type 2 diabetes. Specific endpoints include glycemic control, insulin levels, lipid profiles and free fatty acid levels; and 4) Quantitate the effect of isoflavones when given in the chronic setting to subjects with Type 2 diabetes. In addition to the serum measurements outlined above, the chronic study will utilize glucose clamp techniques and intravenous glucose tolerance tests to further assess glucose utilization and insulin secretion.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Retinal pigment epithelium (RPE) cells and photoreceptor cells re functionally and developmentally closely integrated. Derangements of the RPE are involved in certain retinal diseases. An enhanced knowledge of RPE- expressed genes and of the regulation of expression of these genes will improve our understanding of the RPE. In previous years, we have identified, cloned, and characterized an RPE-specific protein, RPE65, a novel conserved, developmentally regulated 65 kDa protein. Using the cDNA for RPE65, we have isolated and cloned full-length human and mouse genes for RPE65. Comparison of the bovine, human, and partial mouse deduced protein sequences reveals a highly conserved molecule with about 99 percent similarity among species. To study the transcriptional regulation of this RPE-specific gene, we have made promote; constructs driving the bacterial chloramphenicol acetyltransferase (CAT) gene, covering 2.8 kilobases of human 5' flanking sequence, and tested them in transient transfection assays in cultured RPE cells to identify regulatory regions. The mouse 5' flanking region also has been sequenced and compared with the human, revealing conserved blocks, including identified transcriptional factor-binding elements in the putative promoter region. Mouse 5' flanking regions will be used to generate transgenic animals for in vivo analysis of promoter function. In addition to regulation at the transcriptional level, RPE65 expression is also regulated posttranscriptionally at the level of translation. Further characterization of the identified distinct sequences in the 3' untranslated region (UTR) of the RPE65 mRNA that control the stability and the efficiency of translation of the RPE65 message reveals that the effect may be at least partially coding-region-sequence specific because the 3' UTR sequence affecting translational efficiency had no effect on CAT expression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The prevalence of dental caries has been reduced significantly in some segments of the population over the past 25 years. The observed reduction has been attributed largely to increased exposure to fluoride in drinking water and use of fluoridated dentrifrices. During the same time period the use of food preservatives e.g. benzoates, sorbates, propionates, salicylates has also increased dramatically. For example the consumption of benzoate in the U.S. has increased from 1.8 million tons in 1970 to 25.5 million in 1995. Thus humans are exposed to preservatives constantly; persons in the U.S. consuming the average amount of soda ingest 800 mg of benzoate daily. Food preservatives are weak acids and exert their anti-microbial effect in a manner similar to that of fluoride i.e. at low pH values they diffuse undissociated through the bacterial cell membrane and acidify the cytoplasm, rendering microorganisms sensitive to acid. Acid tolerance is a characteristic of cariogenic organisms. Weak acids may also affect bacterial membranes. We have preliminary data which show that benzoates reduces the production of glycosyltransferase by microorganisms without affecting the enzymatic activity. This observation is consistent with an earlier report (Bowen and Hewitt, 1971) which showed that fluoride in growth medium influences the production of glucosyltransferase by mutans streptococci. Data from preliminary studies conducted in rats show that benzoate and non-steroidal anti- inflammatory agents e.g. ketoprofen (weak-acid) enhances the cariostatic effect of fluoride and also many suppress mutans populations. The purpose of the present study is to explore the effects of well-recognized food preservatives alone or in combination with fluoride on caries in our animal model. The outcome of this research could result in identifying a novel method of enhancing the effectiveness of fluoride.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Schizophrenia in adolescents is an understudied area and data on its psychopharmacological treatment are scarce. The goal of this First Independent Research Support and Transition (FIRST) Award proposal is to contribute to the development of a rational foundation for the pharmacological treatment of schizophrenia in adolescents. A program for the evaluation of risperidone will be used as a model. Over a 42 month period a minimum of 70 adolescents who meet DSM IV criteria and who need inpatient hospitalization are expected to complete the study. The primary aims of the study in this population are (1) to assess the short efficacy of risperidone (2) to assess critically the short-term safety of risperidone. Secondary aims are to examine the relationship of risperidone plasma levels to clinical response and side effects, the relationship of serotonin-2 (5-HT2) binding in blood platelets to risperidone treatment response and the effect of risperidone on cognitive functioning. The importance of this research arises from the pressing need for data about the safety and efficacy of neuroleptics in adolescents with schizophrenia; current treatment of this disorder in adolescents is based largely on extrapolation from research on adults even though research with other medications suggests that the response of adolescents to psychotropics may differ from adults. A double blind, placebo controlled parallel groups design will be used. An initial one week washout will be followed by a one week placebo baseline during which placebo will be administered under single-blind conditions. Patients who meet clinical criteria will be randomly assigned to two independent treatment groups, risperidone or placebo, for a period of four weeks. Treatment response will be measured with the Positive and Negative Syndrome Scale for Schizophrenia as well as other widely used measures sensitive to medication effects. Side effects will be carefully monitored with standard instruments. Plasma levels of risperidone plus 9-OH-risperidone will be monitored throughout the treatment period and related to treatment response and side effects. Pre- and post-treatment serotonin platelet will be assessed and related to treatment response. The study will advance knowledge about the pharmacological treatment of schizophrenia in adolescents (1) it will replace clinical anecdote and case reports about this now widely used medication with systematic data regarding efficacy and safety relative to placebo (2) it will provide needed information about the relationship between plasma levels, treatment response and side-effects and serve as guide for minimal effective therapeutic dose in the future (3) it will explore 5-HT2 platelet binding as a predictor of treatment response to risperidone, which may permit the selection of patients most likely to respond (4) it will examine the effects of risperidone on cognitive function. The study will also provide the basis for other innovative grant proposals by the Principal Investigator to study questions relevant to the treatment of schizophrenia in adolescents.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this project is the research and development of biostatistical methods and mathematical models appropriate for the analysis of epidemiologic studies related to cancer control and prevention. The various statistical problems studied under this project are derived from the needs of other activities in the Division. This research includes the development of mathematical models which can be used to predict and evaluate the effect of different intervention strategies. A two-stage model of the carcinogenic process, adapted from a model proposed by Moolgavkar, Venzon, and Knudson for studying the effects of cancer-inhibitory agents such as dietary retinoids is currently being developed and tested. In addition, the Armitage-Doll multistage model is being used to quantify the relationship between cigarette smoking prevalence and lung cancer mortality in the U.S. to estimate the effects of hypothetical changes in the future prevalence of cigarette smoking. The trend of lung cancer mortality is also being analyzed by regression models to project its future course which would be expected without any specific population intervention. Other methodolgic problems under investigation include the modification of logistic regression methods to incorporate knowledge of the sampling rates used in population-basec case-control studies of cancer, and development of a set of interactive computer programs which can be used to analyze the trends of cancer rates over time.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The central purpose of the proposed research is to define the roles which drug metabolism and pharmacokinetic factors play in determining the clinical response to certain commonly abused drugs, including the predisposition for compulsive drug use. Attention will be focussed on d-and 1-amphetamine, administered therapeutically to children with minimal brain dysfunction and to adult volunteers; and methadone, given in the course of treatment for chronic heroin addiction. Integrated gas chromatography/mass spectrometry and immunochemical methods will be employed to measure the concentrations of drugs and active metabolites in blood from human subjects, and the clinical response to the drug will be interpreted in terms of these measurements. Concurrent measurements on other body fluids (urine, saliva, sweat) are expected to lead to empirical correlations which may be of greater practical value in routine patient care and drug control programs. Twin studies are proposed to evaluate genetic factors as determinants of both pharmacokinetics and clinical response. Deuterium-labelled variants of amphetamine and methadone will be used to study changes in the single- dose pharmacokinetics of these drugs as tolerance develops in the course of regular administration. An investigation of the metabolism of amphetamine and related compounds will encompass both clinical and experimental studies in order to evaluate both the factors controlling the rate and routes of metabolism and the pharmacological properties of active metabolites. Significant improvements are anticipated in the sensitivity and accuracy of chemical assays and in methods of quantitative evaluation of clinical response to these drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary: Bile acids are secreted by the liver, stored in the gallbladder, and efficiently reabsorbed from the intestine. This enterohepatic cycling is crucial for maintaining the pool of bile acids and for restricting these potentially cytotoxic detergents to specific compartments such as the liver and Gl tract. Many of the transporters that function to maintain the enterohepatic circulation of bile acids have been identified over the past fifteen years. Notably absent from that list is the identity of the ileal basolateral bile acid transporter. The goal of the studies in this application is to determine the relative roles of the recently identified Organic solute transporter alpha-beta, Osta-Ostp, and the multidrug resistance protein-3, Mrp3, in the basolateral transport of bile acids in the small intestine and colon. The studies in Specific Aim 1 will directly test the hypothesis that ileal basolateral bile acid transport is mediated primarily by Osta-Ostp, with Mrp3 serving only a secondary back-up role. These experiments will use recently developed Osta KO, Mrp3 KO and Mrp3/Osta double KO mouse models. In addition to analyzing the gastrointestinal phenotype of these models, the adaptive changes in hepatic and intestinal expression of genes important for bile acid biosynthesis, transport, and regulation will be determined. The direct effects of these specific gene deletions on intestinal bile acid transport will be determined in vivo by measuring changes in bile acid fecal excretion and pool size, and in vitro using ileal everted gut sacs. The studies in Specific Aim 2 will directly test the hypothesis that Mrp3 and Osta-Ostp play important roles in protecting the colonic epithelium from the cytotoxic actions of bile acids. These experiments will use KO mice fed deoxycholic acid rich diets. The endpoints to be measured include colonic macroscopic and microcopic tissue damage, and cytokine levels as markers for the extent of injury. Relevance: A single layer of epithelial cells separates the gut lumen from the underlying immune cells of the gastrointestinal tract. Transporters such as Osta-Ostp and Mrp3 may play important roles in protecting that critical barrier by preventing cellular accumulation of bile acids and toxins. Since diseases such as colon cancer, inflammatory bowel disease, and colitis have been linked to defects in intestinal defense, it will be important to determine the specific functions of transporters such as Osta-Ostp and Mrp3 and if loss of these transporters contributes to the development or progression of gastrointestinal diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract A major component of many inflammatory diseases of mucosal surfaces, particularly in the intestine, is migration of large numbers of neutrophils (PMN) across the epithelium and accumulation within a lumen. In such conditions, disease symptoms are complex but directly related to leukocyte effects on the epithelial barrier and epithelial cell function. While much has been learned about mechanisms of leukocyte emigration from the circulation, much less is known about the receptors that regulate leukocyte interactions with the intestinal epithelium. In our studies, we have shown that a sequential series of adhesive steps are necessary for PMN to traverse the epithelium in a polarized manner that begins with interactions with the basolateral membrane and ends at the level of the apical or luminal membrane. However, many of the details of the transepithelial migration cascade and functional consequences of such remain poorly understood despite the clear link to many inflammatory diseases. This proposal will focus on functional consequences of PMN interactions with specific receptors at the level of the tight junction and apical/luminal epithelial membrane. We will extend recently obtained evidence linking binding interactions between PMN expressed JAM like protein (JAML) and the tight junction associated coxsackie and adenovirus receptor (CAR) with pro-inflammatory effects on epithelial homeostasis. Our studies also suggest that expression of the prototypic JAM protein family member termed JAM-A on leukocytes and epithelial cells is important in preventing pathologic inflammation in the intestine, however the relative contributions of epithelial and leukocyte expressed JAM-A to regulating intestinal mucosal homeostasis are not defined. Mechanisms and relative contributions of JAM-A in leukocytes and epithelial cells to development of experimental colitis will be explored. We have also determined that at late stages of the PMN transepithelial migration response there are binding interactions between PMN and the apical epithelial membrane mediated by specific glycan epitopes displayed on the transmembrane protein CD44v6. Such binding interactions result in epithelial functional responses that positively regulate epithelial barrier function. Details of the mechanisms behind such PMN interactions with epithelial cells are lacking despite the potential importance of resultant epithelial signaling responses in promoting mucosal health and healing. Experiments in this proposal will provide insights into these important issues as well as new ideas for the development of agents that might be used as anti-inflammatories in a tissue targeted manner as well as for enhanced mucosal wound healing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The main objective of this grant request is to facilitate faculty research that will contribute to the body of knowledge about nursing practice and nursing education. The expertise of current faculty suggest several avenues of probable research activity including: 1) investigations of macro issues that shape the delivery of nursing in various health care agencies, 2) investigations related to quality assurance for nursing, 3) basic and applied studies of current nursing clinical practices, 4) theoretical investigations of bio-psycho-social foundations of health and illness, and 5) explorations of usages of electronic technology in nursing practice and nursing education. The long range goal is to develop a larger cadre of sophisticated researchers actively engaged in investigative studies of relevance to nursing through the establishment of continuing supportive services for research at The University of Michigan School of Nursing, including the development of models for assigning faculty responsibilities that will facilitate and encourage their research activities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Glassware Washing and Sterilizing Facility located on the first floor of the McArdle Laboratory does the glassware washing and autoclaving for all of the investigators in the McArdle Laboratory. Dr. William Fahl is the faculty supervisor of this Facility, and Ms. Karen Schwarz oversees the sen to ten employees in the Facility. Washing and autoclaving procedures are established for each of the research groups. Materials are picked up from the laboratories, washed and/or autoclaved in the first-floor facility, and returned to the laboratories. I general, this involves an average two- to three-hour turnaround for pickup, washing, and delivery of the clean material. This rapid turnaround allows up to minimize the cost and space required for maintaining large glassware inventories in laboratories. In addition, other special procedures, including special cleaning agents, ultrasonic cleaning of pipettes, acid washing, rinsing with deionized water, etc., are performed as necessary, especially for the cell culture laboratories. These are five autoclaves, each of which is devoted to a specific application based upon the sophistication of the autoclave and the type of steam it uses for the autoclave cycle; these applications include autoclaving large numbers of media bottles for tissue culture media, pipettes, selected pieces of laboratory glassware, large volumes of bacterial media, as well as the routine decontamination of tissue culture and bacterial waste.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Thirty years ago, HIV was a death sentence with no vaccine, no treatment, and no cure. Its high morbidity and association with stigmatized sexual and drug-use behaviors led to the rise of HIV exceptionalism, the tendency to treat HIV differently from other diseases. Over time, a culture of HIV exceptionalism has profoundly shaped public perceptions, law, policy, advocacy, funding priorities, and the structure of health service delivery. A potential HIV cure signals a new chapter in this complex global story. The history of HIV shows that the social meaning of a disease - including how it is represented and policies surrounding its treatment and control - changes dramatically when it transitions from an untreatable disease to a treatable chronic condition. A further transformation of HIV from a treatable chronic condition to a curable disease will not only potentially decrease morbidity and mortality and contribute to epidemic control, but also alter how HIV is experienced, perceived and approached. Curing HIV is a strategic priority of the International AIDS Society and the NIH. Although HIV cure research is at an early stage, this is a critical time to conduct social and ethical research in order to inform subsequent research and ultimately guide pilot implementation of programs. HIV cure research will have unintended positive and negative implications that should be proactively explored. Some HIV control strategies such as male circumcision have been implemented with minimal research into their likely social and ethical implications, resulting in suboptimal uptake and missed public health opportunities. HIV cure clinical trials will predictably raise fundamental social and ethical questions. Anticipating the implications of HIV cure requires input from a diverse group of stakeholders who have the capacity to better understand unintended implications in order to prevent negative implications and expand positive ones. Building on strong links to ongoing cure research, social science/ethics research expertise, and global stakeholder links, we propose the following aims: (1) Develop a theoretical framework about HIV cure research and early implementation using historical, conceptual and ethical data; (2) Determine HIV cure stakeholder perspectives on cure research and early implementation in Cape Town, South Africa, Guangzhou, China, and Chapel Hill, USA; (3) Develop and assess the feasibility of an online forum to promote stakeholder engagement focusing on the social and ethical implications of HIV cure research. The inclusion of three sites creates an opportunity for powerful cross- cultural comparisons, helping us to identify cross-cutting themes about unintended implications. This research will provide a strong foundation for subsequent HIV cure research and early implementation in a number of settings across the world.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose that pattern recognition receptors, whose normal function is in innate immune responses, are also activated by stress-generated ligands, such as molecules released by dying cells, normal and oxidized lipids and a variety of so called \"danger\" signals in addition to environmental toxins and pathogen associated molecular patterns. Activation of such receptors either by endogenous- (i.e. host-generated) or pathogen- generated ligands turns on stress-activated protein kinases, such as JNK, p38 MAPK and IKK, that serve as molecular transducers that contribute to development of chronic inflammatory diseases. Importantly, these pathogenic mechanisms allow the integration of environmental factors and genetic susceptibility loci that together contribute to the development of some of the most common chronic diseases, including type 2 diabetes, asthma, inflammatory bowel disease and chronic liver disease. In the previous grant period we have generated strong evidence in support of this hypothesis by focusing on the pathogenic functions of the JNK and IKK signaling pathways. We also generated a mouse model expressing the equivalent of the most common susceptibility allele for Crohn's disease, an inflammatory bowl disease whose pathogenesis is affected by genetic and environmental factors. In the present period we will focus our main effort on the pathogenic function of different classes of pattern recognition receptors as targets for stress - and injury- generated stimuli, as well as continue with our studies on the role of p38 MAPK in liver inflammation and toxicity. More specifically we will examine: 1) the role of protein kinase C isozymes and Toll like receptors (TLRs) in obesity-induced JNK activation and insulin resistance;2) the role of TLRs in toxin-induced liver injury, liver inflammation and liver cancer;3) examine the role of p38 MAPK in toxin-induced liver injury, liver inflammation and liver cancer;4) examine the mechanism by which the intracellular NOD-like receptor NOD2 leads to activation of caspase 1 and IL-lbeta secretion;5) construct a conditional mouse mutant that allows constitutive NOD2 activation and use it along with our previously generated Nod2delta33 knockin mutant to examine effects of tobacco smoke, microparticles and bacterial products on development of NOD2- modulated colonic and airway inflammation. To accomplish these aims we will use a combination of cellular biochemistry, molecular genetics and experimental pathology, an approach that has been proven effective during the previous project period.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "CD8+ T cells are crucial to the control of T. cruzi infection and may also hold the key to the development of clinical disease in the chronic Chagas'disease. The goals of this project are to better understand the mechanisms important in the generation of anti-T, cruzi CD8+ T cell responses, to identify the primary and most important targets of these responses, and to obtain a better understanding of why these responses are slow to develop relative to CD8+ T cell responses to other pathogens. A combination of proteomic, expression profiling and bioinformatic tools and information will be used to identify targets of the CD8+ T cell response and the status of each of at least 50 such selected proteins will be tested in ELISPOT, in vivo CTL and eventually MHC Tetramer staining assays. We will investigate two aspects of the generation of CD8+ T cell responses in T. cruzi that might account for the differences between the response generated in this infection and that observed in other better-studied models: the initial encounter of T. cruzi with DC and the antigen presentation function of these DC, and the effect of epitope density on activation of anti-T. cruzi CD8+ T cells. Parasites will be followed from the point of infection to their encounter and activation of naive CD8+ T cells in the lymph node draining the site of infection. Additionally parasite lines will be produced which vary in the level of expression of model epitopes to determine if epitope density plays a role in the rate of generation and the quality and quantity of the CD8+ T cell response to peptides from T. cruzi. Finally, the potential role of altered peptide ligand antagonism by peptides encoded by members of the T. cruzi trans-sialidase gene family in the regulation of CD8+ T cell responses in T. cruzi will be determined and the biological significance of the effect of this antagonism of the generation and effector function of CD8+ T cells will be determined.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "X-ray diffraction and neutron diffraction are being used to study the packing and arrangement of collagen fibrils in corneal stroma (mainly beef cornea). Diffraction methods will also be used to study the electron-density profile along the collagen fibrils themselves, to try to locate the glycosaminoglycans and glycoproteins in the stroma. Where appropriate comparisons will be made with scleral tissue, in both cases the emphasis is on tissue as nearly as possible in the living state, without the fixation and staining necessary for electron microscopy. The overall objective is to relate changes in these structural parameters to changes in the swelling behaviour and the transparency of corneal stroma. To this end attempts are being made to prepare cornea in which the glycosaminoglycans and glycoproteins have been extracted by biochemical means, but which retain sufficient regularity to act as diffracting structures. The swelling, transparency and structure of such cornea will be compared with the intact tissue. The X-ray facilities being used include the small-angle diffraction camera on the DORIS storage ring attached to the DESY synchrotron in Hamburg, West Germany. Neutron diffraction is carried out on the D11 camera at the Institut Laue-Langevin, Grenoble, France.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Description (Taken directly from the application): In the United States, managed care is growing rapidly. The membership of managed care organizations (MCOs) now exceeds 50 million people and may grow by an additional 50 million by the year 2,000. The most rapidly growing type of MCO is that based on the Individual Practice Association (IPA) model. The absence of data attesting to the effectiveness and cost-effectiveness of comprehensive diabetes management within an IPA model MCO constitutes a major barrier to the introduction of such care in this type of health care system. The hypothesis of this project is that within the context of an IPA-model MCO, the system-wide identification of members with diabetes and their enrollment in a comprehensive diabetes management program that includes comprehensive provider profiling and diabetes health management will improve the process, outcomes, and cost-effectiveness of care. The Specific Aims of the project are: 1. develop and implement a medical information system-based method to identify members with previously diagnosed diabetes in an IPA model MCO; 2. randomize patients by practice to comprehensive versus usual diabetes management, and to compare the processes and outcomes of care in the two randomized groups; and 3. apply an economic model to these data to compare the cost and cost-effectiveness of comprehensive versus usual management. We anticipate that the results of this project will improve the ability of IPA-model MCOs and medical care providers to identify patients with previously diagnosed diabetes. Comprehensive health management for people with diabetes in an IPA-model MCO will address many of the barriers to care that exist in the present medical care system, and will represent a new and potentially cost-effective strategy for diabetes management in a rapidly growing type of health care system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Lung cancer is the leading cause of cancer deaths in the United States for both men and women. Until recently, the major focus of lung cancer control has been smoking prevention and cessation. The recently conducted NCI funded National Lung Cancer Screening Trial (NLST) found that screening with low dose computerized tomography (CT) scans compared to a control of chest x-ray reduced lung cancer mortality among a high risk study cohort. Based on the emerging scientific evidence, clinical guidelines supporting lung cancer screening among high risk populations have been developed by the U.S Preventive Services Task Force (USPSTF) and other professional organizations. However, many questions remain regarding the translation of scientific findings from clinical trials to the practice setting. In this NCI conference grant we propose to hold a symposium entitled, Lung Cancer Screening: A Debate of Practice, Policy, and Science. This symposium will be held as part of Society for Medical Decision Making (SMDM) 2014 Annual Meeting to be held in Miami, FL. The overall theme of the 2014 SMDM meeting is Medical Decision Making among Diverse Populations: Advancing Practice, Policy, and Science. In accordance with this theme, the proposed symposium will debate issues pertinent to the translation of evidence from clinical trials to diverse populations. The Society for Medical Decision Making is an interdisciplinary professional society that has approximately 1000 members. The scientific focus of SMDM members and meeting attendees encompasses both the individual and population perspective with methods drawn from a range of fields including the social sciences (economics, psychology, sociology), quantitative sciences (mathematics and statistics), organizational theory, engineering, communication, clinical epidemiology, and clinical medicine (physicians, nurses and other health professionals). In the proposed symposium, an interdisciplinary panel of experts will gather to highlight and debate key questions regarding emerging lung cancer screening paradigms. The symposium will foster interdisciplinary discussions that include leaders in clinical medicine, evaluation science, economics, health behavior, and bioethics. The symposia will be organized into 3 blocks: 1) translation of scientific evidence to clinical practic, 2) health policy and the guideline development process, and 3) determining the value of lung cancer screening from the patient and societal perspective. Findings from the symposium will inform strategies for research, policy, and implementation and evaluation of emerging lung cancer screening programs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We evaluated the effect of aging on fat and carbohydrate metabolism during moderate intensity exercise. Six elderly (73*2 yr) men and women were studied during 60 min of cycle ergometer exercise performed at 5&6*4.5 % of maximum oxygen uptake. Six young adults (26*2 yr), matched by gender and lean body mass (LBM), were studied on two occasions: during exercise performed at the same absolute and at a similar relative intensity as the elderly subjects. Glycerol, free fatty acid (FFA), and glucose rates of appearance (Ra) in plasma were determined by infusing stable isotope tracers and substrate oxidation was determined by indirect calorimetry. Mean fat oxidation during exercise was lower in the elderly subjects (3.43+0.27 umol.min-1~kg LBM-1) than in the young adults exercising at either the same absolute (4.27+0.96 umol.min-1.kg LBM-1) (P <0.001) or similar relative (4.65+1.02 umol.min-1.kg LBM-1) (P<0.001) intensifies. Mean carbohydrate oxidation in the elderly group (86.2+6.7 umol.min-kg LBM-1) was higher than the young adults exercising at the same absolute intensity (63.4+ 8.2 umol.min-1.kg LBM-1) (p<0.001) but lower than the young adults exercising at the same relative intensity (131.9+ 13.1 umol.min.-1.kg LBM-1) (P<0.001). Average glycerol and FFA Ra's in the elderly subjects (5.09+0.51 and 9.67+0.50 umol.min-kg LBM-1, respectively) were higher than in the young adults exercising at the same absolute intensity (4.01+0.37 and 6.49+0.54 umol.min-1.kg LBM-1, respectively) (P<0.001) but lower than the young adults exercising at a similar relative intensity (6.42+1.06 and 12.87+2.46 umol.min-1kg LBM-1, respectively) (P<0.001). Average glucose Ra during exercise was similar in both groups during exercise performed at the same absolute intensity, but was 25 % lower in the elderly than in the young subjects during exercise at a similar relative intensity (P<0.001). We conclude that fat oxidation is decreased while carbohydrate oxidation is increased during moderate intensity exercise in elderly men and women. The shift in substrate oxidation was caused by age-related changes in skeletal muscle respiratory capacity because lipolytic rates and free fatty acid availability were not rate limiting in the older subjects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "At least 20 million people in the US alone are infected with human papillomaviruses, which are associated with a wide range of pathologies, including external cutaneous warts, genital condylomata and cervical carcinoma. Nonetheless, there are no specific, beneficial antiviral therapies for this prevalent problem. In order to develop novel therapeutic approaches for compounds with antiviral activity against papillomaviruses, we have collaborated with Dr. Elliott Androphy of the Tufts University Medical Sch, who has cloned a novel cellular protein, provisionally named 42A. This protein interacts with the papillomavirus E2 transactivation and DNA replication regulator and appears to be required for activity. We propose experiments to determine the feasibility of inhibiting the E2-42A interaction as a means of developing novel antivirals. We plan to map the domains of the proteins responsible for their interaction and then use this information to design peptides which inhibit the interaction. The peptides will be assayed in systems we have developed to measure E2-dependent regulation and for antiviral and cytotoxic activities. These Phase I studies will thus allow us to determine the feasibility of developing high throughput screens for small- molecule inhibitors of th E2-42A interaction and the likelihood that they would be of use in inhibiting papillomavirus-related pathologies. PROPOSED COMMERCIAL APPLICATION: The proposed research will establish the feasibility of disrupting the HPV E2-42A complex as a means to attain antiviral activity. Results from this work will establish whether a drug discovery program based on this approach should be established to develop novel human papillomavirus-specific antivirals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Blood coagulation is ensured through equilibrium between pro- and anti-coagulant coagulation factors that interact with each other and cells. Factor VIII (FVIII) is a large multidomain protein (~280kDa), which in its active form, Factor VIIIa (FVIIIa) acts as a co-factor to the serine protease Factor IXa (FIXa) within the membrane-bound Tenase complex. Binding of FVIIIa to FIXa onto the platelet surface increases Factor Xa (FXa) and Thrombin generation more than 10^6 times. Mutations in FVIII result in mild to severe Haemophilia type A, a life-threatening blood condition affecting one in 5000 of the male population (Wacey et al., 1996). The sole cure for this condition is intravenous administration of FVIII, whose membrane-bound structure has been studied by Stoilova-McPhie using electron microscopy (Parmenter and Stoilova-McPhie, 2008;Stoilova-McPhie et al., 2008). CryoEM is a unique method capable of defining the structure of proteins in their native environment. For this purpose we have designed lipid nanotubes (LNT) for helical crystallization of FVIII and its complexes, allowing its membrane-bond structure to be resolved by Cryo-EM. Defining the structure of membrane-bound blood coagulation Factor Va and Factor VIIIa attached to lipid nanotubes by Cryo-electron microscopy is the primary goal of this project.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neiserria gonorrhoeae and Chlamydia trachomatis are the two most common bacterial sexually transmitted infections (STI) in the United States, affecting an estimated 4 million persons annually at a cost of over $3 billion. After a period of declining morbidity, the prevalence of chlamydial infection is now rising in some areas of the U.S., while rates of gonorrhea are stable, but no longer decreasing. New public health interventions are needed to decrease morbidity from these infections. Improved partner notification and treatment might be such an intervention. Recent randomized controlled trials have demonstrated that expedited partner therapy (EPT), the practice of treating the sex partners of persons with curable STI without their mandatory prior examination by a clinician, can increase the number of partners of persons with gonorrhea or chlamydial infection treated, and can decrease rates of STI reinfection. Study investigators have shown that a public health EPT program can be instituted in a single city to increase partner treatment, but the feasibility of wider scale public health EPT programs has not been established, and the population-level impact and cost-effectiveness of larger EPT programs is unknown. This application proposes a stepped-wedge community-level randomized controlled trial of EPT for gonorrhea and chlamydial infection in Washington (WA) State. A population-based EPT program modeled after one already in place in King County, WA will be instituted in a randomly selected temporal order in 28 health districts in WA State. Free medication will be provided to patients to deliver to their sex partners, and a new case-reporting form will triage public health partner notification assistance to persons at high-risk for not notifying partners. Free medications will be distributed directly to patients via larger clinics in each health district and through a network of commercial pharmacies contracted to participate in the program. The study will assess the impact of EPT on the prevalence of chlamydial infection among women tested in clinics receiving CDC Infertility Prevention Project funding, and on the incidence of reported gonorrhea among women in the state. The proportion of sex partners treated before and after the institution of the EPT program will be estimated based on interviews with a random sample of reported cases: Investigators will evaluate the intervention's cost- effectiveness both from a societal perspective and relative to other publicly funded measures to control gonorrhea and chlamydial infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To investigate the control mechanisms involved in regulating melanogenesis in a variety of tissues of normal and cancerous origin. Tyrosinase and other enzymic activities are investigated both by spectrophotometric and radioactive assay methodology. Tissues are dissected immediately before use in 0.1 M phosphate buffer,homogenized and fractionated into subcellular components by means of density gradient and differential centrifugation. Samples are then incubated with the appropriate substrates and controls after treatments known to alter enzymic activity. The production of melanin and other reaction products can be followed spectrophotometrically or by liquid scintillation counting of the newly formed radioactive products.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Menisci function to distribution load and increase stability of the knee joint. Both partial and complete meniscectomy have been shown to increase the incidence of osteoarthritis. Meniscal replacement with allograft, synthetic or tissue engineered replacements, could act to reduce the occurrences of osteoarthritis. The success of meniscal replacements depends on their ability to restore normal meniscal function both biologically and biomechanically. Meniscal attachments are critical for proper meniscal function. Since rupture of the horn attachments of the menisci are rare it is likely that a gradient in mechanical properties and biochemical make-up exists through the attachment. Previous computational studies have shown that the material properties of the attachments dramatically affect the contact pressures on the meniscus and without gradual transition in the properties, large stress concentrations at the insertion zones into the meniscus and the bone are evident. Additionally, these stress concentrations are seen mostly in the deeper zones of the tissue, not on the proximal surface. Previous research has only documented the material properties on the proximal surface of the tissue. The goal of this project is to quantify the microstructure and material properties of the meniscal attachment where it inserts into the meniscus and bone, as well as in the deep layers of the tissue. Specifically this project will 1) determine the local fluid pressures in the deep layers of the tissue during both physiological loading (static and dynamic) levels and failure testing using a fibre-optic pressure microsensor, 2) determine the transverse material properties 2) quantify the collagen orientation in the transition zone from fibrocartilage to ligamentous attachment using scanning electron microscopy, and lastly the project will 3) determine the mechanical properties of the transition zones into the subchondral bone at the nanolevel. The results of these aims will then be used to quantify a relationship between the structure and function of the native meniscal attachments and validate the current finite element model of the attachments. This data can then be used to develop, design and evaluate meniscal replacements, including tissue engineered constructs. A successful meniscal replacement will work to prevent joint degeneration following meniscectomy. PUBLIC HEALTH RELEVANCE: Menisci within the knee joint function to protect the underlying articular cartilage from degenerative osteoarthritis. The absence of a healthy meniscus can inhibit this function. The goal of this research is to determine the structure and function of native meniscal attachments such that effective meniscal replacements or therapies can be developed to reduce the incidences of knee joint osteoarthritis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY There is no specific treatment available for the subpopulation of patients with salt sensitivity of blood pressure (BP); unfortunately, the molecular mechanisms underlying salt-sensitivity remain poorly understood. One of the major proposed mechanisms for the development of salt-sensitive (SS) hypertension involves a defect in the ability of the kidneys to excrete salt. Atrial Natriuretic Peptide (ANP) encoded by Nppa, is a hormone known to promote salt excretion and BP reduction, and there are clinical data implicating inherently low levels of ANP in the development of SS hypertension. Among other effects, ANP (via cGMP-related mechanisms) is known to be beneficial for mitochondrial bioenergetics and biogenesis. However, there is a gap in knowledge regarding the effects of ANP on mitochondria in the kidney, especially in SS hypertension. Our pilot studies demonstrated that during a high salt challenge Nppa-/- (ANP knockout) Dahl SS rats exhibit exacerbated salt-sensitivity, reduced sodium excretion, and aggravated kidney injury, which is associated with mitochondrial damage and dysfunction. We also showed that there is dysregulation of renal sodium transporters, in the Nppa-/- rats compared to wild-type controls, and the activity of the Epithelial Na+ Channel (ENaC) is elevated in the collecting ducts. Chronic ANP infusion in wild-type SS rats resulted in a dramatic attenuation of salt-induced BP increase and alleviated organ damage. We hypothesize that in SS hypertension ANP deficiency/reduced sensitivity to ANP is causative to renal mitochondrial dysfunction and associated sodium transport imbalance. To address the central hypothesis of this project, we developed three specific aims: Aim 1. Establish whether increased ANP levels are beneficial for renal salt handling and cardiac function in SS hypertension. Aim 2. Determine whether low renal cGMP level resulting from lack of ANP causes an increase in renal mitochondrial Ca2+ and reactive oxygen species (ROS). Aim 3. Test the hypothesis that disrupted Ca2+ balance and excessive ROS production by dysfunctional mitochondria affect renal sodium handling in SS hypertension. We generated abundant evidence to support these aims, created a rigorous and comprehensive experimental design and established novel cutting-edge techniques to address the hypothesis. We recruited strong collaborative expertise, and will implement a combination of whole-animal studies and in vivo techniques (blood pressure monitoring with drug infusion, metabolic studies and GFR measurements), electrophysiology (single channel and whole-cell patch-clamp of the freshly isolated nephrons and isolated mitochondria), advanced microscopy, mitochondrial spectrofluorimetry and respirometry, and routine molecular biology approaches. The successful completion of the proposed studies will unravel the novel causative mechanisms of salt-sensitivity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The general goal of this proposal is to gain a better understanding of the origin, identity, and biological properties of mammalian cells that when properly stimulated are capable of regenerating the adult and senescent myocardium. This objective is based on the premise that adult and senescent myocardium contains a small population of cells with the characteristics and behavior of cardiac stem cells (CSCs): they are clonogenic, self-renewing and multipotent. The progeny of a single CSC isolated from an aging myocardium can differentiate into myocytes, smooth muscle and endothelial cells. When injected or activated in a post-ischemic myocardium, these cells reconstitute a functional ventricular wall. A decrease in their number and/or function might be one of the causal factors in the altered performance of the old and senescent heart. Nothing is known about the origin of the CSCs. Data from animals and humans show a continuous trafficking of these cells between myocardium and extra-myocardial tissues. The bone marrow contains a population of cells with similar characteristics and a several other extra-cardiac cell types have been shown able to differentiate into cardiocytes. We will test the hypothesis that CSCs appear late in development, reach the myocardium through the circulation, and are constantly replenished to allow a continuous turnover of the normal and pathological heart. Once we have established the origin of the CSCs and determined their cardiogenic potential, we initiate the dissection of the genetic regulators of their differentiation, starting by identifying the role of \"stem cell marker genes\" in the production of the cardiogenic phenotype. We will continue by testing the hypothesis that differentiation of CSCs in large measure recapitulates early cardiac ontogeny and requires the down regulation of the canonical Wnt pathway as well as the upregulation of members of the TGFbeta family of regulators, namely certain bone morphogenetic protein (BMPs) genes. Three Specific Aims will be pursued: 1.- To ascertain the origin and relationship among the different adult cells capable of producing the cardiogenic lineages and to compare their myocardial regenerating potential in the post-ischemic heart. 2.- To elucidate the role of \"stem cell marker genes\" on the myocardial regenerating capability of the CSCs. 3.- To initiate the dissection of the signaling pathways responsible for the maintenance of the differentiated state of the CSCs and their differentiation under normal conditions, in aging and in response to ischemia. The information obtained from these experiments will be used to devise more effective ways of fostering myocardial regeneration through the coaching of the CSCs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal represents a competitive renewal of the Boston University Multidisciplinary Clinical Research Center grant which was established in 2001 and built on a strong foundation of outstanding epidemiology and clinical research. The strong record of accomplishment in patient oriented medical research at Boston University has been not only maintained in the last four years but expanded. What we propose creates new directions and areas of research for this already productive group of investigators who work together closely, and it promises to produce further major insights into rheumatic disease causation and treatment. Led by Director, Dr. David Felson and Associate Director, Dr. Saralynn Allaire, this group of full-time MCRC investigators who work together in shared office space will meet weekly in Methodology Core group meetings to discuss and review both projects proposed here and other funded and non-funded clinical research. This Core will provide support for the proposed projects: Bracing in Patellofemoral Osteoarthritis: A Clinical Trial, David J. Hunter, PI, Risk Factors for Gout Attacks: A Case-crossover Study, Yuqing Zhang, PI, Interplay of Thrombosis and Inflammation in Vasculitis, Peter Merkel, PI. An administrative unit will provide oversight to ensure that these projects are successfully accomplished and that core resources for support of these and other projects are allocated efficiently.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The HIV-1 capsid is an important determinant of viral replication. Early studies demonstrated the intrinsic importance of the capsid in mediating uncoating of the viral core. More recently, the capsid has emerged as an important mediator of virus-host interactions. Cyclophilin A (CypA) is an abundant cellular protein known to bind the N-terminal domain of the capsid protein. CypA can promote infection of some HIV-1 strains in certain cells, but it can also inhibit infection. Despite many years of research, we still do not understand these mechanisms. I hypothesize that CypA modulates HIV-1 infection by regulating the interaction of specific cellular proteins with the HIV-1 capsid. Indeed, we have observed that CypA can regulate the non-human primate capsid-binding restriction factor, TRIM5. However, the mechanism of this activity is poorly understood. I will determine the ability of CypA to regulate capsid-binding restriction factors by i) elucidating the mechanism of CypA-dependent TRIM5 restriction and ii) determining the ability of CypA to alter the capsid interactome using comparative proteomics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have recently characterized 2 types of cardiac myosins and will extend preliminary studies indicating a correlation between the chemical reactivity and enzymatic behavior of the myosin and the contractile properties of cardiac muscle. Initial studies have indicated that thyroid hormone influences the type of myosin found in the heart, and this observation will be studied further. The rate of synthesis and turnover of the heavy and 2 light chains of cardiac myosin will be determined, and hormonal influences on the rates will be studied. The influence of serum and of growth-promoting factors in serum on cell multiplication in rat aorta will also be studied.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Rho is a termination factor and an ATPase that binds to nascent RNA transcript and catalyzes their site-specific release within many gram- negative bacterial and phage transcription units. This proposal outlines a research program for the construction and characterization of a paused transcription intermediate that can be used as a substrate to analyze the thermodynamics and kinetics of the Rho-mediated transcript release reaction. The transcription template will be constructed from a segment of DNA containing the E. coli rho attenuator (tRA), a Rho-dependent terminator that is located in front of the rho gene, and which autogenously regulates the level of Rho in the bacterial cell. To create the paused transcription intermediate, site-directed mutagenesis will be used to install an EcoR1 restriction site at a specific location downstream of the region coding for the Rho-attachment site on the transcript; transcription elongation will be blocked by the binding of a cleavage-defective EcoR1 endonuclease. A kinetic formalism will be used to ascertain the rate-determining steps of the Rho attachment/transcript release reaction. This formalism will be used to evaluate the rates of Rho assembly and transcript release in an effort to sort out the mechanisms of these processes during transcription termination. The kinetics of release and the structure of the Rho-bound ternary complexes and the released products will be characterized in the presence of ATP and ATP analogs to determine how the Rho-ATPase activity is involved in the release process. The location of the EcoR1 blockade will be varied by linker scanning mutagenesis. The relationship between the site of the blockade and the energetics and kinetics of Rho attachment and transcript release will be measured. These studies will provide information on the minimum size for the Rho attachment site, and the significance of sequence context in transcript-template interactions during termination.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study is designed to prospectively examine changes in the insulin secretion in first degree relatives of non-insulin dependent diabetics compared with individuals with no family history of diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this proposal is to study, by newer, direct methods, the pathways for the transport of proteins, polypeptides and lipoproteins across cells and basement membranes. The studies will be done on isolated segments of renal tubules employing in vitro and in situ microperfusion methods and electron microscopy. Functional, quantitative data on the absorption and transport of various macromolecular substances will be directly correlated with fine structural changes at the subcellular level by autoradiographic and histochemical techniques. By microperfusing macromolecular substances at varying concentrations and time intervals and by following fine structural changes in the cells and membranes sequentially, the mechanisms for the absorption, transport or degradation of these substances should be elucidated. Ultimately, it is planned to study transport of macromolecular substances across cells injured by a variety of harmful transport of macromolecular substances across cells injured by a variety of harmful agents. This information would be of fundamental importance in understanding the role of macromolecules and their transport process during physiologic and many pathologic conditions. BIBLIOGRAPHIC REFERENCES: Oparil, S., Carone, F.A., Pullman, T.N., and Nakamura, S. Inhibition of proximal tubular hydrolysis and reabsorption of bradykinin by peptides. Am. J. Physiol. 231(3): 743-748, 1976. Peterson, D.R., Oparil, S., Flouret, G., and Carone, F.A. Handling of angiotensin II and oxytocin by renal tubular segments perfused in vitro. Am. J. Physiol. 232(4): F319-F324, 1977 or Am J. Physiol.: Renal Fluid Electrolyte Physiol. 1 (4): F319-F324, 1977.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Modern health monitoring devices at hospitals and wearable sensors in households generate a large amount of time series data at high rate, capturing the physiological status of patients in a real-lime fashion. The premise is that these technology advances enable a data-driven healthcare system that starts making fast, accurate, objective and inexpensive decisions based upon data, in addition to an individual physician's experience and preference. However, there is a significant gap in the mathematical theory and computational tools to promptly extract actionable information from multi-modal non-stationary time series data in a robust and tractable manner, which has become a serious roadblock to further utilize bigger data for better healthcare monitoring. The goal of this research program is to develop a mathematical framework for extracting time-frequency and geometric representations of multi-modal physiological data, in an online and robust manner, and use them to design machine learning algorithms to improve real-lime health monitoring. Specifically, we hypothesize that the development of time-series and geometric methods for large streaming multi-modal monitoring data will lead to more accurate diagnosis on various physiological monitoring applications, including detection and prediction of rare events such as seizure and arrhythmia, classification of sleep stages for newborns and children, and real-time artifact removal of physiological data. To achieve our goal, we plan to develop novel theoretical and computational tools for analyzing non-stationary multi-modal time series data with noise, corruption and missing data as well as real-time algorithms for filtering and event detection from such data. The tools and algorithms will be applied on clinical tasks at the Nationwide Children's Hospital. In addition, the real-time workflow will be implemented on Hadoop clusters with a mission of public sharing of both data and software. The development from the interdisciplinary team composed of mathematicians, biomedical informaticians as well as the hospital will not only transform the frontiers of mathematics knowledge, but also significantly impact clinical applications, data science education, and the development of the $11 O billion emerging market of wireless health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The he'patitiS'Bvirus (HBV) is a major cause of infectious liver disease throughout the world. There are 1.2 million carriers of HBV in the U.S. and approximately 400 million worldwide. Neonatal HBV infection is rarely cleared and as many as 90% of perinatally infected children become chronically infected. Therefore, in addition to worldwide vaccine programs to prevent new infections, methods for treating HBV chronic carriers will be necessary to eradicate this disease. This proposal is focused on understanding the mechanisms responsible for inducing and maintaining chronic HBV infection and more specifically the role of HBV nucleoprotein antigens, the nueleocapsid (HBcAg) and the secreted non-particulate HBeAg. Armed with this information we are designing immunotherapeutic vaccine candidates for the treatment of chronic hepatitis B infection. The specific aims are addressed through the use of HBc/HBeAg-expressing and HBV replicating tra'nsgehic(Tg) mice and three recently developed HBc/HBeAg-specific T cell receptor (TCR)-Tg lineages and the Various combinations of \"double and triple-Tg\" hybrids. The specific aims are: (1) determine mechanisms of HBeAg-induced t cell tolerance at the cellular and molecular levels;(2) examine the relationship between HBc/HBeAg-specific CD4+ and CD8+ T cells;(3) explore the reasons for \"split tolerance\" between the HBcAg and the HBeAg at the cellular and molecular levels;and (4) develop strategies to reverse or bypass HBeAg-induced T cell tolerance as possible immunotherapies for chronic HBV infection. During the previous years of this project we have developed a number of Tg model systems that will facilitate the pursuit of the current specific aims. For example, four in vivo models of HBeAg-induced T cell tolerance mediate'd by clohal deletion, clonal anergy, clonal ignorance and T regulatory (Treg) cells have been developed. Furthermore, models of in vivo anti-HBc and anti-HBe seroconversion and CD4+ T cell-mediated liver injury have been developed. These model systems should be very useful in elucidating the mechanisms of T cell tolerance that play an important role in promoting viral persistence during chronic HBV infection and in designing candidate vaccines for treating chronic HBV infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The studies outlined in this proposal are aimed at establishing any similarities in the biochemical mechanisms for toxicity of halogenated antibacterial agents, particularly bisphenols, salicylanilides and carbanilides. These compounds all appear to alter the integrity of biological membranes. We plan, therefore, to investigate interactions between three representative antibacterials (hexachlorophene, tribomsalan and trichlorcarban) and proteins and membranes of the nervous system and erythrocytes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Asthma is a complex disorder characterized by episodic airway obstruction and hyper-responsiveness, sometimes accompanied by airway remodeling. Although the underlying causes of asthma remain poorly understood, one contributing factor is exposure to respiratory pathogens. For example, asthmatics positive for Mycoplasma pneumoniae (M. pneumoniae) infection have demonstrated an improvement in pulmonary function after antibiotic treatment whereas patients that test negative do not, suggesting a causal relationship between M. pneumoniae infection and asthma symptom severity. Although there is a strong clinical correlation between M. pneumoniae infection and a sub-set of asthma cases, until recently, the identification of a virulence factor that might play a role in disease pathogenesis had remained elusive. This situation changed, however, when we discovered a 591 amino acid protein with ADP-ribosyltransferase (ART) activity in M. pneumoniae designated Community Acquired Respiratory Distress Syndrome ToXin (CARDS TX). The experiments outlined in this proposal are designed to uncover the structure and action of CARDS TX using a range of biophysical techniques. Using the well-established tools of single crystal X-ray diffraction, we will determine: 1) the three-dimensional structure of CARDS TX; 2) CARDS TX in complex with its NAD", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A psoralen monadducted crosslinkable DNA oligonucleotide probe to a conserved portion of the HIV genome will be prepared and used in an innovative solution hybridization assay. The assay is conducted at the melting temperature of the respective probetarget hybrid using high concentrations of probe. Hybridization and photochemical fixation are carried out concurrently over a period of minutes after which excess probe is removed by a denaturing wash step. The sensitivity and ease of use of this probe will be compared using a cloned target sequence to an otherwise identical unmodified probe used in a traditional blot format. Favorable results will justify assays on actual clinical specimens using novel nonradioactive amplification schemes. The resultant test for HTLVIII could serve as a model for additional tests to other infectious and genetic diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Alcoholic Liver Disease (ALD) occurs only in a subset of alcoholics ( 30%) that develop tissue injury and organ dysfunction. This well established epidemiological and clinical observation indicates that alcohol abuse is required but not sufficient to cause tissue injury and additional cofactors are required. We and other have shown that increased intestinal permeability (gut leakiness) and endotoxemia are two required cofactors. The key question is what additional cofactors are needed to develop gut leakiness and endotoxemia. We hypothesize that altered circadian rhythms are an important determinant in alcohol induced gut leakiness and endotoxemia. All subjects will undergo testing for (1) central circadian rhythm from the timing and phase angle of the dim light melatonin onset (DLMO) and (2) peripheral circadian clock gene expression in the gastrointestinal tract measured from buccal mucosal cells. Aim 1: To test the hypothesis that circadian disruption is present in chronic alcoholics with gut leakiness. We will compare chronic alcoholics with gut leakiness/endotoxemia and compare them to matched alcoholic controls without gut leakiness/endotoxemia. We hypothesize that alcoholics with gut leakiness will have greater central and/or peripheral circadian disruption than matched alcoholic controls without leakiness. Aim 2: To test the hypothesis that circadian disruption in shift workers increases susceptibility to alcohol induced gut leakiness. Non alcoholic shift workers and daytime workers will be prospectively studied before and after moderate alcohol consumption for one week (0.4 g ETOH/kg per day). We hypothesize that shift workers will be more susceptible to alcohol induced gut leakiness than the matched controls on a daytime work schedule. This study will reveal significant new information regarding the interaction of alcohol and circadian biology in ALD pathogenesis that could be used to improve prevention and treatment of ALD. PUBLIC HEALTH RELEVANCE: This study is a collaboration between scientists that study how alcohol damages the gut and scientists who study our body's natural rhythms (waking/sleeping, eating, etc.) called circadian rhythms. The proposed study uses human subjects to look for the first time at the relationships between circadian rhythms and alcohol consumption in the intestine. In this study we will answer two questions: 1) Does chronically drinking alcohol upset circadian rhythms (especially in the intestine) and thus result in intestinal injury that promotes disease? 2) Are patients who work nights at increased risk for damage in the intestine by acutely drinking alcohol (red wine). Answering these questions will result in a significant new understanding of the relationship between alcohol and circadian rhythms and could result in new preventative and therapeutic treatments for alcohol induced disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Aminoglycosides are broad-spectrum antibiotics used for treatment of serious bacterial infections, including the deadly tuberculosis and those accompanying AIDS, cystic fibrosis, and cancer. The emergence of pathogens resistant to these drugs represents a major threat to public health and underscores the need for new antimicrobial agents. In Aim 1, we propose to utilize aminoglycoside-modifying enzymes of the aminoglycoside acetyltransferase (AAC) family in conjunction with a newly developed 6'-N-acylation protecting group-free chemical methodology (i) to generate in vitro libraries of new and more potent N-acylated aminoglycoside antibiotics, and (ii) to develop aminoglycoside probes that will serve as baits for identification of novel therapeutic protein targets/pathways for these antibiotics. Our chemoenzymatic and chemical strategies offer an effective solution to the following problems: (i) there are no existing general synthetic methodologies for the creation of N-acylated aminoglycosides, and (ii) there are no efficient methods to chemically modify specific amine groups on an aminoglycoside that contains a series of chemically identical amines. A few existing examples that use solely chemical syntheses are too demanding on research time and cost and are limited to very specific cases. In Aim 2, we propose biochemical and structural studies of the mechanism of action and inhibition of a major determinant of aminoglycoside resistance in extensively drug-resistant strains of M. tuberculosis (XDR-TB). We expect this work to (i) advance the basic understanding of AG resistance of a variety of pathogenic bacteria, including M. tuberculosis, and (ii) provide a potential solution to overcome the aminoglycoside resistance problem in majority of XDR- TB. Relevance to public health: We expect that this work will contribute to the development of novel antibiotics with a potential to combat existing and newly emerging drug-resistant bacteria.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Airway inflammation plays a fundamental role in asthma and several other diseases of the airways. Although airway hyperresponsiveness, edema, and the presence of granulocytes are central features of asthma, their underlying mechanisms and relationship to the development of the disease are unknown. Airway hyperresponsiveness and edema have been observed in human subjects exposed to various inflammatory stimuli such as viral infection, ozone, and allergens. This proposal will use physiological, biochemical, and morphological methods to examine the mechanisms underlying airway smooth muscle hyperresponsiveness and increased airway vascular permeability following exposure to an inhaled inflammatory stimulus. Previous work suggests that both airway hyperresponsiveness and increased vascular permeability are dependent on the presence of extravascular polymorphonuclear leukocytes (PMNs) in the airways. This proposal will first examine which chemoattractants are responsible for bringing PMNs to the site of airway injury. Animals either 1) depleted of complement, 2) treated with an inhibitor of the activation of the fifth component of complement, or 3) congenitally deficient in specific complement components will be examined for increases in airway responsiveness, vascular permeability, and inflammatory cell influx following exposure to an inflammatory stimulus. To examine the role of lipoxygenase products of arachidonic acid metabolism, these same indices of airway inflammation and dysfunction will be examined in guinea pigs treated with inhibitors of lipoxygenase activity or a monoclonal antibody to LTB4. The source and nature of chemotactic factors will also be studied in isolated cells and tissues in vitro. The second portion of this proposal will determine whether various oxygen reduction species produced by PMNs are responsible for the observed increases in airway responsiveness and airway vascular permeability. Guinea pigs will be treated with selective scavengers or promoters of superoxide anion, hydrogen peroxide, and hydroxyl radical to determine which products of activated PMNs may be responsible for the inflammatory airway damage. In summary, the proposed studies may lead to a better understanding and treatment of inflammatory airway diseases by clarifying the role of PMNs in the control of airway responsiveness and vascular permeability.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this program is to understand how epigenetic mechanisms of gene expression contribute to the development and progression of cancer in a cell-lineage specific fashion. As an initial effort, we have focused on the development of small molecules and chemical tools to profile and perturb the activity of lysine acetyltransferases (KATs). These enzymes catalyze lysine acetylation, a widespread protein posttranslational modification involved in the regulation of gene expression, DNA repair, protein stability, and metabolism. To better understand the role of protein acetylation in cancer, we have taken a multi-pronged approach. First, we have developed a method capable of globally profiling cellular KAT activities. This method is being applied to cell-based models to identify KATs whose differential activity regulates acetylation-mediated reprogramming of metabolism and gene expression observed in cancer. Second, we have developed a separation-based assay for the analysis of lysine acetyltransferase (KAT) enzymes. Currently we are working together with colleagues at the National Center for Advanced Translational Science (NCATS) to apply this assay to identify cell-active inhibitors of KATs known to collaborate with oncogenic transcription factors in leukemia. This assay will also be applied to understand the metabolic sensitivity of chromatin-modifying enzymes. These studies provide the basis for the application of similar approaches to diverse classes of epigenomic regulators, and support the discovery of novel biological mechanisms in cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Clostridium difficile infection (CDI) is the primary cause of antibiotic associated diarrhea and the most common nosocomial infection. Due to frequent infection, potential for serious complications, and high (up to 30%) chance of recurrence, CDI treatment costs are estimated at $1-5 billion per year. Once CDI recurs, many patients get into a vicious cycle of antibiotic therapy and relapse. Although Fecal Microbiota Transplants (FMT) have demonstrated effectiveness in treating CDI (~90% success rate), there are also potential risks. The ability of the microbiota to contribute to chronic diseases such as obesity, diabetes, cancer, and cardiovascular disease in animal models demands that the long-term effects of transplanted communities on human health be better understood. This is especially true in children and young adults, where FMT is increasingly being used not only for CDI but also other ailments including inflammatory bowel disease. Treatment of CDI with defined, minimal communities of pure strains is one alternative to FMT that could negate these risks. Thus far, treatment cocktails of 10, 12 or 33 strains have proven effective in limited human trials. Although promising, these studies were limited by their lack of rigorous pre-clinical testig to demonstrate the effectiveness/necessity of all strains and to probe their long-term impacts on human health. The overall goal of this research is to identify human derived, defined microbial communities that are safe and effective for the treatment of recurrent CDI. To this end, we have developed two pre-clinical models of CDI, a human fecal Mini BioReactor Array (MBRA) and humanized microbiota mouse (HMbmouse) model of recurrent disease. The unique strengths of these models are that human-derived strains can be tested against human- derived microbial communities, either in medium-throughput MBRAs or in the context of a host. Aim 1. Develop defined communities to suppress C. difficile invasion in human fecal MBRAs and the HMbmouse model of recurrent C. difficile disease. In the R21 phase, our MBRA model will be used to rapidly screen many combinations of strains to identify minimal communities capable of combatting CDI. Successful communities will be tested in HMbmice to identify those with the highest likelihood for success. Aim 2. Optimization of communities for efficacy against CDI. Aim 3. Assess the safety and long-term effects of defined community transplantation. In the R33 phase, communities will be optimized for effectiveness, safety, and capability of being produced in industrial settings. The potential hazards of strains will be evaluated through genome sequencing/annotation, antibiotic resistance profiling, and long-term association studies with murine models to determine the impact on host health and physiology. Specific impacts on human tissue will be evaluated with human enteroid models. The final products of this research will be one or more defined cocktails of strains with proven effectiveness in treating CDI, acceptable safety profiles, and proven protocols for industrial-scale production.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Knowledge of the principles that govern activity-dependent neural plasticity is integral to understanding both normal and impaired memory function. The overall goal of the studies proposed here is to examine the prerequisite conditions and underlying mechanisms of homosynaptic long-term depression (LTD) at the commissural-CA1 synapse in the hippocampus in vivo. Homosynaptic LTD is a lasting decrease in synaptic transmission that results from activity in the afferent pathway. Research with formal models of learning and memory has shown that synaptic strength must have the capacity to both decrease and increase in a use-dependent manner for successful simulation of these cognitive processes. Research with forebrain synapses maintained in vitro has demonstrated that homosynaptic LTD does occur, that its induction is favored when excitatory synaptic activation occurs during hyperpolarization or attenuated depolarization of the postsynaptic cell, and that its induction requires elevation of postsynaptic calcium levels. Little is known about the induction and expression of homosynaptic LTD in forebrain in vivo. Recent experiments with the commissural-CA1 synapse in the intact hippocampus have shown that robust LTD is induced at that synapse by stimulation of the commissural afferents with pairs of pulses using an interstimulus interval (ISI) that causes inhibition of CA1 pyramidal cell firing evoked by the second pulse of a pair. No LTD develops when the ISI is lengthened and pyramidal cell firing evoked by the second pulse is not inhibited, or when the calcium- permeable N-methyl-D-aspartate (NMDA) receptors is blocked. To gain further insights into the mechanisms by which paired-pulse stimulation induces LTD, the first study proposed here will assess the degree of correlation between paired-pulse inhibition and the induction of LTD. The second study will explore the mechanism that underlies paired-pulse inhibition; a likely candidate is feedforward and recurrent inhibition from local interneurons, because of the nature of the interconnectivity between interneurons and pyramidal cells. The third study will determine whether or not the contribution of this mechanism is required for the induction of LTD by paired-pulse stimulation. To enable analysis of contributing mechanisms at he single-cell level, the fourth study will serve to develop a protocol for studying LTD induced by paired-pulse stimulation in the hippocampal slice preparation. Using this protocol, the final study will examine whether or not the induction of LTD by paired-pulse stimulation requires postsynaptic calcium, as is suggested by the phenomenon's dependence on NMDA receptor activation. Collectively, the findings obtained from these studies will enhance our understanding of the neural processes that underlie learning and memory.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Masonic Medical Research Laboratory (MMRL) was constructed in 1959 to provide a clinical setting for the study of gerontology, a mandate that was abandoned in 1960 in favor of studies in basic biomedical research. Although no clinical studies were ever done the construction and design of the now designated animal facilities reflect their original intent for patient occupancy and for offices. Provisions for animal housing, including appropriate controls for heat and ventilation to meet the needs of advancements in scientific animal care to meet the requirements of federal and state agencies were not made in the original construction. In the intervening 25 years the MMRL has met these by numerous modifications, and restrictions in the capacity to house and use animals, often at the expense of the interests of the investigators using the research facility. These modifications have now reached the limit to correct the deficiencies. The research interacts of the staff and their ability to compete nationally for research funds has been handicapped as a result of these shortcomings. Recent investigator and conceptual changes in areas of their research interest (particularly in microbiology and ageing research) makes it necessary to update and re-equip our animal facility, particularly that part which concerns the rodent areas which not only need better heat and ventilation controls but also for control of their microenvironment. This proposal would provide for a renovation of the heat and ventilation system as well as for equipment and barrier spaces for the housing of the mice and rat population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT This is a competing renewal of a multi-year conference grant that has supported a forum for researchers to pursue collaborative studies of the molecular genetics of obsessive compulsive disorder (OCD). This application is consistent with NIMH initiatives to facilitate meetings in which researchers come together to establish mechanisms for collaborating in a manner that would facilitate the detection of genes predisposing to psychiatric disorders. At the present time, there are approximately 60 researchers who have attended the meetings of this group. These individuals come from 38 different research centers in 13 foreign countries and the United States. OCD is a serious psychiatric condition that can have debilitating affects on both adults and children. The importance of genetic factors in OCD has been demonstrated by twin and family aggregation studies. Furthermore, recent molecular genetic studies have begun to provide evidence that specific genes may play a role in the manifestation of the disorder. Although these preliminary results are encouraging, several considerations suggest that fully clarifying the genetic architecture of OCD will require large samples and collaborative efforts. It is clear from ongoing studies of the genetics of schizophrenia, autism, Gilles de la Tourette's syndrome, ADHD and bipolar disorder that are consistent with statistical considerations regarding the power to locate genes for complex disorders, that samples collected by individual investigators may not be sufficiently large to provide significant evidence for genetic linkage. Furthermore, once linkage has been demonstrated, even larger samples sizes are needed to identify and characterize susceptibility genes. Thus, it is clear that coordinated collaboration is needed to share findings and pool samples. Cooperative research of this type is often difficult. With the support of the Obsessive Compulsive Foundation and this grant from NIMH, this group of investigators is collaborating on the first Genome Wide Association Study of OCD. At the present time, investigators from 19 different sites have agreed to combine their OCD samples. The total sample consists of approximately 4,500 individuals, 1,800 singleton cases and 800 triads consisting of an affected individual and two parents. It is expected that the results of this GWAS will be published before December 2008. Thus, the aim of this proposal is to obtain funding that will allow this group to continue meeting to further establish collaborative working groups focused on different aspects of the genetics of OCD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is an investigational program project aimed at a better understanding of important biological, biochemical and pharmacological factors influencing the growth of selected human solid tumors and their management by chemotherapy. The research program will focus on the cytokinetics of human solid tumors of the female genital tract, skin (melanoma), head and neck, and lung. Cytokinetics will be analyzed in relation to designing chemotherapy programs with cell cycle specific and cell cycle non-specific drugs for clinical use in patients. Animal studies with athymic nude mice heterogeneously transplanted with these human tumors will be investigated in parallel for cytokinetics and chemotherapy models. Specific studies will be performed regarding the pharmacologic effects of cytotoxic drugs on cytokinetics and drug-directed biochemical pathways in these human tumors in vivo and in vitro. Cell sorting and flow microfluorometry techniques will be used in parallel with classical radioautography in the study of kinetics of patient tumors both in the patient and in the athymic mouse. Utilizing the data obtained from these various investigational approaches, the designed chemotherapy programs will be clinically tested in patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Enzymes containing mononuclear non-heme iron sites catalyze a diverse array of reactions that are significant to medicine and to the environment. This proposal describes plans to study three representatives of the largest, but perhaps least well understood, grouping of these enzymes: The a-ketoglutarate (aKG)-dependent dioxygenase superfamily. TauD catalyzes the hydroxylation of taunne and other sulfonates as a first step in their metabolism. TfdA carries out similar chemistry during catabolism of the herbicide 2,4-dichiorophenoxyacetic acid (2,4-D). Phytanoyl-CoA hydroxylase, a new research direction for this laboratory, is required for utilization of C-3 branched fatty acids; deficiencies of this human enzyme lead to Refsum disease, rhizomelic chondrodysplasia punctata, and ZeHweger syndrome. The specific aims include: (1) Characterize the enzyme mechanism of TauD and TfdA by examining the properties of catalytic intermediates and analyzing the effects of site-directed mutagenesis. (2) Examine the biogenesis of the tyrosyl radical, hydroxy-tryptophan, and histidyl-trihydroxyphenylalanine found in TauD and identify the structures & synthesis of modifications present in TfdA. (3) Obtain high-resolution three-dimensional protein structures of TauD, TfdA, and their variants in their various states. (4) Explore the metallocenter properties of phytanoyl-CoA hydroxylase using the recombinant human enzyme and/or a. more tractable microbial model system. Of particular interest will be studies to test a new mechanism for this enzyme superfamily. Specifically, we propose that these enzymes possess catalytically essential tyrosine residues that are used to resonance stabilize Fe(IV)=O as Fe(III)-OH/tyrosine radical states.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Simple, easy-to-use, low-cost nucleic acid amplification tests (NAATs) are not available for diagnosis of infectious diseases in low-resource settings (LRS). Challenges to easy adoption or modification of existing NAAT technologies for LRS use include time-consuming sample preparation;cold chain requirements for reagent storage;and lack of instrumentation, electricity, and training at the point of care. Unfortunately, there are many infectious diseases rampant in LRS where the lack of appropriate NAATs is a critical barrier to timely diagnosis and treatment. PATH proposes a collaborative effort with the CDC that simultaneously merges scientific and technological capabilities with this clinical need to develop a calcium oxide (CaO)-heated DNA amplification kit that obviates the requirements for electricity and instrumentation at the point of care. PATH has demonstrated the use of CaO for heat and a proprietary engineered phase change material to stabilize an assay mixture within a narrow temperature range suitable for isothermal amplification. The CDC has developed a novel combination of lysis buffer;loop mediated isothermal amplification (LAMP) reaction and fluorescently-labeled primers to enable a simplified, extraction-free and lysis-free workflow. These recent innovations can be combined with a simple two-chamber, high-containment reaction tube prefilled with lyophilized LAMP mixture to create the first electricity-free, instrument-free, and easy-to-use, low-cost NAAT kit. We envision multiple NAAT kits customized and validated for specific strains of infectious diseases such as tuberculosis, malaria, and HIV that will advance evidence-based medical practice at the point of care in LRS. We will develop, integrate, and validate components of this NAAT kit with the sensitivity of PCR, the simplicity of a strip test, and sufficient stability for storage out of the cold chain for long periods of time. Aims 1 and 2 below describe development of the kit components. Aim 3 describes the integration of these components into a stand-alone kit designed for low-infrastructure use. Finally, Aim 4 proposes the laboratory validation of this kit using HIV-1, a highly relevant pathogen afflicting many populations in LRS. \"Aim 1: Specify and optimize the properties of the electricity-free incubator active materials using the existing incubator prototype. \"Aim 2: Optimize CDC's innovations to LAMP HIV-1 assay, establish dry reagent formulations, and define sample. \"Aim 3: Design and fabricate optimized kit hardware and consumables: the isothermal incubator and reaction containment tube. \"Aim 4: Validate the electricity-free and instrument-free assay kit created in Aims 1-3. PUBLIC HEALTH RELEVANCE: We propose to integrate two innovations to create and validate the first electricity-free, instrument-free, easy-to-use, low-cost, nucleic acid amplification test kit for diagnosis of infectious diseases in low-resource settings. PATH has demonstrated the use of calcium oxide and a proprietary engineered phase change material to stabilize the temperature of an assay mixture within a narrow range suitable for isothermal amplification. The Centers for Disease Control and Prevention have developed a novel combination of lysis buffer, loop mediated isothermal amplification (LAMP) reaction and fluorescently-labeled primers to enable a simplified, extraction-free and lysis-free workflow. These two technologies will combine with a simple, two-chamber, disposable, high containment reaction tube prefilled with lyophilized LAMP mixture.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This competitive renewal will build upon our previous work on the role of chemokines and their receptors in airway inflammation. Children that had experienced severe responses to RSV infections often progress into developing long-term pulmonary problems. In addition to pediatric populations, recent evidence has indicated that there is an unknown and relatively unexplored relationship to pulmonary disease in adult populations, including those with asthma and COPD. This renewal application will focus on the role of specific chemokine receptors and their ligands in RSV infection as well as the effects of RSV on exacerbation of cockroach allergen induced disease. Our hypothesis for this proposal is that RSV infection causes airways disease via the activation of CD8+ T cell responses dependent upon CCR1-mediated mechanisms, whereas resolution of disease relies upon the activation of CxCR3-mediated mechanisms. We have designed experiments using 3 specific aims to test our hypothesis and to identify the mechanisms of disease progression. These specific aims include: I. To determine what role CCR1+ T lymphocytes have on RSV-induced disease and in exacerbation of allergic airway disease; II. To establish the mechanism of CxCR3+ and its ligands in the immune response leading to the resolution of RSV-induced disease, and III. To identify the differential role of chemokines for DC subset, pDC vs. cDC, trafficking to the lungs and activation leading to altered pulmonary responses. Our studies will examine both a primary RSV-induced response as well as RSV-induced exacerbation of allergic airway disease. Determining the mechanisms that drive the early responses to RSV and mediate or alleviate severe disease will offer an excellent opportunity to target the early manifestations that have long-term detrimental effects in children, and possibly aid in attenuating progression into severe pulmonary disease. Our models have now been well characterized and allow our studies to address the cell populations involved and the relevant mechanisms that drive the detrimental responses. We will extend our hypothesis to include that CCR1+ CD8 T cells are a significant source of Th2 cytokines, especially IL-13, that lead to exacerbated allergic airway disease. The mechanism of the recruitment of CCR1+ CD8 T cells will center on the induced expression of CCR1 ligands, especially CCL5, within the airways of RSV-infected hosts. We have now also provided novel data that has identified that CxCR3-mediated mechanisms induce a critical anti-viral response via recruitment and activation of important innate cells especially plasmacytoid dendritic cells. The use of cellular transfer experiments with specific animals deficient in targeted molecules will enhance our ability to define the particular cellular mechanisms in vivo during a complex immune response. These mechanisms may be similar to those that are involved in infants, where RSV-infected children often progress into having long-term pulmonary problems and in asthmatics for exacerbated disease. Project Narrative: The coordinated production of chemokines during pulmonary inflammation leads to the recruitment of various leukocytes into the lung interstitium and airway. Identifying chemokine mediators as well as the relevant receptor during allergic and viral disease may be important for identifying therapeutics targets for treating chronic airway disease. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The equilibrium between stability and regeneration is critical to maintaining a functional vasculature. In the mature circulatory system, the endothelium provides critical barrier and regulatory functions by controlling fluid, nutrient and cellular transport between intravascular and extravascular compartments. Vascular injury induces an inflammatory response that stimulates the release of cytokines and growth factors. These factors weaken endothelial cell-cell junctions, stimulate migration and proliferation, and pave the way for remodeling and regeneration. Thus, there is constant tug-of-war within the vascular endothelium between signals that maintain vascular homeostasis/stability and signals that incite regeneration/instability. The central hypothesis for this competitive renewal is that Slit-Robo4 is an endogenous ligand receptor pathway that drives the balance toward homeostasis/stability. If this model is correct, then Robo4 signal might oppose the destabilizing influences of injury, ischemia, and inflammation mediated by a variety of angiogenic and inflammatory cytokines. Specific Aim 1: Elucidate the mechanism of Robo4 signaling. Specific Aim 2: Determine whether Slit-Robo4 is a broad platform for blunting the vascular response to cytokines. At the conclusion of our studies, we hope our contribution will be to demonstrate that Robo4 is a broad vascular stabilization program and to define its downstream signaling cascade. PUBLIC HEALTH RELEVANCE: At the conclusion of our studies, we hope our contribution will be to demonstrate that Robo4 is a broad vascular stabilization program and to define its downstream signaling cascade. In pursuing these objectives we hope to demonstrate that activating this vascular stability program will reduce pathologic endothelial hyperpermeability in a variety of ischemic and inflammatory diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/ Abstract Training with robot-applied forces and virtual reality technologies has potential to restore motor functions in stroke rehabilitation, yet the optimal dosage of robotic therapy remains unclear. A fundamental principle in the motor learning literature is that neuroplasticity can be stimulated using distortions to the visual or mechanical environment where exaggerated mistakes (augmented error-feedback or EA) can accelerate and increase learning. However, robotic training using the error-augmentation methods have achieved only modest gains over conventional therapy. One explanation could be that stroke patients exhibit heterogeneous motor deficits and thus require more focused and individualized robotic rehabilitation. We propose a novel model-based approach that explores joint human-system performance by modeling dynamics for error-guided motor learning and recommends the optimal dosage schedule of EA to best enhance motor skill acquisition and retention. This proposal explores computational models of how various forms of EA affect motor learning and evaluates how the optimal EA dosage could be customized to healthy individuals and to chronic stroke patients. Research indicates that training specificity (i.e., training on the tasks you actually want to improve) is important to recovery of activities of daily living (ADL). Here we also test whether customized visual feedback based on optimal EA dosage can recover functional skills in 3D for stroke survivors. This work utilizes a data-driven modeling method in order to control and customize learning of novel motor skills using robotic technologies. This proposal will contribute to neuroscience knowledge by uncovering relationships between motor learning and error-feedback modulation. This proposal will improve engineering methods to optimize motor skill acquisition for the users operating intelligent-machines such as robotic tools, robot guided neuro-rehabilitation and prosthetics/assistive technologies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Multi-photon microscopy is rapidly becoming one of the most versatile imaging modalities for biomedical research. The ability to image intact tissues to depths of over 500 microns constitutes one of the most groundbreaking developments in in-vivo functional imaging in recent years. At the Martinos Center for Biomedical Imaging, Massachusetts General Hospital (MGH), we strive to develop and utilize breakthrough approaches and our continuing success relies on the integration of new and emerging technologies into our research facility. New modalities enhance our ability to validate and interpret data from our existing imaging resources which include functional and high-resolution magnetic resonance imaging (fMRI and MRI), magnetoencephalography (MEG), positron emission tomography (PET) and functional optical imaging. We are proposing to purchase a state-of-the-art multi-photon microscopy system, with fluorescence lifetime imaging capabilities. The configuration of the microscope platform chosen is optimized for rapid in-vivo imaging, as well as for a spectrum of other microscopy functions. Two-photon microscopy provides unrivalled depth sensitivity and resolution in intact tissue, allowing structure and function to be imaged in- vivo on scales as small as a single dendrite. Fluorescence lifetime imaging (FLIM) exploits both intrinsic and exogenous contrast, and provides dramatic improvement to fluorescence resonance energy transfer (FRET) imaging, which can be used to reveal not only biomarkers, but also interactions between individual proteins. The new microscopy system will be an important resource for currently funded medical imaging research initiatives. Major projects include studies of: cortical neurovascular coupling using calcium sensitive dyes and dynamic blood flow microscopy, the morphological and functional effects of cocaine on the brain, mechanisms of cell death in stroke, and development of molecular probes for cancer and diabetes research. The system will allow versatile imaging of the structure and function of in-vivo, ex-vivo and in-vitro tissues. With the new multi-photon microscopy system and our existing imaging facilities under one roof, we can feasibly perform in-vivo experiments using multiple medical imaging modalities in succession or even in parallel. This system will be an invaluable resource for the many interdisciplinary research projects within the Martinos center, in the surrounding MGH community, and for many others in the greater Boston area. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Molecular Profiling to Predict Response to Treatment (MP2PRT) initiative will implement the Blue-Ribbon Panel?s recommendation by leveraging biospecimens collected from completed trials (with available outcome results) that were conducted by the NCI National Clinical Trials Network (NCTN) Program and/or the NCI Community Oncology Research Program (NCORP) and stored in their associated biospecimen repositories. Appropriate research projects solicited from the NCTN/NCORP Groups with archived biospecimens will be selected for molecular characterization under an NCI-designated, multidisciplinary review process with support from this Task Order. The Task Order will support an integrated approach to the molecular characterization of the archived biospecimens (e.g., tumor tissue/normal tissue, whole blood, plasma, serum) from the selected NCTN or NCORP trials, the comprehensive analysis of the molecular characterization data with trial outcome data including predictive and prognostic modeling, and the provision of data sharing of results by depositing the results in the NCI Genomic Data Commons (as well as having the appropriate NCTN/NCORP Group deposit the clinical trial data in the NCTN/NCORP Data Archive, if appropriate).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Collaborative HIV Prevention Research in Minority Communities (CRMC) program, an NIMH and NICHD funded training program, has the dual purpose of 1) increasing the amount of innovative and scientifically excellent HIV prevention research being conducted in minority communities, and 2) increasing the number of funded investigators from underrepresented minority groups. Since 1997 the program has mentored a diverse group of 37 scientists who have been awarded $23 million in extramural research funding to date. In addition to the productivity of individual participants, the program has created a network of scientists conducting innovative HIV prevention research in minority communities. This network has led to the development and successful funding of several collaborative grants involving program participants and faculty. This application proposes a 2-day conference to bring together program participants, prospective participants, faculty and representatives of funding agencies. The conference will provide a structured environment to disseminate innovative research, promote the development of mentoring and collaborative research relationships, and to explore the current state of science and community practice regarding HIV prevention in disproportionately impacted minority communities. The Specific Aims of this R13 conference grant are: 1)To plan and convene a conference for past and present program participants, past and present program faculty and representative of federal, state and private sector agencies who are funding and setting policy regarding minority-focused HIV prevention research; 2) To evaluate the impact of the conference in promoting networking, collaborative research and mentoring relationships and in affecting funding and policy decisions among conference attendees; 3) To create a list of recommendations regarding research and funding priorities for HIV prevention in communities of color, and; 4)To disseminate the conference presentations and the conference-generated recommendations regarding research and funding priorities for HIV prevention in communities of color to a variety of stakeholders including researchers, community service providers and public and private agencies involved in funding and policy decisions regarding HIV prevention research. Minority communities are disproportionately impacted by HIV and AIDS. At the same time, minority researchers are underrepresented among those receiving federal funding for research in this critical area. The Collaborative HIV Prevention in Minority Communities (CRMC) program has been funded since 1997 to 1) increase the amount of innovative and scientifically excellent HIV prevention with minority communities and 2) to increase the number of funded investigators from underrepresented minority groups. To date we have trained 37 scientists who have gone on to conduct research in this area and to create a productive national network. This conference will bring together former and current program participants and faculty and representatives of funding agencies. This historic conference will produce a list of recommendations for future research and funding that will be widely distributed to academics, community practitioners, funders and policy makers. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Non-aromatic polyketide natural products of marine origin are often characterized by both significant structural complexity and extraordinary biological activities. Because these exciting compounds are not typically available in any meaningful quantities from natural sources, total chemical synthesis is the only means by which sufficient amounts of material may be accessed for the full biological/preclinical evaluation of these compounds. No natural product better exemplifies this class of compound than spongistatin 1. This extraordinarily complex and precious marine natural product has an average IC50 value against the NCI panel of 60 human cancer cell lines of 0.12 pM. The ultimate goal of this proposal is to adapt spongistatin 1 for use in an antibody-drug conjugate (ADC) construct, by way of the design, synthesis and evaluation of a series of analogs of spongistatin 1 to identify appropriate linker sites for bioconjugation and to identify a significanly structurally simplified analog that retains the sub-nanomolar potency of the natural product. Our focus on an ADC approach derives mainly from two considerations: 1) this approach requires far less drug material than conventional approaches, rendering the synthesis of the kinds of amounts required for full clinical evaluation a significantly more realistic proposition, and 2) th low pM potency of spongistatin 1 renders it an ideal candidate for use in an ADC, as so little drug material makes it to the target that extraordinary potency is required for any meaningful clinical efficacy. In order to achieve these goals, we will continue to develop synthetic methods for the synthesis of polyketide natural products that are characterized by unprecedented levels of step-economy, efficiency, and scalability to continue to push the frontiers of efficiency in the chemical synthesis. We will then apply these methods to the development of a synthesis of spongistatin 1 that may easily be adapted for use in the preparation of the designed analogs. Finally, we will identify and synthesize significant quantities of the most significantly structuraly simplified compound equipped with a linker that retains the low pM potency of spongistatin 1.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Perceptual decisions are based on sensory information encoded by populations of neurons over short periods. To identify the most important aspects of the population code for guiding behavior, I varied visual attention, which improves perception of an attended location or feature. The amount of information encoded in a population of neurons is limited by both the variability of individual cortical neurons and by correlated variability that is shared across the population (noise correlations). I found that attentional changes in shared variability are likely the major contributor to the resulting behavioral improvement. These results suggest that studies of single neurons that ignore noise correlations miss perhaps the most crucial aspect of the way that responses of populations of neurons guide behavior, so it is necessary to monitor the activity of populations of neurons rather than record one neuron at a time. The goal of this proposal is to use computational methods and multielectrode recordings in awake primates to understand the mechanism by which inter-neuronal correlations arise and the extent to which they can flexibly adapt to task demands. In my previous experiments I found that attentional modulation of firing rates and noise correlations is linked: the neurons that show the biggest firing rate changes (typically increases in multiplicative gain) show the biggest decreases in correlation. I will use the mentored phase of this award to investigate the origin and flexibility of noise correlations by creating a computational model to test whether a mechanism thought to underlie gain changes in single neurons in many sensory, motor and cognitive processes (sensory normalization) could also cause modulation of noise correlations (Aim 1). The result that modulation of correlations accompanies all gain changes would imply that correlation changes are a major factor in most cortical computations. My goal in the independent phase will be to experimentally investigate how flexible and adaptive correlation changes can be and the impact of correlations on communication between cortical areas. Noise correlations can either severely limit or improve the information available in a neuronal population, depending on the way that neuronal responses are combined. In the task I used in my previous experiments, noise correlations limited population sensitivity, and attention adaptively decreased correlations (as predicted by the normalization hypothesis). In Aim 2, I will use a task in which noise correlations improve, rather than limit, performance to determine whether attention can adaptively increase correlations. In Aim 3, I will investigate the origin of noise correlations by measuring correlations between different cortical areas and varying attention to determine whether correlations depend on the strength of shared functional inputs. Collectively, these studies will have implications for the impact of correlations on population coding and the way sensory information is transmitted from area to area and used to guide behavior. PUBLIC HEALTH RELEVANCE: Many neurological diseases including depression, schizophrenia, and attention deficit hyperactivity disorder (ADHD) are thought to involve networks of cortical neurons, so understanding the way that information is encoded and read out by neuronal populations will be critical for diagnosing and developing drug therapies to treat these diseases. The projects in this proposal will elucidate general mechanisms for population coding, and will in particular enhance our understanding of the neuronal mechanisms underlying attention. Deficits of attention such as ADHD are thought to affect as many as 5% of the children in the United States, and better understanding of the basic neuronal mechanisms related to attention is needed for guiding assessment, diagnosis, and treatment of attentional deficits.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will investigate basic mechanisms of bioelectric information as produced by the actions of sodium channel proteins of excitable cell membranes. We will focus our attention on the voltage-sensing domain of the sodium channel found in human skeletal muscle fibers. Mutations in this domain are responsible for a number of inherited diseases, called sodium channelopathies, and include muscle myotonia and periodic paralysis. In this work we will investigate the molecular means by which the segments of this domain, S1 to S4, interact to control basic sodium channel functions of activation, or opening, and fast inactivation, during which the channel is unable to respond to changes in membrane potential. Our hypotheses target putative interactions of negatively charged amino acids in segments S1 to S3, so-called countercharges, with positively charged amino acids in the segment S4. We will use voltage clamp electrophysiology to test the effects of mutations that reverse the charge of negatively, or positively charged amino acids. These charge-reversing mutations will be compared for effects on activation and for two forms of fast inactivation. Our goal is to identify countercharge interaction with the S4 segment of a given sodium channel domain, that determines a specific function of this asymmetric channel. To do this we will quantify the effects of all significant mutations on activation parameters using the IFM / QQQ inactivation deficient background, and using gating currents to directly test voltage sensor movement. Comparison of charge immobilization and its remobilization will allow a similar quantifiable measure of S1-S3 interaction with S4 segments during two forms of fast inactivation, and during recovery. Finally, we will build models of the voltage sensor domains, insert our mutations in these models, and then run computer simulations of the models in response to the change in membrane potential that elicits their typical function in muscle fibers. Our studies will further our understanding of the molecular basis of voltage-sensitivity in sodium channels and provide a foundation for studies on dysfunction produced by channelopathy mutations of muscle fibers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The University of Chicago has had a major commitment to both laboratory and clinical cancer research since 1930. As part of the clinical translation of that research, the University of Chicago and its Cancer Research Center joined the Cancer and Leukemia Group B (CALGB) in 1985, and has subsequently strongly supported the CALGB both scientifically and clinically. Numerous protocols, committees, and programs are lead by Chicago faculty. Over the past five years the University of Chicago has increased its accrual from 184 patient points to 294 patient points, and in 1996 27 percent of the main member's patient accrual was African American. These accomplishments occurred by disciplined activity at the main member and also by supporting seven dedicated affiliated institutions in northern Illinois, Michigan and Indiana. Chicago is in the process of adding three new affiliates in the next year and remains very involved with the training and quality control of all its affiliate institutions. The goals of this application are: (1) increase the patient accrual to 300-350 patients/year; (2) to lead and assist CALGB scientific activities in the disease-related committees of respiratory (Drs. Vokes and Olak), prostate (Drs. Vogelzang, Steinberg and Vijaykumar), breast (Dr. Fleming), leukemia (Dr. Larson) and GI (Dr. Mani); (3) to lead and assist and participate in the CALGB committees such as psycho/oncology (Marcy List, Ph.D.), pharmacology and experimental therapeutics (Dr. Ratain), transplantation (Dr. Williams), genetics (Dr. Olopade), AIDS related malignancies (Dr. Liebowitz), surgical oncology (Dr. Michelassi) and pathology (Dr. Vardiman); (4) to encourage Chicago faculty to be protocol chairs for future protocols; and (5) to actively assist and participate in the CALGB committees of audit, minority issues, oncology nursing, cancer control, and radiotherapy. The University of Chicago proposes to accomplish these goals by the following methods: (1) to increase accrual from the main member with the assistance of new energetic physicians in colorectal, breast, surgery, leukemia and prostate cancer and by generating new ideas for phase I, II and III protocols; (2) to maintain the high accrual from the affiliate hospitals and increasing accrual by adding new qualified affiliates; (3) to maintain the strong leadership roles of Drs. Vogelzang, Vokes, Larson, Schilsky, Ratain, Williams, Olopade, List, Vardiman and Michelassi within the CALGB; (4) to recruit new young investigators to CALGB leadership roles (Drs. Manni, Leibowitz and Daugherty); especially those with a specific laboratory expertise which can be correlated with clinical treatment or outcome; and (5) to provide volunteers for numerous CALGB administrative committees. The University of Chicago and its affiliates remain firmly committed to serving all members of their respective communities, especially serving the needs of women and minorities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Section on Organelle Biology investigates the global principles underlying secretory membrane trafficking, sorting and compartmentalization within eukaryotic cells. Live cell imaging of green fluorescent protein (GFP) fusion proteins in combination with photobleaching and photoactivation techniques are being used to investigate the subcellular localization, mobility, transport routes and binding interactions of a variety of proteins with important roles in the organization and regulation of membrane traffic and compartmentalization. Quantitative measurements of these protein characteristics are used in kinetic modeling and simulation experiments in order to test mechanistic hypotheses related to protein and organelle dynamics. Among the topics currently under study include: growth and maintenance of endoplasmic reticulum (ER) and Golgi morphology in mammalian cells and in developing Drosophila embryos; the mechanism(s) of secretory protein transport into and out of the Golgi apparatus; membrane binding/dissociation kinetics of trafficking machinery and its regulation; the generation and maintenance of cell polarity; and, organelle breakdown and reassembly during mitosis. We have also recently developed a photoactivatable GFP, whose mechanism of photoactivation is currently being investigated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our broad objectives are to explore new reaction chemistry of three classes of cobalt complexes (propargylium)Co2(CO)6-nLn+BF4-(1), (propargylium)Co4(CO)10BF4(2), and (diene)Co(CO)3-nLn(3) in order to establish their patterns of reactivity with selected nucleophilic coupling partners, especially with respect to stereo- and regiocontrol, and to develop applications of this chemistry in the synthesis of natural products and other biologically active compounds. Studies of the propargylium complexes 1 (L=CO) derived from acetylenic acetals and aldehydes will be directed towards developing new approaches to diastero- and enantioselective coupling reactions with nucleophiles. Utilization of such reactions in the synthesis of chiral building blocks containing 2-5 adjacent stereocenters (precursors to carbohydrate antibiotics) and for the synthesis of macrocycles will be examined. Studies of the novel chiral bimetallic complexes 1b,c will be directed towards examining their structures and assessing their potential for directing stereoselective (and possibly stereospecific) coupling reactions with nucleophiles. A modest effort aimed at the preparation, characterization, and reactivity screening of the unknown cluster derivatives 2 is planned in order to determine if their anticipated unique structural and electronic features will provide new, improved opportunities for stereocontrolled coupling reactions an d a future basis for cluster-based stereoselective reactions in general. Our second major emphasis will be directed towards expanding the synthetically useful chemistry of the diene complexes 3. Primary objectives include: 1) deepening our understanding of the factors controlling the regioselectivity of nucleophilic attack on 3 (and the intermediate allyl-complexes); 2) expansion of the scope of annelative bisnucleophilic additions and illustration of their application in the synthesis of prostacyclin and heterocyclic derivatives; 3) preparation and utilization of chiral, non- racemic 3 in enantioselective syntheses, including carbohydrate derivatives; and 4) development of new reactivity patterns of organometallic complexes as well as valuable new synthetic tools for the stereoselective synthesis of complex and biologically active substances.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neuroblastoma is a common childhood tumor, and approximately 40% of patients have aggressive metastatic disease (stage 4) when diagnosed. With improved therapy, survival has increased to 35-40%. Problem: It is not possible at diagnosis to predict which patients will be long-term progression-free survivors (PFS) and which will succumb to disease. Hypothesis: Molecular \"signatures\" that are derived from microarray analyses of tumor RNA and DNA will define subgroups that have either excellent or poor outcomes. We discovered a 55 gene signature from expression profiles of MYCN gene non-amplified tumors that identifies patients with 79% and 16% PFS. Overall goal: Continue developing therapeutically relevant genomic classifiers for these patients. Specific aims: 1) Determine if RNA expression profiles predict PFS. 2) Determine if DNA signatures based upon loss of heterozygosity and copy number abnormalities predict PFS and if combining DNA and RNA signatures improves accuracy of prediction. Research Design: MYCN amplified and non-amplified tumors will be analyzed as separate groups because they are clinically and biologically distinct. Tumors are available from the Children's Oncology Group and other collaborators with annotation and clinical follow-up. Signatures derived from RNA and DNA microarrays will be used to build validated molecular classifiers that predict the likelihood of PFS. RNA: Initially, expression profiling with Human Exon (HuEx) and standard HG 133 microarrays will be compared to determine if HuEx signatures have similar or better accuracy in predicting PFS. The optimal platform will be used to discover signatures with approximately 337 tumors. Clinically applicable TaqMan(r) Low Density Arrays (TLDA) will be designed for microarray signature genes and tested on the same RNAs to validate their predictive ability. Finally, an independent external set of approximately 210 tumors will be tested with TLDA assays to confirm their validity. DNA: DNA from the same specimens originally used for RNA studies along with paired normal cells will be tested with high density 500K SNP arrays to determine if DNA and DNA + RNA signatures predict PFS. If so, clinically applicable TaqMan(r) DNA assays will be designed from SNP microarray results and tested using the same DNAs. Last, the external validation set of tumors will be tested with TaqMan. DNA assays to confirm their clinical value alone or with TLDA RNA assays. Summary: These are the first and currently only studies aimed at defining subgroups among clinically defined high-risk stage 4 patients. Prediction of long-term PFS using genomic classifiers will facilitate assignment of treatment and development of more effective therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Molecular Cytogenetics Core was involved in the following projects: 1. Detection of IgH translocations in lymphoma (DLBCL) cell lines (collaboration with Dr. L. Staudt). 2. Delineation of complex chromosomal rearrangements and identification of IgH translocation partners by FISH mapping in multiple myeloma cell lines, comprehensive metaphase FISH analyses on advanced multiple myeloma tumors and cell lines (collaboration with Dr. M. Kuehl). 3. Delineation of chromosomal rearrangements produced in clones of monocytic leukemia cell line U937 and ovarian carcinoma cell line OVCAR-8 by RAG- and I-SceI- induced breaks (collaboration with Dr. P.Aplan). 4. Detection and delineation of chromosomal rearrangements (by SKY and FISH) generated by a single targeted DNA double strand break in H2AX deficient mouse cell lines (collaboration with Dr. Aplan). 5. Spectral karyotyping of a set of multiple myeloma cell lines and short term cultures, comparison of FISH and SKY data (collaboration with Dr. M. Kuehl and Dr. A. Protopopov). 6. Delineation of complex chromosomal rearrangements in a collection of mesothelioma cell lines (collaboration with Dr. F.Kaye, Dr. G. Klorin and Dr. O Glebov). 7. Pilot study: translocation between TMPRSS2 and ERG loci in prostate cancer cell lines (collaboration with Dr.L.Cao). 8. Pilot study: verification of flow sorted probes by FISH (collaboration with G. Stone) 9. Determination of ongoing rates of structural and numerical chromosomal instability in selected cancer cell lines using SKY and interphase FISH. Cytokinesis-blocking assay is implemented and tested on cancer cell lines to detect outcomes of asymmetrical mitoses (non-disjunction, chromosome or fragment loss, multipolar mitoses, dicentric chromosomes)(collaboration with Dr. G. Klorin and Dr. M.Kuehl)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to identify and characterize plasma membrane anchorage sites for the cytoskeleton in cultured fibroblasts. The approach will use HAHS, a new crosslinking reagent, which transfers a radioactive moiety from an initially labelled protein to other molecules within reach of a reactive group. Neighboring molecules can then be identified simply and unambiguously by their acquired radioactivity. Two complementary methods will be used. In one, inside-out plasma membranes will be purified by letting cells phagocytose specific ligand-coated beads, then reisolating the beads; then HAHS-labelled actin, fodrin and vinculin will be reconstituted onto the membranes, photolyzed, and analyzed by electrophoresis to identify their sites of anchorage. In the other, labelled proteins will be microinjected into living cells, allowed to incorporate into cytoskeletal structures, then photolyzed and analyzed. Experiments will be done with normal vs. transformed cells, well spread vs. suspended, and rapidly dividing vs. growth arrested cells to identify proteins involved in oncogenic transformation, anchorage to the substratum and growth control. The interaction of cells with fibronectin will also be studied by labelling the fibronectin 11.5 k cell-binding fragment with HAHS, and crosslinking it to the cell surface. Once identified, the location, distribution, regulation, function and structure of these proteins will be investigated. Particularly interesting proteins will be isolated and their structure and function studied further.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this project is to define the catalytic properties of the [unreadable]-subunit of the voltage-sensitive potassium channel (Kv). The Kv channels regulate cell excitability and have been shown to play an important role in oxygen-sensing, volume regulation, memory, and learning. The Kv[unreadable] proteins are ancillary proteins that associate with the cytoplasmic domain of Kv channels, but no clear physiological function has been assigned to these proteins. Structural and sequence analyses show that Kv[unreadable] proteins are members of the aldo-keto reductase superfamily. Our central hypothesis is that the Kv[unreadable] proteins catalyze the reduction of endogenous carbonyls whereby they impart redox sensitivity to Kv currents. Such regulation may be an important feature of oxygen-sensing or metabolic dependence of Kv currents. To test this hypothesis we will measure the catalytic efficiency of Kv[unreadable] proteins with endogenous substrate series consisting of prostaglandins, steroids, metabolites of neurotransmitters and lipid peroxidation products. We will also test whether assembly of Kva-[unreadable] complexes enhance enzymatic activity of Kv[unreadable] and determine the sequence and rate-limiting step in its catalytic cycle (Aim 1). To determine whether the endogenous carbonyls utilized by Kv[unreadable] in biochemical experiments, are also functionally active, we will examine the ability of the most efficient series to alter the voltage-sensitivity and the kinetics of Kva-[unreadable] channels expressed in COS-7 cells (Aim 2). Results of these experiments will allow us to distinguish whether nucleotide binding itself, catalytic cycle, or binding of ligands which can serve as pharmacologic agents modulate Kv inactivation. Taken together, the findings of this study will provide a better understanding of the catalytic and ligand binding properties of Kv[unreadable] and form the basis of a more in-depth project to examine the in vivo role of Kv[unreadable] catalysis and its putative role in regulating surface excitability, oxygen-sensing, or encoding memory. Results of these studies could also form the basis of developing pharmacological modulators of Kv[unreadable]-mediated changes in Kv current. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In order to understand mechanisms by which unit-copy bacterial plasmids are stably inherited, we seek to identify and characterize functions of the host that are directly implicated in the maintenance processes. In the period of this report our recently established laboratory has succeeded in resolving contradictory evidence concerning the participation of the dnaA protein of Escherichia coli in maintaining the 90 kb P1 plasmid. It has previously been found that a well-characterized plasmid replicon isolated from P1 carries two copies of a dnaA protein binding site, but appears unaffected in its replication by a mutant allele of dnaA that does not satisfy the dnaA requirement of the E. coli origin of DNA replication. We report here that this P1 replicon cannot do wihout dnaA protein entirely. On the other hand, a presumably different P1 plasmid replicon, which remains to be isolated and characterized, can integratively suppress a null mutation in the dnaA gene. We suggest that P1 carries both dnaA dependent and dnaA-independent plasmid replicons. The biological significance of the pressence of replicons with differing host-function requirements in so simple an organism as P1 remains to be determined.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Botulinum neurotoxins (BoNTs), from which there are seven serotypes are Gram-positive bacteria. The clinical signature of BoNTs is peripheral neuromuscular blockade and flaccid paralysis, which depending on the serotype can last for months. BoNTs are the most toxic proteins known to man and have been classified by the Centers for Disease Control and Prevention (CDC) as one of the six highest-risk biothreat agents. Despite their toxicity and high potential as a bioterrorist weapon, BoNTs are widely used in medical and cosmetic procedures (i.e., Botox). However, even under a controlled environment unwanted side effects have been reported causing complications for patients and in some cases severe life- threatening disorders. Currently, a botulinum heptavalent antitoxin (BAT) is the only medical intervention for BoNT poisoning and this has limited value since it can only neutralize circulating toxin, being useless once cellular poisoning takes place. At a mechanistic level botulinum intoxication proceeds through a series of three steps to produce its neuroparalytic effects: neuronal membrane binding, internalization, and intracellular poisoning. Our objective is to target the intracellular poisoning stage, which result when the toxins' zinc metalloprotease cleaves neuronal cell SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins. Proteolytic damage to the SNARE proteins prevents fusion of synaptic vesicles with the presynaptic membrane blocking the exocytosis of essential neurotransmitters. Botulinum neurotoxin proteases are unique among endopeptidases in that they require long substrates (SNAREs) for catalytic efficiency due to their recognition strategy. Further complicating their inhibition are allosteric patches, termed exosites that both determine specificity and also remodel the catalytic cleft upon substrate binding. The challenges in designing small molecule inhibitors to the BoNT proteases are recognized through the historic efforts against matrix metalloproteinases (MMPs) where the potential pitfalls of targeting the active site of Zn+2 proteases are well documented. We plan to develop selective molecules against BoNT serotype A (BoNT/A), the most toxic of the BoNT proteases, by targeting the enzyme's exosite. By using this approach metalloactive site redundancy as seen amongst this enzyme class will be avoided as will the use of compounds that chelate zinc. Our proposed research will be centered upon our discovery of the first non-peptide/protein exosite inhibitor, chicoric acid. Our chemistry efforts will be directed toward improving potency while building in drug-like properties. This effort will be assisted by NMR structure guided studies to delineate the BoNT/A exosite structural interactions essential for chicoric acid analogue binding. We anticipate that such studies will identify the key interactions required for exosite BoNT/A light chain protease (BoNT/A LC) inhibition, which in turn will allow additional structures to be prepared and tested in a more rational approach.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY The goals of this proposal are to develop in vitro models of adipose tissue that allow a superior hypertrophic growth of adipocytes and facilitate investigation of metabolic stresses and signaling mechanisms during pathological culturing conditions mimicking those of progressing obesity. Existing in vitro adipocyte culture models are not optimal: 2-D monolayer culture does not represent the 3-D adipose morphology, 3-D cell encapsulation models (e.g., hydrogels) restrict the volume of differentiating adipocytes due to compressive stress and limited porosity, and 3-D ?scaffold-free? models (e.g., hanging drop, non-adherent coatings) do not support long term culture due to spheroid loss during media changes. Consequently, the existing in vitro models result in functionally impaired adipocytes that do not reach their full growth potential, seriously limiting the study of how a full range of intracellular triglyceride (fat) deposition affects adipocyte function in the development of obesity. This proposal directly addresses this technical limitation using a novel tissue engineering approach. Specific Aims to prepare physiologically relevant in vitro models of adiposity are to: (1) Create the stable, surface-tethered 3-D spheroid model of adipocyte culture by using an array of copolymers of biocompatible elastin-like polypeptide (ELP) and charged polyelectrolytes (PE) as coating substrates. Here the positively-charged PEs encourage spheroid formation and ELP encourages stable surface-tethering of spheroids. We will systematically investigate the effect of charge content and chemistry on 3-D spheroid organization. Our overarching hypothesis is that we will achieve superior adipocyte maturation and functionality by this surface modification method that achieves 3-D culture without using a cell-size restrictive encapsulation scaffold and achieves long term culture through surface-tethering of spheroids. (2) Define the mechanism of adipogenesis in 3-D spheroid culture and determine the functionally superior model by comparing against 2-D monolayer and 3-D hydrogel cultures. We seek to define the mechanism of enhanced adipogenesis in the context of morphological cues (cell shape through the mmp14 pathway) regulating PPAR-?, a key effector of adipogenesis. (3) Determine the effects of multiple metabolic stresses (fatty acids and TNF-?) on adipocyte phenotype, viability, and function. The stability of 3-D culture atop our ELP-PE coatings allows a substantially longer culture period. This innovation allows us to expose the optimally developing 3-D spheroid cultures to nutritionally relevant fatty acids at physiological levels. Finally, by comparing the functional and genome-wide responses of the metabolically stressed 3-D spheroids to those of primary adipocytes from obese animal (mice) and human donors, we will recapitulate the effects of metabolic stresses predominant in progressing obesity. We expect the functionally superior 3-D spheroid model to be a clinically relevant in vitro adipocyte model with the potential to invent novel therapeutics by examining drug and nutrient treatments on an in vivo-like mature cell population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project summary This application addresses the urgent need for more and better evidence on the appropriate selection, implementation and use of electronic health (eHealth) systems to support the care of HIV patients in developing countries. eHealth system including electronic medical records (EMRs) are potentially powerful instruments to strengthen health systems, to improve access to and quality of care, and reduce costs. While these technologies show great promise in countries whose health systems face severe financial, infrastructural, technical and human resource constraints, key questions remain for decision makers: (1) How can eHealth facilitate scale up of health initiatives in resource poor environments? (2) How can such systems be more effectively developed, and implemented to ensure high quality information for clinical and programmatic use? (3) Have the large investments in eHealth in resource-poor environments demonstrated clinical impacts such as improving HIV patient outcomes? (4) What are the short term and long term costs of implementing such systems and how can their sustainability be improved? While eHealth is proliferating in resource poor environments, often for HIV care supported by large investments from agencies including PEPFAR, data on their implementation, clinical impact and costs is very sparse. Countries like Rwanda and Kenya whose eHealth plans call for national rollouts of EMRs critically need evidence from rigorous, and generalizable studies. The core issue being addressed in this study is: What is the benefit of using an eHealth system to collect and manage clinical data at clinics that are often in remote sites with limited staff, and infrastructure? It builds on a critical opportunity to assess an open source EMR system, OpenMRS, that is being adapted by a technical team from the Rwandan MOH, and rolled out to several hundred clinics. With 11 sites implemented to date, this is the ideal time to perform a randomized controlled trial with 50 or more sites randomized to receive the system early or late. We will address three related questions: how to optimize systems and ensure they are stable, usable and used, what are the impacts on clinical processes and outcomes, and what are the costs of deploying and sustaining such systems? Clinical indicators will include mean CD4 count and viral load, activities such as ordering of laboratory tests, and rate of loss to follow up of pre-ART patients. The results should inform policy and care in many countries for HIV and other diseases. We will also create a training course in eHealth evaluation at the National University of Rwanda School of Public Health, training evaluators to take a lead in designing and carrying out studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Glutamate activates ionotropic glutamate receptors (iGluRs) that mediate fast synaptic transmission, and metabotropic glutamate receptors (mGluRs) that modulate cell excitability. The iGluRs gate extracellular sodium and calcium entry into the cell, while the Group I mGluRs (1, 5) lead to the release of calcium from intracellular stores. Brain injury such as stroke or ischemia leads to increased extracellular glutamate, uncontrolled activation of iGluRs and mGluRs, and the toxic accumulation of intracellular calcium that is an essential initiator of cell death. Thus therapeutic strategies for the treatment of brain ischemia have focused on the use of iGluR and Group I mGluR antagonists. While iGluR antagonists are neuroprotective in modeled ischemia studies, likely due to inhibition of intracellular calcium accumulation, these compounds have failed in clinical trials. Similarly, Group I mGluR antagonists are neuroprotective in modeled ischemia studies, likely due to inhibition of intracellular calcium accumulation, yet delivery of most mGluR-selective compounds to the brain is difficult. We have begun to explore alternative neuroprotective strategies for brain ischemia using proteomics to identify novel proteins or peptides that modulate Group I mGluR signaling via interactions at the pharmacologically accessible extracellular amino terminal domain. Our first proteomic studies focused on the Group I mGluR subtype, mGluRS, and revealed a novel interaction with a recently cloned extracellular protein that promotes cell survival, ADNP (activity-dependent neuroprotective protein). ADNP contains an eight amino acid peptide sequence (NAPVSIPQ; NAP) that was shown to be the smallest active element of ADNP that can induce neuroprotection. Preclinical experiments show that NAP has potent neuroprotective, memory enhancing and neurotrophic properties. However, the mechanisms that underlie neuroprotection by NAP or ADNP are not known. Our preliminary data suggest that one mechanism of neuroprotection by NAP and ADNP is to regulate Group I mGluR signaling. The research studies proposed herein are important because 1) they begin to delineate the mechanisms of neuroprotection by NAP, an exogenous peptide, and ADNP, an endogenous protein, 2) they provide evidence for the novel regulation of mGluR signaling by peptide or protein interactions at the extracellular domain, and 3) they offer new approaches for therapeutic intervention in brain ischemia. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: (Applicant's Description) The capacity of memory T-cells to traffic preferentially to distinct epithelial tissues has only recently been appreciated. Evidence is accumulating that a skin homing memory T-cell population, identified by E/P selectin ligand expression (e.g., CLA/PSGL-1) and absence of integrin alpha 4 beta 7, mediates antigen specific effector responses in skin, while a reciprocal memory T-cell subset (integrin alpha 4 beta 7 positive, E/P selecting ligand negative) mediates effector responses in the gastrointestinal tract (gut). Importantly, these memory T-cells use these cell surface molecules to initiate the tethering and rolling (under physiologic flow conditions) on endothelial cells in post capillary venules that are required for their ultimate extravasation in both normal and inflamed skin or gut. Such polarized populations of memory cells are thought to emerge during the naive to memory T-cell transition in the specialized lymphoid micro-environment of lymph nodes draining each epithelial surface, respectively. GVHD is a nearly inevitable complication of allogeneic bone marrow transplantation. Much of the morbidity from GVHD derives from the involvement of the two major epithelial interfaces with the environment: the skin and the gut. The present proposal tests the hypothesis that distinct subsets of effector T-cells with similar antigenic specificities mediate skin and gut GVHC. A second testable hypothesis is that these polarized subsets of memory T-cell s emerge in part from transferred graft naive T-cells that are activated by alloantigen in lymph node draining either gut or skin, respectively. A final hypothesis is that the strength of the conditioning regimen influences this process in two ways. First, tissue injury and attendant inflammation from primary cytokine release leads to enhanced egress of dendritic cells form blood, which in to carry damaged tissue antigen via afferent lymphatics to draining lymph nodes. Second, the increased expression of adhesion molecules on endothelial cells in inflamed skin and gut enhance the efficiency of memory T-cell extravasation. Each of these hypotheses will be directly testing using a well-characterized murine model allogeneic BMT (MHC identical). The lethal GVHD that results is dependent on T-cells in the allografted population. Both antibodies and blocking molecules specific for memory and naive is dependent on T-cell homing ligand/receptor pairs, as well as transgenic mice deficient in one or more genes critical to these adhesive interactions (e.g., E/P selectin), FucTVll, beta 7 integrin), will be used to ask precise and directed questions about the cellular and molecular requirements for the evolution of skin and gut specific GVHD, respectively. The capacity to selectively modify GVHD involving one or both of these tissues, while not suppressing ostensibly beneficial alloimmune responses (e.g., graft versus leukemia or global immunocompetence is the longterm therapeutic goal underlying this study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of our Hub for Latin America (HLA) is to generate sustained progress toward Community Health Care for individuals with Severe Mental Disorders (CHC-SMD). We envision CHC-SMD as an integrated system of primary care and secondary mental health services that are delivered as close as possible to the communities that use them;that are accessible to all people with severe mental disorders and their families;that promote full community integration;and that engage the affected individuals, their families, and their communities in shaping the health services offered to them. The Research Component proposes to test a pilot regional randomized controlled trial (RCT) of a Critical Time Intervention-Task Shifting (CTI-TS) intervention. CTI-TS is a task shifting integration, and, at the system level, strengthens the connections between mental health and primary care clinics. The proposed pilots are a necessary preparatory step toward a full-scale regional RCT. We wili conduct pilot RCTs (N=40) of CTI-TS at three sites (total N = 120): Rio de Janeiro, Santiago, and a third HLA site selected during year 1. The study complements our Capacity Building Component, because HLA sites will gain experience with R(5TS, with task shifting interventions, and with coordination of research among sites. The aims of the Research Component then are : (a) to complete a pilot RCT of CTI-TS at three HLA sites;(b) to analyze the data collected to infomi the design of a full scale regional RCT;and (c) to share results and challenges encountered with all relevant stakeholders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. OBJECTIVE: Our objective was to examine the effects of exercise training (EXS) on quality of life (QoL) in highly active antiretroviral therapy (HAART)-treated HIV-positive (HIV+) subjects with body fat redistribution (BFR) in Rwanda. METHODS: The effects of a randomised controlled trial of EXS on QoL were measured using World Health Organisation Quality of Life (WHOQOL)-BREF in HIV+ subjects with BFR randomised to EXS (n = 50;BFR + EXS) or no exercise training (n = 50;BFR + noEXS).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The activity of tryptophan hydroxylase, the rate-limiting enzyme in the synthesis of the neurotransmitter serotonin, was determined in extracts of various rat brain structures after treatments which increase or decrease the availability of serotonin in the synaptic cleft. Drugs promoting the rapid release of serotonin (e.g., p-chloroamphetamine (PCA) and p-chloromethamphetamine (PCMA) produce an acute reduction (onset by 2-3 hours) in the activity of tryptophan hydroxylase in brain areas rich in serotonin nerve endings. In preliminary experiments, inhibition of the serotonin releasing effects of PCMA by chlorimpramine blocked the reduction in enzyme activity. While it is usually hypothesized that PCA itself is taken up into the presynaptic bouton where it inhibits tryptophan hydroxylase, this project will examine the possibility that 5-HT release into functional activity (i.e., onto the receptor) initiates a sequence of neuronal and biochemical events which has as its end point, a rapid compensatory decrease in serotonin synthesis at the rat limiting hydroxylase step. BIBLIOGRAPHIC REFERENCES: Lovenberg, W.: Enzyme changes as an index of neurotoxin specificity. Ann. N.Y. Acad. Sci. In press (1977). Baumgarten, H.G., Klemm, H.P., Lachenmayer, L., Bjorklund, A., Lovenberg, W. and Schlossberger, H.G.: Mode and mechanism of action of neurotoxic indoleamines: A review and progress report. Ann. N.Y. Acad. Sci., In press (1977).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The bioinstrumentation laboratory facility consists of a centralized instrument laboratory, and the electron microscopy facility. The centralized instrument laboratory contains major equipment for support of the various research projects in biomedical sciences. This laboratory houses the HPLC, UV-Vis spectrophotometer, gas chromatographs, electrophoresis apparatus, and other basic instruments for conducting research in the Health Research Center. The spacious laboratory will house all major equipment that will be acquired through RCMI funding.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This study will verify the absence of microscopic evidence of hepatic damage even in the absence of abnormalities on liver function tests one year following distal gastric bypass.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goals of this study are to determine whether 10 candidate genes influence susceptibility to SLE and to two specific disease manifestations: lupus nephritis and the antiphospholipid antibody (APLA) syndrome. The GCRC will be utilized to obtain blood samples for APLA testing, as well as serum and DNA samples for long-term storage (and future use). In addition, during the GCRC visit SLE patients and family members will complete the enrollment process including provision of buccal smear samples and questionnaire information.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. DEVELOPMENT AND APPLICATIONS OF NEW PROBES AND METHODS FOR CORRELATED LIGHT AND ELECTRON MICROSCOPY, LIVE IMAGING, AND FUNCTIONAL ANALYSIS In this subproject, our efforts are directed at (1) extending and generalizing the tetracysteine/biarsenical labeling technology already shown to be a powerful and versatile method for correlated LM and EM, and (2) developing other complementary and unique labeling methods. We are exploring approaches to increase the sensitivity of the tetracysteine/biarsenical tag while minimizing its impact on the host protein normal behavior. We extend the application of biarsenical labeling to chemically fixed tissues and propose the utilization of these methods to explore mesoscale structure and function in complex tissues in virally transduced animals and in transgenic models. In collaboration with the laboratory of Roger Y. Tsien, we will develop a new genetically encoded small Singlet Oxygen Generator (miniSOG) specifically for improved photooxidation for EM labeling. The miniSOG tag can be used alone or in combination with other molecular tags, and shows its applicability to correlated LM/EM studies. In addition, we continue to develop a repository and database of our successful molecular-tagged proteins as a \"cell biology toolkit\" to facilitate new projects using these probes and expand their availability and usefulness to a larger segment of the scientific community. The ultimate objective of this core project is to expand our toolset to label one or more proteins in cells and tissues with minimal perturbation of the native specimen ultrastructure for multi-scale-correlated LM and EM through further development and refinement of tools and methods like the tetracysteine system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary The long-term objectives of the proposed research are to deepen our understanding of the epidemiology of pediatric epilepsy in a geographic area with a racially and economically diverse population. Children in DC are among the most racially and socioeconomically diverse populations in the country, with approximately 66% Black, 18% white, 11% Hispanic and 5% some other race. A total of 29% of all DC children live below the poverty level, and almost 50% of families are run by a single head of household. DC presents a rich population for study to formulate policies and programs to address the needs of a vulnerable and potentially underserved population. To meet our objectives, the following specific aims will be addressed: (1) determine utilization of healthcare resources, responsiveness of health systems and obstacles to care experienced by children with epilepsy and their families, (2) characterize risk factors, outcomes and comorbidities related to prevalent and new onset pediatric epilepsy, and (3) reassess population-based estimates of epilepsy incidence and prevalence over a 2-year period among children living in DC. In support of Aims 1 and 2, we propose creating a cohort of at least 500 DC children with epilepsy. We will use a self-administered questionnaire to obtain data from the parents of the pediatric cases. This questionnaire will ask questions about personal history of epilepsy, types of care sought and received, comorbidities such as cardiac disease and sleep disorders, family history of epilepsy, and the family and social environment. To supplement these data, medical records will be abstracted for treatment schedules, medication use, epilepsy phenotype, and other health measures. Follow- up data collection is expected to occur every 6 months. In support of Aim 3, we will determine whether previously obtained prevalence and incidence estimates for pediatric epilepsy are accurate and stable over time. This will be performed by using an existing health screening tool, the Behavioral Risk Factor Surveillance Survey, used to obtain data on public health in DC and across the country. We will also examine DC-mandated public school immunization records, maintained by CNMC, that also include an indicator for epilepsy, to compare estimates of epilepsy in school-age children with that found in the telephone survey. Children with epilepsy are a socially and educationally vulnerable group. Characterizing epilepsy in the DC school age population will play a role in better defining the resources needed to provide essential health and educational services to an underserved population. As a community with such rich diversity, DC is an ideal geographical area to implement population-based studies of epilepsy. Results from our study may contribute to recommendations for changes in health policy and guidelines used by primary care providers and neurologists for treatment of children with epilepsy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of the proposed research is to extend our information regarding hexose metabolism during aging in Dictyostelium discoideum. Evidence for a pentose phosphate pathway will be investigated, in order to expose unique characteristics of the metabolic network under analysis and to test specific predictions regarding the effects of perturbing this system with exogenous glucose. Those enzymes directly involved in pentose phosphate metabolism will be fractionated, to determine the reaction mechanism and relevant kinetic parameters. The pentose phosphate pathway will be incorporated into the present kinetic model for carbohydrate metabolism in this organism.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Inosine monophosphate dehydrogenase (IMPDH) is the rate limiting enzyme in the synthesis of guanine nucleotides from IMP and has been demonstrated to be a critical determinant of overall guanine nucleotide levels in most cell types. Inhibitors of this enzyme have demonstrated usefulness as immunosuppressive agents and may be of use in the chemotherapy of malignancy. The two isoenzymes of IMDH, I and II, are highly similar in amino acid composition and function, but appear to be regulated differently at the molecular level. The overall objective of this proposal is to elucidate the mechanisms regulating the expression of these two isoenzymes and to develop new approaches to the modulation the expression of the two gene products. In addition, we propose to investigate in greater detail the effects of guanine nucleotide depletion on DNA synthesis and cell cycle progression. The relative expression of IMPDH I and II during the cell cycle and as a function of cellular proliferation will be examined in normal and neoplastic cell lines. Genomic clones containing the regulatory regions for the two genes have been isolated and will be characterized in terms of transcriptional elements important for the constitutive expression of the two enzymes, as well as factors responsible for their up-regulation with pharmacologic stimuli and with cell proliferation. To achieve these goals, the structures of the two genes will be determined, DNase I hypersensitive sites will be mapped to detect potential regulatory regions within the genes, and the promoters will be analyzed for similarities and differences in structure and transcription factor binding sites. We will analyze the two expressed and purified IMPDH proteins for differences in structure that could be exploited in the development of new inhibitors. In addition, antisense oligonucleotides will be used to determine whether the expression of IMPDH I and II genes can be differentially inhibited and whether such inhibition affects T lymphocyte proliferation. Finally, we will attempt to gain new insights into the role of guanine nucleotide depletion in inhibiting cellular proliferation by examining the effects of IMPDH inhibitors on factors influencing the progression of cells from 01 to s phase in the cell cycle, including the expression of p53, the retinoblastoma suppressor protein (pRb), and enzymes directly required for initiating DNA synthesis. We anticipate that the results of these experiments will shed further light on both the mechanisms generating increased levels of guanine nucleotides in proliferating cells and the sequelae of reductions in guanine nucleotides for cell growth. These data should be of use in developing new approaches to immunosuppression and chemotherapy based on more selective manipulation of IMPDH expression and guanine nucleotide synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent advances in molecular biology have shown that the products of the cellular homologs of retroviral transforming genes are important regulators of cell growth and behavior in normal as well as malignant cells. We propose to study the role of these products in vascular endothelial cells. We will initially focus upon c-sis, the cellular homolog of the simian sarcoma virus transforming gene (v-sis), which encodes for one chain of platelet-derived growth factor (PDGF) because endothelial cells in culture synthesize PDGF-like mitogens. In the context of vessel wall, c-sis expression has generated intense interest since PDGF is known to be mitogenic for smooth muscle cells and chemotactic for leukocytes, key elements in the generation of an atherosclerotic plaque. First we will determine the complete structure of c-sis mRNA cloned from human endothelium and elaborate the regulatory mechanisms utilized by the endothelial cell in controlling the production of the c-sis product. These experiments are based upon our isolation and sequence analysis of a 2.1 kb v-sis homologous cDNA clone from human endothelial cells, and provide the first opportunity to study the structure of sis as transcribed by a normal (untransformed) cell. We propose to extend our studies to regulatory sequences associated with the c-sis gene. We also plan to identify and characterize mRNA's encoding the less cis-homologous chain of PDGF. Additionally we will use expression systems for a structure/function analysis of both c-sis regulatory sequences and the c-sis encoded polypeptide. Understanding the structure of PDGF produced by endothelial cells may permit construction of PDGF analogs which may be clinically useful in altering atherogenic or neovascular processes. In a second project, we will investigate the role of other tranforming gene homologs in the control of endothelial cell growth by using pseudotyped amphotropic retroviruses to introduce exogenous retroviral transforming genes into cultured endothelial cells. We propose to correlate modifications of endothelial cell behavior with expression of both exogeneous and endogenous cellular tranforming gene homologs. By comparing the retrovirus modified cells with normal endothelial cells we will infer mechanisms for normal growth and phenotypic conrol, generating specific hypotheses to be tested.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project is designed to evaluate the developmental patterns of mammary glands of mice fed diets containing different levels of saturated and unsaturated fatty acids. In addition, we wished to establish an atrophied mammary gland similar to that of immature animals but in an \"adult\" animal. The subsequent impact of manipulation of hormones and dietary lipid intake on mammary tumor incidence is also to be evaluated. Studies include: 1) Examination of the morphology of mammary glands of mice at various ages after being fed EFA-deficient and control diets; 2) evaluation of the effects of various periods of EFA-deficiency on formation of hyperplastic alveolar nodules (HAN) and ultimately on mammary tumor formation in mice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this proposal is to gain insight into mechanisms of cell-mediated immunity and unresponsiveness to intracellular pathogens in humans. Leprosy provides an extraordinary model to investigate these immunoregulatory processes. First, the disease primarily affects skin and is therefore accessible for study, proving a unique opportunity for investigating the nature of the local inflammatory response in human infectious disease. Secondly, the disease represents a spectrum of clinical manifestations that correlate with the pattern of cytokines produced at the site of infection. Patients with the resistant tuberculoid form (T-lep) express type 1 cytokines in lesions; whereas patients with progressive lepromatous form (L-lep) manifest type 2 cytokines in lesions. Our approach has been to study immune responses in lesions at the site of disease activity. Such studies have provided insight that cannot be obtained from the study of the peripheral blood from these patients. We hypothesize that the outcome of the host response to M. leprae is determined by the nature of the innate and adaptive immune response in lesions. We now propose to: 1) elucidate the gene expression profiles associated with resistance versus susceptibility in the human immune response to an intracellular pathogen, 2) determine the role of leukocyte immunoglobulin-like receptor (LIR) family members in contributing to immune unresponsiveness in leprosy; and, 3) elucidate the mechanism by which T-cell recognition of a major microbial antigen contributes to host defense in leprosy. The studies we propose are intended to provide a comprehensive analysis of the immune response in leprosy, comparing host responses in patients that are resistant to infection vs. susceptible to progressive disease. Such studies should provide new insights into mechanisms of immunoregulation in humans, and, we would hope, would provide the ability to predict disease outcome and also lead to the development of new immunomodulatory treatments for a variety of human infectious diseases. These issues are timely, given the rise in emerging infections and the threat of bioterrorist attacks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Variations in the rates of neuropeptide synthesis and release are to be expected for all neuroendocrine systems. Nerve terminal compartmentation (differential distribution of different aged granules with respect to release) will interact with the kinetics of precursor biosynthesis, processing and release to yield complex variations in the mixture of biologically active peptides secreted. These variations will not be predictable without a knowledge of both the compartmentation characteristics of the nerve terminal and the kinetics of synthesis, processing and release. The experiments proposed here seek a precise definition of these complex properties in order to better understand the possible physiological role of the peptidergic neuron. Two simple nerve terminal preparations will be studied; the neural lobe of the rat pituitary and the sinus gland of the land crab. The major experimental approach will be to use HPLC analysis of the relative proporations of the products of partial proteolysis in the secretory granule to determine the kinetic history or \"age\" of a secretory granule population. The probability of secretory granule release as a function of secretory granule age (compartmentation) will be defined in the neural lobe by determining by HPLC the ratios of the oxytocin neurophysins throughout density gradient granule separations in neural lobes from control, 1, 2, 3 and 5 day dehydrated rats. A similar definition of compartmentation in the sinus gland will be based upon HPLC determination of the partial proteolysis products of the sinus gland H peptide in density gradient separations. Further experiments will define synthesis rates, enzymatic processing rates and the peptide content of release in both systems. Taken together, the data will provide a comprehensive description of factors governing variation in peptide content for two chemically simple neurosecretory systems. The data will then be used to predict variations occurring in more complex systems by mathematical modeling of the effect of compartmentation on variations in relative amounts of released peptides with complex processing situations. The approach is planned to yield an understanding of how the biochemical properties of the secretory neuron are related to its function in the transmission of neural and endocrine information.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our major objective is to demonstrate that heart wall motion abnormalities characterized by early shortening/late lengthening trigger in the epicardium, local inflammation, reactive oxygen species generation, matrix metalloproteinase (MMP) activation and fibrillar collagen degradation leading to adverse chamber remodeling. Wall motion abnormalities secondary to dysynchronous contraction of the left ventricle (LV) as seen in heart failure reduce ejection fraction and promote remodeling. Similar changes occur in animal models of anterior LV wall epicardial pacing whereby adverse remodeling is characterized by thinning at the pacing site and hypertrophy of the posterior wall. Local strain patterns demonstrate early shortening and late systolic lengthening in the early activated areas (anterior wall) and early lengthening and subsequent shortening of the posterior wall. Preliminary data indicates that anterior wall pacing yields localized evidence for epicardial leucocyte (likely neutrophil) mediated inflammation, activation of MMPs, reactive oxygen species generation (ROS) and collagen degradation. We propose that the mechanism responsible for inflammation/injury is the local expression of adhesion molecules in the microvasculature secondary to alterations in coronary venule blood flow patterns. Four aims are proposed. Aim 1 will test the hypothesis that pacing induced early shortening leads to the local expression of adhesion molecules in coronary venules, trapping of leucocytes, generation of ROS, pro-MMP activation and fibrillar collagen degradation in the epicardium. Aim 2 will test the hypothesis that early shortening regions of the LV demonstrate abnormal epicardial systolic tissue volume changes and modified patterns of epicardial venular blood flow. Aim 3 will test the hypothesis that leucocyte generated ROS structurally modifies pro-MMP-9 in vitro and in vivo. Aim 4 will test the hypothesis that long- term LV wall motion abnormalities induce adverse chamber remodeling characterized by epicardial wall thinning in areas associated with early shortening. A variety of cardiac mechanics, intravital microscopy, proteomics, biochemical and histological methods are to be used. Results from the proposed studies should aid in our understanding of how ventricular dysynchrony as seen in patients with heart failure and left bundle branch block leads to the development of adverse chamber remodeling and loss of contractile function. Anticipated outcome from the proposed studies may also aid clinicians in optimizing cardiac resynchronization therapy. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE The proposed research addresses an important clinical problem related to how, when the heart does not contract in unison it develops structural and functional abnormalities that are detrimental in nature. These abnormalities are frequently seen in patients with heart failure and our observations indicate that an underlying inflammatory process may explain these adverse changes. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Abstract The project seeks to understand the function of the cis-regulatory sequences that control animal development, using the embryo of the pomace fly Drosophila melanogaster as a model system. Given the importance of gene regulation in virtually every facet of biology, including many human diseases, it is remarkable how poorly we understand the relationship between the base sequence of the genomic regions that control transcription and their function. Despite a growing catalog of well-characterized regulatory sequences from numerous species, especially D. melanogaster, we still can not reliably recognize regulatory sequences in DNA, determine the expression pattern of a gene from the sequences that surround it, predict the consequences of variation in regulatory sequences, or design regulatory sequences de novo to produce a desired pattern of expression. The early D. melanogaster embryo has long been a model for the study of transcriptional regulation. During the first several hours of its development, the D. melanogaster embryo transforms a small number of crude spatial cues left behind by its mother into intricate spatial patterns of expression of thousands of genes that establish the body plan and tissue identities of the embryo, larvae and adult fly. Technological advances in the last decade have enabled the generation of an increasingly detailed portrait of this regulatory network and the molecular events that underlie it, as well as genome sequences of D. melanogaster and many of its genetic variants and sister species. The central premise of this proposal is that we can infer from these data the molecular logic of gene regulation. In particular, we propose to model three aspects of this system: 1) the manner in which regulatory information is distributed across the D. melanogaster genome, 2) constraints on the evolution of regulatory sequences, and 3) the detailed relationship between DNA sequence and gene expression. Each of these models will reveal not only details of the D. melanogaster regulatory network, but will also illuminate the biophysical and logical principles that unite gene regulation in all animals, including humans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "After a metaanalysis we published in the journal Transfusion suggested the age of stored blood may have a significant effect on survival after transfusion, we intiated a series of pre-clinical studies in our canine model of critical illness. Our initial study showed a significantly worse outcome if critically ill patients were transfused with old (42 day) vs. fresh (7 day) blood. We have also shown that old blood will not worsen a very severely ill or healthy patient but in a patient population with a 50-70% mortality, tranfusion of old blood significantly worsens outcome. We also found the age of blood that becomes harmful is likely between 14 and 28 days. In a therapeutic study, we should that washing stored blood (to wash out any accummilated harmful particles) with saline improves survival when transfusing old stored blood but worsens survival when transfusing washed fresh blood. Ongoing studies will narrow the window when the age of stored blood adversely affects survival and the role of the volume of transfusion. We are also pursuing the role of iron as a mechanism underlying the harmful effects of stored blood. This line of research has resulted in the publication of 5 papers to date.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this project is to use a retrovirus-based mouse model system to understand how inappropriate activation of transcription factors in hematopoietic cells can result in the development of leukemia. We have been studying the myb-ets-containing ME26 virus, which causes a high incidence of leukemia in mice by a novel mechanism involving the inappropriate activation of erythroid-specific genes in hematopoietic precursor cells. We have previously shown that the ME26 viral protein can transactivate the promoter of the erythroid-specific transcription factor, GATA-1, and then cooperate with the GATA-1 protein to transcriptionally activate the erythropoietin receptor and other erythroid-specific genes. This results in the unregulated proliferation of virus-infected cells in the presence of the erythroid hormone erythropoietin. In an attempt to better understand how the viral protein can transactivate the GATA-1 promoter, we engineered and analyzed deletion mutants to determine which sequences in the promoter are crucial for activation by the ME26 viral protein. Our results indicate that sequences in the 3' end of the GATA-1 promoter, which include two CACCC elements, are essential for transactivation by ME26 virus, while other upstream sites contribute to full activation by the virus. Mutation of the CACCC sites abolishes ME26 viral transactivation. The interaction of cell extracts containing ME26 viral protein and the GATA-1 fragment containing the two CACCC elements was examined by electrophoretic mobility shift analysis and the results showed no direct interaction between the two. However, we could detect the ubiquitous transcription factor Sp1 bound to this sequence. These data demonstrate that the CACCC element is necessary for GATA-1 promoter transactivation by ME26 virus and that the viral protein may indirectly transactivate the promoter by binding to Sp1.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The present proposal represents an amalgam of two current graduate training programs at UCSD, one covering the areas of Cell and Molecular Biology, and the other focused on the application of Genetic methods to problems of gene regulation and development. The primary mission of this new combined program is to train graduate students for career as independent research scientists. All entering graduate students complete an intensive, one year core curriculum covering virtually every facet of contemporary biology, with an emphasis on the diverse techniques now available for the characterization of fundamental and conserved cellular processes. During this first year of study, students are exposed to a broad spectrum of research problems through 4 or 5 laboratory \"rotations.\" After successful completion of a rigorous comprehensive examination, and selection of a thesis mentor, each student conducts independent research. Given recent progress in the elucidation of common mechanisms underlying seemingly disparate processes, we place a heavy emphasis on a broad training environment, whereby students engaged in traditionally diverse fields of study are brought together in common symposia, workshops, journal clubs, retreats, and informal tutorials. The proposed training program includes 98 faculty spanning two different departments in the Faculty of Arts and Sciences, the UCSD School of Medicine, and The Salk Institute. Funds are requested to maintain the current level of support (53 graduate students), even though there has been a dramatic expansion in the training program due to the inclusion of Salk faculty in a common graduate curriculum. The proposed training program includes two tiers of organization. First, faculty are organized according to their participation in the formal training of all graduate students, with an emphasis on the traditional disciplines of Genetics, Molecular Biology, Structural Biology & Biochemistry, and Cell Biology. Secondly, the faculty are distributed among four principal fields of contemporary research, including Transcription and Gene Regulation, Signal Transduction, Cell Biology (particularly membrane biogenesis and cellular compartments), and Molecular Evolution.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Selective arteriography (radiographic) of the spinal cord is a diagnostic technique which has proven to be very informative in cases of arteriovenous malformation, tumor, and post-radiation damage of the spinal cord. Radioisotope angiography of the spinal cord offers distinct advantages as a screening method, and in certain types of intraspinal pathology may give information not available by any other diagnostic test.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research intends to design and build an instrument of accurately measure centrally located pathological areas of non-vision, or central scotomas, within the 30 to 60 degrees field of vision. The new device is intended to serve as a supplement to commercially available automatic perimeters, which are known to be inaccurate when retinal defects are confined to the central field of vision. The device used in a novel technique that enables a patient that suffers from a central defect to maintain central fixation, thereby producing measurements of their central and paracentral visual field with unprecedented accuracy. It is anticipated that the device will be able to gather data on patients even when the retinal defect lies in the area required to fixate. The device will consist of both hardware and software, ant the test administered to the patient will be similar to available automatic perimeter, which include computerized algorithms resulting in a rapid test, storage and analysis of data, and a variety of test options. Initial validation of the device will be done through testing a select group of patients with central defects. The measurement results from these patients will be compared with other types of visual field devices used commonly today. It is the intent that the Campimeter be made available to the practitioner, by keeping the cost of the commercial unit inline with comparable ophthalmic instruments. PROPOSED COMMERCIAL APPLICATION: Once complete and developed, the device will be well suited for a clinic or private practice. The inability to measure and map central scotomas has been a long-standing problem with clinicians. Now more than ever, with the aging population and the subsequent increase in macular degeneration, the wide variety of refractive procedures, as well as numerous retinal pathologies, there is a consistent need for the accurate mapping of central scotomas. The ability to measure and map, track changes, and quantify treatment modalities, will be monumental in diagnosing, following and managing macular and retinal diseases. The anticipated moderate cost of the complete instrument will make it available to a wide variety of clinicians, and will not be limited to a research environment. If properly designed, engineered, and tested, the ultimate hope is for wide spread use and early detection and management of central field defects.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The focus of this research project is to elucidate the mechanism of the ring-opening of the azetidine ring. The reactions that will be undertaken will involve the action of a series of meta- and para- substituted phenols on 1-alkylazetidines rand 1-alkyl-2-methylazetidines. The reaction products will be separated and isolated by fractional distillation and high performance liquid chromatography. Characterization of these reaction products will be made by IR, 1H and 13C NMR spectral studies. A second dimension of the project will involve kinetic studies of the ring- opening reactions by a suitable combination of spectral studies. During the previous funding period, research-size quantities of compounds possessing beta-adrenolytic properties (helpful in controlling hypertension, angina pectoris and cardiac arrhythmia) have been prepared by our laboratory using these reactions. During the proposed funding period this project will be aimed at providing a detailed understanding of the mechanism, with the ultimate objective of providing not only a convenient means of preparing new compounds but also new classes of compounds possessing more specificity for the treatment of the organs affected by these diseases--the heart and the blood vessels. The research design will involve: (1) the synthesis of N-substituted azetidines by various methods; (2) isolation of the products by distillation and/or various chromatographic techniques; (3) ring-opening of N-substituted azetidines with substituted phenols; (4) separation, isolation and characterization of the ring opening products; and (5) kinetic studies of the ring opening reactions. Students will participate in every aspect of this project, including preparation of starting materials, performing kinetics, interpreting data from IR and NMR spectra, etc.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Bone marrow transplantation is viewed as a potentially invaluable tool for treating different cancers and immune deficiencies. However, little is known about the immunological functions of most bone marrow cells. This is evident by the fact that bone marrow transplantation is not always fully successful because of host versus graft responses (HVGR) and graft versus host disease (GVHD). The role that bone marrow pre-T cells play in these problems is unclear. In vitro models of GVHD and HVGR using bone marrow cells could greatly improve our understanding of these problems. I have developed an in vitro culture system which demonstrates that murine bone marrow cells can generate high levels of cytotoxic effector T cell activity alloantigens. This response may involve functional differentiation of bone marrow pre-T cells to cytotoxic effector T cells and/or helper T cells. The aim of this proposal is to investigate the cellular basis for this in vitro response and to determine whether or not bone marrow cells can generate in vitro cytotoxic responses against hapten-modified syngeneic cells or against parental hemopoietic histocompatibility (Hh) antigens. The reactive cells involved in these responses will be chracterized according to size, density presence of Fc receptors, adherence properties, and for surface antigens associated with pre-T and splenic T cells. The expression of Thy 1 antigen and the development of cytotoxic T cell activity will be correlated for antigen stimulated Thy 1 negative bone marrow cells. Soluble factor(s) produced by activated T cells will be investigated for a possible role in this differentiation. Informaton obtained from these studies will not only add greatly to our understanding of bone marrow cells but will also provide information necessary for establishing in vitro models of GVHD and HVGR using bone marrow cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is a competitive renewal application for our research program on the role of the Caspase family of proteases, the c-Jun NH2-terminal kinase (JNK), and the phosphatidylserine receptor (PSR) in neuronal apoptosis in brain development and neurological diseases. In the present project, we will extend our research of these signaling pathways with a new emphasis on the mechanism of apoptosis and renewal of neural progenitor cells. Aim 1: We will investigate the mechanism downstream of phosphatidylserine and phosphatidylserine receptor (PS-PSR) recognition leading to the apoptosis of neural progenitor cells. We will express PS on the outer cell surface of neural progenitor cells and examine the signaling events following PS-PSR recognition. In addition, we will test whether the disruption of PS-PSR recognition in the embryonic brain will alter the apoptosis of neural progenitor cells. Aim 2: We will examine the interaction between the JNK and canonical Wnt/beta-catenin signaling in the decision of self-renewal or cell-cycle exit by neural progenitor cells. We will determine whether the canonical Wnt/beta-catenin pathway mediates self-renewal, and whether JNK activation attenuates the canonical Wnt/ beta-catenin signaling to promote differentiation of the daughter cells. Aim 3: We will use a transgenic system to calculate the net increase of adult-generated neurons in the mouse brain. In addition, we will test whether EOF and FGF2 infusion can accelerate adult neurogenesis and attract newly born neurons into stroke damaged brain areas. In summary, the present project is built on our research of JNK, phosphatidylserine receptor (PSR), and canonical Wnt/beta-catenin signaling with an emphasis on the mechanism governing the adjustment of the number of neural progenitor cells (Aim 1), the decision of cell-cycle exit by neural progenitor cells (Aim 2), and the accelerated neurogenesis after brain injury (Aim 3). These studies will enhance our understanding of neural plasticity and functional recovery after brain injury.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Protein sequence homology (i.e., descent from a common ancestral sequence) is perhaps the most widely used tool for annotating the putative functions of genes. Homologous proteins often share functions inherited from the common ancestor, so if the function of one protein has been experimentally determined, the function of its homologues can often, but not always, be inferred to be the same. Homology-based inference allows functional data from experimentally tractable model organisms (such as E. coli, yeast, Drosophila, C. elegans and the mouse) to be applied to other organisms, most notably humans. The past several years have seen a dramatic increase in the amount of structured, computationally accessible data available on the functions of proteins and the genes that encode them, primarily using the Gene Ontology (GO). The most useful of these data have been manually entered (}curated}) by a biologist after reading papers in the scientific literature. The goal of this proposal is to leverage these literature-derived ontology annotations by using them, in a carefully curated and structured manner, as the basis for inferred annotations in other organisms. We will utilize and extend existing software developed in our groups to develop a web-accessible environment for curation of GO terms in the context of evolutionary relationships, and link the data to biological pathway data and data standards. We will integrate the software into current GO term annotation projects, and support a broad data ex- change and dissemination plan across GO and pathway ontology curation efforts and the communities of bio- medical researchers they serve. PUBLIC HEALTH RELEVANCE: The current research project provides a cost-effective, accurate methodology for taking experimentally based information about genes in a wide range of well-studied species, and applying this information to understanding human biology, genetics and disease. The results from this methodology will be made broadly available to both researchers and the public, in formats accessible to both people and computers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Two isozymes of NADP ion-dependent isocitrate dehydrogenase (IDH) are preferentially localized in cytoplasm and mitochondrial, respectively, of various tissues. Genetic variants of the cytoplasmic isozyme have been identified in DBA/2J and in C57BL/6J mice. The purpose of this study is: (1) to provide a simple purification procedure for the isolation of these two isozymes; (2) to compare their structural and biochemical properties, in order to correlate their differential distribution in the tissues or their subcellular localization; (3) to compare biochemically the two cytoplasmic IDH variants from DBA/2J and C57BL/6J mice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite growing interest about osteoporosis in men, there is little data regarding the pathogenesis and risk factors for fracture in men, or effective diagnostic strategies in this population. To address these issues, the ongoing Osteoporosis in Men Study (MrOS) has successfully recruited over 6000 men over age 65 from 6 US clinical centers. MrOS collected extensive baseline measurements of anthropometric and health habits, as well as axial densitometry and lateral lumbosacral spine radiographs. Baseline QCT of the hip and spine, and sex hormones have been measured in subsets. Since the baseline visit, subjects have been followed for the occuence of new fractures, and as of Feburary 2005, 46 hip fractures and a total of 256 non-spine fractures have been reported and centrally adjudicated. Study participants will return to the clinical centers for repeat spine x-rays and densitometry beginning January, 2005, after a mean follow-up of 4.5 years. In this MrOS ancillary study, we plan to use archived serum and urine specimens from baseline and propose a prospective analysis of the relationship between bone turnover and incident vertebral, hip and any non-spine fracture using an efficient nested case-cohort study design. Our sampling scheme will take advantage of the many existing measurements in MrOS, including DXA, QCT and sex hormone levels. In addition, we will analyze the relationship between a new non-invasive biochemical index of bone collagen quality, type 1 collagen isomerization, and incident vertebral, hip and any non-spine fracture. Propective studies among older women suggest that both bone turnover and collage isomerization are independently associated with fracture risk, but such relationships have not been studied in men. Lastly, we plan to study bone turnover and bone loss, and determine which anthropometric, historical and health habit factors are associated with bone turnover. These analyses will address critical questions about the pathogenesis of spine and non-spine fracture in men, and will determine the clinical utility of bone turnover measurements in this population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Survivors of a pediatric malignancy are at risk for serious long-term morbidity and premature mortality related to their treatment. Women who were treated with chest radiation for their childhood cancer have an increased risk of developing breast cancer at a young age. We use the term chest radiation to refer to radiation that includes the following fields: mantle, mediastinal, lung, total body irradiation, or spinal. Models for predicting the absolute risk of breast cancer, such as the well-known Gail model, have been used extensively to advise patients on their individual risk of developing breast cancer and to design prevention trials. The majority of these risk calculators are not immediately applicable to survivors of a previous malignancy who have a risk that is modified by previous treatments. Moreover, there is conflicting evidence as to whether various non-treatment related factors, including the traditional breast cancer risk factors, are associated with the risk of breast cancer in these women. The traditional risk factors include age, age at menarche, age at birth of first live child, number of first-degree relatives with breast cancer, number of previous breast biopsies, and history of atypical hyperplasia. We propose to utilize the unique resources of the North American Childhood Cancer Survivor Study (CCSS) and the Dutch LATe Effect Registry (LATER) cohorts. Our first aim is to develop a risk prediction model that integrates chest radiation dose and volume, treatment-related exposures, the traditional risk factors, and other possible risk factors such as age at menopause, body mass index, the number of years with intact ovarian function after radiotherapy, and use of hormone replacement or oral contraceptive therapy, in order to estimate the individualized absolute risk of breast cancer for women who were treated with chest radiation for a childhood cancer. The model will be developed using the original CCSS cohort in which there are 1,677 female participants who were treated with chest radiation and would be included in this analysis. Of these 1,677 women, 187 women had developed breast cancer by the time of this application. Our second aim is to validate the prediction model on independent data. The validation cohort will consist of about 1225 female participants in the expanded CCSS cohort and 1087 female participants in the Dutch LATER cohort. We estimate that approximately 100 of these women will have breast cancer. The third aim is to create and disseminate a risk calculator that provides computer-assisted risk prediction. Our objective is to present a tool in an easily accessible format and disseminate it for clinical use by physicians and patients. The long-term goals of this project are to provide a means for facilitating conversations between physicians and their patients about appropriate screening and preventive strategies and to help refine screening recommendations for this population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ictacalcin is a calcium-binding protein that is found in olfactory receptor neurons in the catfish. This AREA proposal describes a series of experiments aimed at characterizing the ictacalcin gene and the mechanisms underlying its restricted expression to olfactory neurons. Olfactory signal transduction in vertebrates, mediated through G protein coupled pathways, results eventually in fluctuations of cytosolic calcium in the cilia and soma of receptor neurons. Thus, calcium is thought to be involved in signal generation, adaptation and/or termination of the sensory signal. Ictacalcin belongs to the S100 family of calcium binding proteins, the members of which may play a role in mediating the effects of calcium in a variety of other tissues. Because of its tissue-specific expression in the olfactory epithelium, the regulation of the ictacalcin gene is potentially interesting. In this application, Dr. Porta proposes to isolate the full-length ictacalcin gene and initiate a definition of transcriptional elements in its regulatory regions and of cell type specific transcriptional factors involved in its regulation. In the long-term, the P.I. believes that this work has significance for understanding the regulation of chemosensory transduction and other neuronal processes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Electron spin resonance spectrometry is a widely used technique for the detection of free radicals and paramagnetic centers. There are over 23,000 published biomedical research papers citing this technique, particularly the use of spin labeling and measurement of reactive oxygen species and reactive nitrogen species. However, applications of electron spin resonance also extend into chemistry, biochemistry, physics, radiation dating, and commercial applications in petrochemical exploration and refining, oil condition monitoring, food sterilization and many other areas. Historically, electron spin resonance spectrometers have been very large, very expensive pieces of lab equipment that are therefore not readily accessible to many potential users. What is proposed is the development of a miniature, inexpensive, high sensitivity electron spin resonance spectrometer instrument that will make this powerful analytical technique accessible to a much broader range of users. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE: Free radicals are an important contributor to many disease processes. We propose to make a simple, low cost, portable analysis tool for the measurement of free radicals, particularly in clinical and biomedical research settings. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a revised application for competitive renewal of the NHLBI Short-Term Research Training for Minority Students program which has been in place at the University of Florida since 1999. The purpose of this training program is to offer undergraduate students and students in health professional schools a ten week experience in an active research laboratory, to offer them didactic material covering topics of interest to, and needed by, young scientists. The students recruited into this program will be underrepresented in medical science (under-represented minorities, disabled, or disadvantaged students). The proposed renewal of this training program is planned as a broad- based exposure to research in cardiovascular, pulmonary, and hematologic research in three colleges of the University of Florida Health Science Center. The faculty for this proposed training program spans several departments. We want this program to capture the diversity of interests and approaches to cardiovascular science, to offer a snapshot of professors in three colleges (Medicine, Pharmacy, and Veterinary Medicine) with overlapping research interests but different academic missions, to offer to our trainees a wealth of possibilities in science beyond what are sometimes naive notions of medical practice. Most of the faculty are directly involved in research which is easily identifiable as cardiovascular, pulmonary, or cardiorenal research. The program director is Charles E. Wood, Ph.D., a well-known developmental cardiovascular physiologist who has made fundamental contributions to the understanding of the reflex control of the fetal cardiovascular system. Dr. Wood will direct the relevant activities in the office of the department chairman's office as well as the Office of Minority Affairs (OMA). Selection of trainees is done by a committee consisting of four faculty members, all leading members of the U.F. College of Medicine faculty. Selection and oversight of lectures and training seminars offered during the training program will be done by Dr. Wood in his role as program director. The structure of the program is designed to be an 8-10 week mentored laboratory experience with an overarching weekly lecture series that will is designed to inform students about the structure of academic science and doctoral training programs. The students will also be required to attend a short course in the Responsible Conduct of Research. A plan for evaluation of program effectiveness is proposed that will assess career goals, perceived barriers, knowledge of academic research careers, and knowledge of higher education in research. The evaluation plan will be used as the basis of a report, after 5 years, of effectiveness of this modality of research training for undergraduates. At the end of the program, the trainees are tracked by the OMA. Students are encouraged to apply for a second summer in this program. When appropriate, individual students are encouraged to apply to the various graduate programs at the University of Florida, especially to the Interdisciplinary Program in Biomedical Sciences, but also to Ph.D., M.D., D.V.M., and Pharm.D. programs. We want to make our trainees aware of the power of basic research, especially in the cardiovascular sciences, so that they will be motivated to choose careers in research and to successfully initiate graduate-level training. We will be a success if we improve recruitment and success of under-represented minorities in academic science.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objective of this research is the production of dimer-specific antibodies and the development of immunohistological techniques for screening and identification of high risk, nonxeroderma pigmentosum (XP) individuals especially in families with a higher than normal susceptibility to skin carcinogenesis. Radioimmunoassays (RIAs) will be developed to distinguish between ultraviolet radiation (UVR)-induced thymine-thymine, thymine-cytosine, and cytosine-cytosine dimers. The antibodies specific for pyrimidine dimers will be incorporated into immunohistological procedures that will allow the rates of induction, distribution, and fate of DNA damage to be measured in mouse epithelial cells exposed in vivo to UVR on the presumption that the risk of cutaneous carcinoma in a \"normal\" or non-XP population may be related to the ability to repair damaged DNA. During the first year, we have produced polyclonal antibodies from two rabbits that are specific for UVR-induced DNA damage. Binding of antibodies to irradiated DNA was measurable at antiserum dilutions of 1/10,000. These antisera have been incorporated into an ELISA procedure that is as sensitive as previously reported radioimmune assays for the measurement of UVR-induced DNA damage. (2)", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Development of antiviral drugs for the treatment of human cytomegalovirus (HCMV) infections, particularly in AIDS and other immunosuppressed patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The distinction between healthy aging and pathology is imprecise and is clouded by overlap among normal brain aging markers and neural changes associated with Alzheimer?s disease (AD). Even in the case of beta-amyloid deposition, a key hallmark of AD, 20-30% of healthy older individuals show elevated amyloid, yet it remains unclear if these individuals will necessarily progress to dementia. Therefore, there is significant need for studies designed to investigate novel, lesser-studied mechanisms of brain aging, which may help disambiguate the neural changes that are early indicators of a pathological trajectory vs. signs of normal aging. The present study aims to provide new insight into neurocognitive aging with a multimodal investigation of important and understudied mechanisms of brain aging from molecular changes (iron accumulation and A? deposition) to synaptic changes (neurite complexity) to changes in brain function (BOLD modulation of activation) and their influence on cognition in middle-aged, older and MCI individuals. Critically, we propose to prospectively study healthy aging individuals who systematically vary in risk for AD (cognitively healthy low risk for AD, cognitively healthy elevated risk for AD) alongside individuals with Mild Cognitive Impairment (MCI), who are more likely to be in the early stages of neuropathological development. Through this approach, we aim to help elucidate some of the mechanisms underlying key aspects of brain and cognitive aging in healthy adults and also to distinguish which age-related brain changes may signal preclinical AD vs. non-pathological aging. The first aim proposes to examine brain iron accumulation as an early marker for cognitive and neural decline. We will study the impact of increased brain iron on cognitive performance and fMRI measured brain activation in healthy aging, preclinical AD and MCI. Secondly, we aim to examine the effect of synaptic complexity on cognitive performance and brain activation across risk groups. We will examine the relationship of both iron accumulation and synaptic complexity measures to beta-amyloid accumulation in preclinical aging and MCI to gauge whether these variables may serve as sensitive biomarkers for pathological aging. Finally, we plan to test the hypothesis that differences in neural modulation to cognitive difficulty as measured with fMRI may serve as a biomarker for preclinical AD. Completion of the current study is expected to advance understanding of the mechanisms and predictors of healthy brain aging versus a pathological aging trajectory.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development of bone metastases is one of the most debilitating complications of advanced breast cancer. Patients suffer from severe pain, spontaneous fractures and possible spinal cord compression caused by development of osteolytic lesions. We don't understand how breast cancer cells develop the ability to metastasize and proliferate in bone. As a consequence, therapeutic options at this advanced stage of breast cancer are extremely limited. Breast cancer bone metastatic cells undergo striking changes, expressing genes involved in bone development such as OPN, MMPs, VEGF, all known markers of tumor growth, invasion and metastasis. Importantly, these genes are controlled by the master regulator of bone formation RUNX2, and RUNX2 becomes overexpressed in breast cancer metastatic cells. We identified that the WWOX tumor suppressor protein physically interacts with and represses RUNX2 transcriptional activity. We hypothesize that loss of WWOX expression (a common event in breast cancer) is key for the progression of breast cancer cells towards development of a bone metastatic phenotype. Our current research suggests that loss of WWOX in breast cancer cells leads to unrestrained RUNX2 activity, Osteopontin overexpression and these maybe key determinants for metastasis development. We expect aborting development of bone metastases by restoring function of the WWOX-RUNX2 axis. We have also created a conditional Wwox KO mouse that will allow in vivo study of effects in mammary gland growth, tumor development and metastasis. Therefore we will pursue the following Specific Aims: Aim 1: We will A. determine whether WWOX depletion alters the phenotype of normal mammary epithelial and non-metastatic breast cancer cells and B. determine the effects of restoring WWOX expression in metastatic breast cancer cells. Aim 2: A. To determine whether WWOX restoration suppresses spontaneous bone metastasis and osteolytic bone disease in breast cancer metastatic cells. B. To determine if WWOX depletion induces a metastatic and osteolytic phenotype in non-metastatic breast cancer cells. C. To determine whether WWOX influences the ability of breast cancer cells to crosstalk with the bone microenvironment by affecting osteogenic differentiation. Aim 3: To determine whether the integrity of the WWOX-RUNX2 physical interaction is relevant for impeding bone metastases and associated osteolytic disease. Aim 4: To determine whether targeted Wwox ablation in the mammary gland leads to enhanced mammary tumorigenesis and increases metastasis. The proposed studies will provide novel information on the mechanistic basis of bone metastasis development and identify potential novel targets for therapeutic intervention. PUBLIC HEALTH RELEVANCE: We do not understand how breast cancer cells develop the ability to metastasize and proliferate in bone;therapeutic options at this advanced stage of breast cancer are extremely limited. We hypothesized that loss of expression of the tumor suppressor WWOX in breast cancer cells leads to unrestrained activity of the RUNX2 transcription factor and that this maybe a key determinant of metastasis development. The proposed studies will test this hypothesis, provide novel information on the mechanistic basis of bone metastasis development and identify potential novel targets for therapeutic intervention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The ultimate objective for this program is to develop a hybrid active and passive magnetic bearing based, rotodynamic, continuous flow implantable ventricular assist device. Development and integration of the MiTi hybrid active/passive magnetic bearing with a motor driven Impeller will result in a compact, low power consumption, highly reliable continuous flow blood pump. Key elements of the proposed program include 1) a detailed integrated blood pump and hybrid bearing design; 2) component bearing testing to validate the bearing design; 3) fabrication of up to three prototype pumps; 4) in-vitro testing of the fabricated pump; and 5) preliminary in-vivo studies. In-vitro testing will validate pump and bearing performance, verify that washing flows limit deposition, and verify the lack of hemolysis damage to blood circulated through the pump. In-vivo testing is planned to demonstrate freedom from deposition-related difficulties with the rotodynamic pump. Key features of the blood pump include only one actively controlled magnetic bearing axis for reliability and simplicity; elimination of conventional displacement sensors through the use of self sensing technology to further enhance reliability; large clearance passages surrounding the pump motor and bearing yielding low blood shear stresses (well below the hemolysis damage threshold) and promoting good forward washing flows. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will comprehensively catalogue the molecular alterations present in melanoma stroma in the setting of BRAF targeted therapy. Global transcriptomic, epigenomic and proteomic analyses will generate an unbiased profile in both human and mouse BRAF-mutant melanomas including paired baseline and post-treatment or relapse samples. These integrative and cross-species comparative analyses will define candidate driver stromal alterations that may dictate the response to BRAF targeted therapy. The functional relevance of candidate events will be validated through genetic or pharmacologic perturbation of the targets or their canonical pathways in vitro and in vivo. Further, confirmation of human relevance will be obtained through analysis of a large cohort of annotated human melanoma samples. Beyond discovery and validation, we will elucidate underlying mechanisms with the goal of converting these insights into rational therapeutic combinations designed to neutralize both tumor and stromal targets and avoid/minimize the emergence of BRAFi resistance.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this application is to demonstrate that Interleukin-19 (IL-19), an anti-inflammatory, Th2 interleukin, can drive angiogenesis and improve perfusion of ischemic tissue. IL-19 is a newly described Th2, (T regulatory) anti-inflammatory interleukin which until our work, had been ascribed to be inflammatory cell-specific. We remain the only laboratory to investigate a role for this Interleukin in vascular biology, particularly with respect to EC and VSMC pathophysiology. Both inflammatory and anti- inflammatory cytokines participate in wound healing and neo-vascularization, but the role of anti- inflammatory cytokines in angiogenesis and the cross-talk between these processes remain under characterized. In contrast to our previous work indicating that IL-19 suppresses vascular smooth muscle cells (VSMC) migration and proliferation, we have recently reported the surprising finding that IL-19 has potent pro-angiogenic effects on human endothelial cells (EC). IL-19 is not detected in normal EC but is expressed in EC in capillaries in human angiogenic tissue. IL-19 is mitogenic and chemotactic for EC, promotes cell spreading, and activates MAPK and Rac1. IL-19 promotes microvessel formation in aortic rings, and PECAM1-positive microvessels in vivo. IL-19 can induce angiogenic gene expression in EC. IL-19 effects are independent of VEGF and bFGF, as neither can induce IL-19 expression, and IL-19 cannot induce expression of either. Neutralization of bFGF and VEGF does not affect IL-19 activity, suggesting a novel, Th2-induced pathway to stimulate EC activation and angiogenesis. IL-19 can polarize human macrophage to the M2, wound healing phenotype, and induce angiogenic gene expression in macrophage. IL-19 expression and function is reciprocal to and regulates the pro- inflammatory anti-angiogenic cytokine IL-12. These preliminary and published data have driven the hypothesis that IL-19 is a novel vasculogenic cytokine with multiple effector cells. What needs to be determined is if IL-19 can restore blood flow in ischemic tissue, if the major effector cell is endothelial cells or the M2 macrophage, what soluble factors mediate IL-19 effects, and the molecular mechanisms and mediators of IL-19 differential effects in EC and VSMC. We will determine if absence of IL-19 attenuates, and if over expression of IL-19 promotes angiogenesis and restores blood flow in ischemic tissue, if IL-19 regulation of angiogenesis is facilitated by direct effects on vascular cells, or by macrophage M2 polarization, will identify and characterize IL-19 inducible factors necessary for IL-19- driven angiogenesis in vivo, and test the hypothesis that differential expression of IL-20 receptor subunits account for pleiotropic effects of IL-19 in VSMC compared with EC. This application is potentially paradigm-changing as it will implicate a Th2 interleukin as a novel anti-inflammatory, pro-vasculogenic cytokine expressed by inflamed resident vascular cells to promote neovascularization.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (Applicant's Abstract): The purposes of this research are: (1) to evaluate interpersonal psychotherapy (IPT) as a treatment for postpartum depression; (2) to evaluate the effect of IPT on a) women's adjustment in their relationships with their spouses, children, families, and friends, b) the women's responsiveness to their infants, and c) the infants' affective expression and emotional regulation; (3) to determine the rate of recurrence of depression in successfully treated women over an 18 month period following the end of treatment; and (4) to evaluate a clinical-social model of depression recurrence in successfully treated women. A sample of 104 postpartum depressed women is being randomly assigned to immediate or delayed (a waiting list control) IPT. Treatment will consist of 12 to 16 sessions (over 12 weeks) with a private therapist. Assessment of women before, during and after therapy with several measures of depression and social adjustment will be undertaken. After the treatment trial, all women will be followed for 18 months with assessments of depression and social adjustment occurring every 3 months. Assessments of children will take place at pre-therapy, post-therapy, and at 2 years of age. We expect that women receiving interpersonal psychotherapy will evidence significantly lower levels of depression, higher levels of social adjustment than women in the waiting list control condition, and be more responsive to their infants. We also expect that approximately 40% to 50% of successfully treated women will experience a recurrence of their depression during the 18 month follow-up period. We expect that previous history of depression and other psychiatric disorders will be associated with increased risk of depression recurrence. The ongoing research should prepare for later studies to (a) prevent depression recurrence in women successfully treated for postpartum depression and (b) to improve the social adjustment of young children by reducing their exposure to maternal depression. The final purpose of this proposed research is to evaluate the contribution of maternal depression in the first two years of children's lives to their adjustment at age 2. Finally, we expect that children whose mothers were chronically depressed during their first two years of life will show poorer adjustment at 2 years of age.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Photoreceptor development and maintenance require precisely regulated gene expression. This regulation is controlled by the Gene Regulatory Network (GRN) of photoreceptor transcription factors (TFs) and their target non-coding cis-regulatory DNA elements (CREs). Mutations in either TFs or CREs can cause misregulation, leading to photoreceptor diseases. How mutations in coding sequences cause diseases, such as CRX-linked retinopathies, has been extensively studied, and many such mutations have been identified and can be tested for. However, only a few disease-associated mutations in non-coding DNA regulatory elements are known. One example is X-linked blue-cone monochromacy, a red/green color blindness associated with disruption of the Locus Control Region (LCR), a distal CRE of the Red/Green gene array. Little is known about the location or functional importance of other retinal CREs, presenting a bottleneck for identifying and evaluating disease-associated variants in the largely unexplored non-coding portion (99%) of the genome. Thus, there is an urgent need to catalog functional CREs and understand their sequence logic. To this end, we have identified the subset of CRX bound CREs that are Dependent on the activity of CRX to establish an active chromatin state and promote photoreceptor gene expression. We hypothesize that many of these CREs are essential for rod gene expression, cell fate, and survival, and their sequence features determine how they mediate the commands of CRX and/or other TFs. We propose to test this hypothesis in three specific aims using innovative high-throughput functional genomics approaches. Aim 1 will provide a deep understanding of two previously-identified CREs using loss-of-function studies in mice: By thoroughly characterizing the individual and combined deletions of the two rhodopsin (Rho) enhancers CBR and RER, we will decipher their in vivo roles in regulating endogenous Rho expression, rod cellular function, and health. Aims 2 and 3 will identify the subset of essential retinal CREs using unbiased high-throughput functional tests in mouse retinas: We will first identify activating CREs that are sufficient to enhance the expression of a reporter gene driven by a minimal rod gene promoter using massively parallel reporter assays (MPRA) (Aim 2); We will then identify active CREs that are necessary for rod gene expression and cell identity using a CRISPR-Cas9 genomic deletion screen (Aim 3). Finally, we will combine the information gained from these two complementary approaches to decipher the sequence logic mediating CRE regulatory function. Ultimately, we will identify the most functionally important photoreceptor CREs, gain a deep understanding of their genetic and epigenetic grammar, and be able to predict the effects of specific disruptions to these CREs on rod gene expression and cell fate. The findings will significantly advance our understanding of normal and pathogenic photoreceptor gene expression. It will also provide new resources for genetic screens and functional testing of disease mutations, and new insights into disease phenotype variability.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (adapted from the applicant's description): Experiments proposed here test hypothesis that alterations in the structure, function, and regulation of cardiac troponin I (cTnI) and cTnT contribute to the evolution and end-stage pathology of heart failure. The long term objective is know how myofilament remodeling and covalent modulation play a role in the evolution of hypertrophy/failure in human hearts. Aims #1, #2, and #3 address the questions: Can altered tension and economy of myofilaments from human hearts in end-stage failure be rescued by replacing components of the Tn complex with recombinant proteins? What is the nature of the thin filament alterations in human heart failure? Are there changes in protein phosphorylation of specific sites on cTnI or isoform population of cTnT? How do these changes alter force and ATPase rate in reconstituted preparations? Aim #4 addresses the question: What is the specific role of protein kinase C (PKC) sites on cTnI in the development of hypertrophy and failure in response to hemodynamic stress of pressure overload? The approach to these objectives involves the use of procedures for exchanging thin filament proteins in the intact force generating lattice, and for isolating Tn complex from small heart tissue samples. Recombinant proteins are prepared in an unphosphorylated state and specifically phosphorylated at PKC and PKA sites. Gel electrophoresis, immunoblotting and antibodies that recognize phosphorylated forms of cTnl are used to detect changes in the Tn complex. To test the role of specific PKC sites on cTnI in the evolution of hypertrophy and failure, mice harboring transgenes expressing a mutant form of cTnl (lacking PKC sites at Ser 43 and Ser 45) and slow skeletal TnI (lacking a PKC site at Thr 144 in the inhibitory peptide) are stressed by pressure overload. These experiments provide crucial information on the mechanisms of heart failure and on the potential value of developing pharmacological approaches to the inhibition of the PKC pathway.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The focus of this research involves the analysis of the molecular mimicry of mammalian DNA and protein sequences by the protozoan parasite, Trypanosoma cruzi. We have demonstrated that the parasite genome contains DNA sequences which mimic host sequences and which encode proteins with functional similarity to host immune regulatory proteins. These parasite proteins assist its survival in the mammalian bloodstream by down regulating the host immune response to the parasites.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Oxytocin (OT) functions as a hormone in labor and lactation and as a neuromodulator in the brain. OT receptors are present on OT neurons, which they autoregulate during somatodendritic OT release. During pregnancy and lactation, OT release is pulsatile, not continuous, and this pattern is critical to avoid OT receptor desensitization and insure proper hormone function at target smooth muscle in uterus and mammary gland. Neurons in the supraoptic (SON) and paraventricular nuclei (PVN) undergo remarkable plasticity during late pregnancy and lactation, including a glial-neuronal rearrangement and increases in synaptic activity. We have found enhanced spike afterhyperpolarizations (AHPs) during both periods. AHPs are critical in sculpting the short (~4 sec) high frequency (~50 Hz) bursts from OT neurons underlying this pulsatility, bursts that produce a bolus release of OT into the bloodstream to maximally contract uterine smooth muscle or mammary gland myoepithelium. Reproductive-associated plasticity in the SON and PVN depends on the pattern of ovarian steroid release during pregnancy and on the somatodendritic release of OT. We present the first in vivo evidence showing that central OT receptors are critical for the enhancement of the calcium-dependent AHPs normally manifest at late pregnancy. A specific OT antagonist administered chronically to the third ventricle during late pregnancy blocked this form of plasticity with no effect on VP neurons. In addition, we mimicked this plasticity by applying OT to hypothalamic slices from pregnant (E18-19) rats. The goal of this proposal is to understand the mechanisms of this adaptation made by OT neurons, and how it shapes OT neuronal firing to maximize hormone release and thereby insure normal parturition and lactation for infant development. Understanding the mechanisms of central OT receptor transduction at the OT neuron will have important applications for understanding the wider roles of OTRs in brain function. There are Three Specific Aims: Specific Aim 1. To determine if the enhancement of AHPs in OT neurons by OT is specific to OT neurons and OT receptor activation, whether it is specific to pregnant animals, and whether it is due to a change in the Ca2+ sensitivity of the underlying currents. Specific Aim 2. To test whether OT's effects on AHPs are mediated through a pERK 1/2-MAPK cascade, and whether it is associated with an increase expression of the AHP channels and/or the enzymes CK2 and PP2a, known to modulate AHP Ca2+ sensitivity. Specific Aim 3. To determine how enhanced AHPs modulate OT neuronal bursts, and to determine whether the reinsertion of missing excitatory activity differentially alters OT firing pattern in lactating vs. virgin rats, in an OT-dependent manner.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Tobacco cessation is a global health priority not yet addressed in low and low-middle income countries such as India and Indonesia. An important lesson gleaned from international health is that interventions proven effective in western countries cannot simply be exported to developing nations without significant cultural adaptation. Research is urgently needed to find the most effective means of promoting cessation in local contexts. A first step in gaining public attention about the harm of tobacco use and the need for cessation is to involve the medical community in tobacco cessation efforts. It has been well established that before tobacco use declines in the general population, health care providers must be at the forefront of tobacco cessation efforts. To do so, they must quit tobacco use themselves, routinely ask patients about tobacco use, and advise them to quit. At present, there is little involvement of physicians in tobacco cessation efforts in India and Indonesia. Based on four years of experience during Project Quit Tobacco International (QTI) (under the previous Fogarty initiative), the proposed project unfolds as a four-step process to develop a cohort of tobacco cessation researchers in India and Indonesia, using local medical schools as a hub for research activities in both the health care community and the community at large. Medical schools will become centers for recruiting and training tobacco cessation researchers as well as sites for implementing pilot studies and mobilizing local tobacco cessation research networks. The specific objectives of the project are to (1) increase knowledge about the risks of tobacco use and the importance of cessation through dissemination of a model tobacco education curriculum for medical schools in India and Indonesia;(2) recruit and train tobacco researchers concurrently with introducing tobacco education in nine medical schools in each country;(3) involve partner medical schools in tobacco cessation-related community-based research pilot studies, and (4) build capacity in tobacco-related research in both countries by creating tobacco cessation research networks in the six states (three in each country) where project activities will take place. Developed over the past four years, the QTI tobacco cessation research centers in India and Indonesia have begun to engage the communities in which they are located. In-country researchers have gained the skills and confidence to take the next step toward building research capacity beyond their institutions in these two culturally diverse nations. The proposed project will leverage lessons learned during QTI and provide an infrastructure for training a next generation of local tobacco researchers to meet the challenges of tobacco cessation within their own countries.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Test the relationship of hypovolemia to th eseverity of orthostatic tachycardia and that interventions that perturb volume or autonomic function will improve the hemodynamic profile in these patients. We are beginning recruitment and screening.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Institute of Medicine and others have strongly encouraged research on structural interventions. Outreach HIV/STD testing programs are an example of a structural intervention that have been studied for their individual level effects but not for structural level effects. Nevertheless, some anecdotal evidence suggests that this intervention may indeed have a structural level impact on risk behavior in at least one type of risk environment. Using secondary data analysis, the proposed exploratory study will expand the science of structural interventions by investigating the possible structural-level impact of a local prevention program at a club where men at high risk for HIV infection go and where risk behavior is known to occur. Although this subgroup of men are from a small proportion of the US population, they account for almost two-thirds (63%) of the AIDS cases nationwide. For an on-site HIV testing program to be a successful structural intervention, the critical question is whether the altered social climate results in change to the overall risk behavior in the club, even among those who did not test. Additionally, any observed change must not be due to unintended consequences, such as moving the high-risk men or behavior someplace else. Finally, it would be good to identify evidence that the structural-level changes anticipated are actually occurring in the club's social climate. Thus the Specific Aims of the proposed study are: 1) Compare risk behavior at the club during times with on-site-testing vs. times with no on-site-testing. 2) Assess indicators of unintended consequences (i.e., moving high-risk men and behavior elsewhere). 3) Describe the impact of the on-site testing programs on the club's social climate. This study proposes to conduct secondary analyses using multiple data sources from a single club: 1) Exit Surveys: two independent probability samples of men exiting the club, 2) Club Membership/Utilization Data: collected by club staff as members enter and leave the club, 3) Ethnographic Data: interviews with managers, staff and patrons about the social climate at the club, and 4) Process Data: interviews with managers, staff and patrons from an evaluation of the testing program at the club. The proposed study has the significant advantage of addressing the intervention's impact using multiple data sources and analyses, providing a robust assessment of our research question. The results could support designing a controlled clinical trial to test the efficacy of an on-site HIV testing program as a structural intervention to reduce high-risk behavior in clubs. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Prescribed footwear, such as Removable Cast Walkers, are used to reduce harmful pressures on the bottom of the foot for prevention and treatment of diabetic foot ulceration (DFU). Although a key component to the healing and prevention of DFU, patient adherence with these devices is excessively poor as determined by patient interviews and initial objective assessments. The use of \"off the shelf\" activity monitors for objective monitoring of adherence has provided some valuable information regarding adherence and wound healing, however, the methodology used to date has had several significant restraints and is not suitable for use outside of the research setting. This methodology is limited to the assessment of foot loading during walking and thereby ignores the significant loading of feet that occurs while standing. Also there is no means of identifying whether sensors were worn, allowing for significantly underreported activity/adherence values. Lastly, the calculation of adherence from these devices is quite labor intensive and cannot be done in real time. In this proposal, we develop an innovative and low cost technology that could help improve patient adherence via both an alarm notification to the patient and valuable data logger information provided to clinician. The proposed system combines a widely available inexpensive device with innovative physical activity monitoring technology for improving patients'adherence to wearing prescribed footwear. The device consists of an active RFID tag (Radio-frequency identification) integrated in the subject's footwear, together with a body worn sensor that includes an RFID tag reader and motion sensors based on micro-electro-mechanical systems (MEMS) technology for monitoring spontaneous daily physical activity. The RFID tag will act as a proximity sensor, with the range limited to the length of the patient's body. Therefore for physical activity to be classified as adherent with the prescribed footwear, the MEMS sensor recording the activity must concurrently detect the footwear as being within sensor range (e.g. being worn). The suggested technology is an extension of our work in monitoring spontaneous daily physical activities in real-time using unobtrusive body wearable technology. The body worn sensor measures specific body postures in real-time, such as sitting, walking, standing, and lying down and is also capable of measuring patients'compliance in wearing the sensor by monitoring chest movement due to respiration. The software algorithm is sensitive and specific enough to integrate an alarm, should the subject ambulate unsafely without their RFID tagged footwear. (e.g., walk more than five steps). In case that the RFID tag is out of range (i.e. subject didn't wear the footwear), the sensor unit will notify an alarm (via sound or vibration) and will mark the time of the alarm in the on-board memory system. In the clinical part of the study, two different tests are designed to address its validity as well as its application in (a) quantification of patient adherence as compared to previous measures of adherence to prescribed footwear (e.g. face-to-face interview) while ambulating at home and in the community and (b) investigating changes in patients'footwear behavior in response to commonly used practices, such as addressing patient preferences, patient education;and use of alarm. The suggested technology could make an immediate difference in diabetic foot care management and enable clinicians to better manage diabetic wound care. It may also lead to smart foot ulcer prevention strategies via improving subjects'adherence to prescribed footwear and allowing for dosage of daily physical activity. Lastly this device and the data gathered by it may also serve as a valuable patient education tool. PUBLIC HEALTH RELEVANCE: The use of specialized footwear that limits physical stress to the feet during weight bearing activities is a key factor in the prevention and treatment of diabetic foot wounds (ulcers);however, patients generally wear the footwear for only a small fraction of their daily activities. This project seeks to develop an unobtrusive monitor that will identify patients'use of their footwear during their daily physical activity. This device may be used to provide clinicians valuable information regarding patient compliance and also may be used as a tool to directly improve compliance by providing a warning alert to patients when they are active while not wearing their footwear.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal seeks renewal of an Older Americans Independence Center at the Johns Hopkins Medical Institutions. The focus of this Center is to promote independence for frail older adults through the development of interventions to prevent or ameliorate frailty and the many serious, adverse outcomes that frail older adults are at risk for. To accomplish this, a multidisciplinary group of senior and junior investigators, in multiple departments and Programs at this institution, propose to continue and enhance, through this OAIC, 1. the Leadership Council that will stimulate, lead and sustain a University-wide interdisciplinary research program investigating frailty, its etiology and opportunities for treatment; 2. the infrastructure and methodologic development essential to creating the next generations of research on frailty in older adults; 3. support for the career development of outstanding junior faculty committed to research careers that will lead to treatments for frailty and its prevention; and 4. to cultivate and support pilot studies that will provide the basis for investigator-initiated and funded research towards interventions on frailty. The OAIC would provide the support critical to complement and harness the substantial strengths already at this institution into a unified and potent effort directed at developing interventions for frailty. It is designed to ensure multidisciplinary translation between basic, mechanistic and clinical/population-based research, and of each into development of treatments for frailty. This organized infrastructure and scientific leadership will serve as a source of advice and collaborations to investigators at our own institution and at other institutions in the area of research on frailty in aging. The OAIC will be administratively led by, and housed in, the Johns Hopkins Center on Aging and Health, with strong leadership and participation by all of the institutional programs committed to aging research; it will also seek to expand to incorporate leaders and investigators not previously involved in research on frailty but who have made a commitment to this work through the planning process for this proposal. This OAIC has strong institutional commitment, based on the priority at the Medical Institutions for the unification and strengthening of aging research and spotlighting its importance for the entire institution. Overall, it is designed to spearhead a new depth and intensity of research to improve outcomes and promote independence for frail, older adults. [unreadable] [unreadable] PROGRAM AS A WHOLE [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Despite decades of research, the basic pathogenetic mechanisms involved in inflammatory bowel disease (IBD) remain elusive, but the answers appear to lie in the scientific fields of immunology and genetics and these are the major areas addressed in this proposal. The basic theme of this Program Project relates to the immunologic mechanisms determining chronic inflammation in the intestine, the impact of chronic enteritis on the mucosal immune system, the mechanism of antigen handling in the intact and inflamed colon, and the genetic influences that determine susceptibility to disease, as well as the interplay and interaction among these areas. The focus of these studies will be on newly developed mouse models of colitis including spontaneous colitis in C3H/HeJBir mice, a substrain which was generated during the initial phase of this project. Although no model exactly duplicates human IBD, models can allow important insights into certain disease mechanisms. The mouse has been chosen because of the rich array of techniques and reagents available only in this species in both immunology and genetics. The Program Project will consist of four Projects and two Cores. Project #1, will focus on the enteric bacterial antigens recognized by the immune system during chronic colitis, particularly by T cells, the role that such T cells play in disease pathogenesis, and how such T cells are regulated. Project #2 will study a number of issues of mucosal immunity, oral tolerance, and isotype regulation in the setting of chronic intestinal inflammation. Project #3 will exploit two powerful new technologies, a T cell transgenic mouse, and in situ hybridization techniques, to examine antigen handling and T cell induction in the normal and inflamed colon. Project #4 will be located at the Jackson Laboratory in Bar Harbor, ME. This project will map genes important in susceptibility to colitis induction in inbred mouse strains, including the G3H/HeJBir colitic substrain. These projects will be supported by an Administrative Core which will provide administrative and fiscal support and coordination, and an Animal Model Core which will centralize the production of mice with experimental colitis, maintain strict quality control, and provide a centralized pathologic analysis. These studies should generate new concepts and paradigms to test in man, and possibly new approaches to immunotherapy, as has happened with mouse models of other diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recently, a number of studies have shown that mammalian central nervous system axons can regrow following lesion if given the proper environment (Richardson, 1980; David and Aguayo, 1981; Bently and Aguayo, 1982; Guth et al., 1981; Silver and Ogawa, 1983; Goldberg et al., 1986). These findings have renewed interest in regeneration of central nervous system neurons with much of the attention focused on inducing regeneration in species that under normal conditions exhibit little capacity for recovery of function, e.g., rats. While these experiments have produced dramatic and exciting results, they have not revealed the requirements for, or the mechanisms of, successful regeneration. The experiments described in this proposal will provide a detailed description of naturally occurring regeneration in an adult system, the salamander spinal cord. These studies will complement the mammalian studies of regeneration and provide a basis for future studies of the molcular biology of regeneration. The salamander spinal cord has been a model system for the study of regeneration for several decades because of the salamander's ability to recover function after transection of large ablation (e.g., Piatt, 1955a,b; Holtzer, 1951; Butler and Ward, 1967; Stensaas, 1983). Despite this long history many important questions remain unanswered: 1) what percentage of cells regenerate following ablation or transaction of the spinal cord and what is the origin of these cells, 2) where are the somata that give rise to the axons found in the regenerated spinal cord and how far do regenerated axons grow pass the lesion site into the undamaged spinal cord, 3) do regenerated axons make synapses which are appropriate, and 4) how does the repopulation of the damaged spinal cord by neurons and fiber tracts relate to recovery of function. Horseradish peroxidase (HRP) pathway tracing techniques and 3H-thymidine labeling will be utilized to examine these questions in order to produce a complete description of the events involved in the regeneration of the thoracic and lumber regions of the salamander spinal cord.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We discovered that M. tuberculosis (Mtb) infection causes apoptosis of primary human alveolar macrophages (AMphi) in vitro by a tumor necrosis factor-alpha death signaling pathway. Apoptotic AMphi are present in bronchoalveolar lavage of tuberculosis (TB) patients and in TB lung biopsies. We found that virulent Mtb strains downregulate apoptosis of host AMphi, and we have shown that apoptosis is linked to microbicidal processing. Based on these observations, we hypothesize that lung Mphi apoptosis constitutes a newly recognized mechanism of innate immunity in TB. TB is a major co-infection in AIDS, contributing to increased morbidity and mortality. With declining adaptive immunity in AIDS, the relative importance of innate immunity may be augmented. Our research plan proposes novel use of the murine aerosol TB model to define the role of lung Mphi apoptosis in disease susceptibility and resistance in vivo. The specific aims address three key issues: 1, we will characterize the kinetics and distribution of lung Mphi apoptosis following aerosol Mtb infection; 2, we will examine the relationship between lung Mphi apoptosis and host resistance in experiments comparing Mtb strains of high and low virulence, common inbred mouse strains that are innately susceptible or resistant to TB, wildtype and knockout mice with targeted deletion of genes regulating apoptosis, and Mtb transduced to express apoptosis regulating genes; 3, we will evaluate the impact of lung Mphi apoptosis on cytokine expression and the pattern of leukocyte recruitment to the lung in TB, and it's contribution to the development of delayed type hypersensitivity. These studies will provide a detailed analysis of early events in murine TB, and will be the first to fully characterize lung Mphi apoptosis in vivo and to define it's contribution to the integrated host response to infection. Our model may reflect a mechanism broadly involved in the defense against a variety of other intracellular pathogens (such as Legionella) of immediate relevance to the HIV-infected host.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The existence and biochemical mechanisms of organ-specific toxicity are the subject of heightened interest among toxicologists. Earlier studies in this laboratory described selective necrosis of pulmonary non-ciliated bronchiolar (Clara) cells following administration of naphthalene to mice. No damage to other lung cells was noted and no pathologic changes, as evidenced by histology or enzymic alterations, were observed. The work described in this section describes conditions under which 1,1-dicloroethylene (DC) produces selective damage to mouse lung without morphologic or enzymatic evidence of nephro- or hepatotoxicity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Data will be collected on the safety and efficacy of performing excimer laser photorefractive keratectomy in two animal models. A previously developed 193 nm excimer laser system, the ExciMed UV200, will be used to perform photorefractive keratectomy using two different treatment techniques. The ExciMed UV200 with an electronically driven iris aperture delivery system will be compared to an ExciMed UV200 with a fixed beam diameter used in conjunction with an ablatable contact mask. A comparison will be made of the two methods in rabbits and monkeys. In Phase I, initial development of the contract mask will be performed and a range of operating parameters for performing photorefractive keratectomy with the contact mask will be determined in-vitro. Simultaneously, in-vivo experiments in rabbits and monkeys will begin with the previously developed iris aperture delivery system. During Phase II, in-vivo work will be performed in rabbits and monkeys with the ablatable contact mask. Further development of the contact mask will take place during Phase II based on the results of the in-vivo experiments. Analysis of the data collected in rabbits and monkeys will determine which of the two treatment techniques will be used in further human clinical trials of photorefractive keratectomy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Previous studies have established the feasibility of removing albumin-bound substances such as bilirubin from the blood by in vivo extracorporeal perfusion using albumin-agarose beads. Because a number of substances in addition to bilirubin accumulate in the blood of patients with liver damage, we plan to test albumin-agarose beads and other adsorbents in extracorporeal perfusion of experimental animals with liver damage. At present we are setting up the assays to measure blood levels of some substances known to accumulate in liver failure, amino acids, fatty acids, mercaptans, and the false neurotransmitter, octopamine. We will test the efficacy of various adsorbents for binding these substances.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Crohn's disease (CD) is a chronic inflammatory bowel disease (IBD) that cannot be cured with either surgery or currently available medical therapy. CD is characterized histologically by infiltration of the intestinal mucosa and submocosa with chronic inflammatory cells and clinically by abdominal pain, diarrhea, and weight loss. How these inflammatory cells are attracted to the intestinal mucosa is not known, but adhesion molecules are believed to play an important role in lymphocyte transmigration into the intestine. ISIS 2302 is a 20 base oligodeoxynucleotide designed to specifically hybridize to a sequence of the human intercellular adhesion molecule-1 (ICAM-1) messenger RNA (mRNA), thus blocking translation of this message. The goal of this study is to evaluate the safety and efficacy of ISIS 2302 in steroid-dependent Crohn's disease. We hypothesize that inhibition of ICAM-1 will decrease intestinal inflammation and lead to clinical symptom improvement in patients with Crohn's disease. To date one patient has completed the study and enrollment remains open.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION(provided by applicant: Urease, which catalyzes the hydrolysis of urea to ammonia and carbonic acid is produced by diverse bacterial species including various aerobes, facultative anaerobes and obligate anaerohes Both Gram-negative and Gram-positive organisms are urease producers, as are species of mycobacteria and ureaplasma. Urease plays a significant role in virulence when expressed by urinary tract, oral, and gastroduodenal pathogens. Providencia stuartii and Proteus inirabilis, the two most common ureolytic uropathogens, express urcase only in the presence of urea. This urea-dependent expression is mediated by UreR, a transcriptional activator belonging to the AraCfamily of regulators. Evidence suggests that urea interacts directly with UreR, and thus is the effector molecule for this activator. Urea is Found at concentrations up to 500 mM in the urinary tract, a concentration that is at least 50 fold higher than that observed at other sites in the body. Thus for these uropathogens, urea is a signal molecule, and UreR is acting as a signal receptor, alerting the organism that it is in the urinary tract. UreR bound to urea is active as an transcriptional activator and has a high affinity for DNA. UreR not bound to urea is not active and has a low affinity for the I)NA binding site. Studies in this proposal are to examine the urea-UreR interaction and to determine the conformational changes associated with urea binding that result in active UreR. Two models are proposed, one in which UrcR forms a dimer, and the other in which UreR is active as a monomer. Studies are proposed to prove one of these models. The crucial urca-UreR interaction is examined in Aim #1, and the urea-UreR-DNA interaction in Aim #2. Aim #3 is to determine the X-ray structure of UreR, UreR bound to urea, and UreR bound to urea and the DNA binding site. Structures of mutant forms of UreR will also be generated. This work will elucidate the molecular mechanisms of this important regulator of a urovirulence, as well as extend our knowledge on the AraC-fainily of transcriptional activators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research project is directed toward establishing experimental systems to enable the discovery of cytokine response (CR) genes, and to determine which CR genes function to facilitate or inhibit T cell cycle progression, cellular survival, cytokine production, and surface protein expression. The aims of the project are focused on interleukin-2 (IL-2) and T cells, but the methods to be developed should be applicable to other cytokines and cells, and to other ligands, including transcription factors. The proposed research follows from the hypothesis that IL-2 promotes the expression of specific genes, the products of which regulate lymphocyte proliferation and differentiation. Two Specific Aims test this hypothesis: 1) To create a more rapid and efficient system to identify additional CR genes, by coupling our methods to enrich for CR genes via sulfhydryl labeling and affinity purification (SLAP) to the technique of serial analysis of gene expression (SAGE); and 2) To employ retroviral-mediated gene transfer in T lymphocytes, and flow cytometry-based assay systems to screen for CR genes which affect cell cycle progression, apoptosis, cytokine production, and immunomodulatory surface protein expression. The SLAP method yields a 40-fold enrichment of CR genes, so that approximately 98 percent of constitutively-expressed, \"housekeeping\" genes are removed. The SAGE method to identify genes by sequencing 9 base pair \"tags\" also improves efficiency by 40-fold. Therefore, the combination of these 2 synergistic methods will reduce the time necessary to identify new CR genes from several months to a few weeks. To enable the functional analysis of each CR gene product identified, a gibbon ape leukemia virus- pseudotyped, bicistronic retroviral vector will be used. This system allows co-expression of the CR gene with a selectable marker, green fluorescent protein (GFP), in peripheral blood T cells. Functional analyses of infected cells may then be performed, using flow cytometry with gating on the GFP-positive population. The combination of retrovirus-mediated gene transfer into normal T cells, and sensitive flow cytometry assays for cellular proliferation, apoptosis, cytokine production, and surface protein expression provide a powerful means to assess CR gene functions. Consequently, this research project will contribute to our understanding of the molecular mechanisms responsible for fundamental lymphocyte responses to cytokines, and to the diagnosis and therapeutic manipulation of immune system disorders including immunodeficiency as well as hypersensitivity disease states.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The PI for this project, Dr. E. P. Greenberg has worked on quorum sensing for over 25 years. His long-term goals are to investigate the molecular mechanisms of social activity in bacteria, and to learn about why, when, where and how cooperative behavior exists. This proposal is to continue studies on quorum sensing in Pseudomonas aeruginosa. Prior work on this project has established that P. aeruginosa employs two quorum-sensing signals to govern expression of a battery of genes in a complex fashion. Although it is clear that P. aeruginosa shows substantial diversity as a species quorum sensing has been studied primarily in the context of the single strain PAO1, which was isolated from a human infection. In addition to the two known quorum-sensing signal and receptor circuits, Dr. Greenberg has shown there is a so-called orphan receptor for one of the signals, 3-oxo-C12-homoserine lactone. There are two receptors for this single signal. There are fundamental biological questions about the value of possessing two receptors for a single signal. In addition, the complete quorum-sensing systems are required for virulence of this opportunistic pathogen and the orphan receptor appears to suppress virulence. The proposed research derives from recent work on this project and represents a next logical set of about the biological significance of P. aeruginosa quorum sensing. We will study P. aeruginosa strains other than PAO1 to identify genes universally regulated by quorum sensing in this species and to gain insights about how environment might influence the makeup of the regulon. We will test two hypotheses that each address why P. aeruginosa might possess two receptors for the same signal. The relevance of this research to public health is that P. aeruginosa can cause difficult to resolve and often incurable human infections. Although it is unlikely that quorum sensing evolved to activate functions for human infections it is nevertheless true that quorum sensing is required for full P. aeruginosa virulence in a variety of animal infection models. Perhaps because of the quorum sensing involvement in virulence, cell-cell communication and cooperative behavior in P. aeruginosa are at the forefront of our knowledge of bacterial social behavior. The work proposed here will lend insight into the selective advantage of bacterial communication and cooperative behavior. PUBLIC HEALTH RELEVANCE: This research program has developed a mechanistic understanding of quorum sensing in Pseudomonas aeruginosa, an opportunistic pathogen that causes devastating human infections. Because quorum sensing controls virulence of P. aeruginosa it has become a target for therapeutic development. The proposed research aims to increase our understanding of the elements of the P. aeruginosa quorum sensing system and how this bacterium uses quorum sensing to control its social activities. The findings will provide guidance about how to effectively target quorum sensing in the course of an infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is an amended application 1 R41 AI052921-01. The overall goal of this project is to develop new targeted formulations of antibiotics for prevention and treatment of diseases caused by Category A pathogens, in particular Bacillus anthracis (anthrax), Yersinia pestis (plague), and Francisella tularensis (tularemia). These preparations will specifically accumulate inside and or in the near vicinity of cells harboring such pathogens after exposure to weaponized (aerosolized) biological agents. Availability of such preparations will provide new strategies of disease prevention for those at risk of exposure, and novel effective treatments for those already infected. The proposed new formulations will be based on lymph node-specific nanocarriers developed at Massachusetts General Hospital. These carriers are capable of transporting various drug substances to lymph node phagocytes (primary pathogen-harboring cells) after systemic administration. It is expected that delivery of antibiotics by such carriers will result in much higher drug levels in the infected lymph node tissue than conventional preparations of the same antibiotics. Thus, the primary site of germination and development of weaponized biological agents will be sufficiently saturated with drugs to prevent or stop disease development at the very early stage (before the onset of symptoms). The objective of this project is to test feasibility of previously established nanocarrier technology for loading lymph node phagocytes with two model antibiotics (one of Fluoroquinolone and one of Tetracycline families), and to develop two respective prototype preparations for further characterization and optimization. The specific aims are: (1) develop two prototype lymph node specific preparations (of Ciprofloxacin and Tetracycline); (2) investigate, in a rodent model, the degree of drug accumulation inside and in the near vicinity of target cells, and (3) evaluate efficacy in infected cells. If successful, this project will result in the development of a new class of preparations and a new strategy for prophylactic and early post-exposure treatment of bacterial diseases of Category A. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The chemical synthesis of 2'(3')-0-aminoacyloligoribonucleotides and 2'(3')-0-(N-acylaminoacyl)oligoribonucleotides with nucleotide sequences identical to or resembling those of the 3' terminus of AA-tRNA will be carried out. These compounds will be used as models of the 3' terminus of AA-tRNA or peptidyl-tRNA in order to study their involvement in the various steps of protein biosynthesis on ribosomes and their interactions with elongation factors (EF-Tu, P and G). Modified tRNA containing \"fraudulent\" nucleoside units on the 3' terminus will be synthesized by combination of chemical and enzymatical methods. These modified tRNA's will be used for study of various subreactions of protein biosynthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Broad Institute performs high-throughput large scale biology with academic collaborators from across the country, investigating a wide variety of biological systems. In this proposal the Broad Institute core small molecule screening group is requesting funding for an automated tissue culture system to enhance our large scale biology capabilities. The instrument would be managed and maintained by the Chemical Biology platform screening group. The screening group is a team of professional scientists that has extensive industrial experience and expertise in developing and executing high-throughput cell-based assays for small molecule probe discovery. Large scale tissue culture is already performed by this team to support more than 30 MLPCN and Broad screening projects per year. Cell supply can become a limiting factor in terms of the quality and quantity of cells required. Automated tissue culture would expand our capabilities to support more projects, increase our screening throughput, improve assay quality, and free up scientists to carry out more research. The automated tissue culture platform, CompacT SelecT, grows and maintains cells from multiple cell lines in T-flasks and delivers them into microtitre plates. It is designed to generate assay-ready plates on demand for cell based screening and assay development. A CompacT SelecT would enable unattended operation, up to 24 hours a day, 7 days a week to maintain 130 T-175 tissue culture flasks and deliver 210 384 assay ready plates per day with a 3- fold reduction in manpower. The rate of progress of the large scale biology projects described in this proposal are limited by the researchers ability to scale-up cell culture, propagate multiple cell lines and maintain cell quality. We are in a unique position to offer access to this high throughput cell culture capability to many members of the scientific community. This proposal describes five projects currently funded by the NIH that would greatly benefit from the increased scale, and cell quality automated tissue culture provides. These projects are of significant interest to the biomedical research community, focusing on disease areas in metabolism, cellular differentiation, cancer, and functional regulation. Beyond these projects, the screening group has established various pathways for outreach, forming collaborations, and providing industrial-scale HTS capabilities to academic groups, including numerous NIH-funded investigators. The Broad Institute is a Comprehensive Screening Center in the NIH Molecular Libraries Probe Production Center Network (MLPCN). Automated cell culture would enable us to support many more projects without increasing our headcount.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of our research program is to understand the mechanisms that control sexual differentiation of brain structures, behaviors, and diseases. The developmental process of brain sexual differentiation is critical for the establishment of dimorphism in neural function and behaviors between the sexes. Meanwhile, sex differences in the incidence, age of onset, and severity of neurobehavioral diseases are also well documented. Many studies have established an essential role for sex steroids and their nuclear receptors in these processes. However, the genetic and molecular mechanisms that underlie the actions of sex steroids on differential gene transcription and translation relevat to sex differences are still not clear. Recently, we have identified splicing factor, suppressor of white-apricot homolog (Drosophila) (Sfswap), also known as splicing factor arginine/serine-rich 8, as one of the many candidate genes differentially expressed in the neonatal mouse cortex between the sexes. Thus, we propose the novel hypothesis that sex steroids and their receptors regulate transcription of Sfswap gene in the developing cortex, which leads to sex difference in brain structures and behaviors. To test our hypothesis, the proposed project has three major aims: (1) to characterize expression of Sfswap gene in the mouse cortex during early development; (2) to examine if Sfswap expression in the cortex is regulated by perinatal exposure to elevated testosterone via androgen receptor; and (3) to investigate the physiological roles of Sfswap in the development of sexually dimorphic structures in the cortex and the behavior(s) mediated by this brain region. Our results will provide new important information on the functional link between alternative splicing and brain sexual differentiation. Successful completion of this project will also help the PI establish a new line of research in neuroendocrinology with the enhanced competitiveness and quality of research required to secure NIH non- SCORE funding in the future. PUBLIC HEALTH RELEVANCE: Sexual differentiation during early development has a significant subsequent impact on human health throughout life. Not only are gender differences well known in normal physiological function and behaviors but also in susceptibility to different diseases. Understanding the genetic and molecular bases of these gender differences will lead to better understanding and new treatments for sex-biased diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The principal cause of cancer mortality is the dissemination of cells from the primary tumor to form distant metastases. The potential for a cell to migrate and invade depends on a multitude of signaling pathways, which present novel targets for anti-cancer therapies. Reactive oxygen species (ROS) have increasingly been implicated as modulators of tumorigenicity and many cancer types exhibit elevated ROS levels. An important regulatory enzyme of Redox balance is the mitochondrially expressed manganese superoxide dismutase (Sod2), which catalyzes the conversion of superoxide anion to hydrogen peroxide (H2O2). Elevated Sod2 expression has been described in a variety of invasive cancers including those of breast, brain and bladder. We have shown that Sod2 overexpression, and a concomitant increase in H2O2, can lead to a more metastatic and migratory behavior of fibrosarcoma HT-1080 cells. In this proposal we hope to elucidate how an increase in Sod2 expression and a surge in mitochondrial H2O2 production leads to this pro-migratory phenotype. To address this we will determine the spatial localization and site of action of mitochondrial H2O2, using novel Redox sensing green fluorescing protein probes. This will indicate if ROS can be targeted to specific areas within a moving cell, where they may carry out their role as second messengers. A novel role for ROS in cellular signaling is the oxidation and inactivation of protein tyrosine phosphatases, including those that are involved in the dynamic rearrangement of focal contacts at the plasma membrane by focal adhesion kinase. Using biochemical approaches we will investigate whether expression of Sod2 can significantly enhance pro-migratory focal adhesion kinase signaling via enhanced oxidation/inactivation of phosphatases that normally inhibit this pathway. Finally, we will investigate the role of anti-oxidant molecules in reversing the pro-migratory phenotype and signaling events of Sod2 expressing cells, both in vivo and in vitro. This will determine the validity of antioxidant based therapies for cancers that display an enhanced ROS burden due to elevated Sod2 expression. The studies of this proposal aim to address the hypothesize that elevated Sod2 expression results in the spatial inactivation of protein tyrosine phosphatases via increased mitochondrial H2O2 production, leading to pro-migratory signaling and an enhancement of the metastatic potential of cancer cells. This research will further our knowledge of the intracellular mechanisms that regulate metastasis. Understanding the role of reactive oxygen species in these events will aid in the development of novel antioxidant based cancer therapies. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Esophageal cancer occurs in two major histologic types; squamous cell carcinoma (SCC) and adenocarcinoma (EA) both of which have an extremely poor prognosis. Although these tumors are often treated as one disease, they have distinct differences in epidemiology, risk factors and biological behavior. In Westernized countries the incidence of SCC has been steadily decreasing over the past 25 years, while EA rates have been increasing dramatically (~400%). In fact, EA now has the fastest rate of increased incidence of any tumor in the United States. This is most likely linked to the rise in obesity and the high prevalence of gastroesophageal reflux disease, which is estimated to affect up to 44% of the American population and is strongly linked to the development of EA. Despite this rise, research on esophageal adenocarcinoma is made difficult by the relatively low number of cases per year and the inability of single institutions to acquire large patient cohorts for study. Consequently much less is known about the biology of this disease than about more prevalent tumor types. On a genome-wide scale, very little is known about the genetic aberrations and gene expression alterations that are specific to EA. Partly as a result of these deficiencies in our knowledge, there are currently very few biomarkers for disease diagnosis, progression or prognosis and there are no targeted therapeutic agents indicated for the treatment of this highly lethal tumor. Our research group is in a strong position to address these deficiencies since we have a multi-disciplinary team focused on translational research and our ongoing, R01-funded studies have enabled us to develop large tissue banks with detailed clinical information on all patients. In this proposal, we intend to utilize these resources in order to generate molecular staging tools and to identify novel therapeutic targets to improve the treatment of esophageal adenocarcinoma patients. Specifically, we propose to examine the genome of EA using high density DNA arrays in order to identify genomic regions that are frequently altered in this tumor type. We will then identify all genes within these altered regions and extract their expression levels from Affymetrix U133 arrays. Genes whose expression levels correlate with DNA copy number change will be cross-referenced against druggable target databases in order to identify potential new therapeutic targets for esophageal adenocarcinoma. We will also determine which of the regions and genes are associated with disease stage, metastasis and survival. The association of these regions and the expression of genes within them with clinical endpoints will then be validated in an independent tumor set using quantitative PCR. Finally, validated prognostic markers will be combined to develop a molecular staging algorithm that can be used to guide therapy for esophageal adenocarcinoma patients. Thus, at the end of this study we will have identified, verified and validated novel genetic markers that could be used for diagnosis, staging and treatment of esophageal adenocarcinoma. PUBLIC HEALTH RELEVANCE: Esophageal adenocarcinoma is increasing faster than any other malignancy and typically has a late diagnosis and poor outcome. Despite major advances in high-throughput genomic analysis methods, this disease remains largely unstudied at the level of genome-wide DNA alterations. The identification of specific DNA alterations and the genes within them could lead to markers for diagnosis, progression and prognosis as well as providing novel targets for therapeutic discovery and treatment of esophageal adenocarcinoma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The p53 tumor suppressor is central to human DNA repair, damage checkpoints and many aspects of human biology. Importantly, most cancers are altered for p53 function. Although the guardian of the genome p53, much about its function and its targets remains unknown. Because of its importance as a human tumor suppressor and its other recently discovered functions identifying the universe of p53 direct transcriptional targets is clinically relevant, especially since many of these may have therapeutic value. There is considerable variation in p53 dependent expression across targeted genes leading to differences in p53-mediated biological consequences, due in part to variation in target response element (RE) sequence. We have focused on RE functionality, i.e., the ability of REs to support transactivation by p53 to do that we developed reporter systems in budding yeast for variable human p53 expression and have translated many of the findings to human cells in culture and ex vivo. Several studies have used genome-wide, next-generation sequencing approaches to identify p53 binding sites in the human genome. Recently, we took a rigorous method by reanalyzing all the raw data from the individual ChIP-seq studies and associated gene expression using a single analysis workflow, then combining information using a common set of criteria. Our approach revealed a large p53 genome-wide cistrome composed of >900 genes directly targeted by p53, including the identification of new potential p53 transcriptional targets involved both in the classical roles of p53 function (DNA repair) as well as in processes not immediately related to classical p53 outcomes, for example, the immune response. INFLUENCE OF P53 TRANSCRIPTIONAL NETWORK IN DNA DAMAGE RESPONSE. Using our p53 cistrome studies we have identified nearly 120 potential human target genes associated with DNA Damage Response (DDR). We have validated the p53 transcriptional responsiveness of potential p53 DNA damage response target genes under stressed and unstressed conditions with various human cancer cells that vary in p53 functional status. We identified many putative novel p53 transcriptional targets. We validated by RT-qPCR the p53-dependency and expression profiles of nine of these genes, APTX, ATR, ATRIP, DDX5, LIG1, MSH4, POLD1, POLH and REV3L after the treatment with three p53-inducing drugs: Doxorubicin, Etoposide, and Nutlin. THE INFLUENCE OF p53 ON HUMAN IMMUNE RESPONSES. We have established that p53 plays important physiologic roles in the immune system. p53 upregulates most members of the pathogen sensor TLR family in human cells to consequently enhance TLR-dependent production of proinflammatory cytokines in response to cognate ligands. The transcriptional cofactors ASPP1 and ASPP2 can enhance p53 mediated transactivation of several TLR genes. Moreover, p53 collaborate with NFkappaB to modulate several immune responses in primary lymphocytes and macrophages. Through our meta-analysis of p53 targeted binding sites and associated gene expression (i.e., cistrome genes) following stresses in many cell types, including human lymphocytes, treated with p53 activating drugs, we identified 100 genes target by p53 involved in cellular immune and inflammatory process including several viral/bacterial restriction factors. Our studies emphasized the influence of p53 in modulating the immune system, which defends against external and internal threats as well as tumorigenesis. Among these genes we found several of the human cytidine deaminase APOBEC3 gene family (A3) involved in the innate immune that deal with RNA virus infections are also subject to p53 transcriptional control by WT and mutant p53 in different directions. We observed that RSV, the major cause of respiratory problems in young children worldwide, induced p53 protein activation, resulting in the expression of several A3 genes in a p53-dependent manner and influencing negatively the virus replication. A SNP in TLR8 regulatory region creates a p53 responsive sequence that renders responsiveness and transcriptional control of this gene by p53. We found that this SNP influences RSV disease severity in infants infected by this virus. In a project supported by the Office of AIDS Research (OAR), currently we are investigating the immune role that p53, its polymorphism in codon 72 and SNPs in TLR8 including the p53RE SNP (rs3761624), might have during HIV-1 infection. We recently confirmed in H9 leukemia T cells (p53 null) cells and CD4+ T lymphocytes of healthy people, that activation of p53 results in upregulation of subset of HIV restriction factors genes including A3G, A3H, SAMHD1, TRIM5, HERC5, TRIM5, MARCH2, MX1, TSG101 PRK and ZAP. Furthermore, we have found that activation of p53 with Nutlin in CD4+ human T lymphocytes reduces HIV1 replication. In collaboration with Johns Hopkins School of Public Health, we identified 263 HIV-1 seroconverters in the Multicenter AIDS Cohort Study (MACS) previously genotyped for TP53 SNP rs1042522 and TLR8 SNP rs37646880, which is in high linkage disequilibrium with TLR8-p53RE SNP. A preliminary analysis showed that especially TP53 codon 72 variant, may be associated with a lower risk of developing AIDS but the presence of both protective alleles (TP53 rs1042522=GG and TLR8 rs37646880=A) might convey an overall 65% reduction in the risk of AIDS. CANCER-ASSOCIATED p53 MUTANTS. With inclusion of immune response-related TLR genes into the p53 network, we evaluated the effect of 25 tumor-associated p53 mutants on TLR gene family expression after transient transfection in p53-null cancer cell. Changes in TLR transactivation patterns, including change-of-spectrum, were observed, suggesting that p53 tumor status might be an important factor in adjuvant therapy employing TLR pathways to treat cancer. p53 mutants that induced expression of TLR3, enhanced cytokine and chemokine responses mediated by this receptor after exposing cells to TLR3 ligand poly-I:C alone or in presence of Doxorubicin. We also found that functional rescue of loss-of-function p53 mutants by the p53 reactivating drug RITA, restored TLR gene expression in a mutant p53 cell line, enhancing DNA damage induced-apoptosis via TLR3 signaling. Furthermore, several p53 mutants also altered the expression A3 gene family, particularly, several mutants promoted expression of A3B, which normally is repressed by WT p53, suggesting a clear gain of function phenotype for these p53 mutants. Since TP53 gene mutations occur in many human cancers, it is important to identify anticancer drugs that specifically target p53 mutant tumor cells. We are pursuing the identification of synthetic lethal (SL) genes with genome-wide siRNA-based screens, that when reduced in expression in p53 mutant cells cause death or reduced growth. The SLs are expected to lead to potential targets and provide opportunities for anticancer drug development. We have identified several SL targets for two of the most frequent tumor-associated p53 mutants (R175H and R273H) in the presence of anticancer drug etoposide as well as for the WT p53 and p53 null conditions. Among the SL genes identified, several are related to the DNA damage response including ATR, BRCA2, SOD1, TOP1, ZNF45. We have found that drug mediated inhibition of etoposide p53SL target ATR, impacts Topoisomerase-mediated enzymatic activities. In addition, we developed another SL screen using genome wide sRNAi approach to identify synthetic enhancement of lethality genes that alter responses of p53 mutant cells to ionizing radiation. In addition we studied the mechanism of Doxorubicin resistance in p53 mutant breast cancer cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Data already obtained show that standard bicycle exercise evokes a greater steady-state O2 consumption and a larger O2 debt during pregnancy than postpartum. Continuing studies will extend observations to the total O2 cost of exercise, and measure changes in fetal heart rate associated with maternal exercise, collating them with maternal responses. The objectives are to measure the hemodynamic and ventilatory requirements of exercise, the changes in these requirements during pregnancy and the range of response shown by normal women, women with a disease limiting O2 transport, women with twins, and women whose fetuses show unusual responses of heart rate to maternal exercise, hypoxemia or oxytocin infusion. Subjects are studied at intervals during pregnancy and postpartum. Measurements are made at rest, during exercise and during recovery: they include the rates of O2 consumption and CO2 production, tidal volume, end-tidal PCO2, maternal heart rate and blood pressure, and fetal heart rate. Minute-to-minute changes in respiration will be obtained, measuring gas flow and composition by a mass spectrometer and a pneumotachograph, and alveolar ventilation, maternal cardiac output and respiratory gas exchange will be calculated. Findings will be correlated with stage of gestation, maternal age, weight, height, blood hemoglobin concentration, and with fetal heart rate in labor, the size and condition of the newborn and placental weight. Measurements in pregnant, trained Pygmy goats of maternal cardiac output and its distribution, fetal heart rate and fetal oxygenation (from blood samples drawn through indwelling catheters), at rest and during standard exercise are currently underway and will be continued. Similar studies will be made in goats with blood-loss anemia.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary objectives of this proposal are to evaluate the use of leukocyte-depleted blood products and/or 8-methoxypsoralen (8-MOP) & UV-A pre-treated random donor platelets (RDP) to prevent primary alloimmunization and to investigate the use of HLA- matched vs random single donor platelets and red cells to prevent broad sensitization among patients with prior alloimmunization. the distinction of patients with and without prior alloimmunization is critical because of their different immunological behavior in response to blood transfusions. Prior sensitization will be determined by conventional cytotoxicity and by flow cytometry. for the first objective, non-alloimmunized patients with newly diagnosed acute non-lymphocytic leukemia will be randomized into 3 treatment groups: 1) Group A will receive only leukocyte-depleted RDP and red cells. 2) Group B will receive only RDP with leukocytes inactivated by 8-MOP and UV-A and leukocyte-depleted red cells. 3) Group C will receive regular RDP and red cells as control. Special emphasis will be made on accurate quantitation of leukocytes that remain in the leukocyte- depleted products so that a potentially immunogenic threshold can be established. A novel method to achieve precise quantitation of low numbers of white cells in leukocyte depleted blood is described. Antibodies will be measured weekly by lymphocytotoxicity and flow cytometry. HLA or non-HLA specificity will be determined by neutralization of antibody with purified HLA antigens. For the second objective, patients with prior allosensitization will be randomized into 2 groups. 1) Group A will receive HLA matched single donor blood products. 2) Group B will receive random single donor blood products. Changes of antibody titers will be measured by flow cytometry and by panel lymphocytotoxic antibody reactivity. Retrospective crossmatches will be performed to determine if 10W level anti-donor antibodies consistently induce poor transfusion responses or acceptable increments can still occur in their presence. The results of the proposed studies will not only allow us to evaluate methods to prevent allosensitization but they will also further our understanding about the role of leukocytes in triggering humoral immune responses to HLA antigens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Retinoids have broad effects including development, growth, cell death, and anti-oxidation. Their actions have been extensively studied in the skin, eye, and in many types of cancer. The liver is a major retinoic acid (RA) target site. Surprisingly, the action of RA in the liver has received very little attention. Our data generated in the past 10 years have uncovered many novel roles of retinoids in the liver. We showed that in the liver retinoids and their receptors have a broad spectrum of actions ranging from xenobiotic biotransformation to cholesterol, fatty acid, bile acid, carbohydrate, and amino acid homeostasis. In addition, hepatic retinoid signaling regulates cell proliferation and apoptosis as well as inflammation. Thus, we hypothesize that RA and its receptors regulate liver metabolism and function in general. The objective for the current application is twofold: to provide an unbiased hepatic genome-wide RA receptor target-gene profile in a gender-specific manner and to elucidate the mechanism by which RA and its receptors regulate gene transcription and expression. Among the diverse pathways, we propose to focus on studying the role of RA/receptors in regulating lipid homeostasis because hepatocyte RXR1 (retinoid x receptor 1) deficiency increases and RA treatment reduces serum lipids in vivo. Furthermore, lack of hepatic RXR1 increases the susceptibility to develop steatosis and steatohepatitis. Three specific aims are proposed to study the global effect of RA/receptors in the liver and the underlying mechanisms. Aim 1 determines genome-wide RA/receptor target genes/pathways in the liver in a gender- specific manner using ChIP-sequencing and microarray analysis. The generated data may account for gender differences in liver function and susceptibility to liver disease. Aim 2 studies the mechanism by which retinoid- mediated pathways regulate lipid homeostasis. We propose to study the mechanisms that regulate two clusters of lipid homeostasis genes, which either do or do not respond to RA but both depend on hepatic RXR1 for their expression. The transcriptional machinery, which dictates RA responsiveness, will be elucidated. Aim 3 studies the mechanism by which RA regulates its own signaling. We will test the hypothesis that RA and hepatic RXR1 regulate genes via modification of chromatin structure and epigenetic signatures. The mechanism by which retinoid signaling is controlled at the basal and RA-regulated level in the liver will be addressed at the transcriptional and epigenetic level. The proposed study may be the first attempt to uncover the fundamental effects of retinoid signaling within the liver with an emphasis on lipid homeostasis. The generated data will have a huge impact on cancer, metabolic syndrome, diabetes, and cardiovascular disease as well as toxicology. PUBLIC HEALTH RELEVANCE: The action of retinoids in the liver has received very little attention. We show that retinoids and their receptors have a broad spectrum of actions ranging from xenobiotic biotransformation to lipid, carbohydrate, and amino acid homeostasis in the liver. The proposed project will profile hepatic genome-wide RA receptor target-gene in a gender specific manner and elucidate the mechanism by which RA and its receptors regulate lipid homeostasis and its own signaling. The generated data will have a significant impact on conditions including but not limited to metabolic syndrome, cancer, toxicology, diabetes, and cardiovascular disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A flexible facility for genetic computation, including both ongoing software development and expansion of hardware resources, is essential for high throughput sequencing operations to succeed. As the technology is new and still evolving rapidly, different platforms are introduced two to three times per year, each requiring some adjustment in order to harness the data pipeline. Stable artifacts must be monitored and recorded in order to eliminate most false calls; annotated coding region and splice junctions must be updated periodically; putative discrepancies that change coding sense must be parsed from those that do not; and finally all calls must be validated or excluded from consideration. Once authentic causative or incidental mutations are established, the data must be ported to a permanent repository without the introduction of error by human operators. This repository, built during the first period of funding, is Mutagenetix. A parallel repository will be established for drosophila mutations. All of these tasks will fall within the purview of Core E. In addition. Core E will model mutagenesis to allow optimized use of ENU in somatic cell mutagenesis studies, to be carried out in Project 1, and in future projects. Core E will be supported by hardware that includes a two large (-3,000 node) Linux cluster computers at the Texas Advanced Computing Center (TACC), a smaller dedicated cluster devoted solely to mapping short reads, and servers that support Mutagenetix (now accessed approximately 3,000 times per month by approximately 1,000 independent users worldwide).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hardware and software for computerized transaxial nuclide reconstruction based on transverse section emission data from living patients is currently being developed in many institutions. This is a particularly promising imaging modality for structure-specific labeling by an enormous number of natural and artificial radioactive compounds. Emission imaging potentially permits not only static reconstruction, but quantitative estimates of time-course of labeled compounds through structures deep within the body. Transaxial reconstruction permits isolation of radioactivity in very small body parts that would be obscured by overlying and underlying radioactivity in conventional views.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The research in my laboratory is centered around computer simulations of liquids and proteins. The simulations are being used to gain a better understanding of how solvation affects a variety of chemical and biochemical processes. The current focus of our liquid state simulations is twofold: (a) the development of algorithms for very large liquid state simulations on massively parallel computers which include novel methods for treating long range interactions, and (b) simulations of polarization effects. There are four ongoing protein simulations projects in my laboratory: (a) the calculation of pKa shifts in proteins from simulations with explicit solvent models, (b) characterization of the thermodynamic and structural properties of the protein solvation shell, (c) computer simulations of protein-ligand interactions, and (d) determination of protein structure and dynamics from simulations and NMR data. This is an umbrella proposal for all the projects in my laboratory.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Comparative genomics is a powerful tool for uncovering patterns of mutations within genes targeted for issues in human health. Our research approach utilizes molecular phylogenetic methods and the well-defined evolutionary relationships (based on multiple genetic markers) that unite the 37 species within the cat family Felidae as a reference phylogeny. Current research is focused on the pattern of diversity and substitution in genes located on chromosomes X and Y. The results indicate genes on the Y evolve in a highly precise manner and accurately reconstruct expected evolutionary relationships. Yet we describe an unusual form of recombination exists between X and Y, represented by one-way exon exchange (ectopic gene conversion) from Zfx to Zfy genes, that may be an adaptive mechanism for maintaining function of Y genes located outside the pseudoautosomal region. Further results identify a novel subfamily of SINEs (repetitive selfish DNA) providing the first example of homoplasy (insertion into an identical site unrelated to shared evolutionary history) that warrants additional investigation of possible functional effects of repetitive DNA in genes located in the non-recombining regions of the Y. These and other insights from comparative genomics describe molecular genetic changes taking place in genes of sex chromosomes that may be highly informative in fertility and reproduction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Anti-angiogenic molecules have tremendous potential in the treatment of cancer. Anti-angiogenic therapy has[unreadable] several benefits over conventional chemotherapy as different tumors can be treated with less toxicity. Most significantly, drug resistance is unlikely. Despite these potential advantages, the utility of current anti-angiogenic agents is limited since the mechanism of action of these proteins is unknown. The PI?s long term objective is to study the mechanism of action of antiangiogenic proteins and to identify the cell surface molecules that interact with these proteins. This proposal focuses on Restin, a C-terminal fragment of the NC1 domain in human collagen XV. This fragment possesses antimigratory function against endothelial cells. This proposal plans to understand the mechanism of action of restin with the following specific aims:[unreadable] Specific Aim 1: To define the structure-function relationship of restin.[unreadable] Specific Aim 2: To determine the intracellular signaling events mediated by restin.[unreadable] Specific Aim 3: To identify and clone the cell surface binding partner(s) receptor for restin.[unreadable] We will use molecular biology and biochemical techniques to address these aims. Once the mechanism of restin?s action has been elucidated, we will understand better the utility of restin and similar agents as anti-cancer drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposal is to understand the adaptive mechanisms in the structure, mechanical properties and function of the coronary arterial vessels to altered mechanical forces, including principal and shear stresses, in experimental models of hypertension and flow overload. The specific aims include studies on alterations in the macro-structure including the length, diameter, and branching pattern of all generations of the coronary tree in response to increased mechanical stresses; alterations in the vessel walls and their micro-structure, including the intima, media and adventitia of all generations; to determine the remodeling of the mechanical properties, including the zero-stress state; and to express the structural and mechanical remodeling data mathematically in terms of indicial functions. This project will provide a structural and mechanical foundation for the study of tissue engineering of the blood vessels, to clarify the role of mechanical stresses on coronary blood vessel remodeling, and to clarify the structural and mechanical remodeling that signifies hypertension and hypertrophy as important risk factors for coronary artery disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Autophagy is an ancient evolutionarily conserved pathway designed to maintain cellular homeostasis by degrading long-lived proteins and organelles in the cytosol. It is also used as a survival mechanism under starvation conditions. Recent studies demonstrated that autophagy is utilized by the cells of the innate and adaptive immune systems to combat viral infections. Innate recognition of viruses occurs via two distinct pathways. In professional viral sensors, the plasmacytoid dendritic cells (pDC), recognition of viruses occurs in the endosomes via Toll-like receptors (TLR) 7 and 9. Our recent work has demonstrated that autophagy plays a key role in recognizing signatures of viral infection in pDCs through TLR7. In contrast to pDCs, most other cell types of the body utilize cytosolic sensors of viral replication via RIG-I like receptor (RLR) family. Molecules involved in autophagy have been shown to block RLR signaling. In addition, recent reports indicate that autophagy delivers endogenous viral antigens to the MHC class II loading compartment, allowing activation of CD4 T cells. However, the relevance of such pathways during in vivo virus infection is unknown. In this application, we present preliminary data that reveal the requirement for Atg5, a key molecule required for formation of autophagosomes, in the transduction of signaling through TLR9 leading to the activation of type I IFN genes in pDCs upon herpes simplex virus (HSV; TLR9 agonist) infection. In addition, we show that autophagy negatively regulates RLR pathway in non-plasmacytoid dendritic cells upon vesicular stomatitis virus infection (VSV; RIG-I agonist). Finally, we demonstrate a key in vivo role for autophagy in the processing and presentation of various forms of antigens on MHC class II in dendritic cells upon HSV-1 infection. Building on these preliminary studies, we propose to examine the importance of autophagy in both innate and adaptive immune responses using a variety of molecular and cell biological techniques and using established mouse models of virus infection. In the first Aim, we will determine the mechanism by which Atg5 and/or autophagy mediates signaling through TLR9 upon HSV infection in pDCs through the use of molecular and cellular biological techniques. In the second Aim, we propose to determine how autophagy regulates RNA sensor activation upon VSV infection through proteomics and biochemical approaches. In the final Aim, we will interrogate how dendritic cells utilize autophagy for processing and presentation of extracellular viral antigens in vitro and in vivo in mice selectively deficient in autophagy within the dendritic cell populations. By providing basic understanding of how autophagy orchestrates the generation of innate and adaptive immunity against virus infections, these studies will help to establish important foundation with which to design vaccines and anti-infective measures against a variety of viral pathogens", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cardiac cells are critically important, because heart disease is the leading cause of death in both men and women is US. About 600,000 people die of heart disease in the US every year-that's 1 in every 4 deaths. Heart failure is caused by loss of function and death of cardiac cells. Thus, transplantation of functional and healthy cardiac cells differentiated from embryonic stem cells (ESCs) offers potential treatments for heart disease. Studies have showed that bone morphogenetic protein-4 (BMP4) plays key roles in cardiac differentiation. The binding of just a few BMP4 molecules with its receptors (BMPR) on single mouse ESCs (mESCs) can induce their differentiation, triggering down and up regulation of a few receptor molecules (SSEA1 and CXCR4), respectively. However, their related molecular mechanisms and how to precisely control their specific cardiac differentiation remain largely unknown. We hypothesize that we may be able to more precisely understand and direct their differentiation into cardiac cells if we can tune the inducer with single-molecule (SM) sensitivity and in real-time. Currently, most differentiation studies were performed by culturing ESCs with inducing agents, observing morphological characteristics of ESCs, using PCR to probe gene expression, or using immunostaining assays to detect biomarkers on fixed dead cells. These methods require several steps, which are unable to real-time study their dynamic differentiation in live cells or guide their differentiation as it occurs. The differentiation of ECs often takes days. Currently, fluorescence microscopy using fluorescence probes is the primary tool for live cell imaging. Unfortunately, fluorescence probes photo-bleach within seconds. Thus, they cannot continuously capture the dynamic events of live cells over hours and days. In this proposal, we aim to develop photostable multicolored single-molecule nanoparticle optical biosensors (SMNOBS), and far-field photostable optical nanoscopy (PHOTON) and use them to study molecular mechanisms and kinetics of cardiac specific differentiation of mESCs induced by the binding of BMP4 with BMPR on single live mESCs, while simultaneously and quantitatively imaging individual SSEA1 (undifferentiated biomarker receptor) and CXCR4 (differentiated biomarker receptor) on single live mESCs in real time with SM sensitivity to monitor their differentiation. Thus, the proposed study will offer innovative tools to direct, contol and study the differentiation of single live ESCs into a specific cell lineage, and depict their related molecular mechanisms and pathways in real time with SM sensitivity. The study will also offer new insights into rationally directing differentiation of mESCs into cardiac cells, and lay down the foundation for design of potential ESC-based therapies to treat heart diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Department of Obstetrics, Gynecology, and Reproductive Sciences proposes to continue the Yale Women's Reproductive Health Research (WRHR) Career Development Center with the goal of selecting outstanding novice or more experienced physician scientists for advanced training in basic, translational, or clinical science under the guidance of skilled scientific mentors. The Principal Investigator/Program Director (PD/PI) and Research Director (RD) will monitor the Scholars'academic progress toward research independence. We identify prospective scholars from our residents, subspecialty fellows, and junior faculty, as well as national networking by the PD/PI and RD and advertisements in professional journals, letters to program directors and Chairs and postings at national meetings. Our selection committee chooses WRHR Scholars who exhibit exceptional promise and commitment to a career in women's health research. The PD/PI, RD, and Advisory Committee are directly involved in the training, evaluation and academic development of each WRHR Scholar. Mentors are experienced in career development and are outstanding scientists in fields related to Women's Reproductive Health. Core training in basic lab techniques and molecular biology, along with a didactic program, are required for all WRHR Scholars and include instruction in the responsible and ethical conduct of research. There are three WRHR Scholars in the Department at any given time, with each Scholar assigned departmental laboratory space with access to shared teaching and training cores. In addition to having 75% time directly covered by the Center, in order to pursue career development. Scholars will receive research support for 2-5 years to underwrite costs while they obtain preliminary data and develop grant applications. Scholars enter the Department of Ob/Gyn as Assistant Professors, with appropriate office and support staff. Clinical duties and responsibilities relate directly to research interests. Career development will span two to five years, depending on the individual Scholar's initial level of experience and particular training goals as a Scholar. The Yale WRHR Center aims to foster Scholars'development into established, independent investigators. Yale seeks to retain as faculty Scholars who successfully complete the individualized training program at Yale and who have made significant progress toward independent funding. PUBLIC RELEVANCE: The Yale WRHR Center will train Obstetrician /Gynecologists to develop into independent physician- scientists with research expertise in women's reproductive health, thereby addressing a national shortage of scientists with the appropriate clinical skills necessary to foster translational research discoveries required to advance women's health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Experts in the treatment of depression have emphasized the importance of striving for remission. Studies have shown that depressed patients who respond to treatment but do not achieve remission, currently defined as a lack of symptoms, continue to experience greater likelihood of continued impairment and future recurrence of depression. However, there is much debate as to how to properly define remission and if the definitions currently used adequately represent the construct. If remission is to be the primary goal of treatment for depression, it is important to use an empirically validated definition. There is also a need to develop tools that can comprehensively assess remission yet be feasibly administered by clinicians in their clinical practice to identify remitted patients. The goal of this Phase I project is to develop an instrument, the Remission from Depression Questionnaire (RDQ) that will provide a multi-faceted assessment of remission from major depressive disorder (MOD). This is in contrast to current definitions of remission that rely exclusively on symptom-based indices. The specific aims of Phase I of the current project are: to generate content rendering a more comprehensive representation of factors related to remission through the use of both patient- and clinician-based focus groups; to use the content generated in the groups towards the composition of preliminary items for the RDQ; to assess the relevance and comprehensibility of preliminary items among patients with depression, modifying items based on patient feedback; to assess the practicality of using the scale in clinical practice; and to examine the comparative relevance of the scale in addressing patient goals relative to the most commonly used self-report, symptom-based measure of depression. At the completion of Phase I we will have developed a comprehensive, understandable measure of remission from depression that is practical to use in clinical practice. Phase II will concern a more rigorous examination of the validity and psychometric properties of the RDQ and will establish the prognostic and incremental validity of the measure over existing symptom-based definitions of remission. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal outlines a Pharmacological Sciences Training Program (PSTP) for years 40-44 at Duke University. The goal of the PSTP is to prepare Ph.D. candidates to use Pharmacology in their careers as active research scientists in leadership roles. This is an interdisciplinary training program composed of students from an innovative triumvirate of Departments of Pharmacology and Cancer Biology, Chemistry and Biomedical Engineering. Leadership in the program is provided by a Director, Co- Director, Executive, Admissions, Curriculum Committee and Trainee Review Committees. Participating faculty are selected on the basis of research interest in Pharmacology and enthusiasm for participating in Program activities including advising Ph.D. students, participation on student committees, classroom teaching, student advising and survival skills mentoring. The training includes coursework in four core courses (Essentials of Pharmacology and Toxicology, Interdisciplinary Approach to Pharmacology, Drug Discovery and Biostatistics) and laboratory research related to Pharmacology. Students also receive training in oral and written presentations in multiple venues including core classes, student seminar, the Pharmacology and Cancer Biology department retreat and attendance at national meetings funded by the training grant. This training is integrated with requirements of the students' specific discipline. Training in Responsible Conduct is a required part of the curriculum. The program sponsors a monthly Survival Skills luncheon for all trainees and students are introduced to the wide array of professional development opportunities offered by Duke University. The Executive Committee selects candidates based on interest in Pharmacology, excellent undergraduate record, strong GREs and outstanding recommendations. The program in Pharmacological Sciences has a long-standing, active and effective minority recruitment program which integrates with strong programs from the University and the Biomedical Sciences.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The main objective of this study is to evaluate the role of sympathetic nervous system in neurovascular control in patients with diastolic heart failure. In this protocol, the investigators plan to study muscle sympathetic nerve activity and plasma norepinephrine levels in patients with chronic heart failure due to diastolic heart dysfunction during rest, exercise (handgrip exercise) and orthostatic stress (head up tilt). They also plan to study the effects of exercise on renal blood flow in these patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We shall isolate dimeric intermediates in the genetic recombination pathways of phage lambda. Dimers generated by the Red, Rec, and Int pathways will be separately characterized. The role of DNA synthesis in Red-mediated recombination will be determined by assessing the involvement of synthesis in dimer formation and dimer resolution. Dimers will be isolated by isopycnic centrifugation following infection of DNA-zero cells with dense and with radioactive particles. The isolated dimers will be defined topologically and topographically (following partial denaturation) by electron microscopy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (From the Applicant's Abstract): MRSDA Candidate: The applicant describes a plan to broaden his research experience and skills in sophisticated imaging approaches for cellular neuroscience. A mechanism of structural plasticity on dendrites of CA1 hippocampal pyramidal neurons will be investigated drawing on techniques in serial electron microscopy and immunogold labeling, and training in multiphoton laser scanning microscopy, one of the most advanced techniques for live tissue imaging. The candidate's long-term goal is to become an independent investigator in an academic setting. Environment: The MRSDA sponsor is one of the foremost researchers studying synaptic ultrastructure. The investigator will interact with developmental, cellular, molecular, and systems neurobiologists as well as neuroethologists in a diverse Department of Biology and neuroscientists in other Departments. Two faculty collaborators with expertise in immunogold labeling and optical physics will participate in the MRSDA program. The sponsor's laboratory is outstanding, possessing all of the physical and instrumental resources needed for this MRSDA program and comprised of research professionals, postdoctoral fellows, and technical staff. Research: Most excitatory synapses in the adult brain occur on dendritic spines and changes in spine number or structure and composition can affect communication between neurons. It was thought that spine number would increase to meet the demand of enhanced synaptic activity and vice versa. We have shown on the mature hippocampal neurons that spine number dramatically increases when synaptic activity is reduced or blocked. In contrast, developing neurons appear to require activation to initiate spine formation. The MRSDA specific aims are: 1) Determine under what conditions of neuronal activity dendritic spines form on immature hippocampal neurons. 2) Establish the ultrastructural locations of synapses under conditions of spine formation on immature hippocampal neurons. 3) Determine how rapidly new spines can form on mature hippocampal neurons when synaptic activity is blocked. 4) Compare the composition and detailed structure of newly formed versus previously stable dendritic spines on mature hippocampal neurons. The results will have important implications for understanding how new spines might be generated when overall neuronal activity is low and could be lost during excessive activation such as with epileptic seizures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although natural disasters present serious challenges for residents of affected communities, many questions about their effects on the physical health of older adults remain. The long-term goal is to inform intervention and resource planning efforts designed to mitigate the effects of disasters on older people, reduce health care utilization and expenditures and improve the quality of life of older people. The objective of this application is o identify how trajectories of functional limitations are affected by disaster. Our central hypothesi is that the majority of older adults exposed to disaster are resilient, healthcare utilization and expenditures associated with some functional limitation trajectories in the wake of disaster are significant, and pre-disaster characteristics of individuals and communities can distinguish trajectories of resilience and vulnerability. Our hypothesis was formulated based on our previous empirical and conceptual work and on work by others. The rationale for the proposed research is that, once it is known how disasters affect functional limitation trajectories, interventions ca be developed that will reduce the vulnerability of older adults and increase their resilience in th face of disaster, resulting in new and innovative approaches to improving the quality of life for older people. The hypothesis will be tested by pursing three specific aims: (1) To ascertain the effects of a natural disaster on the functional limitation trajectories of older adults; (2) To contrast the healthcare utilization and expenditures associated with functional limitation trajectories for older people experiencing disaster with those of older people not exposed to disaster; and (3) To identify the pathways by which risk factors and resilience resources influence trajectories of functional limitation. The aims will be addressed by building on a longitudinal panel of 1,977 people aged 65 to 80 when Hurricane Sandy hit New Jersey. Three post-hurricane self-reported assessments would complement data from two prior data collection efforts. These data would be linked with Medicare/Medicaid claims data. This work is significant because it is the first step in a continuum of research expected to lead to the development of interventions that will bolster the capabilities of older people to face natural disasters without increasing functional limitations. The proposed research is innovative, because it: (1) focuses on functional limitations, a linchpin in the disability cycle, (2) examines the effects of disaster ona large representative sample of people living in one of the states hardest hit by a recent hurricane, and (3) builds on data collected before the storm struck, includes people with and without direct exposure to the storm, and has 3 post- disaster assessments, yielding one of the few studies able to disentangle causal relationships attributable to disaster. Knowledge from this project will advance understanding of disaster's effects on older people, and inform interventions likely to improve their quality of life, both before and after disasters strike.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "American Indian/Alaskan native (AI/AN) children have overweight rates that are 18-30% higher than the general American population of children. Adult obesity has early antecedents, and date show that eating and physical activity behaviors are formed and set as early as 3 years of age, through primary socialization within families. Yet there are few, if any, obesity preventionprograms that target children younger than 3 years of age. We propose a community-partneredrandomized study to prevent early childhood overweight in American Indian (AI) children.A birth cohort of about 780 children from six Indian communities born over a period 18 monthswill be randomized by community to either a control (3 communities) or intervention condition (3 communities). The interventioncomprises a community-wide intervention coupled with individualized family counseling to improve nutrition and physical activity in infants and toddlers. Nutrition goals are to increase breastfeeding initiationand sustainability, limit sugared beverages for infants and toddlers, and make healthful choices during weaning and solid food introduction. Physical activity goals include promoting motor development, limitingvideo/TV viewing and creating play opportunities for infants and toddlers. Interventionapproaches and methods used in the pilot study on which this project is based will be combined with new formative data collected at the beginning of the project to enhance the intervention's feeding and physical activity components. Each component in the community-wide interventions will be collaboratively designed with the tribes, and tailored to each community's needs. Trained peer counselors will deliver the family interventions,during a series of 12 cluster visits, each covering a different developmental stage of the infant/toddler. The primary outcome is BMI Z scores at ages 24-30 months.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary The protein tyrosine phosphatases (PTPs), which dephosphorylate specific phosphotyrosine residues in protein substrates, constitute a large family of signaling enzymes, whose activity is ubiquitously misregulated in human diseases such as leukemias, solid-tumor cancers, type I and type II diabetes, and autoimmune disorders. Tools that are capable of inducing target-specific PTP inhibition or activation would be invaluable for delineation of the precise functions of PTPs in cell signaling and for the validation of PTPs as therapeutic targets. However, the common architecture of the conserved PTP-domain fold impedes the discovery of selective PTP inhibitors, and cell-permeable PTP activators have not been identified to date. The broad objective of the proposed research is to develop tools to specifically inhibit or activate PTP enzymes by targeting either engineered cysteine residues (Specific Aim 1) or naturally occurring cysteine residues (Specific Aim 2). Specific Aim 1 (Target-specific inhibition and activation of engineered PTPs with biarsenical small molecules) proposes several complementary means for engineering inhibitor- sensitive and activator-sensitive (collectively, ligand-sensitive) PTPs: engineered phosphatases whose activities are uniquely responsive to small molecules. The proposed inhibitor- and activator-sensitization strategies, which can potentially be applied widely across the PTP superfamily, will afford PTP-signaling researchers an unprecedented level of chemical control with which to study a key family of signaling enzymes. Specific Aim 2 (Targeting naturally occurring cysteine residues for the discovery of Shp2- specific inhibitors and activators) is focused on selectively targeting naturally occurring cysteine residues in a medicinally important PTP, Shp2, with the goal of discovering compounds that have direct implications for drug development. The proposed experiments are aimed at the discovery of drug-like compounds that can target Shp2?s unique allosteric site, potentially providing highly selective Shp2 inhibitors that have not been realized through active-site- directed efforts. Also proposed are small-molecule-discovery efforts targeting two disease- causing Shp2 mutants that contain missense cysteine residues (Y63C and Y279C Shp2, which cause Noonan and LEOPARD syndromes, respectively). Both mutations affect the enzymatic activity of Shp2; efforts to identify compounds that restore wild-type-like Shp2 function by targeting the disease-causing cysteine residues are proposed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Self-renewal and pluripotency are defining properties of an embryonic stem cell (ESC). Although extremely transient in vivo, these cells can be captured from the inner cell mass of the embryo and maintained indefinitely in a pluripotent state or differentiated into essentially any specialized cell in vitro. As such, embryonic stem cells (ESCs) hold great promise for regenerative medicine, which seeks to use ESCs to regenerate or rejuvenate cells and tissues that have been damaged due to injury, disease, or underlying genetic mutations. However, to leverage their full potential, we must understand the mechanisms that control ESC self-renewal and pluriptency to allow (re)generation of any tissue while preventing aberrant growth or depletion of the stem cell pool. Stem cell identity is defined by a gene regulatory network dictated by the master regulators OCT4, SOX2 and NANOG. However, the binding of these transcriptional factors is in turn highly dependent on the accessibility of the chromatin landscape. To access the underlying genomic information, histone proteins must be repositioned or removed, a function that is performed by ATP-dependent chromatin remodeling complexes. In particular, several components of the mammalian SWI/SNF or BAF complex are essential for formation of the inner cell mass and for derivation of ESCs in vitro. BAF complexes are regulated by the combinatorial assembly of homologous subunits from gene families. However, there is currently no mechanistic understanding of how unique combinations of distinct subunits affect BAF complex targeting or function. Using biochemical approaches, a completely novel and previously undescribed form of the BAF complex was discovered in ESCs that contains the acetyl lysine reader, Bromodomain-containing protein 9 (BRD9). Due to the incorporation of BRD9 and other unique subunits, the BRD9-containing BAF complex or 'BBAF complex' is uniquely targeted across the genome and exhibits specific regulatory interactions not found associated with canonical BAF complexes. Moreover, the BBAF complex lacks certain subunits that are critical for ATP- dependent chromatin remodeling, suggesting that it may have alternate function. The goals for the next five years are to determine the mechanism of BRD9-mediated chromatin targeting of the BBAF complex and how that relates to the transcriptional regulation of genes involved in ESC self-renewal and pluripotency. These studies will aid in our understanding of how BAF complex heterogeneity contributes to the precise control of the ESC transcriptional program and provide a framework for understanding the roles of individual BAF subunits in contributing to cell type- and developmental stage-specific function of BAF complexes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Single-molecule Forster resonance energy transfer (FRET) measurements on freely diffusing molecules contain information about conformational dynamics because the rate of energy transfer depends on the distance between donor and acceptor labels attached to a molecule. In these experiments, a molecule diffuses through a spot illuminated by a laser, and the donor is excited. The output of these experiments is a sequence of photons of different colors (some emitted by the donor and some by the acceptor) separated by apparently random time intervals. As described in last years report, we have developed a rigorous theory that describes how statistics of photons is influenced by protein conformational dynamics, the diffusion of the protein through the laser spot, shot noise, etc. It was shown that the exact FRET efficiency and photon counting histograms can be obtained by solving an appropriate reaction-diffusion equation. We have obtained a simple analytical yet rigorous result for the width of FRET efficiency distribution and showed that the shape of the distribution depends dramatically on the bin size.[unreadable] [unreadable] This year our continued efforts have lead to four publications in this area. First, we have written (1) a book chapter where we have applied our general theory of photon counting to systems with fluorescence quenching. Second, we introduced a new and simpler to use method of analyzing single-molecule data (2). Third, we applied (3) this new method to interpret fluorescence resonance energy tranfer in polyproline (in collaboration with the W.A. Eaton's group, LCP/NIDDK). Finally,we answered the question when do single-molecule experiments actually measure the properties of one molecule by quantatively determining the influence of fluorophore concentration on the observed photon statistics.[unreadable] [unreadable] An important advance we made (2) was to show how to extract information about conformational dynamics from FRET experiments on diffusing molecules without modeling diffusion. Starting from a rigorous theory that does treat diffusion, we first examined when the single-molecule FRET efficiency distribution can be decomposed into the measured distribution of the total number of photons and the efficiency distribution of an immobilized molecule in the absence of shot noise. If the conformation does not change during the time the molecule spends in the laser spot, this is possible when (I) the efficiency is independent of the location in the laser spot and (II) the total number of photons does not depend on conformation. This decomposition is approximate when the conformation changes during the diffusion time. However, it does provide a simple framework for analyzing data. This is illustrated for a two-state system where the FRET efficiency distribution can be found analytically for all values of the interconversion rates. [unreadable] [unreadable] As an alternative to the analysis of FRET efficiency distributions, we introduced a simpler procedure that allows one to extract the rates of conformational changes by decoding the pattern of colors in the donor/acceptor photon trajectory (2). This can be done in the framework of statistical inference because the likelihood function, which must be optimized with respect to the model rate parameters, depends only on how the conformation changes during the interval between photons with specified colors. The procedure works even when the photon colors appear to be scrambled (i.e., one cannot identify states by visual inspection of a photon trajectory) because the photophysical properties of the conformers are similar and/or conformational dynamics is on a similar time scale as the photon counts.[unreadable] [unreadable] The above work has played an important role in analyzing recent experiments performed by Dr. W. A. Eaton's group (3). In this project, quantitative analysis of the FRET efficiency distribution of polyproline was performed. The analysis resolved the puzzle about the excess width of the distribution and allowed one to draw important conclusions concerning the flexibility of the all-trans form of polyproline and the distribution of cis isomers. More details are provided in Dr. Eaton's report.[unreadable] [unreadable] Finally, the optimal concentrations for single-molecule measurements of freely diffusing molecules (4) was established. It was found that a concentration as low as 0.1 molecule per observation volume may not be small enough for single-molecule FRET efficiency measurements. This result follows from a rigorous theory that takes many molecules into account. We considered the distributions of the number of photons (photon counting histograms) and showed that multiple-molecule effects are pronounced at large photon counts even at low concentrations. FRET efficiency distributions reveal multiple-molecule effects at large threshold values. This might be misinterpreted as multiple conformational states. Multiple-molecule effects strongly depend on the brightness of fluorophores. A simple test was suggested to determine parameters for which the single-molecule description is applicable.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The tissue lesion of the connective tissue diseases appears to be due to vascular injury induced by immune complexes. The cold precipitable immunoglobulins found in the circulation of many patients with these diseases have all the characteristics of immune complexes. Since cryoglobulins are easily isolated and quantified, we propose to: (1) study their relationship to the activity and vascular complications in RA and SLE, (2) analyze those features, such as complement fixing ability, that could account for their phlogogenic capacity, (3) determine their ability to influence lymphocyte function (especially lymphocyte-mediated cytotoxicity) and, finally, (4) to attempt to isolate antigens from SLE cryoglobulins that might be important in the pathogenesis of that disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The fulminant hepatoxicity caused by halothane has been thought to have an immunological basis because this toxicity occurs most often after repeated administration of halothane and because sera from patients recovering from severe halothane hepatoxicity have been found to contain antibodies that bind to the surface membranes of hepatocytes of rabbits treated with halothane. In order to determine whether halothane's major reactive metabolite, trifluoroacetyl halide (CF3COX) results in covalent binding of trifluoroacetyl (TFA) groups to hepatocytes, we have developed specific and sensitive peroxidase enzyme-linked immunosorbent assays (ELISA) and an indirect immunofluorescence staining method for identifying TFA-hepatocytes. Liver sections prepared from rats at 4 hr after halothane administration were stained preferentially in the centrilobular region with anti TFA antiserum. The specificity of the assay for the TFA group was confirmed by the complete inhibition of the staining by TFA-Llysine. Moreover, treatment of rat with deuterated halothane resulted in significantly less staining than did halothane. Twenty-four hours after halothane administration hepatocytes were isolated and stained by indirect immunofluorescence. The immunofluorescence showed a linear or a granular pattern on the surface membranes. These results indicate that CF3COX either reacts directly with plasma membrane or with other cellular components which become incorporated into the plasma membrane. In order to study further the immunological mechanism of halothane hepatitis, we intend to determine what component is modified by TFA, and how these altered hepatocyte membranes can initiate an immune response, which can result in hepatocellular damage. We also plan to develop an animal model of halothane hepatitis to evaluate the immunological mechanism, and to study this disease in greater detail. The immunological methodologies that will be developed in these studies may serve as a model for investigating the mechanism of other drug induced toxicities which are believed to have an immunological origin. In susceptible individuals these membrane changes may lead to sensitization and damaging immune reactions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This protocol describes a study designed to compare a new sleeping pill, zolpidem with a currently available preparation called tempazepam with regard to \"morning after performance\". This will be a multicenter study involving 15 centers and approximately 630 normal subjects, 42 of whom will be enrolled a the BWH. Subjects will be randomized to three treatment groups such that 3/7 will be in each drug group and 1/7 in placebo group.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Many patients receiving chemotherapy and/or ionizing radiation (IR) develop acute and residual (or long- term) bone marrow (BM) injury that limits the success of cancer treatment and adversely affects their quality of life. Acute myelosuppression is the result of the induction of apoptosis in the rapidly proliferating hematopoietic progenitor cells (HPCs) and to a lesser degree in the relatively quiescent hematopoietic stem cells (HSCs). Its clinical manifestations have been successfully managed by the use of various hematopoietic growth factors. In contrast, residual BM injury has been largely attributed to the induction of HSC senescence. However, neither the molecular mechanisms by which chemotherapy and/or IR induce HSC senescence have been clearly defined, nor has an effective treatment been developed to ameliorate residual BM injury. Recent studies from our laboratory and others provide new insights into HSC damage. First, we have found that exposure of mice to a sublethal dose of total body irradiation (TBI) perturbs the balance of reduction/oxidation (redox) reactions ONLY in HSCs, leading to a persistent and prolonged increase in reactive oxygen species (ROS) production. Second, HSCs are more sensitive to ROS-induced oxidative damage than HPCs and other hematopoietic cells. Moreover, it appears that ROS injures HSCs not by a nonspecific cytotoxic effect as previously hypothesized. Instead, the damage is at least partially mediated by induction of cellular senescence through redox-dependent activation of the p38 mitogen-activated protein kinase (p38)-p16lnk4a (p16) pathway. Based on these novel findings, we hypothesize that chemotherapy and IR cause residual BM injury by SELECTIVELY inducing HSC senescence through ROS-mediated activation of the p38-p16 pathway. Thus, we predict that antioxidants can be used to effectively mitigate residual BM injury. Moreover, antioxidant therapy provides additional benefits to cancer patients by suppressing chemotherapy- and IR-induced genetic instability, a primary cause of secondary tumors and a contributing factor to the development of tumor resistance. Therefore, this strategy offers the promise of significantly improving the quality of life and increasing the efficacy of chemotherapy and radiotherapy for cancer patients. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Addressing the needs of dying children- a benchmark of societal compassion and a gauge of health care quality-rises as a challenge for the 21st century. As deaths due to injury, prematurity, and sudden infant death syndrome are prevented, the remaining deaths may be increasingly associated with chronic conditions. I hypothesize that deaths attributable to complex chronic conditions (CCCs) constitute an increasing proportion of all pediatric deaths, both at a population level and more specifically among patients cared for by children's hospitals, and that the prolongation of life has led to a rising prevalence of these conditions. Furthermore, I postulate that quality of care and support for these dying children and their families deteriorates over the last two years of life, with declining continuity of care and untimely referral to hospice and other social support services. This five-year research plan will profile the needs of dying children specifically aiming to: 1. Determine whether the prevalence of terminal congenital CCCs is rising in the national population. 2. Measure the change in prevalence of children with terminal CCCs in tertiary children's hospitals, thereby assessing the magnitude of need for hospital-based pediatric palliative care services. 3. Test whether the 'intensity' of medical care increases in the 30-day interval prior to the day on which death compared to the preceding 23 month interval, so as to improve clinicians prognostic inferences regarding likelihood of death. 4. Examine two markers of quality of care of children with CCCs by: Testing whether the continuity of care deteriorates in the last two years of life for children with CCCs; and 5. Examining the timing of referral to hospice. This research seeks to improve the care and health policy for dying children in three ways. First, evidence of a 'epidemiological transition' in pediatric mortality-from mostly accidental and sudden death to deaths that occur somewhat inevitably with a longer yet unpredictable dying process-would motivate the redesign of pediatric health care services and physician training at both the regional and national level. Second, this research will produce several techniques to monitor health utilization data for indicators of the quality of care provided to terminally ill children. Finally, the results of these studies will ultimately inform the development and trial of a longitudinal needs assessment program for children with complex chronic conditions. These studies of the epidemiology of pediatric death and the needs of dying children will help enhance the individual care provided to these children and their families as well as broader policy. They will also foster the development of the applicant's research skills so as to become an independent researcher addressing the needs of children with complex chronic conditions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The alveoli of the lung are covered by a thin layer of surface-active material, referred to as pulmonary surfactant. This material, which is essential for normal gas exchange, consists of phospholipids and specific proteins. The best characterized surfactant proteins include SP-A, and two smaller hydrophobic proteins, SP-B and SP-C. We have recently purified a collagenous surfactant-associated glycoprotein from rat lung. This protein, designated SP-D, is different that SP-A and previously described collagenous proteins. Preliminary experiments indicate that SP-D has carbohydrate-binding activity in vitro, and that there is structural homology of SP-D with the C-type (calcium-dependent) lectins. It is our hypothesis that the carbohydrate-binding activity mediates interactions of SP-D with other components of surfactant, and that carbohydrate-mediated interactions contribute to the function of this protein and pulmonary surfactant in vivo. We propose the following specific aims: 1. To further define the structure of SP-D; 2. To characterize its carbohydrate-binding activity; 3. To identify the ligands for SP-D in surfactant; and 4. To explore the possible contributions of SP-D to surfactant activity and metabolism. These studies should contribute important new information relating to the macromolecular composition, organization, and functions of pulmonary surfactant, and to the structure and function of collagenous lectins.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This request for core support is based on the desirability of maintaining and improving a research resource in the fundamental aspects of radiation oncology now in existence at this institution. Although all individual research projects presently associated with the Divisions of Radiation Biology and Radiologic Physics have been subject to peer review and are funded by separate grants, they also have a common primary objective - namely, the study of the basic nature of cancer and its detection and treatment through the use of radiation. A perhaps less important but inseparable objective of the staff associated with these endeavors has been the education and training of individuals specializing in various aspects of radiation effects and ranging from pre-doctoral graduate students to post-doctoral investigators and residents in radiology. Funding aimed at (1) support of common service type mechanical shops, tissue culture facilities, library, and administration, etc., (2) salary support for staff investigators, and (3) modest but critical developmental research support should materially improve the efficacy of the personnel and eventually increase research contributions in radiation oncology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies from our laboratory over the past decade have found the existence of a novel light-dependent insulin receptor (IR) signaling pathway in rod photoreceptors. We have discovered that IR activation is functionally important for rod survival, since its deletion in rods resulted in the loss of neuroprotective survival signaling. This novel pathway uses growth factor receptor bound protein (Grb14), an upstream regulator of IR, and requires photobleaching of rhodopsin for membrane targeting. Grb14 protects light-dependent IR activation in rod photoreceptors against dephosphorylation by PTP1B. Light-activated IR is subsequently associated with phosphoinositide 3-kinase (PI3K), a cell survival factor, and thus regulates the downstream survival pathway. These studies suggest that rhodopsin photoexcitation may trigger signaling events alternative to the classical transducin activation. Recently, it was suggested that the IR signaling pathway is important for cone photoreceptor survival in retinitis pigmentosa (RP) models since systemic administration of insulin delays the death of cone photoreceptors. We found that insulin-like growth factor-1 receptor (IGF-1R) also activates PI3K and Akt survival pathway in rods under light stress. It is our hypothesis that IR/IGF-1R signaling pathways are important for survival and maintenance of rod and cone photoreceptor structure and function. The long-term goal of our project is to gain a greater understanding of the intracellular signaling pathways that provide neuroprotection to both rod and cone photoreceptor cells. The specific objective is to investigate the role of IR and IGF-1R in the regulation of photoreceptor structure and function. To this end, we will determine the mechanism by which Grb14 activates the IR and determine the functional roles of IR and IGF-1R in both rod and cone photoreceptor cells. We will utilize a combination of genetic, molecular, and biochemical approaches to address our specific aims. Results from these studies will lead to a better understanding the roles of IR and IGF-1R signaling in photoreceptor structure, function and survival. PUBLIC HEALTH RELEVANCE: Insulin receptors (IR) and insulin signaling proteins are widely distributed throughout the central nervous system (CNS). Disregulation of IR signaling in the CNS has been linked to the pathogenesis of neurodegenerative disorders such as Alzheimer's and Parkinson's disease. In retinal rods, IR signaling is neuroprotective, and recently it was shown that insulin delays the death of cone photoreceptor in retinitis pigmentosa animal models. Besides IR, insulin-like growth factor-1 receptors (IGF-1R) are also expressed in both rods and cones;however, their functional role has not been elucidated. Our studies suggest that protein tyrosine phosphatase 1B (PTP1B) negatively regulated neuroprotective survival signaling of the IR. Our studies also suggest that a defect in the photobleaching of rhodopsin and mutations in rhodopsin gene enhances the activity of PTP1B and this activated activity could down regulate the IR survival signaling. Controlling the PTP1B activity and activating the IR signaling would provide sustained neuroprotection to both rods and cones. Our new and innovative approaches to target PTP1B will facilitate future translational application of our work, with the goal of applying our findings to the clinical care of human retinal degenerations. Studies proposed in this grant application would help to understand the functional roles of IR and IGF-1R in photoreceptor structure, function and survival.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Type II Diabetes Mellitus is one of the most common diseases in the Western world, with prevalence ranging from 10-50% in elderly populations. An integral component of Type II diabetes mellitus is insulin resistance, i.e. the inability of maximal concentrations of the hormone to stimulate appropriately muscle glucose transport, suppression of hepatic glucose output and other physiological responses. A plausible strategy leading to the development of novel insulin-sensitizing drugs begins with the systematic analysis of insulin signaling. After over 30 years of intensive research in the pursuit of relevant protein kinases, Akt/PKB has emerged as the only serine/threonine protein kinases definitively established as a mediator of insulin's regulation of important metabolic targets. The activity of Akt/PKB depends on the lipid products of phosphotidylinositol 3'- kinase, an enzyme known to be required for virtually all of the metabolic actions of insulin. The Akt/PKB kinases represent a family of three closely related proteins that have been implicated in the regulation of processes as diverse as apoptosis, cell growth, cell cycle progression, differentiation and angiogenesis as well as metabolism. Recent work has shown that mice deficient in Akt2/PKBbeta mimic several features of diabetes, including insulin resistance in liver, muscle and adipose tissue. Interestingly, Akt1/PKBalpha null mice are absolutely normal metabolically, but are about 20% reduced in size. The differences in phenotype between Akt1/PKBalpha and Akt2/beta knockout mice are due to both to isoform-speciflc patterns of expression as well as distinct signaling properties intrinsic to the proteins. The underlying rationale of these proposed studies is that by investigating the biochemical basis of the preferential ability of Akt2/PKBbeta to signal to glucose transport, significant progress will be made in understanding the fundamental mechanism of insulin action. Specificity will be approached using several related strategies: 1) the domains of Akt/PKB conferring isoform specific signaling will be mapped by the use of chimeric proteins;2) the two isoforms will be analyzed for differences in intracellular localization;and 3) a search will be conducted for Akt2-specific substrates. It is anticipated that these studies will significantly extend the knowledge of physiological insulin-independent signaling and ultimately lead to the identification of potential targets for therapeutic intervention in diabetes mellitus.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The spatial distribution of energy from ionizing radiations in cells is a strong determinant of radiobiological response. It is the objective of this research to derive an understanding of this dependence by the use of short-ranged radiation sources. In particular we intend to probe the mechanism(s) whereby chromosomal changes come about, on the understanding that chromosomal changes are intimately associated with cell death, mutation, anj cancer. We have successfully used Al soft X rays for radiobiological investigations and intend to construct a gas target (B,C,N,O,Ne,Ar) for incident protons as a source of a wide range of soft X rays with energy depositions down to about 200 eV with electron tracks of a few nanometers. We will also use soft neutrons which produce recoil Proton tracks of sub-micrometer ranges. This radiation source will provide information of direct relevance to radiation protection, where personnel exposure to soft neutrons carries an as yet undetermined risk. Thirdly, the localized energy depositions from I-125 disintegrations will be used to ask about the basis of aberration formation. Mammalian cells (human and hamster) will be irradiated at different stages of the cell cycle and the frequencies and types of chromosomal changes, cell survival and cytokinetic changes monitored. Low-dose studies emphasizing response and the basis thereof in the initial part of dose-response curves will be stressed. Analyses of responses will be oriented to distinction between models of biophysical action, e.g., \"critical lesion\" versus \"dual lesion.\" The detailed track structures of the radiations will be calculated, and analyses of the biological responses will lead to predictions for other radiations. Stringent testing of models of biophysical action in this way will lead to model acceptance or rejection. In aiding understanding of mechanisms leading to the formation of chromosomal changes and to the biological effects of ionizing radiations, this proposal will impact significantly in areas concerned with optimization of radiation therapy and the determination of standards for human radiation protection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objective of the proposed project is to separate, characterize and localize the acid phosphatases in developing bones and the enamel organ of tooth buds and to determine the effect of parathyroid hormone and vitamin D in vivo/in vitro on the different phosphatases, in order to gain insight into the role of these enzymes in hard tissue formation and function. After solubilization of the acid phosphatases in rat tissue homogenates, separation and purification of the enzymes will be achieved by means of ion-exchange chromatography and polyacrylamide disc gel electrophoresis. The enzymes will be characterized in terms of molecular size, pH optimum, thermolability, kinetic characteristics for various substrates and the influence of various effectors on the kinetic characteristics. Information concerning the function of the various acid phosphatases will be obtained by determining (a) the specificity of the enzymes for potential in vivo substrates; (b) localizaton of enzymes in cells and cellular organelles and (c) by determining the effect of vitamin D and parathyroid hormone on activity and tissue level of specific acid phosphatases in bone and tooth buds in vivo and in bone cultured in vitro. The latter studies will be carried out with vitamin D-deficient suckling rats, or, for the in vitro experiments, with bones from such rats. Localization of the enzymes will be determined by means of (a) biochemical studies on dissected tissues containing primarily one type of cell; (b) cytochemical techniques at the light and electron microscopic levels using specific substrates and inhibitors determined biochemically; and (c) immunocytochemical techniques using antiserum to one or more specific phosphatase.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The project is focused on a neglected area within children's health services research, the relationship between provider-child-caregiver communication during pediatric asthma visits and treatment adherence. Treatment adherence comprises medication adherence, symptom monitoring, and environmental trigger control. Identification of effective communication strategies is important for future intervention research. Thirty-five physicians and their nurses and 360 English-speaking child-caregiver pairs will be recruited at eight pediatric clinics. Children will be eligible if they are between 8 and 15 years of age, have mild, moderate, or severe persistent asthma and have previously visited this clinic at least once before for asthma. Caregivers will be eligible if they are at least 18 years of age and live with the child with asthma. Consent and assent to participate will be obtained from the caregivers and children respectively. Physicians and nurses will fill out a demographic questionnaire at the start of the study. Following enrollment into the study, the children will have their medical visits audio-taped. The children will be interviewed after their medical visits, asked to demonstrate inhaler and peak flow technique, and assessment of their lung function will occur using spirometry. The children's caregivers will fill out questionnaires after these medical visits. A home visit will occur one month after the audio-taped visit. Children will be interviewed during the home visits, asked to demonstrate inhaler and peak flow technique a second time, and have their lung function examined using spirometry. Caregivers will fill out questionnaires. Generalized estimating equations will be used to examine how physician and nurse communication about asthma management, modeling of asthma care behaviors, and engagement of the child and caregiver during medical visits, are related to treatment adherence.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cerebral amyloid angiopathy (CAA), the age-related deposition of cerebrovascular ft-amyloid (AH), is a common cause of hemorrhagic stroke, an accompanying pathology in most cases of Alzheimer disease (AD), and a potential cause of adverse responses to anti-All immunotherapeutic approaches to AD. Studies of CAA in the postmortem human brain are limited by the inability to make observations over time as the disease progresses. We have used in vivo multiphoton microscopy in mouse models of CAA to define the spatial and temporal progression of the disease in living animals. We have shown that cerebrovascular AH deposition occurs predictably with readily measured kinetics and that passive immunotherapy alters the progression of CAA and can lead to clearance of amyloid deposits from vessel walls. The consequences of CAA are associated in part with breakdown of the vessel wall. For this reason, we now propose to use novel methods to define the mechanisms that underlie Ali-induced vascular wall injury, including oxidative stress, activation of matrix metalloproteinases and other proteases and injury to vascular smooth muscle cells. In Specific Aim #1, we will perform serial imaging of CAA progression in mouse models and define the spatial and temporal relationship between amyloid deposition and markers of injury to the vessel wall. The goal of this aim will be a precise understanding of the sequence of events in the pathway that lead to vascular wall injury. In Specific Aim #2, we will examine which of the CAA-induced alterations in vessel wall injury can be interrupted or reversed by clearance of Afi using passive immunization and other methods. Because mouse models are only relevant in as much as they speak to human disease, in Specific Aim #3, we will determine which of our validated set of markers of vascular wall injury from mouse models are also found in human CAA. We will make use of our expertise in the clinical and neuropathologic characteristics of CAA and our access to a wide range of human samples (including sporadic CAA, CAA linked to APP mutations, tissue from the AN1792 trial of the Aft vaccine and familial British and Danish dementias. From these studies we will develop a clearer understanding of (1) mechanisms by which AIS deposition results in vascular wall injury and neurologic injury;(2) points suitable for therapeutic intervention;and (3) risks associated with the CAA in patients undergoing anti-amyloid therapies for Alzheimer disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "7. PROJECT SUMMARY/ABSTRACT Mental health disorders cause immense personal suffering and represent a significant societal burden. Recent research emphasizes the potential of psychiatric electroceutical interventions (PEIs) ? bioelectronic treatments that employ electrical stimulation to affect and modify brain function ? to effectively treat such disorders. Novel PEIs, however, also raise significant ethical concerns. Not uncommonly, they are negatively associated with historically controversial interventions such as electroconvulsive therapy and lobotomy. Moreover, while their goal is to alter the underlying pathophysiology of disease, these interventions can also have unintended effects on cognition, behavior, and emotions. Those effects, in turn, raise ethical concerns regarding autonomy, personal identity, and capacity for informed consent. PEIs also are sometimes perceived to be more harmful than helpful. A large body of scholarly literature addresses ethical concerns about pharmacological psychiatric interventions, yet there is limited research on ethical concerns and related beliefs and attitudes about PEIs among the clinicians, patients, and the broader public. With the attention novel PEIs have garnered, this proposed study addresses that significant gap by (a) examining ethical concerns, beliefs, and attitudes about the use of PEIs among relevant stakeholders, and (b) developing a guide to anticipate future policy challenges regarding PEI innovation and use. Using a pragmatic neuroethics framework together with a responsible research and innovation framework, our goal is to develop a strategy to support responsible and ethical PEI innovation and use. To achieve this goal, our established, transdisciplinary research team, guided by a distinguished Scientific Advisory Board, will answer three questions: (1) How do relevant stakeholder groups perceive different PEIs for the treatment of depression? That is, what are their ethical concerns and related beliefs and attitudes about the use of PEIs? (2) How do PEI characteristics and disease severity shape ethical concerns and related beliefs and attitudes about PEIs? (3) How does the influence of PEI characteristics and disease severity vary across stakeholder groups? To answer these questions, we will analyze data from hypothesis-driven, national surveys containing an embedded factorial experiment (Aims 1-3). We will examine the key factors shaping ethical concerns and related beliefs and attitudes about PEIs among three relevant stakeholder groups: medical professionals (Aim 1), patients diagnosed with depression (Aim 2), and the broader public (Aim 3). These results will inform the development of an analytical map of the ethical challenges and policy choices that ethical concerns, beliefs and attitudes might generate for PEI innovation and use (Aim 4). The significance of this work lies in the in-depth new knowledge it will generate regarding specific PEI characteristics and relevant vantage points most influential in shaping stakeholders? ethical concerns, beliefs, and attitudes toward PEIs. Responding to the BRAIN Initiative imperatives, our evidence-based map will anticipate and guide choices related to concerns and attitudes surrounding PEI innovation and use.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Subproject #1 3D Whole-Cell PALM for Use with Genetically Expressed Proteins. The ability to merge fluorescence microscopy with appropriate labeling technologies has proven invaluable for the cell biologist, providing three dimensional views of protein distributions with high contrast and specificity, while minimizing sample perturbation. Despite these advantages, the optical diffraction limit has historically placed a lower bound of 250 nm on the smallest structures that may be resolved with optical wavelengths. A number of optical super-resolution techniques now allow spatial resolutions down to 20 nm while retaining the advantages of fluorescence microscopy. One such technique, photoactivated localization microscopy (PALM (1)), relies on the repeated stochastic photoactivation of single molecules and their subsequent localization over thousands of widefield images to provide 20-30 nm resolution in 2D and sub-100 nm resolution in 3D. PALM builds up superresolution images literally molecule-by-molecule, so maximizing the number of successful localizations is critical for resolving small structures. This procedure depends on successfully isolating the fluorescent signal emitted from a single activated molecule from the potentially much larger sea of background arising from cellular autofluorescence and extraneous activation and excitation of other molecules. For surface bound systems or thin samples existing techniques may be used to limit background, allowing even relatively dim, genetically expressed photoactivatable fluorescent proteins (PA-FPs) to be utilized. These markers are especially useful in super-resolution imaging, as they offer greater specificity and effectively higher labeling densities than brighter, but exogenously-introduced caged dyes. Imaging thicker, three-dimensional samples is problematic, however, as illumination activates and excites the entire sample, increasing background and generally precluding the use of dim PA-FPs. Furthermore, if out-of-focus molecules are not localized, they are wasted, thus decreasing the effective label density and reducing image resolution. Along with collaborators Mike Davidson (Florida State University) and Alipasha Vaziri (Janelia Farm Research Campus), we developed a technique that mitigates these problems, allowing PA-FPs to be successfully utilized for 3D superresolution in cells, exceeding a depth of 8 microns. We reduce background by confining photoactivation to the focal plane via two photon, line-scanning temporal focus activation of the sample. We have also developed model-free, 3D subdiffractive localization software that is far more tolerant of microscope aberrations than previous algorithms. The combination of these two technical innovations has enabled the imaging of a variety of cellular constructs (the nucleus, endoplasmic reticulum, vimentin, and mitochondria), with the genetically expressed protein pa-mCherry1. A paper describing this work has been published in Nature Methods, and the subdiffractive localization code we have developed is open-source, at code.google.com/p/palm3d/ We are in the process of applying the 3D PALM microscope to problems in biology. To that end, we are collaborating with Kumaran Ramamurthi (NCI) in order to perform 3D superresolution imaging of Bacillus subtilis;and Jan Liphardt(UC Berkeley) in order to perform 3D imaging of the nucleus in thymocytes. Subproject #2 Selective Plane Illumination Microscopy for worm neurobiology and embryology Selective plane illumination microscopy (SPIM (2)) is a technique whereby a sample is illuminated with a thin plane of light from the side, so that fluorescence detection occurs in a direction perpendicular to excitation. Such an experimental geometry has major advantages over conventional 3D microscopy techniques, such as confocal microscopy or 2 photon microscopy. First, acquisition speed is greatly increased relative to point-scanning methods, as the entire imaging plane is detected simultaneously. Second, excitation is confined to the focal plane, so each pixel is imaged only once during each volumetric acquisition. This drastically reduces light exposure and results in far lower photobleaching and photodamage than is possible with conventional imaging techniques. These advantages have been applied to studying whole-animal (zebrafish, drosophila) embryogenesis, and to the measurement of calcium transients in tissue slices. We have recently built a SPIM that we intend to use in constructing the first atlas of neuron positions in the developing nematode C. Elegans, in a collaboration with extramural researchers Daniel Colon-Ramos (Yale University) and Zhirong Bao (Memorial Sloan-Kettering Cancer Center). Due to the greatly reduced light dosage applied, we can already image developmental events at 30x the speed of the best available competing technology (spinning disk confocal microscopy), with equivalent signal-to-noise ratio, which should aid in embryo lineaging. Initial results from this collaboration are under review. On the technical side, we are pursuing various instrumental approaches that should allow us to image with better 3D resolution (a weakness of SPIM compared to other technologies) in living samples. We were also awarded an NIH Director's Challenge Award to use SPIM for visualizing very fast volumetric (<10 ms) dynamics in the developing zebrafish brain. Together with collaborator Harold Burgess (NICHD), we aim to induce a startle response in a live zebrafish embryo, and monitor the resulting calcium transients with SPIM. We are currently developing the tools for this experiment. Subproject 3 Structured Illumination Microscopy with a Swept Field Confocal Microscope Besides PALM, structured illumination microscopy (SIM) is another superresolution technique (3) that offers more modest resolution (2x better than the diffraction limit), but is readily compatible with live samples. SIM, while commercially available, is expensive and remains the province of relatively few labs. Together with Chris Combs (NHLBI), we are developing SIM so it can be performed on a swept field microscope. This should allow the technique to be performed by many more labs, and should offer some optical sectioning ability in addition to the resolution enhancement. (1) Betzig, E. et al. Imaging Intracellular Fluorescent Proteins at Nanometer Resolution. Science 313, 1642-1645 (2006). (2) Huisken, J., Swoger, J., Del Bene, F., Wittbrodt, J. &Stelzer, E. H. K. Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy. Science 305, 1007-1009 (2004). (3) Gustafsson, M.G. Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J Microsc. 198, 82-7 (2000).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will demonstrate a powerful new strategy for diagnosing infectious diseases in minimally equipped laboratories or at point of care (POC). Developed for biodefense and similar low-complexity testing purposes, the RAMP(R) antigen detection system (Response Biomedical Corporation, Vancouver, BC, Canada) will be adapted for POC diagnosis of tuberculosis (TB). The RAMP system has been extensively evaluated by independent testers, and its high sensitivity and outstanding robustness have been demonstrated. Its sensitivity will be enhanced by integration with a novel electrohydrodynamic microtip system designed to selectively concentrate Mycobacterium tuberculosis (MTB) cells in sputum samples prior to RAMP analysis. The microtip system was developed and validated by NanoFacture, Inc., in collaboration with the Nanomanufacturing Laboratory at the University of Washington. The integrated microtip-RAMP test will detect MTB cells with a lower limit of detection of <100 cfu per mL of sputum, comparable to PCR in a 20 min total test time. Specific aims of the project are: 1) To develop a RAMP test for MTB cells in sputum; 2) to develop sample processing procedures that utilize the microtip system to concentrate MTB cells from human sputum; and 3) to evaluate the performance of the microtip-RAMP system in a field trial in Dhaka, Bangladesh. Targeted pre-clinical performance for the 20-min microtip-RAMP test will have sensitivity and specificity of >90% relative to culture. In addition, the hypothesis will be tested that microtip-RAMP is significantly more sensitive than AFB smear microscopy, currently the world's most widely used rapid test to detect MTB cells in sputum.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to understand the molecular structural basis of transport and select inhibition of lipid flippases/multidrug resistance ABC (MDR-ABC) transporters. We will be integrating structure, function, and chemistry to provide a much more unified and comprehensive view of lipid transport coupled to ATP hydrolysis by MsbA. Through this work, we will discover specific and potent inhibitors of MsbA, which has very significant and direct implications for the treatment of cancer and infectious disease. An inhibitor of MsbA will likely disrupt the formation of lipopolysaccharide (IPS) in the outer membrane and kill a broad range of Gram-negative pathogenic bacteria. We have already obtained co-crystals of MsbA with a variety of cancer compounds, which include the MDR reversal compound PSC833. We have recently collected preliminary diffraction data and have initial electron density maps indicating the position of these ligands relative to the structure of MsbA. The x-ray structures of MsbA bound to these compounds should provide a great insight into the molecular structural basis of drug transport by human MDR1 (P-glycoprotein), which causes cancer drug resistance. The co-crystal structures of MsbA with inhibitors could also serve as a close model for other bacterial and mammalian MDR transporters. Finally, we continue to push the frontier of membrane protein structural biology by determining the x-ray structure of a new and distantly related bacterial MDR-ABC transporter from H. influenza. We have already obtained crystals and the x-ray structure determination is well-under way. Our Specific Aims are: 1. Structural studies of MsbA with anti-cancer compounds and nucleotide analogs. 2. Functional studies of MsbA and the detection of inhibitors. 3. Structural studies of MsbA with compounds that occupy the \"ON\" and \"OFF\" sites of MsbA (Class I MsbA inhibitors). 4. Structural studies of MsbA with cyclic inhibitors based on cyclosporin A and other MDR cyclopeptides (Class II MsbA inhibitors). 5. Structural studies of a new MDR-ABC transporter from H. Influenza.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The properties and capabilities are described of an analytical ultracentrifuge, Beckman Optima XL-I, with both interference and absorption optics, that will be used by a group of Purdue research scientists to investigate the interactions between macromolecules in biologically important systems. The acquisition of this instrument is part of a plan to assemble an array of instruments in a center for biophysical studies of macromolecules. This array of instruments currently includes a surface plasmon resonance instrument, an isothermal calorimeter, and a ten-year old Beckman Model XL-A analytical ultracentrifuge. Future development of this center targets the addition of state-of-the-art fluorescence, FTIR and Raman spectrometers, plus a stopped flow rapid reaction instrument. This proposal for the XL-I analytical ultracentrifuge is submitted by a group of six investigators involved in studies of protein-protein, protein-nucleic acid, protein-ligand and protein- membrane interactions. These investigators have expertise in high resolution crystallographic structural studies as well as in biochemistry, biophysics and molecular genetics, and are interested in complementing their structural results with the biophysical measurements that can illuminate the details of assembly of large complexes, changes in interaction states with ligand substrates or effectors or with modifications that mimic cellular signaling states. Investigations of stability of large complexes and search for conditions of monodisperse preparations of membrane proteins are also proposed, as an aid to the crystallization of these difficult systems. Initial studies with the Beckman XL-A have demonstrated the usefulness of analytical ultracentrifugation in a wide variety of systems including: the binding affinity of membrane proteins for their soluble counterparts, assembly of animal viruses, signal transduction protein interactions, interactions between toxins and their cellular receptors as well as the assembly of large protein complexes for transmembrane transport. The XL-I will extend the range of studies into those involving weaker interactions as well as to studies where the optical characteristics of the samples are inappropriate for the absorption optics of the XL-A and will compliment and extend the information obtained by both structural and biophysical methods at Purdue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Heart failure (HF), a syndrome that affects more than five million Americans, is associated with disability, increased morbidity, and decreased quality-of-life. Self-care may be one way of controlling fluid retention and endocrine responses to HF that impair cardiac performance and lead to poor patient outcomes. Although self-care is hypothesized to influence cardiac performance, the scientific basis of this claim has not yet been established. We need further evidence that HF patient's day-to-day self-care practices can influence cardiac performance in order to impress on patients the important role that they play in maintaining optimal cardiac performance and in increasing the quality and years of their health. We intend to use bioimpedance cardiography technology and hormone measurement, along with a strategic procedure of head-down tilt to passively and non-invasively challenge the heart's performance to gain insight into this relationship. A cross-sectional study is proposed to examine the relationship between HF self-care measured using subjective and objective measures and the objective measurement of cardiac performance in response to fifteen minutes of a 15[unreadable] head-down tilt positional challenge. The specific aims of this study are to: 1) describe the relationship between HF self-care and baseline cardiac performance, by measuring A) self-care with the Self-Care of Heart Failure Index, the Medical Outcomes Study Specific Adherence Scale, and 24-hour urine sodium levels to assess adherence to a low sodium diet, and B) cardiac performance with blood levels of atrial natriuretic peptide, as an index of atrial distension, and cardiac hemodynamics with bioimpedence cardiography, including thoracic fluid volume index, stroke volume index, and acceleration index, and 2) describe the relationship between HF self-care and dynamic cardiac performance in response to a fifteen minute, 15[unreadable] head-down tilt positional challenge. Measures of HF self-care will be compared to the relative change in measures of cardiac performance in response to body positioning. This research study examines the self-care behaviors of persons with heart failure and how their heart functions at rest and when stressed. This study will help determine if the day-to-day self-care behaviors of heart failure patients can influence how the heart functions. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this project is to develop improved methods for studying the metabolism of biologically significant materials through improved instrumental methods of microanalysis. Primary emphasis will be on chromatographic methods of separation, including gas-liquid, thin-layer, and liquid-liquid chromatography all used in conjunction with radioisotopes and with electron impact and chemical ionization mass spectrometry. Effort will be directed toward devising new detection methods for the various forms of chromatography that are specific for distinctive chemical groupings and classes of compounds. The objective will be to develop better methods of analysis both for compounds that contain these specific groupings, and for those that can be labeled with these groupings by reaction with specific reagents. We will also undertake the development of improved methods for assaying radioactivity in compounds separated by various forms of chromatography. When appropriate, we will devise specific procedures for the assay of compounds of interest, such as pharmaceuticals in body fluids, that demonstrate the improved capability offered by the new instrumental methods.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The rising prevalence of obesity in the United States is believed to be due to increased exposure to adverse environmental factors, such as food portion sizes and increased dietary variety. Although decreasing portion sizes is a strategy used in weight loss programs, research has not studied the effects of decreasing dietary variety. Cross-sectional studies show a positive association between variety and body weight and in our own studies we have shown that greater reductions in the number of different snack foods (i.e. cookies, chips) consumed predicted greater decreases in overall caloric and fat intake and greater weight loss. Limiting variety may reduce intake through long-term sensory-specific satiety and/or monotony. Reducing dietary variety is a novel dietary approach with the potential to improve long-term weight loss, which has not been studied as a clinical strategy in obesity research. The objective of this application is to conduct a randomized controlled trial of a behavioral weight loss intervention limiting the number of different snack foods consumed. Two hundred overweight and obese participants will be randomized to a standard behavioral intervention (Standard) or to a standard behavioral intervention that also limits the number of different snack foods consumed (Limited Variety). Both conditions will receive an 18-month standard behavioral intervention, using behavioral techniques (i.e., self-monitoring) to change eating behaviors. Participants in the Limited Variety condition will also limit variety in snack foods to only two chosen snack foods throughout the intervention. Measures of weight, snack food consumption and hedonics, and diet satisfaction will be taken at 0, 6, 12, and 18 months. This investigation will determine: 1) if the Limited Variety condition produces greater weight loss than the Standard condition at 18 months; 2) if the Limited Variety condition consumes fewer servings and calories from snack foods than the Standard condition; and 3) if limiting snack food variety produces long-term sensory-specific satiety and/or monotony. Relevance: Experimental studies show that limiting dietary variety profoundly reduces intake. To date, there is no dietary prescription that has been tested that capitalizes on the effect of variety on intake that can be maintained. This will be the first investigation to examine methods of manipulating dietary variety that can be adhered to over time and that influence intake, weight loss, and weight loss maintenance. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. : Peripheral nerve injury frequently leads to a variety of debilitating chronic pain states that are thought to reflect heightened excitability of ascending spinal nociceptive pathways. Attempts to identify the mechanisms underlying the pathogenesis of chronic pain are hampered by a general lack of knowledge of the connectivity and synaptic efficacy of nociceptors with known populations of second-order interneurons under any condition. The overall goals of the proposed studies are to increase our understanding of the anatomical and functional connectivity between physiologically identified cutaneous nociceptors and specific subpopulations of local and ascending interneurons in the superficial dorsal horn under normal conditions and in an animal model of neuropathic pain, and the role of inhibitory interactions in shaping these patterns of functional connectivity. These studies are motivated by the PI's recent discovery of a novel class of polymodal nociceptors with large, thickly myelinated (A[unreadable]) fibers. Unlike most nociceptors, their projections to nocireceptive regions of the dorsal horn appear to be unaffected by nerve injury and thus may constitute the principle substrate underlying various neuropathic pain states. Further, because these afferents in normal animals become active well below pain thresholds, they provide an excellent window for investigations of 1) the physiological mechanisms underlying the regulation of nociceptive throughput to higher centers, 2) how these are altered under neuropathic conditions, and 3) the contribution of inhibitory controls to nociceptive processing and throughput to higher centers under both normal and pathologic conditions. The proposed studies will use a combination of electrophysiological, light and ultrastructural microscopical analyses, and computational modeling to address these objectives through the use of physiologically identified nociceptors and anatomically and physiologically identified subsets of second-order spinal interneurons. Detailed knowledge of the cellular mechanisms underlying the normal balance between excitatory and inhibitory interconnections that regulate neuronal activity levels in the superficial dorsal horn, and how this balance is disrupted in pathological conditions, will be pivotal to the development of future strategies in pain management and the translation of these strategies into effective practice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Type 2 diabetes mellitus and its atherosclerotic complications impose a substantial burden on health of Americans in general and on African Americans in particular. Recent discoveries in molecular genetics have lead to the identification of functional variations in several candidate genes for susceptibility to obesity, insulin resistance, and/or diabetes. These include genes which code for beta-2and beta-3 adrenergic receptors, insulin receptor substrate 1, fatty acid binding protein 2, frataxin, and leptin receptor. If their role as novel susceptibility factors is confirmed, these variants promise to illuminate the pathophysiologic basis of diabetes and diabetes related cardiovascular diseases, accelerate the development of chemopreventive agents, and facilitate the conduct of prevention trials by marking individuals at high risk. Unfortunately many previous association studies of these mutations in human populations have been limited by small, selected study samples; by limited cross-sectional data on behavioral factors and on cardiovascular risk phenotype; and by paucity of data on African Americans. The investigators, therefore, propose to conduct an epidemiologic study of functional variants in 10 candidate genes for susceptibility to type 2 diabetes, obesity, and insulin resistance. The main objective will be to detect modest effects consistent with polygenic nature of diabetes but with better sensitivity and precision than previous association and linkage studies. The study sample will a community based cohort of 3,250 African Americans and 3,250 Whites aged 45 to 64 who are participants in the ongoing Atherosclerosis Risk in Communities (ARIC) study. Supported by NHLBI the ARIC study has assembled an extensive data base including behavioral assessment (e.g. diet and physical activity), anthropometry, laboratory blood tests (e.g. oral glucose tolerance test and serum lipids), and carotid ultrasonography as well as clinical events and mortality. Using race specific case-control, cross-sectional, and longitudinal analyses the investigators will determine if these putative diabetes alleles are associated with incident and prevalent diabetes, with obesity and weight gain, with hyperinsulinemia in non-diabetic individuals with the presence of an adverse cardiovascular risk factor profile, and with atherosclerosis progression and cardiovascular disease incidence over 12 years of follow up. The investigators will assess how behavioral and environmental factors such as obesity, diet, and physical activity, influence the expression of genetically conferred risk. Strength of this proposal include the close collaboration between clinical, epidemiologic, and laboratory researchers, a wealth of prospectively collected data from an NIH sponsored study, and a sample size large enough to detect modest gene effects and to investigate gene-gene and gene-environmental interactions. Most important, this study will provide unique information on the expression of diabetes susceptibility genes in the general population and possibly suggest genetic explanations for the excess prevalence of type 2 diabetes and obesity in African Americans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract The Immunologic Monitoring (IM) Core will serve as a shared resource to assist the target faculty of the COBRE to pursue high quality research by monitoring immune cell functions following treatment with the dietary supplements in naive and experimental models of inflammatory disease. The Core will be useful to characterize the immune status before, during and after treatment of diseased animals with the plant products so as to provide insights into their prognostic and therapeutic effects. In addition, while the individual projects deal with specialized studies related to the effects of plant products on macrophages that play a pivotal role in inflammation, it is critical to know how these would affect the other immune cells and the overall immunity. This is especially important because inflammatory diseases are systemic and involve multiple types of immune cells. Furthermore, such information on the immunologic effects of the plant derivates is critical in the development of bench-to-bedside research. Specifically, the IM Core will 1) offer a wide range of state-of-the-art resources to pursue cellular and molecular immunological assays so as to enable the investigators to pursue cutting-edge research on the projects. 2) provide technological assistance and training in the use of major equipment by the users 3) participate in the design of the experiments, selection of the appropriate assays, trouble-shoot and interpretation of results. 4) develop and standardize new technologies based on the changing needs of the users by optimization and evaluation of sensitivity, specificity and reproducibility. 5) aid in data collection, evaluation and analysis as well as sharing of the data to enhance collaborations. There are two major components of this core. The core will 1) Serve as a resource of multi-user equipment 2) Perform Immune Function Assessment by 1) Evaluation of the general health status 2) Level I immunological testing which includes a variety of assays that examine the functions of T cells, B cells, NK cells, dendritic cells and macrophages. 3) Level II immunological testing of the genetic, transcriptional and epigenetic mechanisms underlying immune cell dysregulation. This functional assessment performed by the IM Core will complement the cell phenotyping and animal imaging studies performed by other cores and thereby lead to comprehensive integration between projects and cores. These shared resources could be used for translational studies in the future. In summary, the IM Core will provide effective and economical shared resources and service to conduct high-quality research that will lead to increased productivity and enhanced interactions between investigators of all the projects resulting in their ability to attract independent, extramural funding, and advancement of scientific careers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research will examine the importance of biological and physical causes of variation in the production of bioactive secondary metabolites by coral reef invertebrates. The research has important biomedical consequences since it will focus on understanding what factors affect the yields of biomedically important marine natural products. First, differences in types and concentrations of secondary metabolites within and among individuals, at different life history stages, and among populations of selected invertebrates will be examined. Facultative changes in chemical defenses will be examined for.sponges, ascidians, and soft corals exposed to: l) increased grazing by fishes and specialist predators; 2) different environmental levels of light and nutrients; and 3) known chemical elicitors of induced defenses. The proposed research will provide information on chemical variation and its causes and the ability and timing of different coral reef invertebrates to change their allocation of chemical defenses. The study will contribute to our limited knowledge of the chemical interactions of marine invertebrates and their predators and the relationship between natural adaptive functions of secondary metabolites and their biomedical potential. Bioactive metabolites will be incorporated into established testing programs (Univ. of Hawaii, UC Santa Barbara, Wayne State University Medical School, Virginia Tech, and Univ. of Mississippi School of Pharmacy) to determine their pharmacological activities in specific assays. This research program will provide educational opportunities for a postdoctoral researcher, a graduate student, and undergraduate students at the University of Guam, a predominantly undergraduate and minority institution.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application proposes a plan for the detailed investigation of renal function in the Wistar-Kyoto spontaneously hypertensive rat (SHR). In addition to renal clearance methodology, the isolated perfused rat kidney (IPRK) technique will be utilized in organs derived from SHR's and controls. The IPRK allows complete characterization of the synthetic medium perfusing the organ, in the absence of unpredictable circulatory, neural and humoral factors. Studies will include the effect of antihypertensive drug therapy upon renal function. Also, the chronology of development of tendencies toward sodium retention, increased renal vascular resistance, and blood pressure elevation will be delineated in SHR's and SHR IPRK's at progressive ages - always in comparison to appropriate control animals and organs. Metablic and physical factors which could promote sodium retention in the SHR will be investigated in SHR IPRK's. The effects of angiotensin II generation and inhibition will be assessed in the preparation, utilizing pharmacological probes. Also, effects of prostaglandins, and the inhibition of prostaglandin synthesis, will be assessed. Effects on kidneys from normotensives also will be studied. In addition, effects of cyclic nucleotides, and sympathomimetic amines will be investigated. The actions - unique and overlapping - of various pharmacologic inhibitors of sodium reabsorption will be determined in the SHR IPRK, as compared to controls. Finally, a possible role of intracellular calcium in regulating renal resistance and sodium reabsorption will be studied, utilizing a rather specific calcium ionophore.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mutagens contribute to the human burden of heritable birth defects and cancer and probably to heart disease as well. Mutagenic DNA damage usually consists of lesions that block DNA replication. Error-prone repair, the major mechanism of damage-induced mutagenesis, occurs when DNA replication bypasses such damaged bases in a poorly templated (and thus highly mutagenic) manner. Replication blocks have been studied in vitro but hardly at all in vivo. We are establishing systems to map, at the nucleotide level, those sites at which mammalian replication forks terminate synthesis synthesis when replicating damaged DNA and to analyze the relationship between DNA blocking lesions and mutation-prone sites.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: This application is for the renewal of an NEI Core Grant in support of the research programs of 16 principal investigators holding 23 active NEI individual research grants and an NEI Institutional Training grant. The previous funding cycle generated 100 publications in the major areas of aging and diabetes, extraocular muscle biology and diseases, ocular immunology and inflammation, and retinal biology and diseases. The current investigators form the nucleus of the Case Visual Sciences Research Center (VSRC), a broader group of 40 vision researchers in 19 different basic and clinical science departments at the Case Medical Center (Case Western Reserve University School of Medicine and University Hospitals of Cleveland). Since our initial Core Grant award in 1997 and its renewal in 2002 with five Modules, these modules have continued to evolve over the current funding cycle due to a changing investigator group and scientific direction, expanded services and new technologies. The five modules that are proposed are: 1) Tissue Culture and Hybridoma, 2) Histology, Microscopy and Imaging, 3) Molecular Biology, 4) Specialized Animal Resource, and 5) Proteomics. Major new functions since our previous submission include hybridoma production, live cell imaging, laser capture microdissection, and a full service proteomics facility to acquire and analyze protein expression in ocular tissues. With the recent renewal of the T32 Training grant, these modules will also continue to serve as an integral resource for pre- and postdoctoral trainees in the Case Visual Sciences Training Program. The Visual Sciences at Case has receives very strong institutional support, including, construction of a centralized facility for Core Grant modules and functions, recruitment funds and supplemental support for technical salaries and supplies in the core facilities. The change in P.I. from Dr. Lass to Dr. Pearlman will ensure continuity in leadership and direction of the Core Grant, which will enable collaborative goals and objectives while enhancing individual programs, and facilitate Case VSRC investigators'continuing efforts toward analysis of fundamental mechanisms in ocular and visual system development, function, and pathogenesis of diseases of the eye and visual system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dr. Cosloy continues to study the genetics and regulation of the heme biosynthetic pathway of Escherichia coli in collaboration with Dr. Charlotte Russell of Area L The heme IX synthesis pathway in E coli has several branches. G1utamyl-tRNA participates in protein synthesis as well as in the synthesis of aminolevulinic acid (ALA), the first committed intermediate in heme biosynthesis. Another branch point occurs at the uroporphyrinogen III step and it leads to sirohydrochlorin or to heme. Sirohydrochlorin is the precursor for the branches leading to Vitamin B12 and to siroheme. Siroheme is a cofactor in the synthesis of cysteine. Heme plays a role in both aerobic and anaerobic respiration, and it has been found that 02 may regulate the production of ALA. These considerations suggest that the control of this pathway requires interesting strategies which are worth investigating.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this project is to understand the function and regulation of mitotic proteolysis through the development and application of novel small molecule inhibitors. The ubiquitin-proteasome pathway plays an essential role in regulating progression through mitosis in all eukaryotic cells. The critical regulated component of this pathway is a ubiquitin ligase termed the Anaphase-Promoting Complex (ARC). Once this ligase ubiquitinates its substrates, they are targeted for degradation by the proteasome. We have identified small molecule inhibitors that block that ability of the Anaphase-Promoting Complex to ubiquitinate its substrates in cell extracts, and also small molecules that inhibit the degradation of proteasome substrates by binding ubiquitin chains. In this proposal, we seek to use these compounds to understand the mechanism of APC-dependent ubiquitination, which remains poorly understood. Furthermore, recent work from our group suggests that the APC ubiquitinates its substrates with novel types of ubiquitin chains, and we therefore want to understand the significance of this form of ubiquitiantion and how it affects targeting of substrates to the proteasome. Finally, we seek to synthesize new versions of small molecules that bind to ubiquitin chains to better understand how these molecules function. Together these studies will provide new insights into how the APC ubiqutinates its substrates and how ubiquitianted substrates are targeted for degradation by the proteasome, as well as providing new chemical tools that may lead to the development of drugs that target this important biochemical pathway. The ubiquitin-proteasome system regulates many essential processes that impact on disease, including neurodegenerative diseases such as Alzheimer's and Parkinsons's disease, and also many forms of cancer. Recently, inhibitors of the proteasome have been approved for treatment of cancers including multiple myeloma. In this project, we seek to use and develop small molecule inhibitors that block the ubiquitinproteasome pathway through novel mechanisms. Recent work suggests that the Anaphase-Promoting Complex may be dysregulated in cancers, and thus finding new methods to interfere with its function may lead to the development of novel agents for the treatment of cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "An ancient legacy of life transitioning from oceans to land is the role of the skeleton and kidney in regulating mineralization and Ca/PO4 homeostasis. Key players in this bone kidney axis are a group of related extracellular matrix proteins (SIBLINGs). These proteins all map to a tightly clustered region on chromosome 4q. A unique protein PHEX, regulates at least two SIBLING proteins (MEPE and DMP1), indirectly or directly. PHEX is a zn-metalloendopeptidase in search of a substrate and may also play a role as an orphan ligand or receptor. Loss of PHEX function results in X-linked hypophosphatemic rickets and a novel cytokine FGF23 (a phosphatonin), plays a major role in the phosphate phenotype. Our data confirms direct binding of PHEX to MEPE, a SIBLING protein expressed in bone, teeth and in the proximal convoluted tubules of the kidney. The binding between these proteins occurs between the PHEX Zn-binding motif and a short COOH-terminal region of MEPE called the ASARM-motif. This short region is exquisitely resistant to proteolysis and conserved across species. Biologically, the released ASARM-peptide is a potent inhibitor of mineralization, inhibits renal phosphate handling and binds to hydroxyapatite. Binding of PHEX to MEPE may serve to regulate release of the ASARM peptide and thus locally control mineralization. PHEX also binds with high affinity to free ASARM- peptide and also inactivates the peptide by hydrolysis. In HYP, as well as a loss of PHEX function, there is a marked up regulation of MEPE and osteoblastic protease activity. This results in degradation of MEPE and DMP1 and release of ASARM-peptides from both these proteins and perhaps other SIBLING proteins (DSPP, osteopontin BSP, enamelin). These protease resistant ASARM-peptides accumulate in kidney, bone, urine and in the circulation. To study the role of SIBLINGs, PHEX and ASARM- peptides we have made transgenic mice (MEPE-TRG) that overexpress MEPE protein 50X in bone and 7X in kidney. Our specific aims are: 1. Characterize bone and renal PO4 phenotypes of MEPE-TRG mice, 2. Determine the bone and renal- calcification roles of processed forms of MEPE and ASARM-peptides in MEPE-TRG, HYP and NPT2a-/- mice, 3. Design PHEX synthetic peptides as tools to probe the role of ASARM peptides and PHEX in health and disease. PUBLIC HEALTH RELEVANCE: A new class of recently discovered bone-teeth and renal proteins (SIBLINGs) play a major role in upholding the health of the skeleton and regulating phosphate. Bioactive processed peptides from these proteins impact on mineralization, bone formation, bone growth and soft tissue calcification in disease and health. The study of these new proteins will help develop therapies for mineral loss disorders (inherited and tumor-acquired), renal osteodystrophy, renal transplantation, ectopic arterial-calcification, renal calcification and osteoporosis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The mission of the Cancer Therapy Evaluation Program (CTEP) is to improve the lives of cancer patients by finding better ways to treat, control and cure cancer. CTEP accomplishes this mission by funding an extensive national program of cancer research and by sponsoring clinical trials to evaluate new anti-cancer agents, with a particular emphasis on translational research to elucidate molecular targets and mechanisms of drug effects. CTEP uses a scientific process to accomplish its mission: * promising basic science findings are identified and translated into clinical research, both by identifying new agents for evaluation and by identifying biologic characteristics of tumors that may be clinically exploited. * novel anti-cancer agents with distinctive molecular targets, mechanisms of action, or properties are identified and introduced into clinical trials, with prioritization of agents based on scientific criteria and therapeutic needs. * the anti-tumor activity of new anti-cancer agents is systematically evaluated in clinical trials. * promising new cancer treatments are rigorously compared to best available treatments in hypothesis-driven clinical trials to reliably define superior treatments for specific types of cancer CTEP attempts to forge broad collaborations within the research community and works extensively with the pharmaceutical/biotechnology industry to effectively develop new cancer treatments. CTEP also seeks to involve outside experts and patients or their advocates in the formulation of research priorities. In the selection of clinical research for NCI sponsorship, CTEP attempts to fill critical gaps in the national cancer research effort and to avoid duplication of ongoing private sector efforts. In further efforts to control cancer, active new anticancer agents are made available as rapidly and widely as possible for patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neurological syndromes have frequently been described in patients with acquired immunodeficiency syndrome (AIDS). Recent reports on the isolation of infectious human immunodeficiency virus (HIV) from cerebrospinal fluid (CSF) and brain tissues of patients with AIDS and the detection of HIV DNA in the brain strongly suggest that HIV is directly responsible for some of the neurological manifestations. The type(s) of brain- derived cells which can be infected by HIV is under active investigation; however, the possibility that a neurotropic variant strain of the virus might be responsible for neural infection remains to be determined. Several HIV isolates from the CSF and brain tissue of AIDS patients have been cultured in our laboratory. In some cases, the HIV from lymphocytes of the same individual were also obtained. When some of the isolates were inoculated onto brain-derived cell cultures, they productively infected glial cells, particularly those with astrocyte markers. Thus, an in vitro assay to study neurotropism is available. We propose to characterize, at the molecular level, two central nervous system (CNS)-derived HIV isolates and one corresponding lymphocyte-derived HIV isolate. These HIV can be distinguished by their replicating properties in different cells. We shall clone their proviral forms from genetic libraries of virus-infected cellular DNA. Molecular clones of these viruses will be subjected to restriction endonuclease analysis and the resulting DNA fragments will be isolated and sequenced. AT the biological level, differences in the ability of these three HIV isolates to infect human cell lines, particularly those derived from brain will be further studied. To map the viral sequences encoding the determinant(s) for cell type tropism, we shall construct recombinant viral DNA genomes using biologically active molecular clones of HIV with different cell tropisms. Infectious recombinant HIV's, recovered from transfection of human lymphoid cells with recombinant DNA's will then be tested for their ability to infect different cell types. Data obtained from this proposed study will help determine (1) the extent of genomic variations between isolates from the brain and lymphocytes of the same individual, (2) whether \"neurotropic HIV\" is a subtype of the AIDs virus, and (3) the region(s) of the HIV genome that governs tropism for different cell types.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT The mission of the Community Outreach and Ethics Core (COEC) is to support the Center in achieving its overarching mission of Building Capacity and Collaborations, inspiring Creativity, and facilitating Communication in environmental health sciences (EHS) research. To do this, the COEC plays a central role in connecting Center investigators, community partners, and other stakeholders in productive EHS-focused dialogue and activities. Our COEC has developed a strong presence in the region and has a reputation as a trusted partner of stakeholders who include educators, tribal nations, public health professionals, government agencies, non-profit organizations, and academia. One of COEC's areas of strength is our strong commitment and expertise to address the ethical, legal, and social implications (ELSI) that relate to Center research. Our aims for the next funding cycle build on our strengths and expertise, support the mission of the Center, promote innovative research, engage stakeholders, and foster the development of the next generation of EHS innovation and networked researchers focused on exposures, diseases, genomics, and the environment. Some proposals in this application build on our ongoing programs that have been in place through several grant cycles and have evolved in collaboration with stakeholders as the COEC has established long-term and meaningful community partnerships. These include The Academy for Teaching About Health and Environmental Associations (ATHENA); Native Tradition, Environment And Community Health (TEACH); Public Health Caf; our partnerships with the Duwamish River Cleanup Coalition and Collaborative for Health & Environment and dissemination of our work on social media through blogs, Twitter, Facebook, and YouTube. New initiatives include cultivating potential new partnerships with participants in the UW Tribal Leadership Summit and the National Conference of State Legislators. In our Native TEACH program, we plan to introduce a Scientific Workshop, career training for Native students, jointly funded collaborations between Center and Native researchers, and to re-establish the inter-COEC Tribal Collaborative Working Group. In addition to our outreach efforts, we will reach inward to our members through monthly Breakfast Club gatherings where research is shared and collaboration encouraged, and offer communications training for members and trainees, including Lightning Talk events and a graduate class in outreach and engagement. COEC is an integrating element in the Center, translating and disseminating Center research to stakeholders and the community, and connecting CRTs and Center members with each other and with our community partners. The COEC will build on its previous efforts and successes as well as expand in new directions as we enter the new grant cycle and the next phase of EHS research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The inaugural meeting of the Cognitive Development Society took place in Chapel Hill, NC, October 8-9, 1999. The 270 + participants included senior researchers and graduate students from the US and abroad who are interested in the study of cognitive development. The present request is for funds to support their second meeting, to be held October 26-27, 2001 in Virginia Beach, VA. The goal of the meeting is to enhance scientific progress in their understanding of cognitive development by updating researchers' awareness of new developments in the field, fostering debate on relevant topics, teaching new methodological and statistical skills, and encouraging broad applications to the health, education, and welfare of children and adolescents. The meeting will include plenary addresses and break-out groups organized as symposia and workshops, and open poster sessions to encourage participation by all attendees at the meeting. The program will be approximately one-half invited and one-half submitted, with a Program Committee in charge of issuing invitations and selecting among submissions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objective of this project is to determine the biological significance of the \"non-communicative\" function of vocalization and characterize the underlying physiological mechanisms. Although vocalization has traditionally been viewed as a mean of communication, recent evidence from my laboratory strongly indicates that communication also function in an integral physiological capacity. Birdsong, especially in passerine species, has long been a favored research system. The issue of learning in development of song and adaptive significance of acoustic communication are among the problems which have been addressed. The primary question of the mechanism by which the male song may influence the female's endocrine response, culminating in egg- laying, has not been studied. However, this issue has been subject of intensive study of non-songbird system. Recent findings in the study of the female ringdove (Streptopelia risoria) reproductive system has revealed an interesting dimension of vocalization which has received little attention. We have found that a bird's vocalization alters its endocrine state. What neural pathways mediate this endocrine response? Can we demonstrate this basic finding in morphological terms? Are there different circuities involved in endocrine consequence of courtship vs other vocalization such as alarm calls? In this application, I will determine and characterize the chemical features of the neural pathways which mediate the conversion of vocalization into endocrine response. In other words, we intend to demonstrate that an increased luteinizing hormone-releasing hormone (LHRH) release can be traced to neuronal activity in the vocal control nuclei associated with the performance of female nest calls. The proposed studies will add useful information about how acoustic inputs, especially those generated from the individual's own vocalization affect endocrine state and hence the well-being of the individual.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (provided by investigator): Talipes equinovarus or clubfoot occurs in approximately one of every 1000 live births. Although it is one of the most common structural malformations, little is known about its etiology. There is strong evidence to suggest that genetic factors play a role in the development of clubfoot. In a separate study population, our investigative team has identified candidate genes for clubfoot in the homeobox signaling, program cell death, insulin growth factor, and n-acetyl transferase pathways. Further, maternal cigarette smoking has been linked to clubfoot in several studies, and this association may be well be modified by n-acetyl transferase genotypes because this enzyme is involved in the biotransformation of the byproducts of cigarette smoke. In the parent study, DNA is collected from baby and mother with Oragene saliva kits. However, the parent study included no specific aims or funding to examine DNA for genetic risk factors. This application aims to study genetic variation in candidate genes, which were previously shown to be associated with clubfoot in family studies. The candidate genes will be identified in a separate study of clubfoot which has currently identified associations between clubfoot and variation in HoxA, HoxD, IGFBP3 and apoptotic pathway genes. In addition, three functional polymorphisms in NAT2 will be evaluated to look for an interaction with maternal smoking and the risk of clubfoot. Saliva samples will be available on over 400 clubfoot cases and their mothers and over 900 control mother-baby pairs. Mothers are interviewed within one year after delivery and detailed information is collected on cigarette smoking. We anticipate >80 per cent statistical power to detect slight differences for polymorphisms and 2.5-fold odds ratios for gene-smoking interaction. This supplement will provide the resources to test this powerful dataset for genetic and environmental causes of clubfoot and will yield important information that will translate into better management of clubfoot. PUBLIC HEALTH RELEVANCE: Clubfoot is one of the most common congenital malformations but its causes are not known. The aims of this supplemental grant are to identify genetic factors, as well as gene-smoking interactions, in relation to risk of clubfoot. Our goal is to understand the etiology and pathogenesis of clubfoot, leading to prevention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To maintain genomic stability, cells have developed an elaborate DNA damage response (DDR) system, which is responsible for sensing DNA damage, halting the ongoing cell cycle, and repairing DNA damage. Failure to detect and repair DNA damage leads to genomic instability, which in turn can drive tumorigenesis. Many human genetic cancer predisposition syndromes are linked to defective DDR. For example, mutations in the BRCA1 gene were found in about 50% of familial breast cancer cases. Because individual tumors often have unique defects in the DDR pathway, insight into the basic mechanisms by which cells repair different DNA lesions could also guide individualized therapy. A promising example is the use of PARP inhibitors in cancers with BRCA1 and BRCA2 mutations. On the other hand, many studies suggest that overexpression of DNA repair factors contributes to resistance to radiochemotherapy. Therefore, studying this pathway has important implications in cancer pathogenesis and cancer therapy. UFM1 is the most recently identified Ub-like proteins. The cellular function of ufmylation remains unclear. Here we show for the first time that Ufm1 signaling is involved in the DNA damage response. We found that the E3 ligase UFL1 is important for ATM activation and Ufl1 deficiency sensitizes prostate cancer cells to radiation. In addition, UfSP2, which cleaves Ufm1 from target proteins, inhibits ATM activation and its overexpression sensitizes cells to DNA damage. Based on these Preliminary Data, we hypothesize that UFL1 and UfSP2 are two new factors in the DNA damage response. UFL1 promotes ATM activation and DDR, while UfSP2 has a opposite function. Because UFL1 and UFM1 are deleted in 20% of prostate cancer, we further hypothesize that defective UFM1 signaling would sensitize prostate cancer to radiation. In this application, we will further explore how UFL1 and UfSP2 regulates NBS1, ATM activation and DDR. In addition, we will examine how UFL1 and UfSP2 themselves are regulated. We will also test the role of UFL1 and UfSP2 in radiosensitivity using clinically relevant models. Our Specific Aims are: Aim 1. Investigate the regulation of ATM signaling by Ufmylation; Aim 2. Study the regulation of UFL1 and UfSP2 by DNA damage signaling; Aim 3. Investigate the role of UFM1 signaling in prostate cancer therapy. Our studies will comprehensively reveal a novel function of UFL1 signaling in the DNA damage response and radiosensitivity. In addition, it will reveal a new therapeutic strategy based on synthetic lethality in treating prostate cancer cells, especially those with deletion of UFL1 or UFM1. !", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although the literature on homeless individualscoping with drug abuse and other mental illnesses has given some attention to lack of resources as a risk factor for homelessness, the argument made is essentially one directional[unreadable]that costs and availabilityof affordable housing are beyond the means of the individual, leading directly to becoming homeless. This argument generally treats housing affordability and availability as static constructs. However, housing also goes through changes over time[unreadable]value and availability vary as the unit ages and as the neighborhood changes. The purpose of this grant is to test two competing constructs of the role that these \"future-homeless\" tenants play in housing over time, specifically examiningthe role played by drug abuse/dependence and other risk factors in housing stock. The first construct is that future-homeless persons represent tenants of last resort. As housing becomes less desirable, any tenant becomes preferable to none, thus individuals at high risk become acceptable. A corollary of this is that pile-ups of risk factors, such as current addiction, increases the likelihood they inhabit this role. The second explanation is that homeless individuals represent the tenants of choice, occupying housing developed specifically for low-income occupants. Methodologically, testing these theories requires longitudinal data on housing value and occupancy, as well as complete data on last stable housingand individual-level characteristics for a representative homeless sample. Using a uniquedata set, Service Use, Needs, Costs, and Consequences of Drug Abuse on homelessness (SUNCODA: R01-DA 10713), we propose to combine the last stable addresses for 400 randomly-sampled homeless with publiclyavailable (but unfortunately low technology) housing stock information, controlling for census-level data for the relevant zip code, in order to test the study hypotheses. The two proposed theories have strong implications for current service use models and development of primary prevention efforts. Because current services focus on housing as a primary outcome, finding that multiple risk factors lead to housing that is likely to become unstable represents critical new informationfor improving current services. Thus, this proposal is responsive to the PA-02-150, Services and Intervention Research with Homeless Persons havingAlcohol, DrugAbuse, or Mental Disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The general significance of the present study is spurred by the fact that the prevalence of communication difficulties increases dramatically with age and that by the year 2030 the elderly population will grow to about one-fifth of the U.S. total population. As an individual increases into and past the sixth decade of life, one of the consequences of aging is the gradual deterioration in auditory sensitivity, coupled with changes in central neurophysiological processes. The goal of this research is to search for neurophysiological mechanisms underlying known deficits in temporal processing. At the single cell level encoding of temporal acuity is mediated in part, by inhibitory neurotransmitters which are known to undergo an age-related decline in the auditory midbrain. The proposed experiments will manipulate the effects of specific inhibitory neurotransmitters which are known to be critical in determining response properties in the inferior colliculus, the major obligatory nucleus for ascending auditory information in the brainstem. It is hoped that the following specific objectives will provide new insight into the auditory deficits associated with presbycusis, eventually leading to effective treatment of sensorimotor problems in the elderly.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed investigation is designed to yield structural and functional information about fibroin messenger RNA and the fibroin gene, as well as specific tRNAs (and their corresponding genes) used in the specialized synthesis of silk fibroin by the silkworm, Bombyx mori. This system is particularly suited to studies of the molecular mechanisms controlling transcription, RNA processing, and translation in a developing eukaryote because the fibroin protein, its messenger RNA and certain tRNAs can be easily isolated from the silk gland. Moreover, sequence data already available for the coding regions of fibroin mRNA and for some the major silk gland tRNAs simplifies the identificaion of structural elements regulating the function of these molecules. Specifically, Gilbert-Maxam techniques will be used to determine the sequence of the functionally interesting 3' terminus of fibroin mRNA by analyzing a phased cDNA transcript originating or the mRNA-polyA junction. Genetic analysis of fibroin protein size variants will be used to investigate the possibility that the repetitive sequences within the fibroin gene undergo rearrangement. In vitro protein synthesis will be used to determine the functional significance of the structural alteration found in a silk gland-specific tRNAAla. Electron microscopic examination and nucleotide sequence analysis of isolated tRNAAla genes will be used to identify structural features regulating the differential expression of these genes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY/ABSTRACT Atherosclerosis is the leading cause of human mortality worldwide. There has been a great interest in developing novel therapeutics for this disease that function via an orthogonal mechanism of action to currently available drugs. One promising strategy is to enhance the function of high-density lipoproteins (HDLs). HDLs facilitate the process of reverse cholesterol transport to transfer excess cholesterol from peripheral tissues to the liver for elimination. The proposed research program, supported by strong proof-of-concept preliminary results, seeks to advance a novel supramolecular strategy for improving HDL function in vivo to combat atherosclerosis. The proposed studies build on the successes of our research program, funded by an NHLBI R01 grant over the past 3 years. Despite substantial progress over the past 30 years of research in the design of apoA-I mimetic peptides, the inherent pharmacological shortcomings typically associated with linear peptides have been a major impediment to advancing HDL-modulating agents through the clinic. The proposed research program seeks to develop a novel class of chemotypes that could recapitulate the functional attributes of helical apoA-I mimetics but without their inherent limitations. We describe here for the first time that appropriately designed eight-residue self-assembling cyclic D,L-?-peptides are effective HDL modulating agents. The abiotic structure of cyclic D,L-?-peptides overcome many of the shortcomings typically associated with linear peptides, such as low serum/plasma stability, lack of oral bioavailability, high production costs, etc. We show that cyclic D,L-?-peptides remodel human and mouse plasma HDLs in vitro, increase the level of pre- beta HDL particles (subspecies of HDLs considered to be the most anti-atherogenic), and enhance cholesterol efflux from cultured macrophages. In vivo, with oral administration, the cyclic peptides increase the level of pre- beta HDL particles, reduce plasma LDL-cholesterol and triglyceride levels, raise HDL levels, and promote anti- inflammatory effects with concomitant prevention of atherosclerotic plaques. This research project aims to develop safe and efficacious cyclic peptides that prevent the development of atherosclerosis by enhancing RCT via improved HDL function. We are following an integrated approach, from medicinal chemistry optimization of our leads to in vivo mechanistic studies, pharmacology, toxicology, and efficacy. Our objectives, building on our preliminary findings, are to explore the in vivo mechanism of action and how it might differ depending on route of administration (Aim 1), optimize and better understand the peptide structure-activity relationship using a series of mechanism-based functional assays (Aim 2), and to determine the oral efficacy and safety of optimized compounds (Aim 3). These studies will advance the development of orally and parenterally efficacious compounds and provide a new chemical framework for developing HDL modulating therapeutics.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mutations in the putative p53 tumor suppressor gene were analyzed in a series of 9 aflatoxin B1 (AFLB1) induced monkey tumors, 6 AFLB1 transformed rat liver cell lines, as well as a group of 19 human hepatocellular carcinomas (HCC) obtained from Qidong, China, and 17 HCCs induced by the food mutagen, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in cynomolgus monkeys. Among the AFLB1 induced monkey tumors only a single point mutation at codon 175 (G to T transversion) was identified in only one HCC by sequencing analysis of the four conserved domains (II to V) in the p53 gene. In 3/17 of the IQ induced monkey tumors the same G to T transversion was detected at codons 159, 175, and 248. Sequence analysis of amplified cDNA by RT-PCR of p53 mRNA from the six AFLB1 transformed rat liver cell lines showed the same point mutation, a transition from G to A in codon 173 (CGC to CAC) which results in a change of the encoded amino acid from arginine to histidine. No wild- type alleles of the p53 gene were observed in any of the AFLB1 cell lines by genomic DNA analysis. The half life of the mutant p53 protein in the AFLB1 transformants was greater than 3 hours versus 30 minutes for normal p53 protein. Analysis of 19 HCCs from Qidong using PCR-SSCP as well as RFLP analysis revealed that 10/19 HCCs (53%) showed p53 mutation. In two tumors single mutations occurred in exons 5 and 6, while in 8 HCCs a G to T mutation at the third position of codon 249 in exon 7 was observed. Loss of heterozygosity (LOH) of the p53 and retinoblastoma (RB) genes was detected in 7/9 (78%) and 3/6 (50%) of informative cases, respectively. No LOH of adenomatis polyposis coli locus was observed in 10 informative cases. LOH of chromosomes 16q and 16p was detected in 10/18 (56%) and 6/13 (46%) informative cases, respectively. These data suggest that the selective mutation and frequent LOH of the p53 play an important role in the development of HCCs from Qidong and at least three additional tumor suppressor genes might be involved in the human hepatocarcinogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "These studies are a continuation of experiments originated in our laboratory, from a previously funded RO1. Since there are no good biomarkers or preventative strategies, infection-associated pregnancy complications represent an important clinical problem. The regulation of Toll-like receptor (TLR) expression and function at the maternal-fetal interface may determine whether a pregnancy succeeds or fails. By understanding the mechanisms that control how trophoblast TLRs function, only then can we move to clinical applications to determine better ways to predict pregnancy outcome and treat women at risk of infection-associated pregnancy complications. Excessive placental apoptosis has been associated with preeclampsia and preterm labor; however the initial trigger and mechanisms involved are not fully understood. We propose that infections represent a potential trigger for placental apoptosis, and that some TLRs can mediate this response. Specifically, our central hypothesis is that some bacterial and viral components, through TLRs, induce trophoblast apoptosis, leading to adverse pregnancy outcome, such as preterm labor or preeclampsia. In our published studies we found TLR2 activation by gram-positive bacterial peptidoglycan (PDG), and TLR8 activation by viral ssRNA, trigger human first trimester trophoblast apoptosis via two distinct pathways: TLR2 directly mediates apoptosis, and this is differentially regulated by the TLR2 co-receptor, TLR6. In contrast, TLR8 indirectly mediates apoptosis by upregulating the cell's production of IFN2. Our objectives are to further characterize the cellular and molecular mechanisms by which TLR2 and TLR8 mediate trophoblast apoptosis in response to bacterial and viral components, and to determine their impact on pregnancy outcome. Within this overall goal, we will address innovative areas in which there are major gaps in our knowledge, such as the: regulation of TLR expression by DNA methylation; regulation of trophoblast miRs by TLRs; function of TLR8 in the trophoblast; and the effect of viral ssRNA on pregnancy outcome. We will also apply translate our in vitro findings into a study to develop better predictive and preventative strategies. Thus, our specific aims are to: 1. Determine the regulation of TLR2-induced apoptosis in response to bacterial components. 2. Determine the mechanism by which TLR8 functions in the trophoblast in response to viral components. 3. Determine the role of microRNAs in the regulation of TLR2- and TLR8-mediated trophoblast apoptosis. 4. Evaluate the role of TLRs in pregnancy. While the link between bacterial infections and pregnancy complications is well established, less is known about how viral infections affect pregnancy. Despite a recent increase in our understanding of the role of placental TLRs in pregnancy, our knowledge about the specific mechanisms involved is still limited, as is the role of TLRs in mediating apoptosis. Our findings will further our understanding of normal placental function, and will lead to a better understanding, prediction and treatment of infection-associated pregnancy complications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Imaging of Angiogenesis Angiogenesis is an important process in the growth and spread of cancer. It is a topic of great interest to scientists and clinicians in the CCR who are utilizing anti-angiogenic strategies for treatment and therefore, it is of importance that the MIP address angiogenesis in its program of targeted tumor imaging. Here, we describe the pre-clinical and clinical aspects of angiogenesis imaging in MIP. Pre-Clinical Research There are two approaches to angiogenesis imaging: physiologic or functional studies that investigate the flow and permeability dynamics of vessels within tumors and a molecular targeted approach. We have investigated both aspects. The MIP has developed expertise in the performance of dynamic contrast enhanced MRI (DCE-MRI)with low molecular weight and macromolecular contrast agents(1-3). This is a seemingly straightforward test in which a bolus of a paramagnetic contrast agent is followed by the rapid acquisition of images. The images are then analyzed and curve fitting parameters are derived that reflect the physiology of the vasculature. Interestingly, while popular, there has been little validation of this method compared to other well known methods of measuring vessel permeability. We have conducted ultrastructure studies in conjunction with Donald McDonalds lab in UCSF(4). We conducted an murine study in which radiolabeled (14C) permeability agent was compared (using Quantitative AutoRadiography (QAR) with DCE-MRI in a brain tumor model(5). We found excellent correlation between the QAR results and the parameter Ktrans which is derived from DCE-MRI. We have also performed validation studies in treatment models involving TNFalpha (6). Over the past year we have developed targeted integrin imaging (optical and radionuclide) using cyclized RGD (cRGD). This work could result in a targeted imaging agent for directly visualizing angiogenic vessels. New targeted imaging agents are being sought to further pursue this work. Additionally, in collaboration with Brad St Croix, we have labeled an antibody against TEM8 which is highly expressed in angiogenic vessels and appears highly specific. This antibody is being developed as a therapeutic agent by a major Pharma company. We have labeled this antibody with both Br76 and Zr89 in order to document its areas of increased uptake. Interestingly, because this is an endothelial marker it is difficult to measure in vitro. Thus, in vivo measurement is a requirement. Preliminary data suggests that the agent is not only sensitive for angiogenesis but it also can be inhibited by therapeutic levels of the antibody. Moreover, to make a more practical imaging agent as a potential companion diagnostic, we have developed antibody fragments with Gary Griffiths lab. Thse should allow more rapid imaging than is possible with full antibodies. Clinical studies The MIP provides a DCE-MRI service to investigators in the Clinical Center. This is now considered a routine study at NIH due to our efforts. One of the clinical fellows in MIP, identifies a target lesion and ensures that the proper study is performed each time the patient returns. The fellow also provides results to the investigators for research purposes. The MIP is providing this service to 5 active protocols by CCR investigators who are looking at various anti-angiogenic agents. Such work, by its nature, is difficult and publications have been slow to accrue, however in the past years two publications have appeared from a breast cancer trial (7, 8). These demonstrate profound changes in vessel permeability in responders to Bevacizumab followed by high dose intensive chemotherapy. The role of DCE-MRI was compared to other biomarkers that were studied and it was found to be among the more useful of the predictive markers. The MIP is also conducting a study with Dr. Wyndham Wilson using a PET agent that targets the AvB3 and AvB5 integrins. This 18F labeled PET agent is made by GE Healthcare and is being offered to NCI for trial purposes. Because we have been able to develop a clinical team (2 nuclear medicine physicians (Karen Kurdziel, Liza Lindenberg), one body radiologist (Peter Choyke), one neuroradiologist (Dima Hammoud) as well as support staff in the form of Nurse Practitioners and Research Nurses, we are prepared to take advantage of this opportunity that would formerly not have been possible. We have initated a proof of principle trial involving 30 patients with this agent, which is named flucilitide. In this trial a single fluciclitide imaging study is performed prior to surgery. The specimen is then analyzed for integrin expression using immunohistochemistry. We have just begun a clinical trial in patients receiving anti-angiogenic therapies (four protocols feed this imaging protocol). In this trial fluciclitide imaging is obtained before and after one cycle of therapy as defined by the treatment protocol. 1. Barrett, T., Brechbiel, M., Bernardo, M., and Choyke, P. L. MRI of tumor angiogenesis. J Magn Reson Imaging, 26: 235-249, 2007. 2. Barrett, T., Kobayashi, H., Brechbiel, M., and Choyke, P. L. Macromolecular MRI contrast agents for imaging tumor angiogenesis. Eur J Radiol, 60: 353-366, 2006. 3. Xu, H., Regino, C. A., Bernardo, M., Koyama, Y., Kobayashi, H., Choyke, P. L., and Brechbiel, M. W. Toward improved syntheses of dendrimer-based magnetic resonance imaging contrast agents: new bifunctional diethylenetriaminepentaacetic acid ligands and nonaqueous conjugation chemistry. J Med Chem, 50: 3185-3193, 2007. 4. Ocak, I., Baluk, P., Barrett, T., McDonald, D. M., and Choyke, P. The biologic basis of in vivo angiogenesis imaging. Front Biosci, 12: 3601-3616, 2007. 5. Ferrier, M. C., Sarin, H., Fung, S. H., Schatlo, B., Pluta, R. M., Gupta, S. N., Choyke, P. L., Oldfield, E. H., Thomasson, D., and Butman, J. A. Validation of dynamic contrast-enhanced magnetic resonance imaging-derived vascular permeability measurements using quantitative autoradiography in the RG2 rat brain tumor model. Neoplasia, 9: 546-555, 2007. 6. Tang, J. S., Choy, G., Bernardo, M., Thomasson, D., Libutti, S. K., and Choyke, P. L. Dynamic contrast-enhanced magnetic resonance imaging in the assessment of early response to tumor necrosis factor alpha in a colon carcinoma model. Invest Radiol, 41: 691-696, 2006. 7. Wedam, S. B., Low, J. A., Yang, S. X., Chow, C. K., Choyke, P., Danforth, D., Hewitt, S. M., Berman, A., Steinberg, S. M., Liewehr, D. J., Plehn, J., Doshi, A., Thomasson, D., McCarthy, N., Koeppen, H., Sherman, M., Zujewski, J., Camphausen, K., Chen, H., and Swain, S. M. Antiangiogenic and antitumor effects of bevacizumab in patients with inflammatory and locally advanced breast cancer. J Clin Oncol, 24: 769-777, 2006. 8. Thukral, A., Thomasson, D. M., Chow, C. K., Eulate, R., Wedam, S. B., Gupta, S. N., Wise, B. J., Steinberg, S. M., Liewehr, D. J., Choyke, P. L., and Swain, S. M. Inflammatory Breast Cancer: Dynamic Contrast-enhanced MR in Patients Receiving Bevacizumab Initial Experience. Radiology, 244: 727-735, 2007.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the proposed project is to clarify the molecular mechanism of both function and regulation of mammalian class VI myosin. Our preliminary studies suggest that Ca 2+and phosphorylation by small G-protein down-stream protein kinases regulate the motor activity of myosin VI. We will first examine whether the specific phosphorylation of myosin VI occurs in cells, and then study the mechanism of Ca 2+ and/or phosphorylation mediated regulation of myosin VI motor activity. A recent study by others and us has revealed that class VI myosin is a processive motor that travels on actin filaments for a long distance without dissociating from actin. However, the mechanism by which myosin VI moves processively along actin filaments is not understood. We will address this problem by using various biophysical and electron microscopy techniques. The best approach to show the processive movement of myosin VI is the use of single molecule analysis. We will employ two techniques, i.e., mechanical measurement with optical trap nanometry, and direct visualization of the movement by total internal reflection (TIRF) microscopy. The rotational motion of myosin VI on actin will be monitored by visualizing the movement of beads attached to myosin VI on actin filament. The conformational changes of myosin VI during the mechanical cycle will be studied by single molecule polarization TIRF microscopy that measures the angular change of myosin head. The overall structural change of myosin VI with 0.1 nm resolution will be monitored by X-ray solution scattering. The structure of the two-headed myosin VI on actin filament will be studied by 3D image reconstitution of the myosin VI decorated actin filaments with cryo-electron microscopy. We will also determine the structural motifs responsible for the processivity and reverse directionality of myosin VI by analyzing the motor properties of the variants in which each structural motif is changed by genetic engineering technology. In order to achieve this goal, we will use recombinant DNA technology to produce engineered myosin VI molecules. Particular regions of the myosin VI molecule that are hypothesized to be critical for the uniqueness and/or regulation of motor function will be modified and functionally expressed. The motor function will then be analyzed by enzymatic analysis, biophysical analysis and in vitro motility assay, ,with a particular emphasis on the single molecule assay system. The itemized specific aims are: 1. To determine the regulatory mechanisms of myosin VI motor function; 2) To define the structural changes of myosin VI during the ATP hydrolysis cycle; 3) To define the mechanism by which myosin VI moves processively along actin filaments; 4) To identify the molecular determinant of the directionality of myosin VI. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Naltrexone and acamprosate have shown efficacy as adjuncts to psychosocial/behavioral alcoholism treatments. A number of studies have suggested that time limited outpatient behavioral approaches may have similar efficacy to more intense and skill based therapies. A multisite study is proposed to compare the effectiveness of combining medications with therapies which differ in intensity and content. A primary main study and a pilot study are proposed in this application. In the primary study 240 alcohol dependent individuals, at our site, will be assigned to eight treatment cells formed by the combination of 4 medication (placebo, naltrexone, acamprosate, naltrexone plus acamprosate) and 2 behavioral (cognitive behavioral (CBT) and modified motivational enhancement [MET-light]) therapies. Subjects will receive behavioral therapy for 12 weeks and medication for 26 weeks and follow-up assessments will occur at 3, 6, 9 and 12 months post-therapy. Standard alcohol consumption instruments, biological markers, and collateral will be used. The measurement of medication compliance (riboflavin as a marker) will be emphasized. In the pilot study 40 alcohol dependent individuals will be assigned to four treatment cells formed by the combination of 2 medication (placebo or naltrexone plus acamprosate) and 2 behavioral (CBT or MET-light) treatments. Subjects will be assessed and treated as proposed for the primary study. However, half of the subjects will receive the medication packaged in blister packs and half in MEMS cap bottles to determine (based on riboflavin measurements) which method has better compliance monitoring capabilities. In addition, one of the subjects will receive \"booster therapy\" at week 20 with retention and medication compliance evaluated in both groups. Patient satisfaction and adverse medication effects will also be evaluated between the two medication groups. If the primary study occurs at eight sites, a total of 1920 subjects (240/treatment cell) should provide enough power to detect medication versus placebo effects in both therapies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have been studying the efficacy of local injections of botulinum toxin for the treatment of different types of focal dystonias. Botulinum toxin injected in small doses directly into muscle, binds to and inactivates the neuromuscular junction. Dystonic contraction is decreased and function improves Treatment is well-tolerated with minimal side effects. We have also been using botulinum toxin to study the physiology of focal dystonias. We have continued the treatment of patients with writer's cramp. These patients are active participants in physiology protocols including transcranial magnetic stimulation, PET, microneurography and somatosensory evoked potentials studies. We have demonstrated that immediate exercise following injection in writer's cramp patients leads to greater loss of strength. We continued to study the long-term effects of botulinum toxin type F for patients who lose response to type A due to antibodies and for patients who never responded to type A despite adequate doses. Two patients who previously responded became resistant to the effects of botulinum toxin F. These responses are being investigated with testing for botulinum toxin F antibodies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Pulaski County Services for Problem Drinking Drivers Program will offer coordination, planning and minimal emergency treatment services to assist clients identified in the Pulaski County Alcohol Safety Action Program to make maximum utilization of treatment and rehabilitation services available in the community. In addition, the Program Staff will identify gaps and strains in the service delivery system and will work with existing agencies or other interested groups to meet these needs or relieve the system's overload that develops. The Program Staff will be housed with a countermeasure unit of the Pulaski County ASAP Program, also sponsored by the University of Arkansas Graduate School of Social Work, for purposes of coordination and client contact. The Program is designed in its coordination phase as an outreach effort concerned with direct assistance to clients in obtaining needed services as identified by ASAP. In addition to the planning function, a limited amount of funds will be allocated to purchase necessary emergency treatment services where no other resources are available.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Differential Proteomics, Inc., is developing a new proteomics technology for comparing the protein composition of biological samples. The technology, Differential Capture Proteomics (DCP), is designed to circumvent serious limitations in the current capabilities of current proteomics technology, notably DCP can identify the protein differences between two body fluid or tissue samples and simultaneously generate an affinity reagent for each identified difference protein. The DCP approach utilizes phage display libraries of peptides from which affinity reagents against the protein components of two samples are isolated. A novel subtraction step removes those affinity reagents against proteins held in common between the two samples, leaving affinity reagents against the difference proteins. These affinity reagents are used to purify the difference proteins, which are then identified using mass spectrometry. DCP will provide a fast, cost-effective process for performing differential proteomic analyses. It will accelerate the pace of biomarker discovery and also simultaneously provide affinity reagents for those biomarkers. These affinity reagents will immediately be available for the development of diagnostic tests for the biomarkers. Thus DCP will play an important role in medicine and biomedical research wherever biomarkers are involved, including disease diagnosis, the monitoring of disease progression, drug discovery and development and the monitoring of drug efficacy in patients. Phase I of this project will build on the success of preliminary experiments and will determine conditions for performing the initial, phage display steps of the DCP process. The format of column chromatography will be used in place of the batch selection protocols currently used for phage display selections. Phase I has 4 specific aims, each of which will optimize a particular step in the phage display portion of the DCP process. Together, specific Aims 1 and 2 will determine the optimal conditions for performing column chromatography of filamentous phage, leading to the selection of phage peptides that bind specifically to model targets (monoclonal anti-peptide antibodies) with the lowest background obtainable. Specific aims 3 and 4 address the novel DCP subtraction step. Here, optimal chromatographic conditions will be determined for the efficient removal of phage that binds to targets present in both of a pair of samples. In addition, the non-specific background will be further reduced in this step. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this work is to develop mathematical models of the blood flow and transcapillary exchanges in capillary networks. An effort is being made to incorporate in the models of the histological structure of capillary networks as well as different flow patterns from available experimental information. In this model the extraction of substrates .with different chemical kinetics at the tissue site will be described. It is expected that this could be used in experimental situations where the extraction of different substrates are measured simultaneously, thus helping to infer the flow pattern features of the microcirculation. In particular a model of the diffusion-consumption of oxygen in striated muscle containing myoglobin (facilitated diffusion) is being developed and pertinent numerical results examined.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A study to evaluate the growth and pubertal development in male rhesus monkeys chronically exposed to methylphenidate. One of the more commonly recognized pediatric behavioral disorders is Attention Deficit Hyperactivity Disorder or ADHD, in which methylphenidate is drug for treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "With mutants, genetic mosaics and electrophysiological techniques it is proposed to analyze specifically the control of aging, flight mechanisms and the neuromuscular control of leg movements in Drosophila. Hyperkinetic 1 and Shaker 5 are neuroloical mutants of Drosophila with reduced life span as compared to their wild-type background stock. The reduction in longevity has been shown to be correlated with an increased activity level and rate of oxygen consumption. The longevity of gynandromorphs has been measured and the data will be analyzed to determine if a limited area or region may be defined as an aging center. Flight mutants exist for which it is proposed to study, genetically and physiologically, mosaic individuals in order to dissect out the genetic control of flight. Various types of mutants will also enlarge our understanding of nerve-muscle interrelationships.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The rapid effects of thyroid hormone on the activity of Kv11.1 channels in rat pituitary cells were recently shown (Storey et al 2006) to be mediated by the classical nuclear receptor for thyroid hormone, TRbeta;, acting at the plasma membrane through the phosphoinositide 3 kinase (PI3K) and the Rac GTPase, which is a well-known effector of PIP3 dependent Rac exchange factors. Signaling was reconstituted in CHO cells by heterologous expression of human Kv11.1 channels and the human TRbeta, but not the TRalpha receptor. We have continued to investigate the mechanism of PI3K stimulation by TRbeta and the consequences of PI3K signaling for the physiological effects of thyroid hormone. We have used a fluorescent PIP3 binding domain from the Akt protein kinase coupled with cyan and yellow fluorecent proteins to detect PIP3 production by fluorescence resonance energy transfer (FRET) in live cells in real time. Fret signals are blocked by inhibition of TRbeta with the nuclear receptor antagonist,1-850, and by wortmannin, an active site inhibitor of PI3K. PIP3 production is also blocked completely by 10 min preexposure to 100 nM TCDD, an environmental toxicant. We have used immunoprecipitation to show that TRbeta associates with the regulatory p85 subunit of PI3K in the absence of ligand but dissociates in the presence of thyroid hormone. We have also observed rapid thyroid hormone-dependent phosphorylation of the Akt protein kinase and recruitment of Rac to the plasma membrane confirming that thyroid hormone stimulates PI3K-dependent effectors. Finally we have discovered that the Src family kinase, Lyn, is part of the signaling complex and identified its binding site with mass spectrometry. Other proteins interact with p85 through two Src homology (SH2) domains which recognize phosphotyrosine. TRbeta but not TRalpha contains a consensus SH2-binding domain with high affinity for p85. Mutating the tyrosine in that consensus site to phenylalanine prevents reconstitution of Kv11.1 regulation by thyroid hormone in CHO cells but not activation of a heterologous transcription of a luciferase gene driven by a thyroid hormone receptor response element (TRE). A second tyrosine that is responsible for Lyn binding has been identified and is also required for PI3K stimulation. Thus, this mechanism provides a direct link for integrating growth signals through thyroid hormone and receptor tyrosine kinases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To determine the magnitude and duration of acetylcholinesterase (ACHE) inhibition in cerebrospinal fluid (CSF) after a single oral 3.0 mg dose of ENA 713 is administered to healthy male volunteers and to correlate these parameters.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Obesity is currently a worldwide epidemic, therefore preventing and reversing obesity is of utmost importance for the general health of the population. Body weight regulation is maintained through a balance of energy intake versus energy expenditure, and targeting appetite or metabolic pathways via obesity therapeutics is a current goal in obesity research. The brain is the center for control and coordination of body weight. Hypothalamic melanocortin system activation reduces appetite, and also sends signals to the nucleus of the solitary tract (NTS) which activates sympathetic nervous system (SNS) innervation of brown adipose tissue (BAT), leading to increased thermogenesis (energy expenditure). The aim of this proposal is to further investigate a family of molecules recently found to be involved in metabolism. The bone morphogenetic proteins (BMPs) are growth factors, and recent studies have demonstrated roles for the BMPs in metabolism and satiety in lower organisms. Our laboratory has also shown that BMP7 in mice is able to stimulate brown adipocyte differentiation and to induce BAT thermogenesis. Additionally, we have shown that BMP7 and its receptors are expressed in the hypothalamus and BMP7 delivery (either i.c.v. or systemically via adenovirus) produces an anorexigenic effect, at least in part through the hypothalamic mTOR pathway and activation of the melanocortin system. However, much is still unknown about the metabolic and physiological roles of BMP signaling in energy balance. Therefore, the overarching goal of this project is to identify how BMP7 impacts whole body energy balance by affecting appetite pathways and BAT thermogenesis. Thus far, we have found that mice with BMP7 haploinsufficiency (BMP7-/+) on a high fat diet (HFD) are more obese and hyperphagic than littermate controls, with decreased energy expenditure (oxygen consumption/VO2) levels. Therefore, our first objective is to determine the physiological mechanism for BMP7's effects on energy balance, with the hypothesis that BMP7 is regulating the hypothalamic-NTS melanocortin system which regulates both central appetite pathways as well as energy expenditure pathways (via sympathetic innervation of BAT). We have also shown that mice with POMC-neuron (ie: in hypothalamus and NTS/brainstem) deletion of BMPR1a (a BMP7 receptor) exhibit hyperphagia as well as increased energy expenditure and BAT thermogenesis. Therefore our second objective is to determine whether the mechanism for POMC-neuron BMPR1a action is through hypothalamic melanocortin pathway inhibition to induce hyperphagia, and through the NTS-SNS pathways to increase thermogenesis in BAT. These studies will utilize novel mouse models for metabolic studies, including an important model linking central BMP signaling disruption with peripheral energy expenditure effects, and will investigate a novel class of appetite factors: the BMP growth factors. Therefore, the research outlined in this proposal could provide important new findings for the field of obesity research, and potentially lead to development of new obesity therapeutics. PUBLIC HEALTH RELEVANCE: Obesity is currently epidemic worldwide, therefore current biomedical science seeks to develop obesity therapeutics and to understand the pathophysiology of obesity in order to implement prevention strategies. This project utilizes two novel mouse models to investigate the role of the BMPs, a family of growth factors, in both nervous system and brown adipose tissue pathways regulating body weight homeostasis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In the central nervous system (CNS), measles virus produces not only acute, post infective encephalomyelitis but also Subacute Sclerosing Panencephalitis (SSPE), a progressive inflammatory disease of children after viral latency of several years. Measles has been implicated in the etiology of Multiple Sclerosis (MS) as well. In these diseases the CNS virus appears to be defective or incomplete and the pathogenesis of them may involve immunopathological responses to the defective, latent infection. A latent CNS infection of immature, or transiently immunosuppressed adult hamsters with hamster adapted SSPE agent has been developed which has many of the virologic, serologic and ultrastructural characteristics of human SSPE. The CNS virus changes from a complete to a defective, cell-associated form coincident with the appearance of host serum antibodies to measles. Using virological, serological, histological, immunofluorescent, electron microscopic and ultrastructural immunoperoxidase methods we propose sequential studies to define more completely characteristics of the latent CNS virus and the host immune response to it. An investigation combining the latent infection with experimental allergic encephalomyelitis has been initiated and will be extended. Study of this chronic, cell-associated CNS viral infection with measles virus will increase our knowledge of SSPE and the findings may also be significant in understanding Multiple Sclerosis. It may also aid our understanding of cellular immune mechanisms as they operate in the CNS and advance the evaluation of new forms of therapy for SSPE and Multiple Sclerosis. BIBLIOGRAPHIC REFERENCES: Raine, C. S., Byrington, D. P., and Johnson, K. P. Subacute sclerosing panencephalitis in the hamster: Ultrastructure of the acute disease in newborns and weanlings. Lab, Invest. 33: 108, 1975. Johnson, K. P., Feldman, E. G., Byington, D. P. The effect of neonatal thymectomy in experimental subacute sclerosing panencephalitis in adult hamsters. Infection and Immunity, 12:1464, 1975.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The TB Academic Award is a mechanism for achieving strong programs of TB education in targeted U.S. Medical Centers. The proposal from Duke University Medical Center demonstrates that our geographic region has an aggregate incidence of TB worthy of attention. Furthermore, we show that TB is a rural-based problem in North Carolina and propose educational programs more relevant to health care providers practicing in a largely rural region such as ours. Finally, we demonstrate that Duke has the infrastructure and commitment to develop and sustain a program of TB education. We believe such a program could serve as a model for states with similar population patterns, healthcare shortages, and incidence of TB. Our goals for the TB academic award at Duke are: a) to improve existing Duke curricula so that clinical trainees at Duke Medical Center will, upon completion of their training: know how to prevent, manage and control TB; know specifically where to turn for continued medical education about TB during their professional lives; and understand the role of local health departments in the management of TB infection and disease; b) to develop curricula and educational interventions that can be customized and used in similar regions of the U.S; c) to improve Duke's effectiveness in teaching practicing clinicians about TB in our region and provide them with effective life- long educational resources; d) to use the coordinated and interdepartmental nature of this educational initiative to improve the care of TB patients at Duke by establishing consistent diagnostic and therapeutic practices; e) to develop and teach computer-based distance learning modalities that are versatile, simple to update and can be integrated into currently developing curricula and strategies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this study is to evaluate acid-base regulation in cardiac and skeletal muscle during combinations of normoxia or hypoxia and metabolic acidosis with or without an accompanying respiratory acidosis. Acute or chronic metabolic acidosis was produced by administering NH4Cl to adult male rats. Intracellular pH was determined by the DMO method. Acute metabolic acidosis. Rats breathing 21% O2 -0% CO2 (normoxia-normocapnia) had significant decreases in cardiac and skeletal muscle pH. The production of hypoxia-normocapnia (10% O2, 3% CO2) or hypoxic-hypercapnic conditions (10% O2, 10% CO2) did not alter the intracellular pH of these two types of muscle with respect to the values obtained under normoxic-normocapnic conditions. Chronic metabolic acidosis. These rats differed from the acutely metabolic acidotic rats in that the extracellular acid-base state was essentially normal. However there was a significant decrease in both cardiac and skeletal muscle pH. Hypoxia and normocapnia produced a further fall in the intracellular pH of these muscles. However, the addition of hypercapnia to the hypoxic state did not result in a further decrease in the pH of these muscles. These data indicate that the production of metabolic acidosis with NH4Cl results in the development of an enhancement of cardiac and skeletal muscle acid-base regulation. The mechanism of this enhanced effective buffering remains to be determined.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The transcriptional regulation of gene expression is pivotal to all biological processes. Each of our ~20,000 protein-coding genes must be expressed at the right place, time and level, as well as under the right developmental or physiological circumstances. Consequently, inappropriate gene expression is implicated in a myriad of human diseases, including congenital disorders, cancer and obesity. Transcription has been studied intensively for decades, resulting in a detailed picture of the basic biochemical mechanisms of mRNA production. However, we know little about gene regulation at a 'systems level', i.e. how TFs function together in complex gene regulatory networks (GRNs) to faithfully orchestrate the expression of large sets of genes. Our long-term goal is to comprehensively characterize the structure, function and evolution of complex metazoan GRNs to gain insights into global mechanisms of gene regulation. It is becoming increasingly clear that textbook explanations of gene regulation in which a TF binds DNA in the genome and upon doing so regulates the most proximal gene are too simplistic because many physical TF binding events lack an apparent regulatory consequence. There are several explanations for this, ranging from technical (e.g. detection limits, attribution of a bindng event to the wrong gene) to biological (e.g. redundancy between TFs, condition-dependent effects). Conversely, regulatory interactions are not necessarily due to a direct effect. For instance, TFs can function in cascades to propagate functional regulation. A major challenge is to combine physical and regulatory interactions to increase our understanding of the mechanisms of gene regulation in the context of complex multicellular organisms. Many GRN studies focus either solely on physical TF interactions, whereas others focus primarily on regulatory interactions. However, integrated GRNs that combine both are scarce and, when available are relatively small in scale. If we had high-quality, large- scale physical and regulatoy interaction data, as well as spatiotemporal and conditional gene expression data, we could build increasingly precise GRNs. Here, we will continue our studies on the nematode C. elegans to map and integrate physical and regulatory GRNs, which will help us to go beyond mapping to understanding the regulatory mechanisms of gene expression at a systems level in a complex multicellular organism.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy in the United States. Thus, there is an urgent need to develop new therapeutic approaches for this disease. EOC is genetically heterogeneous and, therefore, it is imperative that therapeutic strategies be personalized by targeting distinct molecular subsets of EOC. CARM1 is an arginine methyltransferase that asymmetrically dimethylates protein substrates on arginine residues. Notably, EOC is among the cancer types that show the highest CARM1 amplification rates in The Cancer Genome Atlas (TCGA) database. The role of CARM1 in EOC has never been investigated. The ultimate goal of the current proposal is to develop novel approaches to treat and eradicate CARM1-associated EOC. Our unbiased screen show that CARM1-expressing EOC cells are selectively sensitive to the inhibition of EZH2, another epigenetic regulator. Genome-wide profiling suggests that the observed selectivity correlates with upregulation of EZH2 target genes in a CARM1-dependent manner. Inactivation of subunits of the SWI/SNF chromatin- remodeling complex is synthetically lethal with inhibition of EZH2. Interestingly, BAF155, a core subunit of the SWI/SNF complex, is a substrate of CARM1. Indeed, our preliminary data support a model that CARM1 renders cells functionally dependent on EZH2 activity by methylating BAF155. CARM1 expression is typically mutually exclusive with BRCA1/2 mutations in EOC. PARP inhibitor Olaparib is approved by FDA for recurrent BRCA1/2-mutated EOC. Thus, sensitizing BRCA1/2 wildtype EOC to PARP inhibition remains an unmet clinical need. Notably, our preliminary data suggest a synergy between EZH2 inhibitor and PARP inhibitor in a CARM1-dependent manner. Further, EZH2 inhibition enhances CD8+ effector T cell trafficking into tumors by upregulating Th1-type chemokines. This raises the possibility of combining EZH2 inhibitor with immuno-checkpoint blockade such as anti-PD-L1 therapy in CARM-expressing EOCs. Thus, our central hypothesis is that CARM1-expressing EOC can be treated and ultimately eradicated by targeting EZH2 in combination with PARP inhibitor or immune-checkpoint blockade such as anti-PD-L1 therapy. Two specific aims are proposed: Aim 1: To investigate the mechanistic basis underlying the selectivity of EZH2 inhibition against CARM1; and Aim 2: To develop novel therapeutic strategies for CARM1-expressing EOC. The proposed studies are highly innovative because they challenge current research/clinical paradigms, and explore new intervention strategies for CARM1-associated EOC. The research proposed is of high impact because it has the potential to establish the first effective combinational therapeutic strategy for CARM1-associated EOC. Since amplification and overexpression of CARM1 occurs in several other major cancer types, the mechanistic insights gained from the current studies will also have broad implications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Women with triple-negative breast cancer (TNBC), the most aggressive subtype of breast cancer, have the shortest survival times and the highest rate of relapse. These patients have no currently available forms of targeted therapy and most (~77%) succumb to metastases within 5 years. However, the nature of how metastasis relies on signaling from the tumor microenvironment is not well understood. Macrophages have also been linked to human breast cancer invasion and metastasis as well as drug resistance, and recently have been implicated in TNBC. M1-like pro-inflammatory macrophages are thought suppress tumor growth and metastasis, whereas the tumor-associated macrophages (TAMs) are characterized as M2-like pro-metastatic macrophages. However, recent proteomic and RNA studies indicate that macrophage populations are composed of more phenotypic subtypes than previously recognized. Thus, the phenotype of TNBC TAMs and the mechanisms by which they interact with TNBCs are not well understood and require further characterization. In recent studies I have been testing the hypothesis that TNBC recruits a unique subtype of TAMs via CCL5 that drives invasion and metastasis. Using genetically matched metastatic and non-metastatic TNBC tumors that differ only by expression of the metastasis suppressor Raf Kinase Inhibitory Protein (RKIP), I show that RKIP alters not only the number of recruited TAMs but also their phenotype. I further demonstrate that CCL5 expressed by metastatic tumors recruits TAMs that secrete pro-metastatic factors, promote invasion of TNBC cells, are highly expressed in human TNBC patients, and contribute to a prognostic signature for patient survival. Taken together, these findings suggest that TNBC recruited TAMs are both phenotypically and functionally distinct from other TAMs, and M1 or M2 macrophages. I now propose to test the hypothesis that these TAMs can be distinguished by unique surface receptors that regulate expression and secretion of pro-metastatic factors. I further propose that TAMs recruited by CCL5 to metastatic TNBCs drive metastasis through expression of GRN along with other associated factors. Specifically, I will: 1) Characterize the phenotype of pro-metastatic TAMs in TNBC; and 2) Determine the functional role of TAMs & TAM secreted factors in TNBC metastasis. This study is innovative because it seeks to understand the phenotype of a novel TAM subtype identified in TNBC, as well as the in vivo mechanism of TAM driven invasion in TNBC. Together, these results could generate new diagnostic markers and prognostic signatures as well as identify potential therapeutic targets to deplete or reprogram pro-metastatic TAMs in TNBC patients. This work should also lead to a better understanding of the role of TAMs in the poor outcomes of TNBC patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of the proposed research is to characterize the cellular mechanisms that contribute to tuning in the turtle cochlea. By understanding how these cellular mechanisms determine frequency selectivity, we will be able to assess their limitations and applicability to hearing in higher vertebrates including man. The proposed research will study the innate mechanical and electrical basis of tuning and modulation by efferent input. A combination of patch clamp, microelectrode, and confocal imaging techniques will be used on isolated, solitary hair cells and in the intact basilar papilla. Previous measurements of ionic currents in solitary cells will be extended and combined with [Ca2+]i imaging to reconstruct membrane potential resonance. Transduction in solitary cells will be analyzed to determine if tuning due to basolateral conductances is enhanced by active, mechanical processes in the ciliary bundle. Finally, efferent modulation of tuning will be investigated in both solitary cells and the intact papilla. Special attention will be paid to the need to unambiguously determine the ionic basis of efferent action and whether the synaptic conductances are identical to those involved in electrical tuning. These results will be used to construct a complete description of the cellular mechanisms involved in tuning. Experiments will determine whether ciliary bundle motion in turtle hair cells is produced by a change in stiffness or a voltage-dependent force. The site and cellular mechanism underlying the motion will be characterized. In further experiments on the transducer, the channels at sites of transduction along the length of the stereocilia and the apical surface of the hair cell will be inactivated with UV radiation. The currents of solitary cells will be measured with the perforated-patch variation of the whole-cell voltage-clamp technique. Changes in the size and kinetics of the membrane currents in cells of known characteristic frequency will be analyzed using intracellular exchange of monovalent cations. Simultaneous cell-attached single-channel and perforated-patch whole cell recordings will be used to compare the behavior of the single channel and the macroscopic IK(Ca). Changes in [Ca2+]i will be measured simultaneously using confocal imaging of indo-1 fluorescence. The single channel will then be studied in an excised patch, and its voltage- and [Ca2+] - sensitivity assessed. Confocal imaging of indo-1 fluorescence will be used to measure the local variation of [Ca2+]i within a single hair cell in the intact papilla. Confocal imaging of electrically activated afferent and efferent fibers stained with the voltage-sensitive dye, di-4-ANEPPS, will be used to identify afferent and efferent terminals on the hair cell's basolateral surface. Hair cell potential will be controlled with a single microelectrode used in current clamp or in switching-mode voltage clamp. The spatial changes of [Ca2+]i in the hair cell that occur when the cell is depolarized, during mechanical stimulation, or that following direct electrical stimulation of the efferent fibers will be compared with the position of afferent and efferent terminals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION 3 OVERALL PROGRAM CRITIQUE 3 PROGRAM LEADERSHIP 4 PROGRAM AS AN INTEGRATED EFFORT 5 COLLABORATING INSTITUTIONS 5 PROJECT AND CORE SUMMARIES OF DISCUSSION 5 PROTECTION OF HUMAN SUBJECTS 7 VERTEBRATE ANIMALS 8 ADDITIONAL REVIEW CONSIDERATIONS 8 PROJECT 1: NLR in Host Response to Gamma-Herpesviruses 9 PROJECT 2: Regulators of Innate Immune Responses to Gamma-herpesviruses 17 PROJECT 3: Role of Non-Coding RNAs in Regulating Gamma-Herpesvirus-Host-Interactions 26 PROJECT 4: In Vivo Interactions Between a Gamma-Herpesvirus and Innate Immune Responses 34 CORE A: Administrative Service 43 CORE B: Virology Core 46 COMMITTEE BUDGET RECOMMENDATIONS 50 SPECIAL EMPHASIS PANEL ROSTER DESCRIPTION (provided by applicant): Through co-evolution with hosts, herpesviruses have acquired strategies to exploit their hosts, allowing their own life-long persistence while intermittently being secreted and transmitted to naive hosts. By evading host defense mechanisms, herpesviruses can persist in hosts and cause various diseases. Members of the gamma-herpesvirus subfamily (Epsetin-Barr virus, Kaposi's sarcoma-associated herpesviruses and gamma herpesvirus-68) are distinct in their ability to establish latent infections in lymphocytes and cause benign or malignant tumors in infected hosts. It remains elusive how gamma-herpesviruses evade host innate immunity during their acute and persistent infections. Understanding the mechanisms of viral evasion mechanism is essential for developing approaches to prevent or control the virus associated cancers. Innate immunity is not only the first line of defense against pathogens, but is also critical for stimulating adaptive immune responses. The objective is to understand the mechanisms by which cells mount innate defense responses and tumor-associated-herpesviruses thwart components of the defense mechanisms, to establish a foundation for rational design of vaccine candidates that can elicit effective immune responses and reduce associated cancer incidence. There are four projects: 1) Interactions of gamma-herpesviruses with NLRs (Cart Ware); 2) Regulators of innate immune responses to gamma-herpesvirus infection (Sumit Chanda); 3) Role of microRNAs in regulating host interactions (Tariq Rana); 4) Restoring immune responses to gamma-herpesviruses (Ren Sun); coordinated by an Administrative Core (Ren Sun) and facilitated by a Virology Technical Service Core (Ting-Ting Wu). A team of investigators with diverse and complementing expertise has been working together and obtained critical preliminary data as the foundation for the Program. The understanding of the molecular and cellular mechanisms underlying the interactions between virus-host innate defenses will be utilized to develop a new vaccine strategy. This translational research program will provide critical information for vaccine development to prevent cancers associated with gamma-herpesviruses. RELEVANCE: At least ~200,000 new cases of EBV-associated malignances occur annually and similar numbers of Kaposi's sarcoma cases are reported each year worldwide. Vaccines are the most effective and affordable approach to these cancers, especially in resource-limited areas. This Program is to dissect the viral immune evasion mechanism to rationally design vaccine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Studies of the absorption, distribution, metabolism and excretion of two 14C-labeled aromatic amines, p-nitroaniline and diphenyl guanidine, in the rat indicate that clearance rates vary with compound, but that both compounds are readily absorbed, rapidly metabolized and excreted in the form of several metabolites. Very little of either parent compound is excreted prior to metabolism. These compounds do not appear to be accumulated to a significant extent in any tissue, possibly due to the fact that they are rapidly metabolized. Within a few hours after administration most of the dose retained by the body was in the form of metabolites. These compounds were eliminated in both urine and feces, p-nitroaniline primarily in urine, and whole body half-lives for both compounds were less than 1 day. Additional studies of these and other aromatic amines will provide a better understanding of the importance of chemical structure to metabolism, disposition and persistence of aromatic amines.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Post-translational processing of proteins generates structural and functional diversity. Many examples of post-translational modifications have been described in the literature. Of those described, proteolytic processing of precursor proteins remains a dramatic case of post- translational modification. This processing generates a vast array of peptides, growth factors, and hormones which serve as important intercellular messengers. One particular pathway of proteolytic processing is that one that leads to the generation of alpha-carboxamidated peptides. Our efforts have been directed to developing chemical assays for the alpha- carboxamidated peptides and their immediate precursors, the glycine- extended forms of the peptide. Employing thiohydatoin derivatives of these peptides, two assays have been developed which detect aminoacyl-alpha- carboxamides and carboxy-terminal glycine. These assays have been employed in screening tissue extracts of neuroendocrine tissues, known to be rich sources of alpha-carboxamidated and glycine-extended peptides.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DMetrix is developing a line of innovative array-microscope slide scanner instruments to generate virtual slides. Our core technology is an array of miniature objectives, each measuring less than 2 mm in diameter and 7 mm in length, slightly staggered in the array, to produce a single, seamless sweep of a microscope slide. The array microscope concept relies on imaging in parallel in order to significantly reduce the time necessary to acquire digital, high-resolution microscope-slide images. We propose here to develop an epi-fluorescence configuration of the array-microscope concept. The proposed epi-fluorescence array-microscope system will include 100 miniature microscope objectives designed and optimized for fluorescence imaging by DMetrix. This array microscope will be based on a novel \"many-to-one\" optics configuration that allows efficient and simultaneous illumination of the 100 miniature objectives' fields of view. The long-term goal is to create a high-throughput, highly automated fluorescence microscope. The array-microscope virtual-slide scanner will allow digitization of fluorescently labeled specimens at rates at least 10 times faster than conventional single-optical-axis optical systems. Applications of such an epi-fluorescence virtual slide scanner are to be found in molecular pathology, pharmaceutical research, case archiving, and telepathology. The high scanning speed of the proposed instrument will enable either a high throughput measured in slides/day or the acquisition of multi-layer (\"z- stack\") and/or multi-spectral image sets in minutes per slide. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Methylmalonic acidemia is a rare metabolic disorder. This is a case study to evaluate nutritional needs during maternal adaptation to pregnancy, assess the effects of the pregnancy on the underlying disorder and the effect of methylmalonic acidemia on the fetus.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The major objective of our research is to identify recurrent chromosomal alterations and to isolate new genes that are relevant to neoplastic development and may serve as targets for therapy. Our successful utilization of the advanced molecular cytogenetic procedures for multicolor spectral karyotyping (SKY), comparative genomic hybridization (CGH) and high resolution fluorescence in situ hybridization (FISH) led to the identification of novel genetic changes and isolation of genes in solid tumors and hematological malignancies. Continuous progress has been made in molecular genetics and cytogenetics of liver cancer through the identification of new recurrent genomic alterations and isolation of new genes relevant to the initiation and progression of neoplasia and possibly useful markers for prognosis and diagnosis of the disease. In both human and mouse hepatocellular carcinoma (HCC), at several sites of recurrent genomic changes, we have identified new genes and detected alterations of known cancer-related genes. Several regions of DNA copy number gain were identified by comparative genomic hybridization (CGH) and two regions of high level DNA amplifications on chromosomes 1p32 and 11q13 were detected by representational difference analysis.Ten amplification fragments, showing a 5- to 50-fold increase in DNA copy-number were cloned and used to screen a human lambda genomic DNA library to obtain probes for FISH mapping of the amplified fragments. Several clones were localized to chromosome 1p32 and 11q13 at regions that are frequently amplified in several types of cancer including HCC. The sequence of one amplification product matched that of the EMS1/cortactin gene, which is located in the 11q13 amplicon, and encodes a protein that is a substrate of SRC kinase. Two amplifed fragments were isolated and mapped to chromosome 1p32, a region showing copy-number gains in HCC cell lines. Both fragments were present in partially and completely sequenced chromosome 1 BAC clones. The sequence of one fragment is located about 195 kb from r1f, a zinc finger protein gene that is co-amplified and fused with L-MYC in some lung cancer cell lines and it lies in the middle of a 23-kb interval between two novel genes. One gene encodes a new member of the zinc finger protein family, and the other gene encodes a protein of unknown function that contains a zinc binding domain homologous to those identified in ARF GAP (ADP-ribosylation factor GTPase-activating protein) family proteins. The ARF GAP-like gene appeared to be ubiquitously expressed, while the zinc finger protein gene transcript was undetectable in normal human tissues but was over-expressed in HCC cell lines and is a strong candidate proto-oncogene. The cytogenetic changes in HCC from transgenic mice closely evoked those that occur during the progression of spontaneously human carcinomas. Gain of chromosome 19 is a recurrent alteration in both mouse HCC primary tumors and derived cell lines, as well as in preneoplastic liver lesions induced with diethylnitrosamine. Chromosome 19 carries susceptibility genes for liver tumors and possibly a proto-oncogene involved in hepatocarcinogenesis. In addition to the gain of chromosome 19 in the majority of tumor lines, in one tumor line we identified double minute chromosomes (DM) derived from chromosome 19. DM, small pairs of acentric chromatin bodies and homogeneously stained regions (HSR) are cytological manifestations of DNA amplification. A probe generated by PCR from the microdissected DM was localized by FISH on normal chromosome 19 at two sites separated by seven or eight medium-size G-bands. At one site of DNA amplification (19131.1-131.3) the ms-MYC gene and mouse RelA gene recently characterized in the Laboratory of Experimental Carcinogenesis were localized, while the second site, 19 D1-D2 harbors the MXI1 gene. Both ms-MYC and MXI1 are known to interfere with the myc gene, while the Re1A gene may promote cell survival by inhibiting apoptosis, and allow c-MYC to drive proliferation of transformed cells. We are using gene targeting to generate DLC-1 knockout mice to establish an animal model for studying the biological functions of DLC-1, a candidate tumor suppressor gene that we have isolated. The mouse DLC-1 gene was isolated and used to construct a targeting vector that was successfully employed to disrupt the gene by homologous recombination in embryonic stem cells. Chimeric mice derived from ES cells carrying the disrupted gene have shown germline transmission of the mutant DLC-1 allele. The DLC-1 (+/-) mice will be bred to produce mice homozygous for the targeted gene. The effect of DLC-1 deficiency on viability, development, and reproduction will be analyzed. The susceptibility of the DLC-1 (+/-) and (-/-) mice to hepatocarcinogenesis can be studied using the liver cancer models developed in LEC. A new constitutive translocation involving a common fragile site was characterized by FISH and molecular markers. In a family in which multifocal clear cell renal carcinoma (RCC) segregates with a balanced constitutional chromosome translocation, t(2;3) (q33;q21), possibly with translocation positions similar to those of the renal cell cancer-associated t(2;3) (q35;q21) reported in another family. YAC and BAC contigs encompassing the 2q and 3q breakpoints were constructed and BACs crossing the breakpoints were partially sequenced. All known regional markers, genes and ESTs were mapped relative to the contigs, as well as to the breakpoint sequences. Two single ESTs map within the 2q breakpoint BAC, while the repeat rich 3q breakpoint region is gene poor. By FISH, we determined that the 3q break is in 3q13, possibly near the border with 3q21 and 2q breakpoint is closely telomeric to the 2q31 FRA2G site. Characterization of full length cDNAs for the ESTs near the 2q break will determine if a gene (s) is altered by this familial RCC-associated chromosome translocation. Deletion of the short arm of chromosome 5 was the sole structural alteration identified in a case of testicular lymphoma and it may harbor a tumor suppressor gene implicated in this malignancy. In the past year two studies with profound implications on the mechanisms of malignant transformation and drug action were completed. In a human colon carcinoma cell line rendered resistant to ecteinascidine, a potent anticancer agent, we identified a cytogenetic abnormality that affects the locus of the gene implicated in the hereditary disease xeroderma pigmentosum. As a result, it has been discovered that the killing mechanism of Et743 is mediated by nucleotide excision repair. We also demonstrated that the malignant transformation of normal human mammary cells required nonrandom chromosome changes involving the MYC gene, which is commonly altered in spontaneous breast cancer. Thus, it was demonstrated for the first time how a limited, definable number of genes and nonrandom chromosome changes results in the generation of a tumorigenic phenotype. Several genes isolated in our laboratory and by others were mapped by FISH and include the murine mitochondrial aldehyde dehydrogenase gene involved in ethanol metabolism, sca-1, an antigen commonly used for purification of murine pluripotent hematopoietic cells; and a member of the Ly-6 family and pancreatic phospholipase A2 that plays an important role in pancreatic and extrapancreatic tissue. In human 13 UDP glucunosylransferase genes encoded at the UGT1 complex were mapped on chromosome 2.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Marijuana is the most widely used illicit drug in the United States. Heavy marijuana users who attempt to stop using marijuana frequently report sleep disturbance, restlessness, nervousness/anxiety, increased aggression, and appetite changes. We believe that sleep disturbance may pose an enormous hindrance to successful cessation of heavy marijuana use. Surprisingly, despite several subjective reports of sleep disturbance in newly abstinent heavy marijuana users, there have been no studies using objective polysomnographic (PSG) measures validating these subjective reports. Therefore, this study is highly significant and very innovative because in addition to collecting subjective measures of withdrawal symptoms, we will obtain repeated objective measures of sleep architecture using standard PSG procedures. We will determine if there are objective polysomnographic findings of sleep disturbance in marijuana users (N=16) abstaining from drug use for 14 days in an inpatient setting at NIDA-IRP. Sixteen non-drug using controls, will be included for comparison and matched to the marijuana group on age, gender, and sleep-wake pattern. The control group will reside on our GCRC for 3 days. Baseline sleep-wake patterns will be estimated in both groups with actigraphy and sleep log recordings for 5 days prior to withdrawal/admission. The marijuana users will be transferred from NIDA-IRP to the GCRC core sleep lab for PSG recording. We will obtain PSG sleep measures of sleep onset, sleep maintenance, and restlessness at three separate times over 14 days of withdrawal and compare these to PSG measures obtained from the comparison group. Moreover, changes over time will be determined in the marijuana users. We will also determine if marijuana users have delayed sleep phase syndrome, a possible cause of sleep onset difficulty, by measuring dim light melatonin. The long-term goal of this research is to determine if heavy marijuana use is associated with objective sleep abnormalities as measured by polysomnographic procedures. Such findings could lead to new treatments for alleviating the unpleasant symptoms of marijuana withdrawal. For example, delayed sleep phase syndrome, one cause of insomnia, could be treated with light therapy, the same treatment used successfully for treating seasonal affective disorder. New treatments for alleviating the unpleasant symptoms of marijuana withdrawal would likely increase the number of heavy marijuana users who successfully complete treatment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is intense disagreement in the medical community regarding the value of population based PSA screening for asymptomatic men, and the efficacy of aggressive treatment of screen-detected disease. In the midst of this controversy, professional groups are increasingly advocating fully informing men of the pros and cons of screening and allowing them to make a personal decision regarding whether to obtain screening. Although informed decision-making is rapidly becoming the \"standard of care\" in this area, there is a paucity of information regarding how screening decisions are made in real world settings, and the best ways to promote an informed decision. This information gap is especially acute for ethnic minority men, who have not been systematically studied with respect to prostate cancer screening. Therefore, we propose to conduct a randomized clinical trial to evaluate the effect of an intervention designed to increase informed decision-making regarding prostate cancer screening in a community sample of Hispanic men. Hispanics constitute very large proportions of the Los Angeles and California populations, and prostate cancer is the most common form of cancer and the second leading cause of cancer death in this group. We will recruit 900 Hispanic men from multiple community-based organizations in the Los Angeles area, and administer an in-person baseline interview. Following this, subjects will be randomized to the two study arms. The Intervention Group (N=450) will be invited to attend a small group discussion session aimed at informing men about the nature of prostate cancer, pros and cons of screening, the controversy surrounding treatment of early stage disease, and the importance of making an active and informed decision regarding obtaining a PSA [prostate-specific antigen] test. They will also receive culturally tailored print materials summarizing the information presented in the intervention session. The Control Group (N=450) will be given a pre-existing, publicly available pamphlet, that mentions the availability of screening tests for prostate cancer but does not include details regarding the pros and cons of screening, or other elements of the intervention. Subjects in both groups will be re-contacted 12-months post baseline and administered a telephone interview to assess the impact of our intervention. The two main outcomes will include a composite measure of informed decision making (multidimensional construct including knowledge of prostate cancer and the pros and cons of screening, communication with physician or others regarding screening, and active participation in the screening decision) and receipt of PSA in the past 12 months.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective is to determine the three-dimensional structures of a set of protein neurotoxins that interact with sodium channels of excitable membranes. The structures will be determined using the techniques of protein crystallography. Major emphasis will be placed on structure-function studies of protein neurotoxins from the scorpion, Centruroides sculpturatus Ewing, including toxins that affect the inactivation of sodium channels as well as ones that alter activation kinetics. These studies will include high-resolution refinements of the variant-3 toxin; refinement of two crystalline forms of the variant-2 toxin; determination of the structure of Toxin V; and crystallographic studies of toxins III and IV. The results of these studies will be used to better understand the mechanisms by which scorpion neurotoxins affect sodium channels; the structural features that may account for the different effects that these purified toxins exert on activation and inactivation kinetics; structural features that may account for large differences in the toxicities of individual toxins; and structural factors that may eventually help account for differences in species specificity displayed by scorpion toxins. The crystallographic studies will be extended to include sea anemone neurotoxins, which are small proteins that compete for scorpion toxin binding sites. Comparisons of scorpion toxin and sea anemone toxin structures should be of value in identifying those structural features that may account for the neurotoxic activities of these proteins.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A herpesvirus, referred to as LHV-4, was found to be associated with a disease affecting over half of the 55 rabbits on a farm near Anchorage, Alaska. About 28% of the affected animals died or were euthanized because of the severity of the illness. Extracts from the skin of infected rabbits produced syncytia and cell lysis in cultured rabbit skin and kidney cells, and in Vero cells. PCR using degenerative primers to conserved herpesvirus genes was used to amplify sequences from purified viral DNA. Sequencing of these products allowed the design of specific primers so that complete sequence data for a number of genes could be determined. Analysis of this sequence data indicates that the virus is a member of the simplex genus of herpesviruses and is most closely related to bovine herpesvirus 2, HSV-1, HSV-2, and cercopithecine herpesvirus. Experimental exposure of New Zealand White rabbits to the isolate resulted in severe eye disease and necrosis of the spleen and lymph nodes. In addition, viral DNA was identified in a variety of tissues by PCR during the acute infection (within 5 days post- infection), but was only detectable consistently in trigeminal ganglia (TG) after 14 days post- infection. Experimental exposure of Swiss Webster mice to the isolate at 1X103 PFU/eye resulted in mild eye disease and fever. Viral DNA was detected in spleen tissue during the acute infection, and was detectable in the TG after 20 day post-infection. One of the unique features of alphaherpes virus is that they become latent in the TG with no gene expression, except for the latency associated transcript gene (LAT). The role of LAT during latency is still not well understood. The LAT gene may be the key to understand the mechanism of HSV-1 or HSV-2 latency reactivation. The purpose of the research outlined in this proposal is to investigate LAT gene of this novel herpesvirus. To accomplish this we will sequence and annotate the genomes of LHV-4 and characterize the LAT region. Because of its close relationship to the primate virus lineage, and its ability to infect rabbits, information on latency in this virus may provide a natural host model system for understanding latency in herpesvirus infections of humans. PUBLIC HEALTH RELEVANCE: In this research application we propose to sequence the genome of a recently discovered Simplex herpesvirus pathogenic for rabbits that is closely related to HSV-1 and 2. The goal of this research is to characterize the latency associated transcript gene (LAT) region and investigate the role of LAT during latency in a natural host model system. .", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pancreatic cancer (PC) is an aggressive malignancy with one of the worst outcomes among all cancers. This could be partly due to the ability of PC cells to orchestrate in \"turning-on the switch\" for migration, invasion, angiogenesis and metastatic processes during the early course of the disease. Therefore, there is a dire need for the development of novel strategies by which pancreatic tumor progression could be prevented. The constitutive activation of EGFR and Akt signaling, commonly seen in PC, is known to activate NF-:B, which transcriptionally regulates many genes contributing to aggressive tumor growth, angiogenesis and invasion, resulting in tumor progression. Moreover, recent studies have shown that the FoxM1 and Notch-1 signaling pathways are also activated in PC and appear to crosstalk with NF-:B (please see our preliminary results). However, how FoxM1 and Notch-1 crosstalk with NF-:B and regulate their downstream genes are not fully understood. Our preliminary data clearly suggest that the inactivation of FoxM1 and Notch-1 signaling causes down regulation of NF-:B, which contribute to the inhibition of cell growth, induction of apoptosis and inhibition of tumor cell invasion and angiogenesis. Based on our preliminary data and because of the lack of molecular understanding of the regulation and crosstalk between FoxM1, Notch-1 and NF-:B signaling, we hypothesize that further understanding of the molecular crosstalk between FoxM1, Notch- 1 and NF-:B, and their down-regulation by a novel agent could be an effective approach for designing better strategies for the prevention of pancreatic tumor progression. We will test our hypothesis by accomplishing the following specific aims. We will (i) determine how FoxM1 and Notch-1 crosstalk with NF-:B and regulates their downstream genes, and determine the consequence of down regulation of FoxM1/Notch- 1/NF-:B in PC cell growth, apoptosis, tumor cell invasion and angiogenesis. Next, (ii) we will test whether the down regulation of FoxM1/Notch-1 signaling by our novel approach (such as the use of genistein) could not only inhibit invasion and promote apoptotic cell death but also sensitize PC cells to an EGFR-tyrosine kinase inhibitor (erlotinib) and gemcitabine-induced killing. We will also test whether the chemo-sensitizing effect of genistein is mechanistically associated with the down regulation of FoxM1/Notch-1/NF-:B signaling. Finally, (iii) we will conduct in vivo experiments (using both orthotopic mouse model and transgenic mouse models of PC) to recapitulate our in vitro findings by testing (a) whether genistein-induced down regulation of FoxM1/Notch-1/NF-:B signaling could sensitize PC cells to erlotinib and gemcitabine induced killing, and (b) whether the inhibition of tumor progression could correlate with the down regulation of FoxM1/Notch-1/NF-:B signaling in animal tumor tissues. The results of our research will aid in designing a novel and targeted approach for the prevention of tumor progression, which would be highly relevant to public health in general and especially for saving lives of patients diagnosed with this deadly disease. PUBLIC HEALTH RELEVANCE: This project is focused on elucidating the mechanism by which a chemopreventive agent could prevent pancreatic tumor progression. Therefore, this grant is related to the prevention of gastrointestinal malignancy. We hypothesize that genistein down-regulates FoxM1 and Notch-1 signaling, which in turn down-regulate NF-:B and its downstream genes, resulting in the inhibition of tumor progression. We will test our hypothesis by three specific aims using molecular approaches in vitro and by using animal models in vivo (both orthotopic mouse model and transgenic mouse models of PC).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Integrating large-scale genomics data has huge potential to accelerate the identification of disease genes in human. Three major challenges lie in the current integrative approach for predicting disease genes. First, previous integrations in general limit genomic data input to one species at a time, while disease datasets are often generated in multiple model organisms. Second, public functional genomic datasets are dominated and biased by certain data types and accessible tissues, which can be addressed by expert curation of input datasets. Third, when multiple tissue-specific networks have been generated, a mathematical formulation is lacking to prioritize among these competing networks for the specific disease under consideration. This collaborative proposal aims at addressing the above challenges by exploring a prototype of bioinformatics tools to integrate multiple relevant global and tissue-specific networks across mammalian species targeting a specific disease, here ataxia. This proposal is based on our preliminary data in developing both global and cerebellum-specific networks to prioritize ataxia associated genes, and on the two PIs' complementary expertise in genomic data integration and experimental ataxia gene confirmation. We will 1) use domain-specific and multiple species data to establish global, brain, cerebellum, related tissue, and ataxia-specific networks, and develop web tools to explore these networks; and 2) develop multiple kernel learning algorithms to weigh and integrate multiple networks to predict ataxia-associated genes. Although the algorithms will be developed targeting ataxia only, we envision that this expert-driven integrative approach will be adaptable to other disease gene identification scenarios.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Myotonic dystrophy (MtD) is a dominantly-inherited multisystem degenerative disease with features of premature aging. Although the genetic basis of MtD is now known to be a (CTG)n repeat expansion in non-coding sequence of the myotonin protein kinase (MtPK) gene, the disease mechanism remains unclear even in its general outline. There is little experimental support for the initial hypothesis that a change in the amount or function of MtPK peptide explains the phenotype. Alternative hypotheses that the disease involves an effect on the expression of flanking genes, or a gain-of-function by (CTG)n repeats in MtPK mRNA, are the focus of this proposal. The extent (8-12 kb in most tissues) and character (strongest known natural nucleosome positioning element) of the CTG repeat expansion suggest its potential for altering chromatin structure and expression of other genes in the region. The recently discovered homeodomain gene DMAHP is a candidate for cis-acting suppression by the CTG expansion and for involvement in MtD pathogenesis because of its proximity to the repeat expansion and its sequence homology to transcription factors involved in myogenesis and connective tissue patterning. Studies are proposed to compare the levels of DMAHP mRNA in myoblasts from MtD patients, disease controls, and healthy subjects using allele-specific RT/PCR and ribonuclease protection assays. MtPK expression patterns and tissue pathology in MtD are strongly concordant. The clearest immediate consequence of the CTG expansion is to increase the amount of (CUG)n repeat-containing RNA in cells that express MtPK. Preliminary data from RNA folding algorithms, thermal melting, gel retardation, and gel mobility shift experiments are consistent with the idea that long (CUG)n RNA tracts form stable folded structures that bind to PKR, the dsRNA dependent protein kinase which plays a central role in translational control. Activation of PKR by (CUG)n repeat RNA tracts could explain the reduced muscle protein synthetic rate and myofiber atrophy in MtD. Structural characteristics and PKR binding/activation of (CUG)n repeats will be studied by gel retardation, nuclease mapping, gel mobility shift, and in vitro phosphorylation experiments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aim of this research is to elucidate the control of the differentiation of a multipotent stem cell, the interstitial cell (i-cell) of hydra. The approach is to alter the cell composition of intact animals and study the i-cell division and differentiation behavior. The cell composition can be altered by the selective deletion of cell types in intact animals and with a cell aggregation technique. This involves dissociating hydra into single cells and forming cell aggregates which develop into normal animals. Separation of cell types will permit construction of aggregates (hence animals) of selected cell composition. The pattern of i-cell behavior will be analyzed quantitatively as methods have been developed for quantitatively measuring all cell populations in hydra, as well as examining quantitatively the behavior of a cohort of isotopically-labeled cells with autoradiography. The objects of the proposal are to: (1) study the basis of position-dependent differentiation of interstitial cells; (2) study the role of nerve cells in interstitial cell division; (3) study the complexity of the interstitial cell system; (4) contrive to develop cell separation procedures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Pathologists have long recognized that a significant percentage of the mass of a tumor consists of non- transformed cells infiltrating the malignancy. Nowhere has the fascination with these responding host elements been greater than with B cell lymphomas. Mechanistically, the infiltrating cells have been implicated as mediating tumor promotion and survival, or alternatively resisting tumor progression through immune surveillance. Recent progress in profiling gene expression in certain lymphoma subtypes has revealed strong correlations between host immune signatures and clinical prognosis, providing compelling evidence that cancer-extrinsic factors can have a profound impact on cancer biology. While high overall T cell frequencies found within some tumors portend a favorable outcome, the relative abundance of regulatory T cells (Tregs) correlates with short remissions and decreased survival. Although several cell types are capable of suppressing tumor-specific immunity, a subset of CD4+CD25+foxp3+ T cells appears to be among the most potent inhibitors of immune effector function. There are several features of Treg biology that are especially relevant to lymphoma pathogenesis. Lymphomas have been shown to be rich in cytokines such as IL10, IL13, and TGF2, which promote Treg induction or amplification of their suppressive function. Some of the cytokines elaborated by Tregs may serve as survival factors for transformed B cells. Finally, as lymphoma cells constitutively express MHC class II antigens, they are capable of activating CD4+ Tregs directly, blocking the local execution of effector function of a systemically generated (e.g. vaccine induced) T cell response. The central focus of this proposal is to fully characterize the interactions between Tregs, T effector cells, and B cell lymphoma using animal models, with the objective of identifying cellular and molecular targets for therapeutic manipulation. Specifically, we will: 1. Use a mouse model of B cell malignancy to define the influence of Tregs on disease progression, and the features of the tumor microenvironment that support the induction and/or amplification of Tregs. 2. Define the role of direct MHC II antigen presentation by lymphoma cells in mediating CD4+ Treg dependent inhibition of anti-lymphoma effector responses at the site of tumor. 3. Examine the impact of drugs and antibodies targeting Treg depletion, inhibitors of Treg function, and compounds seeking to block Treg reinduction/recruitment once depletion has been achieved. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE: Lymphomas are cancers of the cells of the immune system. Normal immune cells can become alerted to lymphoma and attempt to eliminate it. However one class of cells (so-called regulatory T cells) can inhibit this. Understanding how these cells work will enable strategies to bypass this suppression. Lymphomas are cancers of the cells of the immune system. Normal immune cells can become alerted to lymphoma and attempt to eliminate it. However one class of cells (so-called regulatory T cells) can inhibit this. Understanding how these cells work will enable strategies to bypass this suppression. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Myoclonus refers to sudden, involuntary jerking of a muscle or group of muscles. Dystonia is defined as a syndrome of sustained muscle contractions, frequently causing twisting and repetitive movements, or abnormal postures. Dystonia is generally believed to be a disorder of the central nervous system. Inherited myoclonus- dystonia (M-D), previously referred to as hereditary essential myoclonus or hereditary (alcohol-responsive) myoclonic dystonia, is an autosomal dominant dystonia with incomplete penetrance. Zimprich and colleagues first identified loss-of-function mutations in a gene coding for ?-sarcoglycan (?-SG; gene name: SGCE in human and Sgce in mouse). SGCE is widely expressed in embryonic development and in adults. SGCE is expressed in almost all brain regions tested. We have established a Sgce knockout model of DYT11 dystonia. We found that the Sgce knockout mice showed myoclonus, motor deficits indicative of dystonia, anxiety, depression, and changes in the dopaminergic and serotonergic systems. Furthermore, we have developed a monoclonal antibody specific for ?-SG. While the knockout mice replicated most of the DYT11 symptoms, the function of ?-SG and the role of its mutated forms in causing M-D are largely unknown. Specifically, at molecular level, whether ?-SG forms a complex in the central synapse, if yes, the nature of this complex is not known. These unknowns hamper efforts to adequately understand the pathogenesis of M-D, thus preventing the development of effective therapeutic strategies for patients. Further detailed analysis of the ?-SG complex should provide a unique opportunity for clarifying the functional role of ?-SG and the pathophysiological roles of SGCE mutations. Detailed understanding of pathophysiology of DYT11 dystonia would also accelerate the drug discovery process to develop treatment for myoclonus-dystonia and related dystonia and myoclonus disorders. The broad, long-term objective of our research is to use transgenic mice to determine: 1) the functional role of ?-SG in vivo 2) how the loss of 5-SG protein leads to M-D. The objective of this application is to further characterize ?-SG protein complex using the Sgce knockout mice and monoclonal antibody that we have already made that will enable us to answer these questions. We hypothesize that ?-SG interacts with other proteins and functions at the synapse. Identification of these interacting proteins may elucidate the function of ?-SG in the synaptic transmission. We further hypothesize that ?-SG interacts with other proteins and loss of ?-SG leads to the reduction of the complex at the synapse. The reduction of the complex leads to altered synaptic transmission and plasticity at circuit level, and myoclonus, motor deficits indicative of dystonia, or both, at system level. The rationale for the proposed research is that once the roles of SGCE in causing dysfunction of movement control in the brain are determined, possible interventions to correct M-D can be developed. We are particularly well prepared to undertake the proposed research because we have created a line of Sgce mutant mice that mimic the DYT11 patients. Furthermore, we have developed other genetic and antibody tools that can critically test the above hypothesis. Our other strength is the multidisciplinary approaches we are able to use, which include molecular, genetic, anatomical, biochemical, neurophysiologic, and behavioral techniques. The work will be conducted in a research environment with many NIH-funded investigators and shared NIH-funded core resources that are focused on using animal models of neurological disorders. We plan to test our hypothesis with the following Specific Aims: Specific Aim 1: To identify the interacting proteins, we will prepare the synaptosomal fractions from WT and Sgce KO mouse brains. Immunoprecipitation of the ?-SG complex will be performed using specific monoclonal antibody against ?-SG followed by separation of these proteins by gel electrophoresis. The bands only appeared in the WT mouse brain samples should be ?-SG and the interacting proteins and the bands appears in the Sgce KO mouse brain samples should be false positive bands that non-specifically precipitated in the immunoprecipitation experiment. Protein bands will be isolated and sequenced by Mass Spectrometry. Specific Aim 2: To analyze the effect of loss of ?-SG on the expression levels of the interacting proteins, we will prepare synaptosomal fractions from WT and Sgce KO mouse brains and analyze the levels of the interacting proteins by Western blot with antibodies against the identified proteins. Specific Aim 3: To analyze the effect of loss of ?-SG on the synaptic transmission and plasticity, we will determine input output relationship, paired pulse ratio, and long-term potentiation of hippocampal CA1 Schaffer collateral pathway. The successful completion of the above Specific Aims will produce a list of candidate synaptic proteins that interact with ?-SG and the effects of loss of ?-SG on their expression levels and synaptic transmission. The results should help us to determine the function of ?-SG in vivo and how the mutation of Sgce causes M-D. The results should significantly increase our understanding of the pathophysiology of M-D, which can ultimately aid the development of therapeutic treatments for M-D patients and other dystonia and myoclonus patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "[unreadable] In response to PAR-04-122, Trudeau Institute submits a revised proposal to request matching funds to expand and improve its research core facilities. Trudeau Institute is a small, not-forprofit research institute focused on identifying mechanisms that regulate immunity to infectious disease, autoimmunity and cancer in young and aged individuals. The research aims of Trudeau investigators directly address the national goals of improving human health and increasing our ability to respond to emerging infectious diseases and bioterrorist threats. Trudeau Institute is part of a Regional Center of Excellence for Biodefense research and holds numerous PHS grants, totaling almost $7,000,000 in direct costs. Research at Trudeau Institute is dependent on state-of-the-art core facilities, including the Animal, Molecular Biology, Flow Cytometry and Animal Technical Services cores. However, these cores must be expanded and improved to meet the current and future research needs of the investigators. For example, the ABSL-2 and ABSL-3 facilities are 40 years old and require major updating and expansion. In addition, the Molecular Biology, Flow Cytometry and Animal Technical Services cores need to be consolidated and expanded. Therefore, we propose to construct a new building that will house mice requiring ABSL-2 (Aim 1), ABSL-3 (Aim 2) or quarantine level containment (Aim 3). The new building will also provide animal facility infrastructure and equipment needed to conduct infectious disease research (Aim 4). Finally, we propose to renovate the space currently used for housing infected animals in order to provide expanded, consolidated and contiguous space for the Molecular Biology, Flow Cytometry and the Animal Technical Services Cores (Aim 5). [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Public prejudice and discrimination toward mental illness have a profound effect on the lives of people with serious mental illnesses (SMI), including self-stigma, damaged self-esteem, social isolation, impaired vocational and social functioning, and reduced community participation. However, effective interventions have not yet been developed that reduce these negative effects. This project will evaluate the effects of a 10-week, peer-developed and peer-led group program, Combating Prejudice and Discrimination through Photovoice Empowerment, which blends Photovoice, a novel public health participatory action research methodology, with psychoeducation and teaching proactive coping strategies to empower individuals with serious mental illnesses to resist public prejudice and discrimination, and participate in the community as equal members. The Photovoice program has been manualized, and its feasibility and potential benefits have been established in a pilot randomized controlled trial (n=82) conducted at the Center for Psychiatric Rehabilitation at Boston University, in which participants showed significantly greater reduction in self-stigma and increase in proactive coping with public prejudice and discrimination. The proposed project will extend previous research on the Photovoice program by evaluating it in the context of a large public mental health agency (Riverside Community Care, Inc.), and determining whether improvements in self-stigma and proactive coping also lead to improved community functioning and integration, and psychological well-being and growth. We will conduct a randomized controlled trail with a total of 192 individuals with SMI (50% schizophrenia-spectrum), comparing the Photovoice program to services as usual (SAU), enhanced by the provision of a peer-led educational session about understanding and confronting public prejudice and discrimination. Assessments will be conducted at baseline, post-treatment, 3- and 6-month follow-ups. The primary hypotheses are that the Photovoice program is more effective than the SAU at reducing internalized stigma, enhancing proactive coping with public prejudice and discrimination, and increasing community functioning and integration. A secondary hypothesis will examine the impact of the Photovoice program on psychological well-being, personal growth and recovery. The Photovoice program is the first peer-led program for people with SMI that specifically targets empowerment to resist and confront public prejudice and discrimination in social situations. The proposed research will shed light on the effectiveness of an innovative, peer-led program that has already shown promise for addressing the pressing problem of prejudice and discrimination toward people with SMI.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project utilizes an adoption design to examine the interplay between genetic, prenatal, and postnatal environmental influences on early pathways to conduct, anxiety, and depressive problems by (a) conducting a psychiatric assessment of adopted children between 6.0 and 7.5 years of age and (b) conducting a psychiatric assessment of adoptive parents. Psychiatric assessments of the birth parents have already been conducted and these data will be utilized in the proposed analyses. In addition, DNA is being collected on all participants and will be available for analysis in the present study. The present study builds on a longitudinal prospective adoption study conducted during early childhood: two cohorts of adopted children and their birth and adoptive parents were recruited and assessed across 3 - 6 waves from infancy through early childhood (Early Growth and Development Study; EGDS-Cohort I, R01 HD42608 and EGDS-Cohort II, R01 DA020585). Combined with the rich existing data, the proposed data will allow us to better characterize early emerging emotional and behavioral symptoms in children that have been shown to predict later psychiatric problems. Because we will have DNA samples and data from birth and adoptive parents, we will be able to disentangle how prenatal, genetic, and rearing environment factors are related to early emerging emotional and behavioral symptoms, as well as to later psychiatric symptoms. To our knowledge, no other existing dataset or resource exists that can disentangle and examine the interplay among genetic, prenatal, and postnatal environmental influences on the development of symptoms in young children. The proposed study will allow us to achieve the following specific aims: SA1: Identify early emerging behaviors and emotions in children that predict onset of psychiatric symptoms in young children; SA2: Examine prenatal exposure to substances and prenatal stress in exacerbating genetic risk for psychiatric symptoms; SA3: Consider the mediating and moderating role of the rearing environment on prenatal and genetic risk factors on psychiatric symptoms in childhood; SA4: Assess how adoptive parents' stress and their own psychiatric symptoms impact the rearing environment and mediate and/or moderate genetic and prenatal influences.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The purpose of this study is to determine whether IL-2 activated donor natural killer cells combined with fludarabine and cyclophosphamide is a possible treatment for ovarian cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The general purposes of this research are to understand the physical and chemical bases of the properties and function of proteins, with particular reference to the fibrous protein, collagen. The aims of this program are to elucidate the way in which the crosslinks of collagen participate in its biologic role of imparting form, strength and support to the tissues. The experimental approach includes labelling of the crosslinks and their precursors in collagen by specific chemical means followed by their isolation and the elucidation of their structure. The location of these compounds is then established by isolating portions of the protein which contain them. As the structural information becomes well delineated, its relationship to the biology of maturation, to phylogenetic variations, and possible alterations in the hereditary and acquired diseases of connective tissue will be explored. By this approach, a correlation of the biologic functions and chemistry of collagen should be possible, providing insight into the activities and properties of an important natural polymer. BIBLIOGRAPHIC REFERENCES: Fujii, K., M. L. Tanzer, B. Nusgens and C. Lapiere. 1976. Aldehyde content and crosslinking of type III collagen. Biochem. Biophys. Res. Commun. 69:128-134. Fujii, K. and M. L. Tanzer. 1976. Osteogenesis Imperfecta: Altered crosslinking of bone collagen. Clin. Orthoped. Rel. Res., in press.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have recently developed a unique stable isotope tracer method for studying the absorption, metabolism, and biokinetics of physiological oral doses of beta-carotene and its primary metabolites directly in human subjects. The approach is based on use of per-labeled 13C-beta-carotene and a new generation of high precision gas chromatography-combustion-high precision gas isotope ratio mass spectrometer. This technique typically achieves 24 orders of magnitude more sensitivity and precision than stable isotope techniques relying on lower isotopic enrichments and conventional quadrapole mass spectrometry. 13C-beta-carotene, greater than 8% 13C, has been biosynthesized in significant quantity and purified by crystallization and HPLC. In preliminary studies with a doses as low as 0.5 mg, we have followed 13C-beta-carotene absorption, intestinal conversion to 13C-retinyl esters and 13C-retinol, and the subsequent plasma kinetics of all three 13C-analytes in the absence of changes in total analyte concentration in plasma. At present we are the only laboratory capable of performing such studies in humans at non-perturbing doses resembling those of typical daily intake. We now propose to use this technique to examine several fundamental aspects of the metabolism of beta-carotene in humans. A series of single-dose biokinetic studies in healthy adults are proposed to (a) fully describe and model the fundamental phenomena of beta-carotene absorption, metabolism and distribution; (b) determine the degree of within- and between-person variability in these parameters; (c) determine the effect of beta-carotene dose (1 mg v. 46 mg) on extent of absorption and intestinal metabolism; (d) determine the effect of prior dietary intake of beta-carotene (deprivation or supplementation) on the metabolism of a subsequent dose of 13C-beta-carotene; and (e) compare the absorption and metabolism of 9-cis-beta-carotene to that of the all-trans isomer. A model intestinal mixed lipid micelle system will be used to probe the effects of bile salts, fatty acid unsaturation, cholesterol, phytosterol, and alpha-tocopherol on the capacity of mixed micelles to incorporate beta-carotene. We will then determine if factors shown capable of altering micellar beta-carotene incorporation in vitro also alter 13C-beta-carotene absorption or metabolism in vivo in human subjects. Finally, a rat model will be used to initiate investigation of the effect of iron deficiency on beta-carotene metabolism, in light of intriguing preliminary data showing such an effect.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Osteoarthritis (OA) affects over 200 million people worldwide including 27 million Americans, and it is estimated that the US spends $128 billion dollars each year on OA related health care (1, 2). Current therapies for OA focus on decreasing pain while improving joint movement (3). As none of these treatments can restore joint structures damaged by the progression of OA, joint replacement is often required, with more than 600,000 joint replacement surgeries performed yearly in the US (4). As such, strategies aimed at ameliorating OA represent a highly significant opportunity to contain health care costs while enhancing the quality of life for many Americans. The role of bone morphogenetic proteins (BMPs), in the pathogenesis of OA remains controversial despite many years of study. BMP activity is necessary for initiating chondrogenesis during skeletal development and is required for formation of synovial joints (5-11). BMP signaling is also involved in the maintenance of functional synovial joints after birth (12). Paradoxically, too much BMP activity is also associated with OA (18-20). In this proposal, we focus on BMP2 and examine its role in maintaining postnatal synovial joint function. Our decision to target BMP2 is based on the phenotype of mice lacking BMP2 expression in mesenchymal cells of the early limb (Prx1cre; 21, 22). These mice lose normal skeletal function as they age, as evidenced by spontaneous fractures throughout the limb skeleton and an ability to initiate fracture repair (22-24) and the progressive development of OA of the knee, elbow, wrist and ankle by 28 weeks of age (Fig 1). The development of OA in BMP2Prx1Cre mice is analogous to disease progression in human OA where there is a long delay between the event that initiates the disease and the development of an arthritic joint. If loss of BMP2 does indeed precede the pathological joint changes seen in OA, maintaining BMP2 at normal physiological levels may be a means to slow or halt joint changes that characterize OA. However, Bmp2Prx1cre mice have a variety of developmental limb defects that may predispose synovial joints to develop OA (25, 26) and these confounding factors must be sorted out in order to determine the importance of changing levels of BMP2 signaling in articular cartilage in the pathogenesis of OA. We propose to directly assess the role of BMP2 in OA by tracking BMP2 expression and BMP signaling in adult synovial joints (Aim 1) and determining if loss of BMP2 specifically from articular cartilage leads to a progressive OA phenotype (Aim 2). To perform these experiments we combine the use of BMP2 reporter mice with BMP signaling reporter mice and mice that drive deletion of BMP2 in articular cartilage. We also employ two state-of-the-art technologies to assess OA progression: ultra high resolution micro CT that allows for submicron 3-dimensional examination of both mineralized and soft tissues (27, 28) and atomic force microscopy -based nanoindentation (AFM-N), a method that allows for the quantification of the local mechanical properties of irregularly shaped, small volume musculoskeletal tissues (29). These technologies allow us to obtain precise images and corresponding functional assessments of joint tissues in the transgenic mice we plan to study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "SUMMARY Sleep is pervasive, universal, irresistible, tightly regulated, and its loss impairs performance and cognition. Sleep is thought to perform essential restorative functions for the brain, and increasing evidence suggests that a key function may be to rebalance cellular changes triggered by plasticity during wake. This evidence is consistent with the hypothesis that sleep and wake may occur, be regulated, and perform their functions at the level of individual neurons. Recently, using multi-array recordings in freely moving rats, we have obtained direct evidence that sleep can occur locally within a group of cortical neurons, while the rest of the brain remains awake, and that such local sleep increases with the duration of wake. In this proposal, we will use multi-array recordings to establish whether OFF periods during wake (local sleep) occur in mice, whether it increases with wake duration and whether it is associated with slow/theta waves in the local EEG and impaired performance, providing the rationale for the use of high density EEG in humans in Project III. We will then test whether local sleep, like sleep proper, is regulated by intense synaptic plasticity (tiredness) or instead by mere activity (fatigue). To do so we will first establish if local sleep increases with locally induced intense plasticity. We will induce local tiredness in one sensorimotor cortex using both a learning task (reaching) and synaptic potentiation through electrical stimulation, and compare the effects on local sleep on the ipsi- vs. contralateral side. Next, we will determine the effects on local sleep of local opto-pharmacogenetic activation (intense activity with little plasticity), expected to lead to fatigue, during both wake and sleep, again comparing the results to the contralateral side. Finally, we will use SBF-SEM to test whether there are ultrastructural signatures that distinguish between neurons that have been kept awake and those that have slept, and between intense plasticity (tiredness) and intense activity (fatigue). Altogether, these studies in mice will complement those in flies in Project I, which use similar methods. Together, they will establish if sleep and wake are regulated homeostatically at the single neuron level, and if they leave ultrastructural signatures that reflect their consequences and functions for individual cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY Speech and swallowing impairments occur in 90% of individuals with Parkinson's Disease (PD) and aspiration pneumonia constitutes the leading cause of death in this patient population. While current pharmacological and surgical interventions are effective in alleviating general limb motor symptoms of PD, they have failed to provide significant benefit for cranial motor functions controlled by corticobulbar tracts such as speech and swallowing. This suggests that cranial motor and limb motor deficits are mediated by different underlying neural pathologies in PD, however the nature of these differences are not currently understood. The central goal of this proposal is to investigate differences in the neural mechanisms mediating corticobulbar versus corticospinal impairments in an animal model of PD. Using intracorticial microstimulation and a comprehensive behavioral testing battery consisting of both cranial motor and limb motor tasks, the proposed studies will (1) determine the differential effects of unilateral versus bilateral striatal dopamine depletion on cranial and limb motor function and corticobulbar and corticospinal circuits; (2) determine the differential responses of cranial and limb motor function in PD to targeted motor rehabilitation and dopamine replacement therapy; and (3) determine the differential effects of targeted motor rehabilitation and dopamine replacement therapy on the integrity of corticobulbar and corticospinal circuits. The results have the potential to guide the development of neurobiologically informed therapies that specifically target cranial motor impairment that can be translated to the human patient population. More effective treatment strategies of cranial motor dysfunction in PD will improve patient quality of life, reduce individual health care cost, aspiration pneumonia and ultimately morbidity in this disease population.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "c. Imaging and Tract Tracing Core - Co-Directors: Tim Ebner, Glenn Giesler and Paul Mermelstein Overview and Significance: Purpose of the Core: The Imaging and Tract Tracing Core is designed to allow researchers from diverse experimental backgrounds to take advantage of modern imaging techniques. The Imaging and Tract Tracing Core is intended to strengthen the research of the investigators by providing access to experimental approaches, equipment and skilled personnel that would not normally be readily available. The Core provides researchers with a diverse array of imaging techniques, allowing investigators to probe, at both cellular and systems levels, normal functioning as well as genetically altered changes in brain function. Importance of Core Services: There are three major Divisions in the Imaging and Tract Tracing Core: Confocal Imaging, Tract Tracing/Histology and Activity-Dependent Optical Imaging. The Confocal Imaging Division allows studies ranging from the imaging of structures in single layers of cells to the examination of deeper tissue, in fixed slices and in vivo. The Division provides investigators access and training in single and multiphoton microscopy. The Tract Tracing/Histology Division allows investigators to examine genetic changes in the nervous system at both the cellular and systems level using anterograde and retrograde tracing methods. The Activity-Dependent Optical Imaging Division provides the equipment and technical expertise for in vivo imaging of neuronal activity in mice. Optical imaging of activity-dependent signals includes flavoprotein autofluorescence, voltage sensitive and calcium dyes, pH imaging, and the hemodynamic intrinsic signal. Necessity of the Core: The visualization of nervous system structure and function is one of the most powerful approaches to understanding questions being raised in neuroscience today. Individual laboratories at the University of Minnesota have expertise in specific imaging techniques, but no laboratory has the capability to apply all of these methods in a cohesive manner. In addition, while there are several confocal microscopes on campus, access is limited due to heavy use and for the vast majority of investigators, training is unavailable. The readily available expertise and equipment of the Core allows NINDS investigators to test questions in an integrative fashion, providing greater depth to their findings and driving their research forward.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "AFDO Application for RFA-FD-16-042 - PROJECT SUMMARY The Association of Food and Drug Officials (AFDO) proposes to collaborate with national food safety training partners to establish a coordinated effort for expansion and development of the Integrated Food Safety System (IFSS) National Curriculum Standard (NCS), including continued development of manufactured food (including Animal Feed, Dairy Processing, Game, Meat and Poultry, Manufactured Food, and Shellfish) competency and curriculum frameworks, further development and validation of manufactured food competency statements, and updates to and development of manufactured food training courses to meet the needs of manufactured food regulatory agencies, which is required for a successful, national IFSS. The end goal of this proposed project is to significantly increase training opportunities for manufactured food regulatory agencies, leading to long term improvements in the knowledge, skills, and abilities of regulators as part of an IFSS. AFDO will recruit manufactured food subject matter experts (SMEs) to create a pool of qualified instructors who can assist in the development of the manufactured food curriculum, frameworks, competencies, and courses. AFDO will also assist with the development of train the trainer systems to assure that all SMEs who will deliver United States Food and Drug Administration (FDA) manufactured food courses will first complete instructor skills training (ISTs) and demonstrate their training capabilities through the satisfactory completion of course specific instructor training (CSIT) or an audit assessment conducted during the pilot testing of training courses or programs developed. Achieving all project objectives will require extensive cooperation and coordination between training partners. AFDO will build upon its existing broad network of resources, including its community of federal, state, local, territorial, and tribal entities built over more than 120 years of the association?s existence and experience. The outcomes of this project will be the advancement of a national IFSS through the establishment of a well-coordinated NCS, an increase in the number of quality manufactured food courses available to regulators, and an increase in the number and quality of trained and vetted SME trainers.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In breast cancer, tumorigenesis occurs through an accumulation of genetic and epigenetic changes in epithelial cells involving both the activation of oncogenes and the inactivation of tumor suppressor genes. The Syk protein-tyrosine kinase, known best for its roles in the immune system, has recently been identified as a tumor suppressor in breast cancer; its expression being inversely correlated with malignant cell growth and metastasis. This is an unusual property for a tyrosine kinase, enzymes more typically found as the products of oncogenes. Thus, an understanding of how Syk functions to suppress the malignant phenotype of breast cancer cells is of considerable importance to our understanding of the critical pathways involved in growth control in breast epithelial cells and to the identification of possible, novel therapeutic targets. Preliminary studies have led to the hypothesis that Syk regulates two important aspects of of epithelial cell function: 1) cellular responses to tumor necrosis factor (TNF), a factor that regulates cell death/cell survival decisions and 2) cell adhesion and motility. These hypotheses are based on the identification of Syk- interacting proteins that participate in each of these pathways. The research proposed in this project will 1) characterize the physical and functional interactions between Syk and components of the TNF-signaling pathway with an emphasis on the characterization of a novel Syk-interacting protein thought to participate in this pathway; 2) characterize the critical structural features and mechanisms by which Syk, through its interacting partners, regulates cell adhesion and motility; and 3) characterize through phosphoproteomics studies the substrates and binding partners important for Syk's regulatory functions in breast epithelial cells. Methodologies to be employed include 1) genetic, biochemical and microscopic evaluations of protein- protein interactions and the consequences of their disruption and 2) state-of-the-art analyses of the phosphoproteome of Syk-expressing cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In inflamed ainvay, changes in epithelial architecture, matrix, glands, and vessels promote obstruction and hypersecretion in chronic bronchitis and asthma. This project tests the hypothesis that peptidases control resolution of remodeling and other aspects of chronic ainway inflammafion. It uses novel approaches such as targeting peptidases in living cells with acfivity-based probes, probing immune cell interactions with real time imaging in mouse models of chronic infection, and exploring contribufions of genetic variation in mast cell peptidases to human asthma. Aim 1 is to determine roles of the ainway epithelial peptidase prostasin in chronic infection. Epithelial integrity and hydrafion is essenfial for defense against microbes and toxins. Failure of barrier funcfion leads to chronic infection and remodeling, as in cystic fibrosis. Aim 1 studies test the hypothesis that prostasin support of ainway ion flux and integrity is controlled by membrane anchoring, acfivafion, inhibition and shedding. Aim 2 is to determine roles of mast cell peptidases in resolving inflammafion. Mast cell products can Inflict harm, but mouse studies suggest they also promote survival from septic peritonitis and pneumonia. By helping to resolve infection, their overall effect can be anti-inflammatory. Aim 2 studies test the hypothesis that mast cell pepfidases promote resolufion of chronic inflammation. Aim 3 is to determine impact of mast cell tryptase deficiency on chronic ainway inflammation. Tryptases are implicated in airway remodeling in allergic and autoimmune inflammation and in defense against bacterial infection and thus have the potential to produce as well as to resolve inflammation. Our recent work reveals that dysfunctional human tryptases are common and that Individuals and populafions vary strikingly in number of active tryptase genes inherited. By exploring connecfions between genotype and asthma, a disease associated not only with chronic inflammafion but with exacerbation by infection, the proposed studies test the hypothesis that differences in tryptase genotype contribute to inherited variafion in asthma severity and suscepfibility. Overall, the proposed studies are expected to identify mechanisms that suggest previously unexplored strategies to prevent or reverse the pathology of chronic airway inflammation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Survival rates for metastatic breast and other cancers remain pitifully low and, barring a discovery to prevent cancer or cure it, the challenge of our decade is surely to find ways to prevent cancer cells from disseminating. Divested of this aspect, cancer ceases to be life-threatening particularly for the breast. Compelling evidence suggests that trauma or physiologically unfavorable 'hostile' environments promote metastasis. In this proposal we will use state of the art noninvasive magnetic resonance (MR) imaging (I) and spectroscopy (S) methods to test the hypothesis that a hostile physiological environment with low pH, oxygen and glucose promotes invasion and metastasis of human breast cancer cells. Studies will be performed on three human breast cancer cell lines (MDA-MB-435, MDA-MB-231 and MCF-7) and a hyperplastic human breast cell line (MCF-12A) preselected for differences in invasive and metastatic behavior. A specially designed invasion assay ('Metabolic Boyden Chamber' assay) adapted for noninvasive MR studies will be used to (i) probe metabolism during carefully controlled modulation of the physiological environment and (ii) simultaneously measure invasion or migration of cells through reconstituted basement membrane. A hostile environment will be induced within orthotopic solid tumors derived from the human breast cancer lines (MDA-MB-435, MCF-7) to determine if incidence of metastasis increases after physiological trauma and to identify vascular, metabolic and physiological changes associated with the trauma which play a role in this promotion. To stimulate a clinical situation, the stromal bed around the mammary fat pad will be irradiated before inoculation of breast cancer cells. The impact of tumor bed irradiation on the physiology/metabolism of the stromal tissue, the vascular characteristics of tumors growing in the pre-irradiated bed, and the increase in the incidence of metastasis will be determined. Cells and tissues typically respond to injury with inflammation which results in the secretion of eicosanoids which play a role in cell invasion and metastasis. The ability of anti-inflammatory agents to attenuate or eliminate the invasive metastatic behavior of breast cancer cells under normal and stressful conditions, and the effects of these agents on cell physiology and metabolism will be determined. The proposed studies will identify physiological and metabolic characteristics which promote invasion and metastasis and open avenues of treatment to prevent metastasis. Several existing forms of treatment induce hostile environments and it is imperative to determine if these treatments promote subsequent metastasis by promoting invasive behavior of residual cancer cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Mouse adenovirus type 1 (MAV-1) provides an ideal animal model system for studying the molecular basis of adenoviral pathogenesis, which cannot be pursued using the species-specific human adenoviruses. Genetic and biochemical approaches are being used to examine the viral and host factors involved in MAV-1 pathogenesis, which is characterized by acute and persistent infections. The first MAV-1 viral mutants have been isolated and are being used to investigate the contributions of specific viral genes to the infectious process in mice. There are three specific aims: 1. Characterize the host response to MAV-1 infection, particularly components of innate and adaptive immunity. The specific cell type(s) in the spleen which are infected by MAV-1 will be identified. Mice with deficiencies in immune system components will be tested for their susceptibility to MAV-1 infection. The effects of two key cytokines, tumor necrosis factor-( and interferons, on viral infections will be investigated. The sensitivity of MAV-1 infected cells to natural killer cell lysis will be determined. 2. Determine the contributions of MAV-1 early region 1A (E1A) and early region 3 (E3) gene products to viral pathogenesis. Various virus-host interactions identified for E1A and E3 of human adenoviruses implicate these regions in pathogenesis of acute and persistent infections, and experiments show that MAV-1 viruses with mutations in E1A or E3 have altered lethality for adult mice. Organ or tissue tropism of mutant viruses and expression of their viral gene products in mice will be compared to that of wild-type virus. The ability of wild-type and mutant viruses to produce persistent infections will be examined. Specific cellular proteins which interact with the E1A and E3 proteins will be identified. 3. Investigate the host genetics of susceptibility to MAV-1 infection. Mouse strains with different susceptibilities to wild-type MAV-1 infection have been identified. Additional inbred strains of mice will be tested for susceptibility to MAV-1. The strains chosen will enable determination of whether H-2 haplotype is a factor in susceptibility, and will form the basis for genetic studies to map genes for susceptibility to MAV-1. The combined approaches of characterizing the host response to wild-type MAV-1 infection and investigating virus-host interactions using both viral and host genetics will contribute to the understanding of mechanisms involved in infectious disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "One of the main obstacles inhibiting HIV eradication is a population of latently infected T cells. Although small, the latent population is extremely stable and resistant to current antiretroviral therapies. Upon withdrawal of antiretroviral therapy, HIV derived from the latent reservoir is able to rekindle infection and renew progression to acquired immunodeficiency syndrome (AIDS). Consequently, viral eradication will be dependent on therapeutic strategies to specifically target and clear the latent reservoir. However in order to develop these approaches, a greater understanding of factors that control viral latency is needed. Efforts in this regard have been hampered by the lack of a suitable in vitro primary T cell model of latency. The studies proposed herein will utilize a new in vitro model to characterize multiple aspects of HIV latency including defining viral and host factors that influence viral latency, and to identify agents that may prove useful in an \"activation/elimination\" strategy to purge latent reservoirs. To accomplish these goals, we propose the following Specific Aims: 1. To further characterize, optimize and adapt a primary cell in vitro model for HIV latency to study activation and elimination of latent virus; 2. To develop a high-throughput screening method to identify additional agents that activate or inhibit activation of latently infected primary cells; 3. To determine the role of viral accessory proteins in activation or formation of latent virus; 4. Determine the role T regulatory cells play in viral latency. These studies will define host and viral factors influencing the latent T cell reservoir, and develop strategies to eliminate this reservoir.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Tumor angiogenesis, vascularization, blood flow and oxygen consumption will ultimately determine the nutritional status of and the oxygen delivery to solid tumors essential for their growth. The oxygen concentration within clonogenic tumor cells exhibits both inter- and intra-tumor heterogeneity and is an important determinant for treatment response to radiotherapy and some chemotherapy. There is no standard procedure in clinical use today for measuring this tumor property. For practical reasons, a non-invasive assay which uses equipment available in most cancer centers would be preferred. Some bioreducible drugs may provide for that attractive option. Nitroaromatic compounds are enzymatically reduced within living cells to activated intermediates which become covalently bound to cellular molecules at rates inversely proportional to intracellular oxygen concentration. When labeled with appropriate isotopes, adducted marker can be detected by various imaging procedures. Nitroimidazole-based markers have been used to detect hypoxic cells within rodent and human tumors by autoradiography and immunofluorescence assays of tissue sections and by nuclear medicine imaging techniques. This research project has now synthesized and characterized novel, second-generation nuclear medicine markers with increased water solubility and improved marking potential. Even better hypoxic markers of the azomycin-nucleoside, azomycin-aromatic and azomycin-chelate classes can now be designed and synthesized. Optimal markers of each class will be validated in appropriate tumor models by independent measurements of their physiology (their P-31 NMR spectra), their oxygen levels by microelectrode and their intrinsic radioresistance. In addition, the sensitivity of optimal hypoxic markers for measuring oxygenation change in individual tumors will be determined. This research will define improved nuclear medicine hypoxic markers for human cancer investigations. The optimal marker(s) will be used to 1) determine the role of initial tumor oxygenation in predicting treatment response, 2) measure reoxygenation kinetics in human tumors during treatment and 3) identify subsets of radioresistant tumors for which hypoxia-targeted therapies are indicated.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Strenuous physical activity in most children with reversible obstructive airway disease results in exercise-induced bronchospasm. The possible triggers and mechanisms that produce this acute, reversible airway obstruction are not understood, and the management of EIB consists mostly of pharmacological intevention. The objectives of this research are to investigate the complex, interacting mechanisms causing EIB and to develop means of non-pharmacological management of EIB. This research proposal will investigate the effects that inspired air temperature, humidity, and hypoxia exert on the severity of EIB provoked by a standardized exercise test. The possibility that heat and/or water loss from the respiratory tract might be related to the mechanisms of EIB will be investigated. High altitude exposure appears to aggrevate EIB, and studies exploring the relationship between increased inspired hypoxia and worsened EIB severity will be examined. Routine encouragement of regular physical conditioning and participation in athletes is part of the present therapy for asthmatics, but only suggestive evidence exists for this recommendation. An evaluation will be made of a twelve-week interval training program and its effectiveness in reducing EIB severity. These studies hopefully will demonstrate the extent to which physical factors can modify EIB, clarify the components and operations in some of the mechanisms for EIB, and evaluate plus develop non-pharmacological methods such as physical conditioning for the prevention and therapy of EIB.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The research proposed is the first stage of a program focused on the development and critical evaluation of quantitative predictive models for gas and aerosol deposition efficiency in lung airways in humans and animals under a variety of respiratory modes and rates. The initial objective is to develop realistic physical models of the tracheobronchial airways of humans and experimental animals. The second objective is to use the models to examine the intrabronchial dose distribution of both gases and particles. Such measurements will provide basic experimental data on pollutant mass transfer. The complexity of flow profiles in both human and animal airways precludes useful predictions of mass transfer based on current fluid dynamics theory. The physical models to be made will permit examination of particle deposition in small bronchial and bronchioles for the first time. The ultimate objective is to develop predictive models for deposition using both theory and experimentally derived coefficients, including models that account for the pronounced nonuniformity of epithelial depositions on the airways. We propose to make single path hollow airway casts of human and dog tracheobronchial airways, including a cast composed of all major branches, a cast composed of minor branches, and a cast containing approximately equal numbers of major and minor branches. Flow distributions will be measured prior to cast pruning, and each branch removed will be replaced with an equivalent flow resistor. The casts will be fabricated so as to permit repeated assembly for experimental deposition measurements. Aerosol deposition in the small airways will be examined for the distinctly nonuniform deposition patterns that have been previously observed in the major human airways and, more recently, at alveolar duct bifurcations in rodents. The utility of the casts for quantitative detailed gas dosimetry will be demonstrated using NO as a model air pollutant and triethanolamine (TEA) as an absorbing surface. Overall deposition efficiencies at each airway segment and the pattern of deposition within airways will be defined for various constant and cyclic flow rates. Comparative measurements in these realistic flow systems with the NO2-TEA system and with nondiffusing, noninertial particles will provide basic flow profile information needed for predictive dose estimation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Characterizing the relationship between the structure of the human brain and its function is one of the most important goals in neuroscience today. Medical imaging has been used to gain significant new insights into this relationship through the use of both anatomical and physiological imaging methods. Despite significant recent advances, the current methodology is still limited by the lack of automatic methods for the detailed segmentation, geometric analysis, and labeling of the cerebral cortex. The major goals of the proposed research are to develop fully automated methods to find and mathematically represent the cerebral cortex in volumetric magnetic resonance (MR) images and to automatically identify and label the major sulci and gyri on the cortex using a detailed statistical analysis of cortical geometry. Specifically, we propose to 1) develop and validate methods to find and mathematically represent the cerebral cortex from volumetric MR images; 2) develop and validate methods to calculate regional measures of cortical shape and volume; 3) develop and validate automatic labelling of sulci and gryi; and 4) conduct studies of cortical variability and volume changes in normal aging. All methods will be extensively validated using both computer phantoms and manual in vivo truth models. The methods we develop to automatically represent and label the cortex in large numbers of subjects should also be useful in 1) the development of a description of normal versus diseased cortical geometry, 2) automatic landmark generation for deformable atlas registration, 3) statistical correlation studies of structure/function relationships, and 4) the analysis of morphological changes in ontogenesis, phylogenesis, aging, and disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Short interfering RNAs (siRNAs) are double stranded RNAs of ~19-29 nucleotides designed to suppress the expression of homologous genes by a process known as RNA interference (RNAi). In this study we have characterized several short interfering RNAs to reduce (knockdown) the expression of genes related to the dopaminergic system of the ventral mesencephalon. We report here effective suppression of all targeted genes, tyrosine hydroxylase (TH), the transcription factor Nr4a2 (Nurr1) and the GDNF receptor moiety cRet, by co-transfection of plasmids expressing gene specific siRNAs under control of the human U6 promoter with a reporter plasmid coding for firefly luciferase and fused in tandem to the cDNA for the gene of interest. The effective suppression of Nr4a2, cRet and tyrosine hydroxylase suggest that the U6 expression cassette could be used to deliver siRNA to mammalian cells in vivo and actively suppress the expression of dopamine related mammalian genes. The characterization of highly effective siRNAs for DA phenotypic ma", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The HIV-1 epidemic has resulted in ~2.7 million new infections in 2007 for a total of ~33 million people living with HIV/AIDS. Clinical trials have shown that HIV-1 infection cannot be prevented by immunization with monomeric recombinant forms of viral envelope (Env) proteins. However, it is clear that the HIV-1 Env contains epitopes that can induce neutralizing antibodies and that such antibodies can protect primates from infection. The HIV-1 Env is a transmembrane glycoprotein. Both the external subunit (gp120) and the membrane- proximal external region of Env (located within the gp41 subunit) contain epitopes that are the target of broadly neutralizing monoclonal antibodies (mAbs) isolated from infected patients. Considerable effort has been devoted to creating soluble forms of the Env trimer. The improvements in immunogenicity of these molecules relative to monomeric gp120 are limited at best. Another approach to creating improved Env-based immunogens is to produce virus-like particles (VLP). VLPs are multivalent and often very immunogenic. The full-length HIV-1 Env protein can be presented on the surface of VLPs composed of Gag protein and cellular membrane components. These VLP structures have the potential to represent true mimics of the Env trimer spike. Several challenges must be overcome to create HIV-1 vaccines based on VLPs. They must be produced at high levels. The number of Env molecules on each VLP must be maximized. Processing of the gp160 Env polypeptide must take place, without dissociation of the gp120 subunit, to create a functional form of the Env trimer. It might also be necessary to minimize the immunogenicity of the variable sequences. In preliminary studies on the creation of VLPs carrying HIV-1 Env proteins, we have begun to address the challenges outlined above. The preliminary results are encouraging and provide a basis for more detailed work on preparing VLP-based immunogens as candidates for HIV-1 vaccines. The objective is to create VLPs that carry the Env protein in a form that most resembles the current vision of the functional Env trimer of the virus. The Specific Aims of this Phase I SBIR proposal are as follows: (1) prepare optimized Env constructs for VLP production;(2) prepare VLP constructs with different Env sequences;and (3) evaluate the ability of various VLPs to induce neutralizing antibodies in rabbits If the VLP-based Env immunogens prove to be superior to soluble gp120 or gp140 constructs in their ability to induce neutralizing antibodies, then additional studies will form the basis of a Phase II SBIR proposal. PUBLIC HEALTH RELEVANCE: The HIV/AIDS epidemic has resulted in 2 million deaths and 2.7 million new infections in 2007, for a total of nearly 33 million people living with HIV/AIDS. Development of a vaccine is considered to be an essential component of the public health measures needed to slow the epidemic. This research proposal is designed to create a vaccine that can induce antibodies capable of preventing infection by the HIV virus.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Fragile Families and Child Wellbeing Study is a longitudinal birth cohort study of approximately 3700 unmarried parents and 1200 married parents and their children. We seek funding to: 1) perform assessments at age 9 of children's health, language ability, academic achievement, behavior problems and social relationships; 2) conduct interviews with children's biological parents, to update information on family resources and parental relationships; 3) collect saliva samples for genotyping from primary caregivers and children; 4) Interview social parents who live with the child, and; 5) collect information from children's teachers on child outcomes and school characteristics. These data will allow researchers to answer the following questions: How do children develop over time, and how do family resources influence children's health and development? How do the resources of unmarried parents evolve over time, relative to those of married parents? How do children's genetic endowments interact with their environments to influence their outcomes? What role do social (non- biological) parents play in the lives of children? How do school environments influence children's social and academic outcomes? The study includes a large number of children and adults who are: poor, members of racial and ethnic minority groups, immigrants and disproportionately affected by welfare, child support, child care, and incarceration policies. The children have diverse and often unstable family structures and child care arrangements. Thus, the sample is well-suited to study how multiple sources of disadvantage influence child health and development. The data are also unique in containing detailed information on mothers, fathers, and their relationships with each other. The information on unmarried fathers, in particular, allows us to follow a group of men that is often under-sampled in national surveys. [unreadable] [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this research proposal is to dissect the molecular events that allow the catalytic A1 polypeptide of cholera toxin (CT) to enter the cytosol of intoxicated eukaryotic cells. CTA1 ADP-ribosylates and irreversibly activates the stimulatory alpha subunit of the heterotrimeric G protein at the cytoplasmic face of the eukaryotic plasma membrane. The resulting downstream signaling events induce the life-threatening watery diarrhea seen in over one million cases of cholera since 1995. After it is secreted into the extracellular milieu by Vibrio cholerae, CT binds to the plasma membrane of eukaryotic cells and is transported in retrograde fashion to the endoplasmic reticulum (ER). CTA1 dissociates from holotoxin in the ER and then crosses the ER membrane to enter the cytosol. It is hypothesized that the ER-associated degradation (ERAD) pathway facilitates CTA1 translocation from the ER to the cytosol. ERAD is a quality control mechanism that recognizes misfolded proteins in the ER and exports them to the cytosol for ubiquitination and proteosomal degradation. A hydrophobic region in CTA1 is thought to trigger ERAD activity and stimulate CTA1 translocation to the cytosol; degradation in the cytosol is presumably avoided because CTA1 has a paucity of the lysine residues that serve as ubiquitin attachment sites. This project will test and elaborate upon the ERAD/CTA1 translocation model. Previous methods to measure CTA1 translocation were based on the downstream cytosolic effects of CTA1 activity, but this work will instead utilize a recently developed biochemical assay that directly monitors the CTA1 translocation event. The assay will be used to (i) identify the structural features of CTA1 that are required for translocation to, and persistence in, the cytosol; (ii) identify the putative ERAD factors that interact with CTA1 during the translocation process; (iii) delineate the physiological parameters required for the ER-to-cytosol transfer of CTA1, and (iv) establish a yeast-based system to study CTA1 translocation. These research activities will meet the short-term goal of establishing a productive research lab with the capacity to train undergraduate, graduate, and post-doctoral personnel. The proposed work will contribute significantly to the understanding of cholera pathogenesis, will generate important observations relating to ERAD function, and will produce information relevant to other toxins that follow the CT trafficking and translocation itinerary.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Brain development requires the orderly generation of numerous distinct types of neurons, establishment of complex cell-cell connections, and remodeling of neuronal processes in response to experience. In order to investigate the molecular mechanisms underlying brain development and plasticity, I would like to study how a tiny larval brain of the Drosophila grows enormously and evolves into a much more sophisticated brain after large-scale remodeling of its neural circuits. Understanding how neuronal progenitors can give rise to distinct types of neurons in a functional brain is important for future cell replacement therapies in human brains. Identification of molecules involved in remodeling of neuronal processes may further elucidate how to manipulate neural circuits in our brains. I propose to study the post-embryonic development of the Drosophila central nervous system from three distinct angles. In the first aim, I will conduct extensive genetic screens to identify genes required for development of the mushroom bodies that form the insect learning and memory center. Mushroom body neurons that are homozygous for a random mutation will be created and analyzed in otherwise wild-type organisms. This approach has led to identification of the ultraspiracle (usp), which encodes one subunit of the ecdysone receptor, as an essential gene for pruning of larval-specific processes during neuronal remodeling. In the second aim, I will directly test the hypothesis that the insect hormone ecdysone orchestrates various aspects of the post-embryonic neuronal development. Mosaic approaches will be used to examine roles of USP in both post-embryonic neurogenesis and morphological differentiation of adult-specific neurons. In the third aim, I would like to identify molecular markers that are specific for distinct types of neurons and their precursors during the post-embryonic development, which will greatly facilitate future molecular genetic studies in the complex nervous system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Pets are recognized as an important facet of the family system, yet research on adolescent?pet relationships has been limited in several key ways. Youth?pet relationship quality is a potentially critical factor in predicting outcomes of youth human-animal interaction but the nuances of how adolescent?pet relationship quality may influence the effects of pet ownership have not been fully explored. In particular, there is a significant need for longitudinal designs to unravel if and how quality of relationship remains stable, and how any change patterns are linked to youth outcomes. Additionally, human?animal interactions exist as part of complex family systems, but the majority of youth human-animal interaction (HAI) research focuses on assessing individual dyadic relationships. There is relatively little known about how parents may directly or indirectly socialize the pet into the family system as an intentional (or unintentional) way of creating a positive environment for their children. Currently, little data exist on how parents view adolescent?pet interactions and the potential benefits or risks as associated with these relationships. Exploring parental attitudes and behaviors is an important facet of understanding the processes involved in youth HAI. The specific aims of this project are 1) To longitudinally assess if youth?pet relationship quality is predictive of healthy adolescent behaviors, and 2) to assess how parents influence the ways in which pets are integrated into the family system. To achieve these aims, we will utilize data from a larger ongoing longitudinal survey (1R15HD094281-01) of students aged 11-15 (n=773) and their parents (n=228), and will recruit a subset of parents/guardians to participate in a follow-up exploratory interview study (n=30) about socialization processes focused on how family pets can contribute to the healthy development of early adolescents. The results from this proposed study will reduce gaps in empirical knowledge by focusing research attention on the quality and consistency of human-animal interactions between adolescents and their pets using a robust longitudinal approach. In addition, this study will provide nuanced mixed-methods data that will have significant implications for understanding the diverse ways in which parents have socialized the adolescent-pet relationship.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Varenicline for the treatment of cocaine dependence: A phase II clinical trial. Cocaine dependence remains a major public health problem, even after over two decades of steady attempts to develop effective behavioral and pharmacological interventions for treating cocaine dependence. Due to the health costs that come directly from cocaine use (e.g., coronary and vascular problems), and those that are more indirect (e.g., spread of AIDS, HCV from exchanging sex for drugs), it is imperative that efforts to develop efficacious treatments continue. To that end, this is a proposed double-blind placebo-controlled clinical trial (n = 200) of varenicline for the treatment of cocaine dependence that utilizes contingency management to promote treatment attendance. In a recently completed pilot trial (n = 37) using the same design, we found that subjects randomized to varenicline showed greater decreases in cocaine use as well as reductions in how valuable or reinforcing they found cocaine (Plebani et al., revision under review). In that study, we used a variable schedule of reinforcement to promote high rates of attendance. That reinforcement led to dramatic increases in attendance rates, and led to far less missing data than usually plagues such trials. Due to these promising results, we now seek to replicate that study in a larger sample in order to have the power to more fully determine the efficacy of varenicline as a treatment for cocaine dependence.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This exploratory study aims to develop and test a generalizable process for measuring the relationship between long-term work in a particular occupation and the onset of chronic disease later in life. A secondary aim is to test the utility o that process for assessing the relationship between long-term exposure to various job characteristics and activities (e.g., handling materials and objects) and subsequent chronic disease outcomes (e.g., arthritis). The proposed project is based on a conceptual model that transcends traditional notions of the distinction between work-related and nonwork-related illness. We hypothesize that aggregated long-term work experiences over several decades in particular occupations with characteristic job requirements can potentially affect the propensity for workers to develop specific chronic conditions later in life. This will be perhaps the first sudy in the U.S using long-term longitudinal data to estimate the relative risks of chronic disease by occupational category. The study will help answer questions about whether the development of common chronic health conditions among workers 45 years and older, such as arthritis, asthma, and diabetes, may be related to long-term exposure to work in particular occupations and job activities. The proposed analytical process has broad relevancy and can potentially be applied to numerous occupations, job activities/characteristics, and chronic conditions. The study has practical relevance for the establishment of workplace-based chronic disease prevention programs and chronic disease surveillance. The proposed study uses publicly available de-identified data from the National Longitudinal Survey of Youth-1979 (NLSY79) and the Occupational Information Network (O*NET), and applies occupational codes from the U.S. Census and the Standard Occupational Classifications (SOC) coding systems. O*NET contains ratings (on a scale of 0 to 100) of 277 descriptors characterizing each of 974 occupations. We will conduct two demonstration studies to test the efficacy and utility of using these O*NET codes and ratings to estimate the relative risk of subsequent chronic disease. Study #1 has two components: a) an analysis of the association between aggregated exposure to handling and moving materials (one of the 277 O*NET job descriptors) and diagnosis of arthritis later in life (one of nine chronic conditions outcomes assessed by NLSY79) and b) a subanalysis of the data limited to arthritis outcomes initially reported in surveys years 2000 to 2010. In Study #2a, we will analyze the association between aggregated years in an occupational classification and the risk for arthritis (and potentially any of the other eight chronic conditions assessed by NLSY79: diabetes, asthma, hypertension, heart disease, non-skin cancer, chronic lung disease, emotional and psychiatric disorders, and general health limitations). Additionally, in Study #2b, we will assess arthritis outcomes in five broad occupational groupings compared to a referent group of white-collar occupations (e.g., clerical and sales). By extension, this same process could then potentially be extended to any of the other chronic disease outcomes. PUBLIC HEALTH RELEVANCE: The rising prevalence of chronic disease among people over 45 years old is perhaps the greatest health challenge facing America. Insufficient attention has been paid to investigating possible connections between long-term job history in specific occupations and the risk of eventual chronic disease. This study will greatly advance knowledge in this field, and help target appropriate interventions (e.g., workplace surveillance for chronic conditions) in high-risk occupations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The innate immune system is a first line defense mechanism that relies on pattern recognition receptors (PRRs) to detect foreign pathogens by recognizing pathogen associated molecular patterns (PAMPs). Toll-like receptors (TLRs) comprise one family of membrane-targeted PRRs that respond to a variety of PAMPs. One of these receptors, TLR4, recognizes lipopolysaccharide (LPS) present on cell walls of gram-negative bacteria. TLR4 engagement initiates immediate, but regulated, signaling cascades leading to activation of Nuclear factor KB (NF-KB) and Interferon Regulatory Factor (IRF) proteins and induction of inflammatory cytokines, as well as interferons. Attenuated immune functions, including deregulated or chronic inflammatory states, are often coincident with increasing age. Intriguingly, aged peritoneal macrophages are sensitized to inflammatory stimuli, including LPS. However, the molecular mediators that may contribute to age-related immune disorders remain ambiguous and largely undefined. Here, we propose to employ a systems-based approach, which will integrate a series of functional genomics analyses, to identify novel molecular factors that contribute to increased LPS sensitivity in aged peritoneal macrophages. Specifically, we will optimize and execute genome-wide RNAi screen using a macrophage cell line stably transfected with an NF-KB-luciferase reporter. The cell line will be treated LPS to induce TLR4-dependent pro-inflammatory cascades, and activation of the pathway will be monitored by assessing luciferase activity. Putative hits generated from high-throughput screening will be subsequently validated, and will represent a subset of the genome affecting pro- or anti- inflammatory states in macrophages. In order to determine if genes modulating TLR4 signaling are also alternatively regulated in aging, microarray analysis will be used to generate transcriptional profiles of young and aged peritoneal macrophages, both in the absence and presence of LPS stimulation. A subset of genes which are identified to both regulate TLR4 signaling and are also differentially expressed in aged macrophages will be further characterized at a mechanistic level. These studies will enable the translation of systems-level analyses towards mechanistic and physiological understanding of macrophage response to lipopolysaccharides and age-related chronic inflammation. PUBLIC HEALTH RELEVANCE: The studies proposed here will promote global insights into the molecular bases of innate immune response, and provide novel therapeutic strategies to address age-related immune and inflammatory diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This program is for predoctoral training of biological science PhD students for research careers in Cellular and Molecular Biology. This interdisciplinary program involves students and 40 faculty members from the Divisions of Biology, Chemistry, and Engineering. It is a continuation of a training program supported at Caltech for the past 31 years by NIH. Subjects of special emphasis within Cellular and Molecular Biology include genetics and genomics, regulation of gene expression, signal transduction, eukaryotic cell biology, synthetic biological circuits, biopolymers, and protein and cell structure. Interaction between the Divisions is evidenced by students who, although earning their PhD in one Division, carry out their thesis research mentored by a faculty member of another Division;by joint courses;by a less-formal interaction including research collaborations, and by interdisciplinary graduate programs in BioEngineering and in Biochemistry &Molecular Biophysics. The major components of the training activities are: 1) each student's individual research program, guided by faculty members and carried out within a group of other students and postdoctoral fellows having related interests;2) core graduate courses including courses in bioinformatics and writing;3) preparation for candidacy examinations;4) formal and informal an seminars and group meetings;5) a course in responsible conduct of research, and 6) a research seminar during which CMB students present their own research. Predoctoral trainees are admitted to graduate study in each option based on highly selective admissions criteria, especially high quantitative aptitude and strong motivation for research. Trainees will be selected from admitted students, and will be those who have a primary interest in research in Cellular and Molecular biology. Trainees are expected to pursue research careers that require training in Cellular and Molecular Biology;the superb record of our past trainees supports this expectation. Facilities are located in a complex of adjacent buildings. Multi-user facilities include cell sorting, biological imaging including cryoelectron microscopy, NMR and mass spectrometry, monoclonal antibody production, high throughput DMA sequencing, animal care and production of transgenic mice. RELEVANCE: Cellular and Molecular Biology will continue to underlie the major advances in understanding of human health and disease that can be expected in the next decades. Young researchers trained in this area will make substantial contributions to human welfare. We will help train the next generation of cell and molecular biologists, those who make fundamental, mechanistic insights using both classic and cutting edge methodology and technology, borrowing appropriately from a variety of scientific and engineering disciplines.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Sarcopenia is a major public health problem among the rapidly elderly population expanding elderly population in our society. Disabilities directly related to muscle weakness, and indirectly related to changes in body composition and metabolic dysfunctions, are causing a staggering toll in disability and health care costs. Osteopenia occurs almost simultaneously with sarcopenia in the elderly population and muscle weakness increases the risk for falls and, therefore, fractures. Although these issues have been separately addressed in several studies, an integrated investigational approach to better understand the pathogenesis of sarcopenia and other age-related metabolic abnormalities and to investigate the potential role of androgens have not been undertaken in a comprehensive manner. The program contains four independent research programs, each representing different research disciplines, and four separate cores supporting the four projects. The main focus on the project is to determine the effect of the replacement of testosterone in elderly men and DHEA in elderly men and women and to compare these effects with placebo treatment over a two-year period. Project 1, \"Effect of Androgen Replacement on Muscle Metabolism\" will specifically determine whether these interventions have a different effect on size and quality of muscle in terms of strength and metabolic functions. Project 2, \"Effect of Androgen Replacement on Bone Metabolism,\" will determine the effects of this intervention on bone mineral density and markers of bone turnover. Project 3, \"The Effect of Androgen Replacement on Carbohydrate Metabolism,\" will determine whether the age-associated decrease in circulating androgens contributes to the alterations in carbohydrate metabolism that are commonly observed in the elderly and on insulin action, insulin secretion, and glucose effectiveness. Project 4, \"Effect of Androgen Replacement on Fat Metabolism\" will determine whether changes in fat distribution that occur with aging could result from differences in regional fatty acid uptake and systemic fatty acid kinetics, and whether these determinants of fat distribution are altered by the interventions. The data emerging from these studies will be integrated to determine the intervention of sarcopenia with other metabolic change and hopefully will contribute to a better understanding of the relationship between sarcopenia, hormonal changes, and many associated metabolic dysfunctions of muscle, bone, carbohydrate and fat metabolism. This study will hopefully form the scientific basis for future trials of androgen replacement in the elderly.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Core C will provide all animals required in the 4 Projects. Lovelace Biomedical and Environmental Research Institute (LBERI) has outstanding animal facilities and unique experience in veterinary care. This is a critical premise for studies of physiological myocardial aging in small and large animal models. Additionally, this Core will be responsible for delivering 14C-labeled thymidine to wild-type and transgenic mice, and dogs. Importantly, animals will be maintained at LBERI until shipment to the investigators' laboratories.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "For evolution to occur, there must be modifications in developmental process. However, the relationship between development and evolution is only beginning to receive experimental investigation. The studies proposed here experimentally address the nature of changes in gene expression that underly dramatic shifts in timing and cell fate determination that accompany replacement of typical larval development by direct development in a sea urchin. The use of this system allows us to test mechanisms of change in development at the level of cell lineage determination and behavior at the level of gene activity. We are able to directly examine the mechanistic underpinnings of one of the key concepts in the evolution of development, that is heterochrony, or changes in relative timing of developmental events. The experimental system is a molecular and developmental comparison of a direct developing sea urchin versus its dosest typical developing species. We have already demonstrated that homologous cell lineages can be identified, and we have observed heterochronies at the cellular and molecular levels in these species. We also have begun tracing cell lineages and cloning cell lineage-specific expressed genes. The major questions to be studied include the following: Are cell fates modified by autonomous determination or by modified patterns of induction? What are the mechanisms for the observed heterochronies in the evolution of direct development? How are gene expression programs changed in modified cell lineages? How distant are typical and direct developing species, and what are their precise phylogenetic relationships? The study of the evolution of developmental processes adds a new and potentially powerful means of understanding the underlying processes of developing systems, because closely related organisms that differ in specific developmental processes provide us with natural variants in these processes. The details of process modification by which these variants have come to differ provides a way of probing basic developmental processes and their controls.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall goal of this proposal is to develop appropriate rank based tests for clustered data when the cluster size is potentially informative and apply the resulting methods for various marginal comparisons (e.g., average condition of teeth before and after treatment) using existing dental database resources, specifically as obtained from the Piedmont 65 + Dental Study and Iowa Fluoride Study. Informative cluster size arises when the number of units in a cluster is non-constant/random and in correlation with the outcome of interest. In the context of dental data, all teeth belonging to an individual will form a cluster. Since tooth loss (in adult) is correlated with two of the diseases we are planning to study, namely, periodontal disease and dental caries, we have potentially informative cluster sizes in the Piedmont data sets. It is a methodological challenge to adapt a classical rank test to such situations. As for example, the two sample Wilcoxon rank sum test has difficulty maintaining the correct size/significance level under informative clustering even if it is adjusted for cluster dependence through appropriate variance estimate. This proposal has a goal of developing proper classes of rank based tests (and related R estimators) and studying their statistical properties for three classical problems adapted to marginal inference under cluster dependence with informative cluster size. These are the so called one sample location problem (Aim 1), the regression problem (Aim 2) and the association problem (Aim 3). In each of these problems, we will obtain a class of test statistics using general score functions that maintain proper asymptotic size under the informative cluster size scenario. We will also study the properties of the related R estimates of marginal parameters. Multivariate extensions of the first two problems will also be considered (Aim 4). Another signification component of the proposed research will be to extend these procedures to handle missing data where the missingness mechanism can be modeled using observable covariates (Aim 5). Finally, when the cluster size is not informative, as in the case of Iowa Study which comprises of children only, we will be able to increase the power of our tests by incorporating cluster specific weights in the construction of our test statistics (Aim 6).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal is a continuation of our ongoing studies of lipid metabolism, focusing on the action of a lipid mediator: 1-0-alkyl-2- acetyl-glycerophosphocholine (platelet-activating factor, PAF). We have found, in corneal epithelium, that PAF activates the expression of the early genes c-fos and c-jun and then the expression of collagenase type 1, the metalloproteinase that degrades interstitial collagen, a major protein of the corneal stroma. A PAF antagonist blocks this effect. We will test two hypotheses: a) that PAF is involved in the mechanism of corneal remodeling and ulcer formation by acting as a lipid second messenger in the transcriptional activation of c-fos, c-jun and the collagenase l gene, and b) that lipoxygenase metabolites formed by the activation of phospholipase A2 have a modulatory effect on PAF. We propose to investigate the PAF signaling pathway and the role of lipoxygenase metabolites in the transcription of collagenase type 1. PAF antagonists and lipoxygenase inhibitors will be evaluated to determine their sites of action and to correlate their biochemical effects with the clinical evolution of corneal ulcers. Another goal of this proposal is to investigate the PAF receptors in the cornea in order to define the sites of action of PAF antagonists in the gene cascade. Powerful analytical procedures such as high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry will be used. Quantification of specific mRNAs will be accomplished by using Northern blots with storage phosphor-imaging. The results obtained will define the involvement of PAF and lipoxygenase metabolites in corneal remodeling and ulcer formation. The new mechanism of action of PAF also can be important in a number of tissue disorders in which normal control of the degradative activity of collagenase appears to be lost, e.a. rheumatoid arthritis, leading to pathological tissue destruction. The action of antagonists can be useful to define new therapeutic tools for control of collagenase type 1 activity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Familial Mediterranean Fever (FMF) is a rheumatic disease caused by a single autosomal recessive gene. Patients with this disorder experience acute attacks of fever, arthritis, abdominal pain, and/or pleurisy; some develop amyloidosis as a long-term complication. The biochemical lesion in FMF, as well as the chromosomal location of the FMF gene, is presently unknown. This project is designed to find the chromosomal location of the FMF gene, and ultimately the gene itself. We have identified 55 Israeli families in which more than one individual suffers from FMF. Blood samples have been obtained from 347 individuals from these families, and Epstein-Barr virus-transformed lymphoblastoid cell lives have been established for each individual. Using cell line DNA from a particularly informative subset of these families, initial mapping studies have been undertaken by probing Southern blots with highly polymorphic DNA markers. To date we have excluded approximately 10% of the human genome as a potential site for the FMF gene. Large areas of exclusion have been established on chromosomes 1 and 9; smaller exclusionary area have been established on chromosomes 4 and 22. More importantly, we have found a DNA marker which shows modest evidence for linkage to the FMF gene (LOD=1.3 for Theta=0.1). From the data obtained thus far, the odds in favor of linkage to this marker are 20:1; a 1000:1 ratio is required for proof of linkage. Additional studies are in progress to determine whether this marker is in fact linked to the FMF gene.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project 4 will focus on understanding the role that social determinants play in the link between obesity and cancer at the population level across the lifespan by developing a multi-cohort simulation model of otiosity and non-Hodgkin's lymphoma (NHL). Aim 1: Develop a multi-cohort system dynamics computer simulation model of obesity and NHL population level incidence, treatment toxicity and survival trends. This will extend the obesity modeling from childhood to adult populations, develop a population level model of NHL, and based on emerging individual level analysis from the VHA cancer registry database, integrate these two models over the life course. Aim 2: Analyze the resulting model to identify how social determinants influence obesity and NHL population level incidence and outcome trends. This analysis will identify dominant social determinants, delayed effects, and temporal relationships across the life course. Aim 3: Design guidelines along with their implementation strategies to identify the most effective way to reduce the impact of social determinants of NHL population level outcomes. This will generate different guidelines (e.g., policy, prevention, screening, treatment, and survivorship care) along with potential implementation strategies to determine the best combination of guideline-implementation strategy for reducing the burden of NHL. This project is transdisciplinary. It will complement existing and separately funded work to develop an innovative system dynamics model of childhood obesity by extending the model into adulthood across the lifespan and develop a NHL model that will be combined with the extended obesity model. Once established, additional cancers can be added to the model from this or other TRECs. This project is significant. It will not only provide rigorous conceptual models of how social determinants for obesity and NHL might interact over time, but also help identify key areas for future transdisciplinary research that have high potential for population level impact. Involvement of the expert panel in the modeling process will facilitate the development of transdisciplinary knowledge. The model will also be one of the first to link efforts from the NIH funded Cancer Intervention and Surveillance Modeling Network (CISNET) and the NIH/RWJF funded Comparative Modeling (CompMod) Network for obesity prevention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Toxicity and carcinogenicity of laboratory animals (mice) for Hexachloro-1, 3-butadiene.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The recent Agricultural Health Study among 25,814 farm women conducted by the researchers from NIEHS, NIOSH, and NCI reported a highly significant association between any use of pesticides in farms and atopic asthma in 282 subjects. This association was strongest among women who grew up on a farm. Growing up on farms and application of pesticides were jointly associated with higher overall risk for atopic asthma in contrary to the protective effect of farm environment (as expected based on the Hygiene Hypothesis);which was in the same study demonstrated for women who grew up on farms and never applied pesticides. Another recent study showed that the production of several cytokines induced by endotoxin was suppressed in pesticide treated mice. Exposures to toll like receptor (TLR) ligands in farm environment including MAMPs (microbe-associated molecular patterns) from microorganisms and microbial products have significant influence on the suppression of allergic response. This suppressive response was implicated to the induction of regulatory T cells (Treg) by recent revisions of the Hygiene Hypothesis. Therefore we propose to explore and determine how combined airway exposures to TLR ligands and pesticides influence immune responses associated with atopic asthma in farm environments. In the first specific aim we will characterize exposures of environmental TLR ligands (endotoxin, peptidoglycan, and bacterial DNA) and two conventional pesticides relevant to atopic asthma in 10 farms of Ohio before and after pesticide application. A state-of-the-art bioaerosol sampling methodology will be used. It includes an inhalable sampler and a NIOSH-developed size- selective sampling system with two-stage cyclone (three particle size fractions: <1.0 5m, 1 - 1.8 5m, and >1.8 5m), for the collection of airborne TLR ligands and pesticides and a novel microbial source strength tester for the collection of aerosolizable TLR ligands and pesticides from soil surface. We will determine the respiratory deposition potential of particulate TLR ligands and pesticides for male, female, and children subjects using the LUDEP ICRP respiratory deposition model. Results from the first specific aim will thus allow us to understand the real world simultaneous exposure levels to inhalable TLR ligands and pesticides in farms. In second specific aim we will determine how unpurified but clinically relevant air and surface sample extracts from above-mentioned farms analogous to real world exposures influence host susceptibility towards ovalbumin induced hypersensitivity and induction of Tregs. The TLR4 dependence of these influences will be determined by using ovalbumin-sensitized wild type and TLR4 -/- C57BL/6 mice. The findings will provide vital preliminary information on the relationship between adverse effects of pesticide exposure versus protective effects of TLR ligand exposure in the development of atopic asthma among agricultural workers. PUBLIC HEALTH RELEVANCE: Pesticide usage can be significantly associated with atopic asthma among the farmers and therefore farming environment may not provide the protective effect predicted based on the traditional Hygiene Hypothesis. Wide range of immunostimulatory materials and pesticides are present in farming environments and traditional investigations with purified materials and pesticides may not provide precise information on relationship between adverse effect of pesticide exposures versus protective effect of TLR ligands in developing atopic asthma among farmers. In this R21 project we will explore immunological activities of unpurified but clinically relevant environmental samples collected in farms (before and after pesticide application) in ovalbumin allergen sensitized mice.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term objective of this research is to develop integrated approaches to understanding drug use and abuse. Conceptually, this objective relies on (a) development of a biocultural anthropology approach to drug abuse, and (b) integration of the biocultural approach with more traditional theories of drug abuse, such as the biopsychosocial approach. Methodologically, this objective relies on (a) ethnographic research on ambivalence and drug use and abuse among adolescents in Bogota, Colombia, and (b) psychophysiological research testing a preliminary theoretical approach to drug abuse among these adolescents. This preliminary theory contains three aspects-wanting, frames, and self. Wanting refers to the implication of dopamine systems in drug abuse, frames to the role cognitive processes play, and self as an encompassing construct around wanting and frames. The psychophysiological research is the main emphasis of the present proposal, and relies on integrating physiological responses of the sympathetic nervous systems to verbal statements emphasizing \"wanting\" and \"self\" respects of drugs. Greater sympathetic response by drug abusers is predicted to wanting and self stimuli that emphasize drugs, as well as quicker response to a change (in frames) between these stimuli.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "While magnetic resonance imaging (MRI) is clinically very valuable, current imaging methods are subject to blurring and artifacts in the presence of physiologic (e.g., respiratory and cardiac) motion, as well as of arrhythmias, thus limiting the practical application of MRI in many patients. The currently used MRI methods are also limited in their ability to study the effects of free breathing and arrhythmias on the heart. The proposed research will further develop and evaluate a new approach to imaging in the presence of physiologic motion, which parameterizes such motions with a variable that is treated as an additional dimension to be reconstructed. This would not be practically feasible with conventional methods, due to the additional associated data acquisition that would be required. However, with the use of sparsity-based image reconstruction methods, the high degree of correlation of the images along these additional dimensions permits good quality image reconstructions, even with heavily undersampled imaging data. We already have made successful initial implementations of this new method for 2D cine imaging and 3D MR angiography. In the proposed research, we will further improve these initial implementations, and we will extend them to include implementations of our methods for other MRI sequences, particularly fully 3D cine data acquisitions. We will evaluate the image quality achievable with these new methods in the presence of free breathing and arrhythmias, as compared with conventional clinical imaging methods, using both numerical phantom simulations and clinical cardiac function analysis in pediatric patients to test the performance. We will also evaluate the potential for extracting new kinds of functional information from these multidimensional image sets, including the effects of free breathing and arrhythmias on the heart, using analysis tools that we will be developing. If this research is successful, these new methods will provide significantly improved MR image quality in the presence of free breathing and arrhythmias, as well as providing potentially valuable new kinds of information on the function of the heart. They may also be able to be used for performing MRI in the presence of exercise, which could be useful for both cardiovascular and musculoskeletal applications, as well as in combination with other kinds of imaging, such as with integrated PET/MRI systems. This research should thus further increase the clinical utility of MR imaging for many patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract Iron is an essential element for normal physiological functions. However, excess it can cause extensive tissue damage and participates in numerous ocular pathologies including cataractogenesis and retinal degenerations such as age-related macular degeneration. The study of ocular iron metabolism has been a focus of this laboratory for many years. We have made recent novel observations about iron's physiological role. We found that iron regulates synthesis and secretion of the neurotransmitter glutamate by ocular tissues and neurons. This is of fundamental clinical relevance since iron and glutamate are both dysregulated in neurodegeneration. In high quantities, glutamate can be excitotoxic in the central nervous system as well as the retina. Additionally, in retinal pigmented epithelial cells (RPE) and lens epithelial cells (LEC) iron regulates the activity of the transcription factor, hypoxia-inducible factor, which in turn regulates the synthesis of dozens of proteins. Our preliminary data indicate that hypoxic conditions stimulate glutamate release, another critically important observation since hypoxic conditions occur in stroke and retinal ischemia. Furthermore, there are profound changes in the structure of the iron storage protein ferritin in lenses that occur with age, cataractogenesis and differentiation. We will continue to explore how these changes affect iron storage in ferritin and the protection against iron damage such storage provides. Unfortunately, little is known about how iron levels are regulated in the eye which is isolated from the systemic circulation by blood ocular barriers (BOB). The proposal's hypothesis is that intraocular tissues have unique and independent systems for regulating iron uptake into and efflux from the eye across the BOBs. Their polarized location and iron-regulated quantity within ocular tissues allows for proper control of intraocular iron levels. Hypoxia, hemorrhage and inflammation significantly impact iron uptake storage, utilization and efflux. The resulting dysregulation of iron metabolism plays a critical role in ocular pathology. We will use an innovative integrated approach to determine how the BOB's regulate iron levels in intraocular tissues. The two specific aims utilize normal and pathological human eyes as well as normal canine eyes and tissue cultures of cells which form the BOBs, e.g., RPE and CE. Additionally, the lens will be used to assess how iron handling strategies adapt for survival in a normally hypoxic environment. We will utilize a state-of-the-art live-cell imaging quantitative fluorescence microscope with total internal reflection fluorescence for quantifying events at the plasma membrane and allow for measurement of dynamic processes underlying these complex interactions in four dimensions (4D) in living cells. It is the goal of this proposal to determine how intraocular iron levels are controlled and the specific role(s) iron has in ocular pathology in order to provide a basis for development of therapeutic modalities needed for prevention and treatment of ocular disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Factors affecting hard-of-hearing (HOH) listeners' use of formant transitions in speech will be investigated in three experiments. Listeners will be selected on the basis of: 1) hearing loss:(a) moderate-to-severe or (b) severe-to-profound, 2) audiometric configuration: (a) flat or (b) sloping, and 3) amount of auditory experience: (a) extensive or (b) limited, as defined by age of hearing loss onset, years of amplification use, and home/school mode of communication. The three experiments will use different approaches for the study of formant transition use by HOH listeners. These are: 1) investigation of listeners' use of formant transitions for glide and voiced stop consonant identification as a function of hearing loss and auditory experience. Spoken stimuli will be modified so that formant transitions are the only available acoustic cues for consonant identification. Transition audibility will be determined based on threshold measurements for each of the transition onset and offset frequencies. An audibility metric will be developed and applied to describe the effects of transition audibility on transition use. The relationships between use of formant transitions and the listeners' auditory experience will also be examined. 2) comparison of listeners' frequency/temporal discrimination for transition-like stimuli with transition use for consonant identification. The transition-like stimuli will simulate the transition characteristics found in the glide/stop identification stimuli. Relationships between transition discrimination and transition use will be described. 3) investigation of HOH listeners' perceptual weighting of formant transitions compared to other acoustic cues to the stop/glide contrast. Both synthesized and natural speech stimuli will be developed that vary along two acoustic continua (formant transition and amplitude onset). Analysis of the perceptual importance of the two acoustic cues, auditory experience, and degree of hearing loss will be completed. In all experiments, listeners with normal hearing thresholds will be used as controls.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (adapted from the Abstract): The Undersea and Hyperbaric Medical Society (UHMS) is a professional, non-profit scientific and medical association with a mandate to expand and disseminate the body of knowledge of diving and hyperbaric medicine. It was founded in 1967 and incorporated in 1972. It is a professional membership association of approximately 2800 physicians and researchers. Its purpose is to provide scientific and medical information to protect the health of sport, military and commercial divers and to improve hyperbaric oxygen research and treatment protocols. The Society also provides medical guidelines for physicians and scientists in the fields of diving and hyperbaric medicine. One of its most valuable resources is the Charles W. Shilling Library, which is the repository of the most complete collection of literature relating to these subjects. The long-term objective of this project is to provide wider access to the unique holdings of this special library. The specific aims are: 1) To organize and catalogue the collection for the on-line use of researchers and doctors working, worldwide, in the fields of Diving and Hyperbaric Medicine. 2) To update and systematize the bibliographic database in preparation for on-line accessibility. 3) And, ultimately, to make the entire library collection, including the database, available through the UHMS Internet Website. Reference work is done by the librarian who responds to phone, e-mail and fax requests. Researchers rarely come into the library in person. The librarian will catalogue the collection, including detailed subject indexing and assignment of classification numbers. She will work with a part/time data entry clerk who will assist her in preparing the database for on-line access. In order that the members of the UHMS are familiar with and understand what will be available on line, a training class conducted by the librarian will be offered at the next UHMS Annual Scientific Meeting. Additionally, full instructions for the use of the database will be available on line.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The primary thrust of the Specialized Population Research Center is to investigate mechanisms concerned with regulation of gonadal function. Most of the fundamental studies are directed towards questions of relevance to work dealing with the development of contraceptives. Applied studies deal directly with investigations of the feasibility of using hormonal preparations for male contraception and with problems related to diagnosis and treatment of gonadal disorders. To approach the question of contraception in a scientifically valid and clinically safe way a body of fundamental knowledge concerned specifically with mechanisms involved in regulation by the gonadal steroids of gonadotropin synthesis and release, with the role of gonadotropins and steroids in gonadal function and with the biochemical mechanisms of gonadotropins, androgen, and estrogen action needs to be accumulated. The studies proposed in this application will be directed towards acquisition of this knowledge and elucidation of these mechanisms. In addition, studies dealing with the kinetics of population and the role of androgens in it from physiologic and behavioral viewpoint will be conducted. These studies should place our entire research effort in a broader perspective through an inquiry into the models of natural mechanisms of population control operating in lower species.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary P-glycoprotein (P-gp) is an ATP-dependent efflux transporter that plays a critical role in drug and xenobiotic distribution, drug-drug interactions, and drug-nutrient interactions. Efforts to modulate P-gp activity to control cellular drug resistance or to modulate the action of existing drugs have been only modestly successful. A barrier to progress in the design of P-gp inhibitors has been the uncertainty about its catalytic mechanism. Different types of drugs or ligands elicit different behaviors, wherein some stimulate ATP hydrolysis and are transported, while others stimulate ATP hydrolysis but are not transported. Other drugs inhibit P-gp without stimulating ATP hydrolysis. The mechanism by which different drugs elicit different behaviors is unclear. Specifically, the conformational changes that mediate communication between the nucleotide binding domains (NBDs) that hydrolyze ATP and the transmembrane helices (TMHs) that bind and release xenobiotics remain unknown. One aim of this proposal is to map by H/D exchange mass spectrometry the ligand-dependent conformational changes in the NBDs and the TMHs. This will be performed with Pgp incorporated into lipid bilayer nanodiscs of defined lipid composition. By monitoring the nucleotide-dependent and drug-dependent changes in solvent exposure and dynamics of specific peptides in the sequence of each protein, with inhibitors, substrates, uncouplers and allosteric modulators, the conformational changes that correlate with each behavior will be identified. A second aim of these studies is to measure the on rates and off rates of drug binding to and dissociating from P-gp in varying conformational states. There are currently no data concerning these rates, which are likely to define ligand behavior, as a substrate vs. inhibitor vs. uncoupler. These measurements will be made via surface plasmon resonance and fluorescence correlation spectroscopy with P-gp nanodiscs. In order to correlate the conformational mapping and off rate information with physiologic behavior, cell based transport activity will be measured for representative drugs with different behaviors. Finally, this proposal aims to explore the methodological advancements offered by nanodiscs with an related drug transporter, BCRP. These studies will add methodological infrastructure to the larger transporter field, increase our fundamental understanding of P-gp, further inform pharmacokinetic models, and facilitate drug design aimed to modulate P-gp.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Learned associations between environmental contexts and experience are the basis of decision-making and allow organisms to guide behavior towards advantageous outcomes. Dysfunction in the neuronal processes that regulate these associations, especially in the nucleus accumbens (NAc), is a critical factor in the pathology of addiction. The NAc is a heterogeneous region primarily composed of two opposing cell types: D1 and D2 medium spiny projection neurons (MSNs). Optogenetic stimulation of these cells results in divergent behavioral outputs; thus, it is important to study these populations in isolation to understand the cell-type specific signals that underlie NAc-mediated learning processes. Under the primary mentorship of Dr. Eric Nestler and Dr. Paul Kenny at Icahn School of Medicine at Mount Sinai in New York, the Pathway to Independence Award will provide the opportunity to build on my expertise in cocaine self-administration and synaptic function while simultaneously developing my training and expertise in in vivo calcium imaging and optogenetics. In the mentored K-phase of this grant fiber photometry calcium imaging will be paired with cocaine self-administration in transgenic mouse lines that express Cre-recombinase in D1 or D2 MSN populations. These mice allow for cell-type specific expression of molecular targets, such as calcium indicators (GCaMP6f) and opsins (ChR2; NpHR). By expressing GCaMP6f in D1 or D2 MSNs, the temporally specific signals that mediate cue-induced cocaine seeking will be determined. Further, optogenetic stimulation and inhibition will allow for direct manipulation of these cells and the associated seeking behavior. The innovative combination of these tools will enable the mapping of how D1 and D2 MSNs encode cue information and concurrently establish causality. In the independent phase (R00), these cutting-edge techniques will be combined with the inducible ArcCreERT2 mice which express constructs (GCaMP6f/Opsins) selectively in cells that are activated by environmental stimuli during a temporally specific window. This will allow for the recording and manipulation of neuronal ensembles that are activated by cocaine or cocaine-paired cues to determine their role in drug seeking. Together, these data will elucidate the underlying neural processes that control associative learning and how cocaine exposure dysregulates MSN signaling to drive relapse following abstinence, which will expand our basic understanding of addiction and may lead to the development of novel therapeutic avenues. In summary, the research proposed in this Pathway to Independence Award will elucidate the neural mechanisms involved in addiction while simultaneously preparing me to develop a fully independent research program capable of integrating a wide range of circuit based and behavioral approaches to dissect the neurobiology of addiction. .", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Avian-human influenza A reassortant viruses that contain the human influenza hemagglutinin and neuraminidase genes and the six \"internal\" genes of the A/Mallard/78 (H2N2) avian virus were attenuated for monkeys and man. A similar reassortant derived from the avian A/Pintail/119/79 and the human A/Washington/80 (H3N2) viruses was also attenuated in monkeys and man. The M gene of the A/Mallard/78 virus, which is capable of effecting attenuation by itself, was sequenced and significant divergence from the corresponding gene of a human influenza A virus was observed in the M2 cistron. the RNA 1 and NS genes of the A/Mallard/78 virus, in combination, also contributed to attenuation of the A/Mallard/78 reassortants viruses for monkeys. Vaccinia-influenza HA recombinants were immunogenic and protective in hamsters.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Proposed is a continuation of a 20 year cohort study of the epidemiology of injection drug use and HIV risk among HIV-uninfected injection drug users (IDUs) in Baltimore, MD known as the ALIVE-2 study. The ALIVE cohort is unique in that it comprises a community-based IDU population of both genders who are largely out of drug treatment, with significant representation of African-Americans and those with limited access to appropriate medical care;these populations have been underrepresented in research on persons at risk for HIV infection. ALIVE-2 has provided critical insight into the dynamics of infection and risk behavior while serving as a comparison group to a parallel cohort of HIV positive IDUs (ALIVE-1, DA04334). Continuation of the ALIVE cohort will allow us to characterize current trends in the incidence of blood-borne infections and changing patterns of morbidity and mortality among our unique cohort of aging IDUs. Further, we will consider the broader contextual determinants of risk behavior, disease incidence and mortality. Building upon 20 years of follow-up and experience in studying the health effects of injection drug use, our specific aims are to: 1) Examine temporal trends in risk of HIV and other blood-borne infections (e.g., HCV) among injection drug users in Baltimore, MD;2) Assess mortality and morbidity indices among IDUs according to the burden of blood-borne infections and the spectrum of drug use;3) Characterize the effects of macro-level determinants (e.g., urban redevelopment and drug treatment policy) on HIV risk behaviors, blood-borne infection incidence and morbidity and 4) Continue to serve as a platform for independently funded collaborative investigations of HIV and drug use, including interventions to prevent morbidity and mortality associated with drug use, HIV and HCV infection. To achieve these aims, we will continue follow-up of a cohort of HIV negative IDUs (1000 in active follow-up) with semiannual visits involving interviews and collection of biological specimens for HIV antibody testing and repository. We will implement standardized assessments of chronic disease morbidity that include annual testing of early biological markers of disease as well as detailed medical record abstraction. Further, we will supplement the rich history of individual-level risk data with external data on changes in Baltimore's population, economic situation, and health and drug treatment policies so that we can characterize the effects of macro-level processes on risk behavior and the pathways through which they operate by mapping individuals to their neighborhoods of residence. Multiple external sources including the US Census and the Baltimore Neighborhood Indicators Alliance will be used. To enrich our analyses, we continue a parallel protocol of HIV-infected IDUs (ALIVE-1) to facilitate distinction of the effects of HIV and drug use on our outcomes of interest. The proposed aims will inform HIV prevention and structural interventions and will help to project future health care needs for aging IDUs with a changing spectrum of morbidity. PUBLIC HEALTH RELEVANCE: Injection drug users (IDUs) are at high risk for HIV and hepatitis infections and the long-term complications of these infections as well as drug use itself. The findings from this study will inform HIV prevention and structural interventions and will help to project the future health care needs for aging IDUs with a changing spectrum of morbidity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "In 1992, The University of California San Diego initiated this annual international research symposium in San Diego. Since 1998, the program [now jointly sponsored by the University of Heidelberg], alternated locations so that it is held every other year in Germany. The 2003 program is scheduled in San Diego, CA, where participants receive CME credit. This meeting flourishes because it is based on the exchange of innovative ideas shared among scientists and clinicians from academia and the biomedical/industrial community. Stem cell technology has evolved, with hematopoietic cell transplantation becoming standard therapy for various malignant and hereditary diseases. In the past, most successful transplants were conducted in patients under the age of 55 years. Current advances have significantly broadened the safety of these procedures, so that non-myeloablative transplants can be performed with relative safety in patients up to 75 years of age. Recognizing the importance of the immune system in controlling malignant disease, major advances continue to be reported in immunotherapy. Hematopoietic stem cell therapy provides a platform on which to demonstrate its efficacy. Non myeloablative transplants may permit innovative approaches to stem cell transplantation including tandem autologous transplantation to achieve cytoreduction followed by non-myeloablative allogeneic transplants to achieve the immunotherapeutic effect of the allogeneic cells. An exciting development in the field of stem cell biology has been the observation that hematopoietic stem cells may be capable of trans-differentiation into other tissue types. Reports of hematopoietic stem cells differentiating into hepatocytes, myocytes, neurons, and epithelial and endothelial cells have stimulated intense interest internationally. These findings suggest that hematopoietic stem cells may be important tools for the treatment of a variety of disease conditions by aiding in the repopulating of damaged tissues. Given the current national debate on stem cell research and clinical applications, the potential role of hematopoietic stem cells in regenerative medicine has profound importance. Advances in supportive care, including the diagnosis and treatment of fungal disease and graft vs. host disease, contribute greatly to the increasingly successful outcomes of stem cell transplantation. These issues and others form the major focus on the 11th International Stem Cell Transplantation symposium to be held in May 2003. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Regionally-advanced and metastatic carcinoma (Ca) of the breast is amenable to a variety of therapeutic modalities. Combination chemotherapy can produce response with improvement in survival. Radiotherapy can provide local control but patients (pts) still die of metastasis. This approach utilizes combination chemotherapy to shrink primary masses and treat overt or micrometastases. Seventeen pts (8 stage III, 9 stage IV) were treated with combination chemotherapy including 5 with inflammatory Ca. After tumor regression, 15 of the 17 underwent surgery, achieving local control. All 5 with inflammatory Ca had elimination of the inflammatory component. A variety of combinations are available for recurrent or metastatic disease, but there are few prospective trials which compare the various treatment regimens and stratify for the prognostic variables. This program randomly compares 3 active regimens for response rates and duration of survival, and evaluates the effect of MER immunotherapy. A group effort is necessary to stratify for the variables. Interim evaluations of the 335 pts entered show that prior postoperative adjuvant radiotherapy is a detrimental factor for determining response. MER immunotherapy also had a detrimental effect upon response rate (p equals .003) and survival was shorter. Further randomization to MER has been discontinued. Interim evaluations of the treatment arms revealed slight but not significant differences among the chemotherapy regimens.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The neutral crest is a population of migratory cells that arise from the ectoderm of vertebrate embryos and give rise to a diverse range of cell types, including most of the peripheral nervous system, melanocytes and the craniofacial skeleton. It has been classically assumed that the neutral crest is a segregated population in the early ectoderm, lying between the neutral plate and presumptive epidermis. However, our recent studies on avian embryos show that individual precursor cells within the \"neutral folds\" can form neutral tube (central nervous system), neutral crest (peripheral nervous system) and epidermal derivatives. This led us to explore the interactions that impart the potential form the neural crest. Interestingly, we found that neural crest cells are generated when epidermis and neural plate are juxtaposed-a classic type of embryonic induction. The proposed experiments aim to characterize this inductive interaction that leads to neural crest formation and to examine the plasticity of early ectodermal derivatives. To continue our studies on the mechanisms responsible for genesis of the neural crest, we will further characterize the molecular nature of the inductive interaction and will test the function of some candidate inducers in vivo and in vitro. We will examine the ability of ventralizing signals to compete with induction of the neural crest, using both grafting and ectopic expression paradigms. Finally, we will examine the lineage relationships and plasticity between neural crest cells and other ectodermal derivatives by challenging their prospective fates via transplantation. Much of the experimental design will involve in vivo experimental manipulations coupled with cell marking techniques, molecular biological approaches, as well as in situ hybridization to examine patterns of gene expression. Specific experiments will: 1. Characterize the molecular nature of the inductive interaction underlying neural crest formation. 2. Determine the ability of ventralizing signals to repress neural crest formation. 3. Determine the competence of ectoderm and neural plate to assume a neural crest fate. 4. Determine the competence of neural crest cells to assume a neural tube fate after transplantation into the ventral neural tube.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a 48-week, open-label, AUC-controlled, multicenter study. Sixty children will be enrolled. All subjects will receive DMP 266 and nelfinavir. Concomitant use of nucleoside reverse transcriptase inhibitors (NRTI) will be permitted, but they will not be supplied through this protocol. The initial target AUC for DMP 266 will be between 190 and 380 (M(h. The initial starting dose for children will be 600 mg adjusted for body size. The starting dose for subjects will be adjusted on the basis of the tolerability and DMP 266 plasma concentrations of the first six subjects receiving that dose for two weeks. The target AUC is considered to have been achieved if the dose was tolerated and at least 4 of 6 subjects reached the target AUC. Enrollment of subjects will be ongoing and those begun on a given starting dose will continue on that dose until individual subject dose adjustments are needed according to their individual pharmacokinetics evaluations. Baseline and study efficacy evaluations will include plasma HIV RNA levels, CD4 counts, and viral genotypic and phenotypic resistance analyses. Baseline and study safety evaluations will include the monitoring of adverse experiences, clinical laboratory tests, physical examinations and vital signs. At the end of 48 weeks, subjects will be given the option to continue DMP 266 off-study through a DuPont Merck protocol.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract (Administrative Core) The Administrative Core will oversee coordination and integration of all U19 Program functions; guide and facilitate interactions between the Project and Core leaders, Principal Investigators, and research staff; provide rigorous and regular fiscal oversight of the research projects and cores; ensure that the U19 TeamBCP maximizes the utilization of existing and established resources; and facilitate data, technology and information sharing and collaboration with key BRAIN Initiative stakeholders and the greater neuroscience research community. The core will be lead by our Team Directors, Rui Costa and Tom Jessell, and have an Internal Advisory Committee, a Data Science management sub-committee, an External Advisory Committee and a dedicated staff Program Manager. The core will a) provide overall coordination and support for the TeamBCP U19 research activities, b) foster growth of the Motor Control research within and beyond our research team, c) oversee and ensure resource dissemination and outreach, and d) fiscal and administrative management of cores and research projects. The Internal Advisory Committee will meet monthly to discuss all administrative issues and to provide scientific direction for the program. Drs. Costa and Jessell will co- chair these meetings. The needs, usage and effectiveness of the Cores will be assessed at these meetings and any obstacles to progress will be identified. Each Core, including the Administrative Core, will present a status update on any outstanding scientific, administrative or budgetary issues that require immediate attention. These regular meetings will ensure that the program can address problems in a timely manner. They will also serve to make decisions about the resources, priorities, efforts and data access for the whole program, and to resolve potential conflicts. The Internal Data Science Subcommittee will meet at least once a year, with more frequent ad hoc meetings with individual advisory members to review Data Science Core specific issues and report back to the IAC.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Primary immunodeficiency can be caused by reduced immune cell number or function. Lymphocytes are essential components of the adaptive immune system. Mutations in DNA repair factors compromise lymphocyte development and contribute to three of the eight classes of human primary immunodeficiencies recognized by the International Union of Immunological Societies. XLF (also called Cernuous or NHEJ1) is a member of the non-homologous end joining (NHEJ) family of DSB repair genes. Human patients with loss of function mutations in the XLF gene develop a primary immunodeficiency characterized by age-progressive lymphocytopenia and occasional myeloproliferative diseases. The NHEJ pathway as a whole is required during lymphocyte development at the step of V(D)J recombination that assembles the antigen receptor gene products in developing B and T cells. While the role of XLF in V(D)J recombination, was thought to explain the lymphocytopenia in XLF-deficient patients, we and others reported that XLF is NOT required for chromosomal V(D)J recombination in developing murine lymphocytes, underscoring the need for an alternative mechanism. In this proposal, we investigate an alternative and novel mechanism that might contribute to the XLF related immunodeficiency. Specifically, we hypothesize that XLF-mediated NHEJ is essential for the maintenance of genomic stability in hematopoietic stem cells (HSC). XLF-/- HSCs recapitulate the phenotype of aged HSCs - i.e. loss of self-renewal function, impaired lymphocyte (versus myeloid) differentiation and a relative expansion of the myeloid compartment. The lymphocytopenia observed in XLF-/- mice and in human patients is therefore secondary to HSC dysfunction and is accelerated with age. The deficiencies in HSC function and differentiation in the XLF-deficient patients or mice could be due to defects in the HSCs (cell autonomous) or the microenvironment (cell non-autonomous). HSCs are known to reside in bone marrow niches defined by osteoblasts, mesenchymal cells and endothelial cells, all of which regulate HSC function. Notably, prior studies by the co-investigator- Dr. Siddhartha Mukherjee, revealed that genetic alterations in osteoblasts can impair HSC differentiation. To test of hypothesis, we will determine (Aim 1) the cell autonomous function of XLF and DNA repair in HSC maintenance, function and B cell differentiation and (Aim 2) how XLF deficiency in the bone marrow niche environments affects HSC renewal, function and B cell differentiation. In particular, the later study will provide information on the long term therapeutic effects of bone marrow transplantation, the only curative therapy that is currently available for patients with primary immunodeficiency associated with DNA repair defects. The completion of this proposal will establish a link between HSC and primary immunodeficiency that will have general implication in other genomic instability syndromes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The research will develop improved measures of social connection through coordinated analyses of intensive ethnographic data and a longitudinal demographic data about the same population in a rural area of South Africa. A great volume of research conducted in various contexts demonstrates that social connection is an important determinant of well-being measured by outcomes such as education, nutritional status, employment prospects, access to health care, and support of the aged and infirm. The most common measure of social connection in population and public health research is co-residence, which has critical limitations. Failure to attend to the full range of social relationships limits our ability to understand the social context of health and well-being. To redress this failure, we will use existing ethnographic data and longitudinal demographic data from rural South Africa to develop new measures of social connection that can be used in social surveys. The ethnographic data is from an intensive fieldwork investigation of children's networks of support in the Children's Well Being and Social Connection project (CWSC) which we conducted between 2002 and 2004. The quantitative data is the result of fourteen rounds of a population-wide census in a sub-district population of 70,000 people conducted by the Agincourt Health and Demographic Surveillance System (AHDSS). The project has three specific and coordinated aims. First, we identify meaningful and efficient indicators of social connection by using all the available data from the CWSC. This is done by identifying instances of resource transfer and behavior that directly impact children's well-being, analyzing these instances to determine the relevant social relationships, individual, household and community characteristics, and spatial distribution of the actors and resources involved, and classifying the coded instances and reduce the list by selecting the most frequent and important patterns. Second, we determine which indicators of social connection identified in Aim 1 can, and which cannot, be measured by manipulating the data from the AHDSS by linking new data files to the AHDSS database, adding links between existing files, and deriving new variables. Third, we assess the value of the new indicators of social connection added to the AHDSS database in Aim 2 by comparing the explanatory power of models that do and do not include these variables in predicting measures of child well-being such as education and mortality. This research will increase the utility of the AHDSS, and potentially, of other censuses and surveys, in explaining child well-being outcomes. By integrating ethnographic, spatial, and longitudinal population data it will contribute to the development of interdisciplinary methods for studying population and public health. PUBLIC HEALTH RELEVANCE: Resources, such as money, food, protection, opportunities, care, and emotional support that flow directly from person to person are vital for the well-being of children in poor countries that do not have strong market or government institutions. To understand why some people thrive and others do not, and to direct policy interventions effectively, we must understand the web of social connections that surrounds and supports them. This project will produce improved measures of social connection by integrating ethnographic data and longitudinal demographic data about the same population in a poor, rural area of South Africa.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Administrative Core of the Program will be responsible for theoverall oversight, organization, and management of the scientific aspects of the COBRE grant. In addition to the scientific oversight, the Core will also provide a number of support services to Program-associated investigators, including fiscal/budgetary management, preparation of progress reports, scheduling of research meetings for project-associated investigators, coordinating travel arrangements and meetings of the external science advisory board, and organizing a regular scientific seminar program, liver club, \"school of hard knocks,\" amid various work sessions as described in the mentoring section.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cancer has been the leading cause of disease-related deaths in human beings, yet, its major non-surgery treatments have been chemotherapy and radiotherapy, both of which are quite toxic and cause severe side effects. Also, the survival rates of cancer patients have had little improvement. Thus, it still remains remarkably important, though challenging, to develop more potent and specific molecule-targeted therapies. The inactivation of the most important tumor suppressor p53 is one highly cancer-related molecular alteration, as its gene is mutated in ~50% of all types of human cancers while its activity or leve is often markedly reduced in the remaining 50% of cancers that harbor wild type TP53. The p53 deactivation is primarily due to the negative feedback regulation by its two chief suppressors, MDM2 and MDMX, which are over expressed in cancers and form a complex that mediates p53 ubiquitination and degradation as well as inhibits p53 activity directly. This negative regulation s further facilitated by SIRT1, which is highly expressed in some cancers, as the deacetylation of p53 by this deacetylase favors MDM2/MDMX-mediated ubiquitination of this protein. Thus, re-activation of p53 in cancers by targeting SIRT1 can be utilized to screen small molecules for the development of an anti-cancer therapy. Indeed, our recent work has identified a small molecule named Inauhzin (INZ) that inhibits the activity of SIRT1 and induces p53 acetylation, level and activity, leading to p53-dependent apoptosis and senescence in p53-containing human lung and colon cancer cells and suppressing the growth of xenograft lung and colon tumors. Our further studying INZ surprisingly reveals another target, IMPDH2, which is also highly expressed in human cancers. Our previous study shows that inhibition of this enzyme causes ribosomal stress by reducing the level of nucleostemin (NS), which is essential for rRNA processing. Consistent with this, INZ also binds to IMPDH2 and reduces NS levels, leading to the activation of the ribosomal stress-p53 pathway. In light of these interesting findings, I hypothesize that INZ can activate p53 by simultaneously targeting SIRT1 and IMPDH2 and thus eliminate cancer cells via a p53-dependent mechanism. We will test this hypothesis by addressing two specific aims. 1. To determine if INZ induces ribosomal stress and activates p53 by inhibiting IMPDH2 and downregulating NS. Since we have recently reported that INZ inhibits SIRT1 activity, here in this aim we will determine if INZ targets IMPDH2 by verifying INZ as a specific inhibitor of IMPDH2, consolidating if inhibition of IMPDH2 by INZ reduces NS levels and induces consequent ribosomal stress, and determining if RPL11 and RPL5 are critical for INZ-induced p53 activation. 2. To determine the role of INZ-14, a more potent INZ analog, in p53 activation and tumor suppression. In this aim, we will test our newly synthesized INZ derivatives, particularly potent INZ-14, by further modifying it, characterizing it in our established biochemical, cellular and animal tumor model systems, and testing the cooperative effect of INZ-14 with doxorubicin or -irradiation in xenograft and orthotopic tumor model systems.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Oral administration of low doses of antibiotics (chlortetracycline and penicillin) to rats results in substantial changes in bile acid patterns, especially in the lower bowel. Hyodeoxycholic acid formation is suppressed, and omega-muricholic acid becomes the dominant bile acid. These changes persist after discontinuation of treatment, and may lead to reduced fecal bile acid output. In time certain animals revert to a \"normal\" fecal bile acid pattern, but in others a predominance of omega- muricholic acid has been found to last up to three months. To extend these preliminary observations, we propose to study the effect of oral administration (via drinking water) of chlortetracycline, penicillin and neomycin to adult conventional rats. Two dose levels will be used: low (5.00, 5.00 and 15.00 micron M resp.), and high (250.00, 250.00 and 750.00 micron M resp.), the latter within range of therapeutic application. Treatment will last 10 days. Intestinal and fecal bile acids will be determined at various time intervals after discontinuation of treatment. Those treatments which result in the most pronounced and persistent changes will then be used to investigate possible alterations in cholesterol absorption and catabolic turnover of cholesterol, presumably starting 30 days after discontinuation of treatment. Germfree rats will be used to detect any systemic influence of the antibiotics on cholesterol-bile acid metabolism. Conventional and germfree gerbils will be included later in the program. The projected study will probe the influence of routine antibiotics administration on bile acid and related cholesterol metabolism. The investigation aims especially at the long term effects of such treatment as depicted by persistent shifts in bile acid metabolism as they may relate to cardiovascular disease, and possibly to cancer of the lower bowel.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The vascular smooth muscle cell (VSMC) provides dynamic regulation of contractile tone during homeostasis, and is often dysfunctional in diseases involving vascular obstruction and hyper-proliferation of the vessel wall. VSMC modify their gene expression and contractile proteins in response to a variety of external stimuli, particularly during disease progression. The Notch signaling pathway is critical for proper cardiovascular development and is implicated in the pathogenesis of vascular disease. Human mutations in the JAGGED1, NOTCH2, or NOTCH3 genes lead to syndromes with cardiovascular structural defects and susceptibility to stroke. In addition, dysfunction of the Notch pathway is associated with vascular obstructive disease, cerebral vascular disorders, angiogenesis, arteriovenous malformations, and vasculitis. Despite the high significance of this pathway to human vascular disease, there is limited understanding of signaling mechanisms mediated by distinct Notch ligands and their corresponding receptors. Because components of the Notch pathway are therapeutic targets in cardiovascular disease and cancer, it is critical to clarify these signaling pathways. Our studies previously defined an important role for Jagged1/Notch signaling in promoting VSMC differentiation and suppressing proliferation. Part of this mechanism is via the transcriptional activation of miR143/145, which are important for VSMC maturation and function. In addition to Jagged1, a second Notch ligand, Dll1 is also produced during vasculogenesis and remodeling in response to vascular injury. Studies with human primary cells showed that Jagged1 and Dll1 activate distinct pathways consistent with Jagged1 regulating early commitment and differentiation of VSMC, and Dll1 promoting late maturation. These activities correlate with inverse regulation of miR143/145. We propose that the selective ligand effects are mediated through differential transcriptional complex assembly and gene regulation, leading to ligand-specific protein signatures. In addition, we discovered a novel function of Jagged1 signaling via Notch2 to regulate p27kip1 and suppress VSMC proliferation. Thus, the coordinated activities of Jagged1 and Dll1 regulate all phases of the VSMC life cycle from embryonic development to re-establishment of homeostasis following vascular injury. The aims of this project are to: 1) Elucidate Jagged1- and Dll1-induced differential pathways and functional outcomes on VSMC recruitment and differentiation, 2) Discover novel components of differential Notch-mediated Jagged1- and Dll1-induced transcriptional complexes and their impact on gene expression and protein signatures, and 3) Identify mechanisms by which Jagged1 suppresses VSMC proliferation to maintain the contractile phenotype. These studies will provide novel insight into molecular signaling of the Notch pathway, which is a potential therapeutic target for the treatment of cardiovascular diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Laboratory of Epidemiology and Biometry National Institute on Alcohol Abuse and Alcoholism and the National Institute on Drug Abuse is in the planning stage of a large (N=46500) national survey, the National Epidemiologic Survey on Alcohol and Related Conditions III, that will collect both environmental and biological data (i.e., saliva samples). The assessment battery includes extensive questions on sociodemographic characteristics and risk factors for alcohol and drug use and alcohol and drug use disorders (i.e., abuse and dependence) and their associated physical and mental disabilities. These include detailed measures of alcohol consumption and drug use for ten substances, along with their abuse and dependence measures. Major physical disorders (e.g., liver cirrhosis) and mental disorders (e.g., major depressive disorder) highly comorbid with alcohol and drug use disorders are also measured. Risk factors include sociodemographic characteristics, physical and mental impairment, discrimination, acculturation, stress, childhood adverse experiences (e.g., sexual abuse), needle use and HIV, stigma due to substance use disorders, objective measures of weight and height, sexual preference and behaviors, traumatic experiences, tobacco use and dependence, treatment utilization for alcohol and drug use disorders, permanent and temporary disability, academic functioning and achievement, and cognitive functioning.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A basic requirement of all cells is the ability to sense and respond to changes in the environment. Osmolarity is one of the most basic conditions to which cells must respond. Despite the ubiquitous nature of osmosensing systems, the molecular mechanism by which osmotic pressure is sensed is largely unknown. Studies involving microbial model systems have played an important role in identifying the sensors and signal transduction pathways that respond to changes in osmolarity. We plan to use the yeast osmotic stress sensor, Slnlp, as a paradigm for eukaryotic osmosensors. Changes in osmotic conditions regulate Slnlp causing changes in phosphate flux between the individual proteins comprising the two-component signaling system. The fundamental question addressed by this proposal is how Slnlp activity is regulated in response to changes in osmolarity. Using computational, genetic, and biochemical techniques, we have identified a coiled-coil dimerization domain that plays a key role in mediating the stimulus-activation step. It is located in the linker region between the membrane and the kinase domain. The specific objective of this application is to perform a detailed structure-function analysis of the coiled coil (CC) region of the yeast Slnlp osmosensor to test the hypothesis that the unusual composition of the HK CC contributes to the regulation of the HK family of sensor kinases. The specific aims of the proposal are to (1) Genetically dissect the Sin1 CC domain, (2) Determine the structure of the CC domain and CC mutants, and (3) Develop membrane-based assays for Sin1 function. The elucidation of the unique structural and mechanistic features of the two-component type coiled-coil domain in Slnlp will serve as a model for this class of signaling molecules and may lead to the development of histidine kinase inhibitors for antifungal therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent advances in human genetics have enabled the identification of various mutations responsible for disease. Such advances include collections of families and populations presenting clinical phenotypes, genome sequence and polymorphism databases, mathematical algorithms and computer programs for computational purposes, and novel applications of pedigree and linkage disequilibrum (LD) analyses. LD is the non-random association between alleles at different loci. By tracking genetic markers in LD with a disease phenotype, a genomic region harboring a causal variant can be localized. With association studies, it is necessary to determine the haplotype structure of any given region. Recent studies have demonstrated that many regions of the human genome are characterized by segments or blocks of limited haplotype diversity due to high levels of LD between genetic markers. Chemotactic cytokines are known to direct the migration of specific subsets of leukocytes to sites of infection and inflammation. Further, the natural chemokine receptors CCR5 and CXCR4 are HIV-1 coreceptors. About 40 human chemokine genes are found at eight different chromosomal locations, with major clusters on chromosomes 4 and 17. We are focusing on 16 Chemotactic cytokines (CC) genes located at 17q12-21. Nine of these genes (CCL2, CCL7, CCL11, CCL5, CCL14, CCL3, CCL4, CCL3L1, and CCL4L1) have been implicated in HIV-1/AIDS pathogenesis based upon tissue culture, cellular immunological and virus infection assays. In fact CCL3L1 has the strongest binding affinity of all ligands for CCR5, and is thus an excellent candidate for an HIV-1 entry inhibitor. A comprehensive haplotype analysis is being carried out using 80 single nucleotide polymorphisms (SNPs) covering 400 kb in 200 individuals from each of three racial groups: European Americans, African Americans and Chinese. Approximately 40 'tagged' SNPs will be selected and subsequently genotyped in 4000 subjects enrolled in HIV-1/AIDS cohorts. We have already identified and published genetic influences on HIV-1/AIDS for two gene regions, one containing CCR5 (RANTES) and one containing CCL2-CCL7-CCL11 (MCP-1, MCP-3, and EOTAXIN). In addition, SNP CCL3 459 C/T is associated with an increased rate to AIDS-87 in European Americans (p=0.003), and SNP CCL4 662 C/G is associated with apparent resistance to infection in African Americans (p=0.01). Subjects are also available for lung cancer, skin cancer and Hodgkin's disease studies, and are being developed for hepatitis and nasopharyngeal carcinoma. Once the studies 'tagged' SNPs have been identified they will be genotyped in all disease populations.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Nevi (moles) are important precursors and risk markers for melanoma. Current knowledge, derived largely from cross-sectional studies, indicates that adolescence is a critical period for the appearance and evolution f nevi. There is further compelling evidence that nevus phenotype is largely genetically determined with a significant modifying effect of sun exposure. The primary objective of this study is to evaluate specific genetic and environmental factors as risk factors for nevus development and growth in early adolescence. A secondary aim is to document the clinical and dermoscopic evolution of individual nevi in this age group. We will apply a combined cross-sectional and longitudinal study design to the cohort of all consenting 5th graders in the Framingham, Massachusetts school system (estimated n=735) to address three aims. Aim #1 utilizes a cross-sectional study of the 5th grade students to test the hypothesis that germline melanocortin receptor (MC1 R) variants, intense childhood sun exposure, and lack of sun protection in childhood are associated with increased numbers of nevi and large nevi in early adolescence. Aim #2 applies a longitudinal study design in the same student population followed through 8th grade to test the hypothesis that MC1R variants, ongoing intense sun exposure, and ongoing lack of sun protection are associated with increased numbers and increased size of nevi during adolescence. Aim #3 utilizes digital photography and dermoscopy, a recently developed imaging technique, to document the clinical and subsurface appearance and evolution of common nevi in the cohort under study. We will use parent and child surveys conducted at baseline and repeated annually to ascertain sun sensitivity, childhood sun exposure and current sun exposure. We will conduct examinations of the skin of the back including high resolution overview photography and close up digital photography and digital dermoscopy of four index nevi at baseline (5th grade) and repeated in 8th grade to document pigmentary phenotype, the prevalence of nevi by size, the incidence of new and changed nevi, and the clinical and dermoscopic features of individual nevi. Mouthwash derived DNA collected at the baseline examination will be used for MC1R genotyping. The insights into nevus risk factors and evolution gleaned from this study will have significant implications for reduction of melanoma mortality through improved risk stratification and more informed prevention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The current work is concerned with investigating the role of opiate peptides in the brain. Since there are multiple systems containing naturally occurring opiates, we have chosen to focus on the so-called 31K system - a neuronal pathway within brain containing beta-Endorphin, ACTH, alpha-MSH and related peptides. Our overall goal is to characterize these peptides as neurotransmitters or neuromodulators by studying their anatomy, storage, release, receptor binding and behavioral effects. The techniques being employed include immunocytochemistry, autoradiography, receptor binding, extraction of peptides from brain, blood and CSF, radioimmunoassays and HPLC. Furthermore, we employ behavioral techniques and microinjection procedures in an effort to study the in vivo function of these peptides. It is expected that these studies will shed light on the role of endogenous opiates in pain control, drug addiction and motivated and affective behavior.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Tobacco use causes about one of every five deaths in the United States and is the single most preventable cause of death and disease in our nation. At least 70 percent of smokers visit a physician each year, but most are not advised or assisted in any attempt to quit. While more than 90 percent of children visit a pediatrician annually, little is known about the prevalence of smoking prevention counseling. Few practicing physicians are prepared to prevent smoking or help patients stop smoking and a majority of medical school graduates are not adequately trained to counsel families, treat nicotine dependence, or minimize smoking initiation among youth. Shortcomings in tobacco control curriculum in medical schools is a well-recognized deficit. Since 1996, Boston University has developed a curriculum with 10-15 new hours of tobacco control curriculum for students graduating in the year 2000, interwoven into major preclinical and clinical courses, Boston University?s effort follows successful integration of tobacco related curricula at the University of Massachusetts (since 1990). Other Universities have begun tobacco control education with many opportunities to test and evaluate new curriculum in multiple contexts. In this project, II medical schools from throughout the United States, with a wide range of depth and breadth of tobacco curricula, will collaborate with Boston University to develop, refine, and integrate new modules, train medical school faculty, evaluate teaching content, assess opportunities for diffusion to and adoption by other schools, and disseminate teaching guides, We anticipate that exemplary universities of tobacco teaching can be developed and serve as regional and national role models. We are aided in this effort by national representatives of major primary care prance organizations, preventive health specialists, medical student organizations, and cancer control advocates. With expertise in medical student education, curriculum development, faculty training, and evaluation for tobacco prevention and cessation, we have the following specific aims: Aim1 - Assess current curriculum and convene a national conference Aim2 - Develop new modules, plans for integration, and faculty training Aim3 - Conduct trial implementation Aim4 - Conduct a comprehensive, formative, process and impact evaluation Aim5 - Disseminate teaching guides to other medical schools", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Caloric restriction (CR: reducing caloric intake 30-40 percent below ad libitum levels) has been repeatedly shown to extend lifespan, reduce the incidence and delay the onset of age-related disease, enhance stress protection, and attenuate functional decline in mammals. The NIA longitudinal study of CR and aging in rhesus monkeys is in its 29th year and 25% of the animals remain on study. In 2012, we reported findings of improved health; however, unlike findings from a similar study at the University of Wisconsin, the NIA CR monkeys did not have improved survival compared to the controls. Both studies are ongoing and we will continue to monitor the monkeys for age-related changes in many health parameters. Specifically we have ongoing studies assessing behavior including locomotor and motor performance. We have identified traits traits consistent with macular degeneration. We are analyzing samples to identify markers consistent with intestinal cancers that might be similar between monkeys and humans. Monkeys are currently being trained for learning and memory tasks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Reactive oxygen species (ROS) and related oxidative stress are linked with various diseases including cardiovascular disease, cancer, chronic lung diseases, and diabetes mellitus. Interventions favoring the scavenging of ROS to attenuate the oxidative stress may prevent oxidant stress associated diseases. Recent studies suggest that various fruits and vegetables contain high concentrations of antioxidants. Quercitrin is a glycosylated form of flavonoid compounds, widely distributed in nature, and is ubiquitous in plants, fruits, seeds, and vegetables. Our preliminary studies show that this compound displayed a stronger antioxidant activity than that of ascorbic acid over the same concentration range. It blocked TPA-induced neoplastic transformation in JB6 P+ cells. Pretreatment of JB6 cells with quercitrin down-regulated activation of AP-1, NF- :B induced by UVB or TPA. In the skin of AP-1-luciferase transgenic mice, topical treatment of mouse with quercitrin markedly blocked the TPA-induced AP-1 activation. Further studies indicated that these inhibitory actions appear to be mediated through the inhibition of MAPKs phosphorylation, including ERKs, p38 kinase, and JNKs. In addition, quercitrin stimulated the activation of NF-E2-related factor (Nrf2) and GST ARE- luciferase activity. Comet assays showed that quercitrin could block DNA damage induced by UVB. These preliminary studies indicate that quercitrin may function as a potential chemopreventive and chemotherapeutic agent. The overall hypothesis of this application is that quercitrin functions as an antioxidant and protects UV- induced carcinogenesis. The goal of this proposal is to test this hypothesis. Aim 1 will investigate the antioxidant properties of quercitrin in both non-cellular and cellular. We will study whether quercitrin scavenges free radicals or inhibits their generation and determine reaction rate constants of the reactions between quercitrin and oxygen radicals. Moreover, we will detect the UV-induced free radical generation from UV- irradiated skin of living animal using in vivo electron spin resonance (ESR) spin trapping and identify the radicals generated. Aim 2 will investigate the inhibitory effects of quercitrin against the UV-induced oxidative damage to lipid, protein, and DNA, and UV-induced tumorigenesis and cell proliferation in SKH-1 hairless mice. This study represents first detection of free radicals generated by UV-irradiated skin of living animals and may potentially open a new avenue to evaluate the properties of antioxidant against free radicals generated in living animals. Another significance of the study is the identification of qucertrin as a preventive agent against UV-induced skin cancers. PUBLIC HEALTH RELEVANCE: Reactive oxygen species (ROS) and related oxidative stress are linked with various diseases including cardiovascular disease, cancer, chronic lung diseases, and diabetes mellitus. Interventions favoring the scavenging of ROS to attenuate the oxidative stress may prevent oxidative stress associated diseases. This proposal will test the hypothesis that quercitrin, a compound found in blackberries and other foods, functions as an antioxidant and protects UV-induced carcinogenesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "CENTER-DRIVEN RESEARCH PROJECT 1: 3D HIGH-CONTENT SCREENING A. INTRODUCTION &SPECIFIC AIMS High-throughput cellular screens interrogate more biologically relevant processes than do cell-free screens, but they grow in complexity proportionally to their ability to mimic the in vivo state. While most high-content (cellimage- based) screening (HCS) is performed on standard cell culture models, it is well known that many normal and malignant cells lose key phenotypic and functional characteristics when grown in monolayer culture. Introduction of organotypic 3-dimensional (3D) culture and screening systems into mainstream small-molecule and drug discovery processes is increasingly discussed but severely limited by complex methodological requirements and a lack of sophisticated biological model systems, miniaturized screening methods, and 3D image analysis and instrumentation (21, 22). Burnham's Center-driven component will apply our multidisciplinary expertise in cell biology, cellular imaging, and high throughput microscopy (HTM) algorithm and instrumentation development to address this fundamental unmet need in chemical genomics and drug discovery. The broad objectives are to: (1) develop validated 3D model cellular systems for screening;(2) develop software analysis and instrumentation tools for 3D visualization and screening of the model systems;and (3) utilize the 3D culture protocols and 3D image analysis tools to perform screens and disseminating the tools and screens to the scientific community. Realization of these 3D culture models and HTM screening tools will fill the gap between traditional monolayer cultures and in vivo animal models, allowing all MLPCN users to generate their own 3D-HCS primary and secondary assays to enhance and expand the development of biologically meaningful Probes. We will pursue these goals through the following Specific Aims: Aim 1. Develop a toolbox of model organotypic 3D systems for screening. We propose to develop a wide variety of 3D culture systems, including spheroids, multi-layered cultures, and co-cultures of interacting cell types, which are of high interest to the scientific community, in 384- and 1536-well formats amenable to medium and high throughput screening. Aim 2. Create image acquisition and analysis tools for screening of 3D models. A range of image acquisition and analysis tools will be developed to enable primary and secondary 3D-HCS ~ from fixed endpoint 3D imaging for fast primary screens to high-resolution 3D time-lapse analyses for in-depth secondary screens. The following approaches will enable completion of this Aim: 2.1. Development and validation of 3D image acquisition using existing confocal and non-confocal HCS instruments, including deconvolution protocols. 2.2. Development and implementation of 2D assay read-outs from optical sections of 3D images. 2.3. Development and implementation of 3D assay read-outs directly from the 3D images. 2.4. Development and implementation of parallel confocal microscopy instrumentation to speed 3D image acquisition. 2.5. Implementation of 3D time-lapse image acquisition and analysis tools for living cells. Aim 3. Integrate and validate tools from Aims 1 and 2 for primary and/or secondary 3D-HCS screens. Here we will integrate the tools developed in Aims 1 and 2 into assays that will be screened within the MLPCN, thus validating the technology for wider use in the scientific community.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Increased expression of the Fibroblast growth factor 8 (Fgf8) gene plays an important role in the progression of both breast and prostate cancer. To understand how abnormal Fgf8 expression affects cell function, we are studying its normal role during vertebrate embryogenesis, using the mouse as a model system. Fgf8 is expressed in a variety of regions of the embryo that may be termed \"organizers\": regions that are a source of signals that pattern and thus \"organize\" the surrounding tissue. An allelic series generated at the Fgf8 locus (Meyers et al. 1998 Nature Genetics 18:136), as well as Cre-mediated tissue-specific knockouts (Lewandoski et al. 2000 Nature Genetics, 26:460; Lewandoski 2001 Nature Reviews Genet. 2:743) has revealed a role for Fgf8 in organizers that control gastrulation, limb, and brain development. Recently we have produced a valuable mouse line (TCre) that expresses Cre throughout all embryonic mesodermal lineages but not ectoderm/neuroedctoderm, thus allowing us to control gene expression in these lineages. This line is useful to bypass the embryonic lethal phenotypes of genes that affect early development, yet allows the study of the role of such genes throughout much of the embryo. Inactivation of Fgf8 with TCre has revealed that Fgf8 plays a central role in cell survival and gene expression during kidney development. Another surprising insight emerging from these studies is that Fgf8 is not require for several mesodermal signaling centers where it was thought to play a role. To investigate this, we are studying mutants in which Fgf8 and each of the other five Fgfs expressed in these regions are simultaneously inactivated. One of the intriguing insights that has emerged from these studies is that at different stages of embryogenesis FGF signaling plays different roles in cell migration, proliferation, patterning, and survival. How is this diversity of response achieved? To answer this question, we are studing downstream targets of FGF signaling. One set of such target genes is the homeobox genes Gbx1 and 2. The role of the mouse Gbx2 gene during neurulation and particularly in defining the mid/hindbrain organizer has been well documented (Wasserman et al. 1997 Development 124:2923). We are currently extending this analysis by studying a hypomorphic (partial-loss-of-function) Gbx2 allele, which has revealed that Gbx2 is required at certain threshold levels for different parts of the brain. Compared to Gbx2 relatively little has been reported about Gbx1. We recently described the cloning and embryonic expression pattern of Gbx1 and defined regions of potential molecular redundancy with Gbx2. (Waters et al 2003 Gene Exp. Patterns. 3:313). We are currently generating mice with an allelic series at the Gbx1 locus to study it's function during development, including its role in FGF signaling.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is proposal to study how the frequency and complexity of pretend play among 30, 3- and 5-yr-old black girls vary as a function of age and toy structure (high vs. low). High structure toys are defined as those which have highly specific, readily identifiable, and conventional functions (e.g., tea set). Low structure toys are defined as those which have less specific, relatively ambiguous functions (e.g., piece of cloth, cardboard box). Data for this study have already been gathered. This proposal requests support for processing and analyzing them. Same-age girls were randomly grouped to contsitute 10 triads, each of which was observed in a playroom for four 30-min. sessions. High structure toys were present in two of the sessions and low structure toys in the remaining two. Several dependent indices of frequency and complexity of pretend play will be examained. The significance of this study derives from the fact that the interactive effects of age and toy structure on spontaneous pretend play will be examined in one design, dissimilarity between high and low structure toys is increased, providing a more powerful test of their effects, threats to validity present in previous psychological studies are eliminatated, the range of dependent variables is increased, and a previously neglected population is the focus. This study will lay groundwork for more comprehensive research in which comparative data for children who vary on specific population parameters (e.g., social class) will be generated to test hypotheses about the interactive effects of toy structure and individual differences. The theoretical and practical implications of such research are many. Empirical research and theoretical formulations suggest that pretend play facilitates or is associated with basic social and cognitive skills and mental health. Hence, research is warranted to examine situational and ecological factors with a view toward clarification of conditions conducive to pretend play. Results of this study could identify at least one way in which socializers may structure the play environment, with minimal intervention, to ehhance children's pretend play.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will apply the technique of hyperspectral imaging, via the development of an acousto-optic tunable fiber, to enhance endoscopic imaging for the discrimination of tumors, malignant, and normal tissue. With this system, tissue identification will be based not only on a spectral ratio but on the entire spectrum, thus enhancing the system's sensitivity to very small spectral variations. This system will aquire spectral images of drug or laser induced tissue fluorescence, and will perform near real-time mapping for tumor localization.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: Demyelinating diseases resulting from different etiologies may share common secondary pathogenic mechanisms. The overall objective of the proposed research is to advance the understanding of the role of cytokines in the pathogenesis of demyelinating diseases. This project will focus on one demyelinating disease, globoid cell leukodystrophy, which has several advantages for experimental investigations: 1) it has an authentic genetic animal model called the twitcher mouse, 2) it has an inflammatory response that is simple by comparison to other demyelinating diseases such as multiple sclerosis and adrenoleukodystrophy, and 3) it can be treated with bone marrow transplantation (BMT). The rationale for the proposed research is based on the findings that TNFa and IL-6 expressions are elevated in twitcher mice, and twitcher/IL-6 deficient mice have a different pathological course than twitcher mice. If twitcher mice are given BMT, donor macrophages enter the brain and slow the development of pathological changes. However, donor macrophages and endogenous glial cells are still capable of producing cytokines, which may affect the therapeutic course of BMT. Based on these facts, two central hypotheses are advance: 1) IL-6 modulates the pathogenic course in twitcher mice and it promotes the therapeutic effectiveness of BMT, and 2) activated TNF receptors negatively affect twitcher mice by promoting pathogenesis, and limiting the therapeutic effectiveness of BMT. These hypotheses will be tested by the following specific aims: 1_ produce double mutant mice that carry both the twitcher and IL-6 mutations, and use them to analyze the effects of the IL-6 mutation on the clinical and pathological course in twitcher mice with and without BMT, and 2) produce double mutant mice that carry both the twitcher and TNF-receptor(R) 1 mutations or the twitcher and TNF-R2 mutations, and use them to analyze the effects of the TNF-R1 or TNF-R2 mutation on the clinical and pathological course in twitcher mice with and without BMT. The proposed studies should provide insights about the role of TNF receptors and IL-6 in the pathogenesis of globoid cell leukodystrophy, which should be relevant to other demyelinating diseases in the CNS. The long range objectives are to utilize this information to design treatment strategies that modulate cytokine expression in demyelinating diseases in general and to enhance the therapeutic effectiveness of BMT in Leukodystrophies. The project will utilize the following techniques: production of double mutant mice, immunohistochemistry, pathological and clinical evaluations, electron microscopy, PCR, ELISA and BMT.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research is planned to study the functional development of sympathetc innervation of the heart directly in puppies and indirectly in newborn infants. Experiments are carried out in open chest newborn puppies up to 6 weeks of age. After bilateral dissection of the sympathetic cardiac nerves, refractory period changes are determined during maximal stimulation of the individual nerves. The findings from different age groups including adult dogs are compared and functional maturational changes as well as differences between newborn and adult dogs are found. Serial electrocardiograms are obtained in full term premature infants during the first three months of life, starting 24 hours after birth. The data will be processed through a computer and any evidence of sympathetic nerve imbalance such as ST-T waveform abnormalities and tachyarrhythmias will be identified, followed and correlated with maturation. Finally, the results of the electrocardiographic studies in newborns will be correlated with results of the animal studies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "As a part of a continuing study of the neurophysin-peptide hormone system, a cell-free translation system is used to obtain the radiolabelled precursors of neurophysin I and II. The principal technique to be used involves passage of the translation product through two affinity ligand columns, methionyl-tyrosyl-phenylalanyl-aminohexyl agarose and lysine vasopressin agarose, shown by previous studies to specifically bind neurophysin I and II. The interaction of the precursors with the affinity ligands should yield information on the expression of the neurophysin hormone binding site. In the event that the translated precursors do not recognize the ligands, two enzymatic preparations are known that may produce modified forms of the precursors. Microsomes from dog pancreas are reported to convert the pre-pro-neurophysin to the pro-neurophysin form. Treatment of the precursor with trypsin reportedly produces a neurophysin-like species. Differences in the interaction of these precursors with the affinity ligands may suggest a processing scheme wherein the initially translated precursor is converted to a functional neurophysin through a series of proteolytic cleavages.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research concerns the attempt to discover the brain substrates, both anatomical and pharmacological, that are responsible for intracranial self-stimulation (ICSS). ICSS is the phenomenon seen when an animal can be trained to perform tasks for no reward other than the electrical stimulation of its own brain. Because of the hypothesized link between this behavior and certain psychopathological states, a better understanding of the central mechanisms responsible for it is needed. Proposed experiments include: 1) the identification of all neural systems in the area of ICSS electrodes, making use of silver impregnation, fluoresence, and orthograde as well as retrograde axonal tracing techniques; 2) the experimental elimination of these systems with lesion or neurotoxins to assess their contribution to the behavior; and 3) the administration of psychoactive drugs such as the stimulants and neuroleptics before and after the lesions to assess the contribution of various systems to the facilitatory or inhibitory effects of those drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Ricin, a natural product of the castor bean (Ricinus communis) and a Category B toxin, is significant as a biological weapon because of its heat stability, worldwide availability, and ease in production. It can be disseminated as an aerosol, a likely route that terrorists may use. The ricin toxin is a lectin consisting of two polypeptide chains linked by a disulfide bridge and cellular entry is required for toxicity. The ricin B-chain (RCB) facilitates entry of the toxin into the cell and the ricin A-chain (RCA) possesses RNA N-glycosidase activity that attacks a specific site on the 28S rRNA, preventing polypeptide chain elongation, thereby inactivating ribosomes (ribotoxic) and leading to cell death. Since the ricin toxin is free in the circulation for only a brief period of time (hours to days) before cellular internalization, the greatest clinical benefits will be derived from therapeutics capable of blocking RCA enzymatic activity at intracellular sites. We have recent evidence that a high dose of a novel primatized anti-ricin A-chain antibody (43RCA IgG) administered directly to the lung can offer 83% and 75% survival when administered at 44 hr and 54 hr post-ricin challenge, respectively. The primatized 43RCA IgG was generated by fusing the V domains from macaque-derived 43RCA scFv to the constant regions of the human IgG1. The 43RCA scFv sequence is very similar to that for human IgG germline genes, with 90% sequence identity for the VH and VL regions, thereby increasing the potential for this human-like antibody to be used as ricin antidote in humans. In the case of human exposure, whether it is accidental or deliberate, it is anticipated that there will be delays in post-exposure treatment. It is, therefore, imperative to develop antidotes with a therapeutic window beyond 2 days to allow sufficient time for treatment of exposed individuals. We therefore hypothesize that the therapeutic index and/or window for post- exposure treatment can be improved and extended by cytosolic delivery of neutralizing anti-RCA antibodies to block intracellular ricin activity. To achieve our goal, we will develop cell-permeable monoclonal antibodies (or TransMabs) by conjugating antibodies to transport peptides or cell-penetrating peptides (CPPs) and the therapeutic effects will be evaluated as described in the following specific aims. Specific Aim 1: To Ascertain the Protective Effects of Cell-Permeable antibodies against Ricin Cytotoxicity in Cell Culture Models. Specific Aim 2: To Ascertain the Efficacy of Cell-Permeable Antibodies for Post-Exposure Treatment of Ricin-Induced Lung Injury and Lethality using the Lung Aspiration Model. Specific Aim 3: To Validate the Efficacy of Cell-Permeable Antibodies for Post-Exposure Treatment in an Aerosolized Ricin Mouse Model. PUBLIC HEALTH RELEVANCE: Since there is currently no treatment for the ricin toxin, development of a specific antidote for the treatment of ricin after exposure will contribute significantly to the protection of our civilian and military populations. Additionally, the availability of an effective antidote may also significantly reduce the threat of the use of ricin as a biological weapon.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "More than half of all human proteins are glycosylated, and the physiological significance of glycosylation is exemplified by the numerous instances in which variable glycosylation compromises protein function and causes developmental defects and disease. Despite this, the factors that control which glycans are assembled on proteins are not well understood. Polysialylation is a striking example of a protein specific modification that can dramatically change protein function. Polysialic acid (polySia) is best known for its ability to block neural cell adhesion molecule (NCAM)-dependent cell adhesion and signaling, and for its roles in cell migration, axon guidance, synaptic plasticity, an nervous system development. PolySia is also upregulated on damaged peripheral neurons and facilitates their regeneration, and on the surface of several different types of cancers where it promotes their growth and invasiveness. Remarkably, polySia is found on only five proteins in addition to the polysialyltransferases (polySTs) that modify their own N-glycans. The recent identification of two of these polyST substrates, SynCAM 1, a synaptic adhesion molecule, and neuropilin-2 (NRP-2), a semaphorin and VEGF co-receptor, suggests that the roles of polySia may be more extensive than previously thought, and raises the question of how the polySTs recognize and modify these distinct substrates. Our long-term objectives are to determine the mechanism of protein specific polysialylation, what factors regulate the polymerization of polySia chains on specific substrates, and how polySia modulates the functions of the proteins it modifies. In this proposal we will test the hypothesis that the polySTs recognize common amino acid and structural features of their substrates and that this interaction allows an initial polymerization of the polySia chain on a substrate's glycans, and that this is followed by a polyST-polySia chain interaction that promotes further chain elongation. To do this we will evaluate the domain and sequence requirements for polyST recognition and polysialylation of NCAM, SynCAM 1 and NRP-2, and determine whether residues in a conserved polyST polybasic region mediate substrate protein and/or polySia chain interaction to promote protein specific polySia chain polymerization. We will also test the hypothesis that changes in the length of the stalk regions of SynCAM 1 and NRP-2, generated by alternative splicing, alter their alignment with membrane- associated polySTs and control the polysialylation of these proteins in a cell- and tissue-specific manner. We anticipate that these studies will allow us to identify points in the polysialylation process that are subject to physiological regulation, and that will b amenable to experimental and therapeutic manipulations to control substrate polysialylation and function during development, repair, and disease. PUBLIC HEALTH RELEVANCE: Polysialic acid is a developmentally regulated sugar polymer found on a small group of adhesive and signaling proteins. It is critical for nervous system development, and plays roles in cell migration, synaptic plasticity, neuronal regeneration, and the growth and invasiveness of a wide variety of cancers. The proposed work will elucidate the mechanism of polysialylation, and identify points of physiological regulation that can be experimentally and therapeutically manipulated to control the functions of polysialylated proteins during development and disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Overall: Summary/Abstract Over the past four decades, MIT has had a focused effort in cancer research, first in the form of the MIT Center for Cancer Research (CCR) and, since 2007, as the Koch Institute for Integrative Cancer Research at MIT. This effort has been continuously supported by a Cancer Center Support Grant (CCSG) from the National Cancer Institute (NCI), providing the designation as an NCI-designated Cancer Center at MIT. By supplying infrastructural support for Core Facilities and other organizational components of the Koch Institute as well as funds for faculty recruitment and pilot projects, this CCSG is a critical resource for cancer research at MIT. From the establishment of the CCR in 1974 to the transition to the Koch Institute and continuing to the present, the NCI Cancer Center designation has had a strong influence on the MIT administration, leading to significant institutional support over this entire period. The investment in construction of the Koch Institute building (opened in late 2010) is a recent indication of this support. The building brings together 27 cancer scientists and cancer-oriented engineers to form a highly inter-disciplinary and collaborative research environment. The building is also the hub of cancer research on the MIT campus, with a nearly equal number of Members of the Center having their laboratories in other research buildings nearby. The 56 Center Members are drawn from eight academic departments at the School of Science or School of Engineering at MIT. Beyond the discovery research and technology development being pursued by the Members of this Center, significant emphasis is placed on translational research in the form of collaborations with clinical centers and industry partners. Research in the Koch Institute is organized into three Programs. Each of these Programs has made significant advances over the current grant period. Program 1: Genetic & Cellular Programs in Cancer is co-led by Drs. Phillip Sharp, J Christopher Love, and Eliezer Calo. Program 2: Cancer Biology & Immunology is co-led by Drs. Richard Hynes, Dane Wittrup, and Stefani Spranger. Program 3: Systems and Engineering Approaches to Cancer is co-led by Drs. Michael Yaffe, Scott Manalis, and Angela Koehler. These Programs function to stimulate new research initiatives by their Members as well as to foster intra- and inter-programmatic collaborations. In aggregate the 56 Members of this Center have published 1006 cancer-related articles over the past grant period. Of those, nearly 18% have involved multiple Members. The Center has a cancer-related funding base of $58,596,507 TDC (see Data Tables 2A/2B).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The identification of biomarkers that enable the early detection and prognosis of disease, or that facilitate measurement of the efficacy of response to a specific therapeutic intervention, holds great promise in advancing the capabilities of individualized medicine. Recent technological advances, particularly in the development and application of sensitive and robust mass spectrometry approaches, have enabled large scale biomarker discovery efforts to be initiated, including within the emerging field of lipidomics. Lipids are a diverse group of compounds, including fatty acyls, sterols, glycerolipids, glycerophospholipids and sphingolipids that play key biological roles as the main structural component of cell membranes, in energy storage and metabolism, and in cell signaling. A large number of studies have demonstrated that the disruption of lipid metabolism or signaling pathways can play a key role in the onset and progression of human disease. Thus, a comprehensive comparative analysis of changes in lipid profiles that occur between normal and diseased cells, tissues or organs, may enable the identification and characterization of specific lipids that can serve as effective biomarker signatures of disease. Here, we propose to (i) critically evaluate and optimize extraction and sample handling procedures associated with the isolation of lipids from specific tissue types and (ii) develop a comprehensive strategy, based on the use of electrospray ionization and complementary tandem mass spectrometry approaches, coupled with automated data analysis and principle component analysis methods for lipid identification and quantification. Then, we will apply this lipid biomarker identification approach to examining changes in lipid profiles that are observed between normal tissue and the end organs of two common human disease models in which lipids play a major role, namely diabetes (retina) and hepatocellular carcinoma (liver), and to correlate these changes with those occurring in the blood fractions (plasma, erythrocytes and leukocytes) of these subjects for use as potential clinically relevant biomarkers for early disease diagnosis. PUBLIC HEALTH RELEVANCE: The successful completion of these studies will lead to the development of robust and sensitive bioanalytical mass spectrometry methods for comprehensive lipid analysis, and for the identification of biomarkers of diabetes and diabetic complications, and hepatocellular carcinoma. The results from this work will therefore provide information leading to the development of effective clinical methods for the early detection and monitoring of these diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this protocol is to determine whether staphylococcal toxins (superantigens) contribute to the T cell activation found in atopic dermatitis (AD) and study the role of S. Aureus infection in the pathogenesis of this common skin disease. Specific Aim One will determine whether AD is associated with a selective expansion of T cells in their skin lesion. Specific Aim Two determines whether the selective stimulation of T cells in AD is clonotypic or diverse by cloning and sequencing the individual T cell receptor (TCR) transcripts amplified from AD skin lesions. The final specific aim investigates whether S. Aureus growing on the skin of patients with an exacerbation of AD due to infection, produce toxins known to act as bacterial superantigens E.G. Staphylococcal enterotoxins (SES) and correlate these findings with the TCR VB gene usage of the t cell infiltrate from skin biopsies of the culture site. In addition, nonlesional skin of patients with AD and normal controls will be patch tested with staphylococcal exotoxins to determine they can induce eczematoid skin lesions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Classic antipsychotics treat schizophrenics's hallucinations, but not their social withdrawal and flattened affect. These drugs also produce extremely unpleasant motor side effects, because these compounds were originally selected based on their ability to produce motor disturbances in rats. Clozapine is an atypical antipsychotic which treats both types of symptoms with very few motor side effects, but it can also produce a potentially lethal side effect, agranulocytosis. Thus, a tremendous demand exists to develop safer and more effective atypical antipsychotics. In order for a compound to show promise as an antipsychotic in preclinical trials, it must have appropriate behavioral and biological actions. The goal of this research is to develop a better method for detecting atypical compounds by testing the hypotheses that atypical antipsychotics will differ from classic antipsychotics in terms of their effects on PCP-induced social withdrawal with repeated administration. In particular, atypical antipsychotics are predicted to be more efficacious at alleviating PCP- induced social withdrawal with repeated administration. In particular, atypical antipsychotics are predicted to be more efficacious at alleviating PCP-induced social withdrawal. Atypical antipsychotics are also predicted to produce greater tolerance to the initial increases in extracellular dopamine in mesocorticolimbic structures, while having a limited effect on striatal dopamine, compared to typical antipsychotics. This hypothesis will be examined using simultaneous sampling from pairs of structures using in vivo dialysis coupled to HPLC. The results will provide scientists with: 1) a profile of social behaviors in a rodent model for candidate atypical compounds to restore in order to be effective at treating both types of symptoms in schizophrenia; and 2) a neurochemical model with limited effects on striatal dopamine and only transient elevations in mesocorticolimbic structures. These studies would improve the manner in which compounds are selected for human clinical trials and enhance our understanding of biological mechanism involved in the atypical antipsychotic effect.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Several diverse metabolic events become compromised when mammalian cells are made deficient in essential amino acids or when charging of their tRNA is blocked by amino acid analogs. This rapid general demise of cell function can be due to inhibition of phosphofructokinase (PFK) by uncharged tRNA. It has now been demonstrated that when tRNA is added to PFK in any assay dependent upon the reassociation of inactive, dissociated enzyme subunits, nanomolar concentrations cause complete inhibition. The model for control suggests that charged tRNA becomes associated with EF-1, which is specific for aminoacyl-tRNAs and is present in sufficiently high concentrations in cells to sequester the charged forms from an inhibitory role. Support for this model include: (1) the rapid onset of inhibition of glycolysis and glucose uptake upon amino acid deficiency; (2) the unique role of the product of PFK activity, fructose-1,6-diphosphate, in reactions of peptide chain initiation, particularly its role as a co-factor for purified eIF-2B, the GDP/GTP exchange factor; (3) the correlations of this interaction with the cellular and molecular lesions of insulin insufficiency; (4) the recognition that the anomalous role of high concentrations of cAMP as a stimulant of peptide chain initiation in energy depleted or gel-filtered cell lysates correlates with its stimulatory action on PFK as an analog for the positive effector, adenosine-5'-monophosphate; and (5) the role of fructose-1,6-diphosphate in the formation of glyceraldehyde-3- phosphate, a substrate for synthesis of ribose-5-phosphate via the non- oxidative portion of the pentose phosphate pathway, which, as precursor of phosphoribosylpyrophosphate, is essential for nucleic acid synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY/ABSTRACT The project ?Expansion of Training and Program Components to Maintain Manufactured Food Regulatory Program Standards? is designed to further advance the manufactured food program in the State of Wyoming. Wyoming Department of Agriculture, Consumer Health Services (WDA/CHS) has made great strides in improving the manufactured food program statewide with the assistance of the Manufactured Food Regulatory Program Standards (MFRPS). In October 2017 WDA/CHS was elated to learn that after the 60 month Program Assessment Validation Audit (PAVA) the program was in conformance with the MFRPS. WDA/CHS takes great pride in providing a quality program for our inspection personnel and industry to be part of. This project will address four (4) goals. The first goal is to adequately train our inspection personnel and continue to meet MFPRS requirements. This training will equip inspection personnel with the knowledge and ability to conduct quality inspections and further advance WDA/CHS to be part of an Integrated Food Safety System. The training received will also be used to provide education to industry. The second goal of this project is for WDA/CHS to continue to show incredible growth in outreach to our clientele. Through this project WDA/CHS will develop and produce a guide for Manufactured Food processors addressing 21 CFR 117 and Food Safety Modernization Act (FSMA) regulations and publish the Wyoming Food Safety Rule revision. This project includes outreach to establishments required to have a Preventative Controls Qualified Individual by providing FSPCA Preventative Controls for Human Food Training. The third goal of this project is to adequately equip our inspection personnel to complete the inspections. Additionally, WDA/CHS plans to expand our routine sampling program. All of the goals listed above are incorporated into the final goal of this project, which is to advance the manufactured food program and continue to conform to the MFRPS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "OBJECTIVE To determine whether mRNAs are expressed in the rhesus placenta which could encode a soluble MHC class I molecule. RESULTS An unusual feature of both the human placental MHC class I molecule HLA-G and the rhesus molecule Mamu-AG is expression of multiple protein isoforms and splice variants. Splice variants also exist for HLA-G which encode a soluble protein but the presence of a mRNA encoding a soluble rhesus MHC class I molecule has not previously been detected. We cloned a fragment of the Mamu-AG gene from rhesus monkey genomic DNA to define the sequences of intron 4 and 5 and noted that the sequence of intron 4 was highly homologous to that of HLA-G (88% identical), which is similar to exon 4 homology with HLA-A sequences (~91%), but higher than that for intron 5, suggesting that evolutionary drift of intron 4 has been restrained and implying selection for functionally important sequences. We then conducted RT-PCR analyses with placental mRNA using an upstream primer specific for Mamu-AG exon 4, and a downstream primer which would amplify through the 3U-untranslated region and allow diagnostic identification of Mamu-AG transcripts via the premature stop codon homologous to HLA-G. RT-PCR products were amplified from placental RNA, cloned and sequenced. A stop codon (TAG) is present in the exact codon location of the TAA stop codon in the soluble HLA-G reading frame of intron 4. The predicted amino acid sequences of the carboxy-terminal end of the 4th exon and the intron 4 peptides indicate identity with the human sequence at 16/21 residues, and conservative substitutions (e.g., G to S) at the other 5 residues. Further studies cloned and defined the sequences of the other introns of Mamu-AG. FUTURE DIRECTIONS These results suggest that the rhesus monkey will be a model for addressing the physiology of soluble Mamu-AG. We will evaluate 1) the expression of soluble mRNA in the fetal membranes, 2) whether antibodies against soluble HLA-G recognize the rhesus molecule, and 3) rhesus amniotic fluid and fetal serum for the presence of soluble MHC class I molecules. KEY WORDS maternal-fetal immune tolerance, placenta, amniotic fluid, splice variant FUNDING NIH HD 26458, HD34216", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hematopoietic stem cell transplantation (HSCT) is now the second most frequent major organ transplant in the US and is used as the treatment of a variety of malignancies and bone marrow failure disorders. An estimated 45,000 transplants are performed each year, 2000 in patients under 20 years of age. There are more than 90,000 survivors of this procedure in the US. HSCT recipients and their families are extremely vulnerable as a result of the physical and psychological demands of the treatment, the geographic dislocation, and physical and social isolation. During the transplant recovery process, care shifts back from the transplant center to the child's home and local treatment center with the parents assuming the primary responsibility for the child's care. Traditional hospital-based interventions have focused on the peritransplant period, but given the prolonged and demanding period of recovery (6-12 months), alternative interventions are needed. To address these issues, we are collaborating to develop a highly transportable transplant-specific module, adapted from the well-established Comprehensive Health Enhancement Support System (CHESS), HSCT-CHESS. This module will be an interactive, web-based, health information and support system for pediatric HSCT parents and families. Through a randomized controlled trial of 190 parents at four pediatric HSCT centers across the US, we will evaluate the impact on health-related quality of life (HRQL) of HSCT-CHESS, comparing it to standard care over a nine-month intervention period. Information about the impact of pediatric HSCT on the family will also be collected over the same time period. This proposal sheds new light on the application of interactive health communication systems to a new population, by formally exploring the mechanism of action and identifying who may benefit most by this approach. The study also directly investigates the link between interactive health communications and HRQL. The results of the proposed family-centered evaluation will have important implications for family adaptation in other intensive cancer treatments. This intervention has the potential to serve as a model in other clinical situations in which complex care is shared by health care providers with different expertise in geographically distant sites, particularly those in which the patient and family must be actively engaged in care coordination and disease management. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Leveraging Family Data to Identify Genetic Variants for Sleep Apnea PROJECT SUMMARY (ABSTRACT) Obstructive Sleep Apnea (OSA) affects more than 10% of the population, especially Hispanic- and African- Americans, and is associated with profound cardio-metabolic morbidity. Through the Cleveland Family Study (CFS), a genetic epidemiological study of rigorously phenotyped families ascertained through probands with OSA, we have established that OSA has a strong genetic basis and have identified promising areas of linkage to inform genetic association analysis and sequencing efforts. To meet the objectives of this RFA, we intend to efficiently utilize existing data from th CFS as well as newly available genotype and sleep phenotype data from major NHLBI cohorts (Sleep Heart Health Study cohorts of ARIC, CHS and Framingham Heart; MESA; MrOS- Sleep, Starr County Health Study, and the Hispanic Community Health Study). Our primary phenotype is the continuous trait, the apnea hypopnea index (AHI), derived from sleep studies rigorously analyzed and archived at our central Sleep Reading Center. In toto, the sample includes 1200 CFS family members and ~11,000 members from NHLBI cohorts, including admixed populations at high risk for OSA and likely to harbor rare variants. Capitalizing on the power of family designs combined with focused sequencing efforts and modern statistical tools, including methodological advances by our research team of leading genetic statisticians, geneticists and sleep epidemiologists, we propose to use complementary strategies designed to identify common as well as low frequency and rare variants associated with OSA. We will: 1) leverage information from areas of linkage to the AHI to prioritize genes for further testing and for targeted exon sequencing; and 2) perform whole exome sequencing in individuals selected from our most informative families and extreme OSA phenotypes, a sample likely enriched with rare variants. Our hypotheses and aims will be addressed using advanced gene-based analysis (SNP-set kernel association tests) and meta-analysis in a multi-stage design consisting of discovery and validation phases. Linkage information will be incorporated into association analysis to improve the chance of true discovery. Weighted methods and bioinformatics approaches (using gene networks) will be used to increase analysis power for detecting rare variants. Analyses will control for covariates including population stratification using principal components, local ancestry, and family relatedness. Multiple comparison adjustments will be carried out to control for overall type I error. Additionally, we will develop novel statistical approaches to meet the challenges of this study as well as other studies supported by this RFA, including methods for analyzing family and unrelated samples when multiple rare and common variants contribute to phenotypic variation. RFA HL-07-012 provides a critical opportunity to leverage the family data from the CFS, newly available data from large cohorts, statistical advances and advanced sequencing technology to together fill a major need to discover and replicate functionally important variants for OSA that may serve as targets for novel therapies for this serious health condition.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Vitamin D deficiency affects between 31% and 82% of the U.S. population, depending on race/ethnicity, and is associated with hypertension, as well as other cardiovascular outcomes. Several randomized clinical trials of the efficacy of vitamin D supplementation in reducing blood pressure have been conducted with mixed results. These inconsistent clinical trial findings may be explained by the substantial inter-individual variabiliy in the ability of a given vitamin D dose to raise circulating 25-hydroxyvitamin D [25(OH)D] concentrations and, thus, to influence vitamin D-related outcomes. This variability is influenced by genetic variants. However, these variants have not been used to guide the dose of vitamin D supplementation. Our long-term goal is to use Genotype-gUIded vitamin D supplEmentation (GUIDE) to improve vitamin D-related health outcomes. The objective of this application is to conduct a randomized, double-blind, placebo-controlled clinical trial of daily oral vitamin D3 for three months in 558 healthy participants between the ages of 30 and 70 from an existing population-based study. The vitamin D3 dose will be determined by the number of risk alleles in two vitamin D- related genes. The specific aims are to determine the efficacy of GUIDE in 1) achieving 25(OH)D concentrations of 20-50 ng/mL and 2) lowering blood pressure.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We described 3 patients with hypertrophic cardiomyopathy who after septal myotomy-myectomy had severe mitral regurgitation. All 3 patients had complete relief of the left ventricular outflow obstruction by the myotomy-myectomy operation but nevertheless, severe symptoms of cardiac dysfunction persisted post-operatively. At reoperation, ruptured mitral chordae tendineae were found in 2 of the 3 patients and severe posterior leaflet systolic anterior motion in the third patient. It is likely that the mitral valve damage was done at the time of the myotomy-myectomy operation. Thus, this potential danger must be kept in mind at the time of myotomy-myectomy in these patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Immunoperoxidase studies are designed to determine the presence and intracellular location of measles virus polypeptides in CNS cells of chronically infected hamsters. Virus isolated from acutely and chronically infected hamsters will be compared by polypeptide composition. To maintain the characteristics of neural cell growth, all virus will be maintained in reaggregated brain spheroid cultures prior to biochemical characterization. Immunoprecipitation methods will be used to measure the serum antibody response to acute and chronic SSPE virus infection in the hamster. Defined measles polypeptides will be used to \"vaccinate\" hamsters of various ages prior to exposure to intracerebrally inoculated measles virus. Following this, animals will be analyzed to determine the protective value of individual antibodies to specific polypeptides or the ability of such antibodies to induce \"antigenic modulation\" of virus leading to persistent infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Role of cytoplasmic and nuclear skeletal structures in the function and metabolism of RNA will be further investigated. The apparent association of poly(A) with the cell skeleton will be pursued and the apparent solubility of Friend cell polyribosomes will be investigated and compared to HeLa. A new set of small RNA molecules located in the nucleoplasm and apparently synthesized by a polymerase of one-type activity will be further studied. These RNAs show a profound change in different animal species and are possibly related to the state of cellular differentiation. Macromolecular metabolism will be further studied in anchorage dependent cells suspended in methocell and reattaching on plastic surfaces. Profound changes in protein synthesis and messenger processing are noted and will be characterized. Changes in message composition have been detected and will be measured. Changes in plasma membrane composition of suspended anchorage independent cells will be measured. Evolution of phenotype between anchorage dependence and independence can be seen at the biochemical level and will be studied with cell lines having intermediate phenotypes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "OBJECTIVE: To conduct a research program on the biology and treatment of murine colon carcinoma. APPROACH: The general goal of our colon tumor project is to obtain improved treatment methods (mainly through the use of chemotherapy or surgery in combination with chemotherapy) for colon tumors in mice. These methods will then function as leads for clinical trials against the human disease. The general method of research involves the testing of various chemotherapy agents and combinations of agents against transplantable colon tumors in mice. We are currently using four transplantable colon tumors that were developed in these laboratories. One of these colon tumors is highly metastatic and surgical removal of 1/2 g tumor growths will not cure the animals because cells are shed prior to surgery through the blood and lymph to distant organ sites. We are testing active single agents and combinations of agents following surgical removal of the primary tumors in an effort to determine the curative potential of the various drug treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY It has long been known that methylation of genomic DNA correlates with gene expression. However, the structural mechanisms that underlie these observations remain obscure. In this project, we will pursue several innovative strategies for studying how methylation affects transcription factor (TF) binding. First, we will use Methyl-SELEX-seq ? a novel experimental method developed in the previous cycle of this grant that uses barcoded mixtures of methylated and unmethylated DNA ligands ? to create detailed maps of the effect of methylation on binding affinity for a broad panel of human transcription factors from various structural families. Second, we will perform detailed computational analyses and follow-up experiments to test the hypothesis that methylation causes local changes in DNA shape, which in turn modify TF binding affinity. We have shown that adding a methyl group in the major groove changes the geometry of the minor groove and enhances the electrostatic interaction between negative charges in the DNA minor groove and positively charged amino- acids in the TF. We will extend these analyses to other DNA modifications, as well as a wider range of DNA shape parameters and associated flexibility parameters. By building interpretable TF-DNA recognition models that integrate base, shape, and flexibility features using a powerful new machine learning framework developed in the previous funding cycle, we will make specific predictions regarding sequence and methylation readout mechanisms, and validate these using SELEX experiments with mutated TFs. To assess to what extent our quantitative models for binding to naked DNA built from SELEX data are predictive of binding to genomic DNA in the context of the living cell, we will perform detailed parallel analyses of SELEX and ChIP- seq data for Hox proteins and other TFs. Finally, to study the relationship between DNA binding and gene expression control in human cell lines, we will exploit Survey of Regulatory Elements (SuRE-seq), a novel massively parallel reporter assay that provides unique information about the autonomous transcriptional activity for each of >108 overlapping genomic fragments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal seeks to support and enhance Down Syndrome research capabilities via the DS-CONNECT registry by leveraging PCORnet, the National Patient-Centered Clinical Research Network. Since 2014, the Patient-Centered Research Institute?s investment in creating PCORnet has resulted in a diverse national network covering over 100 millon lives: providing curated, electronic health record data and claims as real world evidence along with heighted patient/clinician/health system engagement across participating organizations. We will link PCORnet to DS-CONNECT and test capability in three dimensions: 1) increasing DS-CONNECT registry enrollment, 2) extract clinical observations, treatments, and outcomes from PCORnet for DS-CONNECT patients, and 3) conducting cognitive assessment via DS-CONNECT in the PCORnet population. People with Down syndrome and intellectual and developmental disabilities experience poorer health-related quality of life than people without a disability. For people with Down syndrome, person factors, such as communication ability and social skills can also impact health in terms of access to quality health care, opportunities for social participation, and challenging behavior. Self-determination which is defined as people with disabilities engaging in goal-directed actions that enhance quality of life has been shown to be a critical factor in improving employment and health-related outcomes. Specifically, the relationship between sleep quality and cognition may play a significant role in community living and employment for adults with Down Syndrome. We will invite PCORnet DS-CONNECT participants to complete the Self-Determination Inventory (SDI) and link SDI scoring of problem solving and goal setting cognitive deficits for Down Syndrome patients with Obstructive Sleep Apnea (OSA) diagnoses recorded in PCORnet. This will be one of the first studies to robustly integrate data regarding the relationships between health and community participation for people with DS, which is made possible linking health data from registry participants in PCORnet using DS-CONNECT and the Self-Determination Inventory (SDI) System Data Dashboard. As such, our proposed research will have far reaching implications for supports and services for people with DS and the influence of health on participation. The addition of self-determination will help researchers better tailor interventions to support specific aspects of cognition that interfere with health and participation outcomes and create a framework for exploring other associations by integrating data to inform intervention development and future clinical trials.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent evidence suggests that H2O2, produced in response to various extracelluar stimuli, functions as an intracellular messenger. H2O2 is also a precursor of hydroxyl radicals that cause irreversible damage on cellular components. This dichotomous function of H2O2 predicts that the production timing and local concentration of H2O2 be stringently regulated. The thioredoxin peroxidases, TPxI and TPx II, are members of the peroxiredoxin (Prx) family that participates in growth factor and cytokine signaling by modulating intracellular concentration of H2O2. In an effort to understand how TPx enzymes cooperate with different agonist signalings, the distribution of TPx proteins has been studied in subcellular fractions obtained by differential centrifugation. TPx I and II were detected in organelle and membrane fractions, as well as cytosol. When the membrane pool was further analyzed on Nycodenz-density gradient, TPxs I and II showed wide distribution throughout light and heavy membrane structures. Furthermore, we detected a part of TPx proteins in Triton X-100-insoluble membrane fraction. Immunofluorescence staining showed an obvious punctuate pattern of TPx II protein in HeLa cell, similar to endosomal staining. These results indicate that TPxs I and II may localize in various membrane structures, including endosomes and lipid raft. Interestingly, treatment of HeLa cells with EGF or H2O2 induced the accumulation of TPx II in perinuclear region, suggesting a stimulation-dependent translocation of TPxII. We also observed that TPxs I and II, but not other Prx isoforms, are present in nuclei fraction purified from HeLa and NIH3T3 cells. The amount of TPxs I and II in nuclei fraction increased in response to serum stimulation and decreased in response to treatment with H2O2 and UVB. TpxI and TpxII contain a site [TP(K/R)K] of phosphorylation by cyclin-dependent kinase (CDK). CDK2, CDK4, and CDK6 immunoprecipitated from cell extracts phosphorylated wild type TpxI and TpxII, but not mutant Tpx proteins with the Ala for Thr substitution at the putative phosphorylation site. The threonine phosphorylation was also confirmed with CDKs co-immunoprecipitated by antibodies against cyclin A, D, and E. Taken together, Tpx enzymes appear to be mobile protein that respond to the local need by translocation. Either translocation to nucleus or their catalytic activity in nucleus is likely regulated by CDK-dependent phosphorylation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Large inhalable particles exist in the workplace and can constitute a substantial amount of inhaled exposure (50% or more). Particles >20 m primarily deposit in the oro-nasal cavities. Symptomatic health effects (e.g. acute or chronic rhinitis, chronic pharyngitis, chronic sinusitis, nasal cancer, chronic laryngitis and gastro- intestinal diseases) occur in many industries; such effects may be due to the presence of large inhalable particles in these workplaces. However, measurement capabilities for counting and sizing particles between 20 and 100 m are limited. Although large particles settle through air quickly (terminal settling velocity of 100 m particle = 0.3 m/s), these particles are also capabl of delivering very high doses to affected tissues (a 100 m particle weighs approximately 1 g). Thus, a real-time instrument is needed to capture the timing and magnitude of such exposures. There are two specific aims for this project. The first aim optimizes the design of a portable inhalable particle spectrometer to segregate inhalable aerosols as a function of particle size and incorporates optical detection for real-time measurement of particle size and concentration. The second aim evaluates the sampling efficiency, cutoff curves, and other performance measures in calm air and low-wind speed environments. The career development plan outlines the activities that will be taken during the K-award period. This plan describes training in three specific skillset areas: engineering design, aerosol optics, and experimental techniques. The career development plan also describes the specific training the mentors will provide, as well as important milestones, steps, and a timeline for achieving the short and long term career goals. One-on-one mentoring (for both research and career development) is described.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A multi-center study of oxacillin resistant S. aureus is in place and should be completed within a year. Phage types have been, and will continue to be used to fingerprint strains of the oxacillin resistant species. We have evidence that strains with oxacillin resistance not reversible by clavulanic acid are few in number and that oxacillin resistant S. aureus is represented in many U.S. hospitals by a single predominant strain. S. aureus in which oxacillin resistance is reduced in the presence of clavulanic acid, the hyperbetalactamase producers, have another, distinct phage pattern. We also are studying loss rates of oxacillin resistance after the strains are cultured from the treated patient. This phase will include initial sensitivity tests done directly from clinical samples, followed by tests done during serial subcultures. This has important bearing on sensitivity testing of these S. aureus strains, since they begin to lose resistance immediately, on initial cultures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This investigator-initiated new R01 application, in response to PA-10-067, proposes to reduce depression and cancer-related emotional distress, while also improving health behaviors in adult hematopoietic stem cell transplantation (HSCT) survivors using an internet-based intervention in a multi-center randomized controlled clinical trial (RCT). Among adult long-term cancer survivors, those who received HSCT live with some of the highest risks for life-threatening chronic health conditions including cardiovascular disease, diabetes, osteoporosis, and recurrence or second cancers along with other risks. Even more prevalent in HSCT survivors are psychological symptoms that inhibit quality of life, specifically depression and cancer-related distress. Our previous internet intervention has demonstrated efficacy in reducing depression and distress with survivors at a single site, and has pilot tested the efficacy of providing a survivorship preventive care plan. With the proposed improvements and the extension of the intervention to multiple transplant centers we expect to strengthen both the reach and efficacy of the intervention while we advance the science of internet-based providing of health behavior change models to cancer survivors. Specific aims of the project are to: 1) determine whether 2-10 year HSCT survivors with elevated depression or cancer-related distress who are randomized to receive access to a tailored internet-based program report reduced depression and distress when compared with control group survivors who receive a survivorship care plan by mail and delayed access to the internet site (enhanced usual care), and 2) determine whether survivors with low survivorship preventive care adherence (PCA) who are randomized to receive access to the tailored internet-based program, including a survivorship preventive care plan, report increased PCA when compared with control group survivors. The study will examine main effects of the intervention as well as moderating (age <40, chronic graft versus host disease, rural residence, lower income) and mediating effects (health self-efficacy, knowledge of survivorship needs) per Protection Motivation Theory. Further analyses will identify risk factors for disparities in survivorship preventive care adherence in HSCT survivors. The five-site multi-center RCT will enroll N=1200 adult HSCT survivors with internet access. Assessments will occur at baseline, 3 and 12 months. Participants without internet access will be invited to complete the baseline assessment for participation in the risk factor analyses. To our knowledge, this study will be the first to explore the efficacy of a tailored internet-based intervention, enhanced with options for alerts and reminders through individualized SMS messaging, mobile applications and social networking media, to target health behaviors in a cancer survivor population. The program is cost-effective and readily tailored. If effective, the program could be disseminated nationwide for long-term HSCT survivors in all transplant centers and would provide a framework for similar survivorship care models in non-HSCT cancer survivor populations. PUBLIC HEALTH RELEVANCE: Among adult long-term cancer survivors, those who receive hematopoietic stem cell transplantation (HSCT: also called blood, marrow or umbilical cord blood transplantation) live with some of the highest risks for life-threatening chronic health conditions including cardiovascular disease, diabetes, osteoporosis, and recurrence or second cancers. Also prevalent in HSCT survivors are symptoms that inhibit quality of life such as depression and cancer-related emotional distress. In a study with five sites across the nation, this research is designed to test whether a tailored internet-based survivorship program with methods for managing health behaviors improves depression, emotional distress and adherence to health care guidelines in adult HSCT survivors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Normal tissue complication probability (NTCP) models can be used to individualize radiation therapy treatment planning, potentially by guiding the design of the dose distribution. Our goal is to produce improved NTCP models, based on dose-volume, patient, and disease characteristics, for head and neck and lung treatment complications. Under the previous grant, we developed a software system, which enables the construction and convenient analysis of databases of 3-D treatment plans, including datasets from multiple institutions. Using data thus obtained, we will construct predictive models using multi-metric logistic regression methods, which include dose-volume terms as well as other patient and disease-related factors. The robustness of variable selection will be tested with bootstrap methods. For lung treatment plans, we will recompute lung and esophagus dose-volume histograms using a novel Monte Carlo-based technique, to improve the consistency and accuracy of the database dose distributions. Under Specific Aim (SA) #1, Improvements in post-RT late pneumonitis/fibrosis NTCP models, we will: (a) expand the currently available Wash. Univ. dataset (from 166 pts. to an estimated 450 in 4 years), (b) study the inclusion of new factors such as spatially-varying sensitivity and pretreatment pulmonary function tests, (c) test and refine our model using the RTOG 93-11 dataset (113 pts.), and (d) test and refine our model against data contributed by Duke University and the Netherlands Cancer Institute (an estimated 550 pts.). Under SA #2, Improvements in acute esophagitis NTCP models, we will: (a) accrue more patients (from 166 to an estimated 450 in 4 years), (b) incorporate new factors such as partial-circumferential irradiation and other metrics based on the shape of the high dose region, and (c) test and refine our model using new data contributed by the Netherlands Cancer Institute (an estimated 300 pts.). Under SA #3, Improvements in post-RT parotid salivary function/xerostomia models, we will: (a) test the effect of spatial placement of high-dose regions, (b) use the model to analyze the radio-protective effect of Amifostine on salivary function in an ongoing intensity modulated radiation therapy trial, and (c) test/refine our model against the University of Michigan xerostomia dataset. In addition, we will establish publicly archived databases with convenient and freely available software tools. We hypothesize that this research will result in a significantly improved ability to predict, on an individualized basis, the risk of xerostomia, pneumonitis, or esophagitis, and could thereby lead to improved radiation therapy treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Atherogenesis in the adult-onset diabetic is difficult to explain satisfactorily on the basis of existing knowledge concerning insulin, glucagon and growth hormone. Additional controlling hormone(s), particularly those involved with the mobilization of depot fat, would seem to be required for a more nearly complete description of the mechanisms involved in diabetogenesis and in the concomitant or subsequent atherogenesis. The existence of such hormones, which are probably produced by the pituitary or hypothalamus, has been clearly indicated by previous investigations, but they have not been isolated and characterized and their relation to various pathophysiological states established. We propose that the over-production of such hormones leads to increased levels of plasma fatty acids which can then lead to diabetes because of insulin resistance and to accelerated atherogenesis because of hyperglyceridemia. These hormones occur in the urines of fasted, normal adults and of diabetics. The hypothesis will be tested by: (a) isolating the active materials from these sources using chromatographic procedures, (b) determining their amino acid sequences, (c) developing radioimmunoassays for their quantitation in plasma, and (d) using such procedures to ascertain the circulating levels of the active materials in pre-diabetics, diabetics, in persons at risk for atherosclerosis. Such measurements will also be carried in persons in other pathophysiological states with the objective of ascertaining the factors which control the release of these materials from their gland of origin.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The overall objective of this research is to understand the mechanism for the enzymatic assembly of cholesterol, the control of this process, and the relationship of this process to lipoprotein assembly in the liver. Specific objectives include the isolation and purification of Sterol Carrier Proteins and delineation of the relationship of these substances to lipoprotein assembly. Other objectives include the determination of chemical structures of intermediates in cholesterol biosynthesis, as well as the mechanism of inhibition for hypocholesterolemic drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Alzheimers disease (AD) is the most common late-onset neurodegenerative disease. It is known to have strong genetic influences. Over the last decade using advanced high-throughput genotyping/sequencing technology, researchers have identified nearly 30 AD susceptibility genes/loci. However, these loci account for only a portion of AD heritability. Furthermore, almost all of them have been identified for the risk of developing AD, where disease status is the primary outcome of interest. In our paper (Li et al. AJHG 2002) we highlighted the hypothesis of genetic predisposition for age-at-onset (AAO) of AD, estimating the heritability for AAO of AD to be ~42% and reporting linkage to new genetic quantitative trait loci for AD AAO. Recently, two genome wide association studies for AAO of AD confirmed findings in the APOE gene, but otherwise, there was little overlap. Overall, much remains to be discovered about genetic factors for AAO of AD. If AAO genes can be identified, they will contribute new knowledge about the genetic modifiers of AD, provide excellent intervention targets for delaying the onset of AD, and improve our ability to predict AAO in an individual. The increase in large genome- wide genetic datasets for AD, provides an excellent resource for the comprehensive investigation of the genetic basis of AAO of AD through a well-conceived statistical analysis plan and methods development. Our central hypothesis is that genetic variants exist that regulate the variation of AAO and/or one's age-associated risk. To test this hypothesis, we will use datasets from the Alzheimer's Disease Genetics Consortium (ADGC) and Alzheimer's Disease Sequencing Project (ADSP). These two large datasets will allow us to search for common and rare variants for AAO of AD. Our analysis strategy is unique from previous studies, because we will not only analyze AAO as a quantitative trait in a case-only design as used in the past, but we will also treat AAO as a censored trait by applying novel survival-analysis methodology. The survival-analysis approach will include unaffected subjects whose AAO was censored at the age of enrollment, and allow us to identify genetic variants that are associated with AD risk in an age-dependent manner. To uncover novel genes tied to AAO and develop predictive models of AAO of AD, we propose the following aims: (1) Conduct discovery analysis in the ADGC dataset to identify common and/or rare variants associated with AAO of AD; (2) Replicate AAO findings in the independent ADSP dataset and additional cohorts; (3) Develop a suite of analysis programs to support the proposed analysis in Aims 1 and 2 and accommodate the different data structures included in the Discovery and Replication datasets; and (4) Develop a polygenic risk score model to predict AAO of AD. This proposal promises to provide targets with the potential to delay onset of AD and to advance the future development of personalized medicine utilizing individual genetic burden to predict AD AAO. Furthermore, it will provide innovative tools for data analysis and risk modeling to the genetic research community.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We plan to study two groups of 5-8 adolescent chimpanzees, each group living in a 1 1/2-acre enclosure partially simulating a natural habitat. The initial focus will be on the adaptation of each group to its new environment. Thereafter the principal focus will be on changes in behavior that occur during adolescence and beyond, noting sex differences and individual differences. We will especially focus on changes in behavior in the sphere of aggression, sex, and attachment, and hormonal correlates of these behavior patterns. We expect to delineate these patterns, to measure changes in their frequency and duration over the years of adolescence, and to search for endocrine correlates. We will differentiate such behavior patterns by age; sex; phase of menstrual cycle; hormonal condition as occurring spontaneously and as manipulated experimentally; and environmental conditions. Our semi-natural chimpanzee facility will permit repeated endocrine measures over several years; the administration of hormones for experimental purposes; continuous, close-range observation of behavior; a simple social system focusing on peer relations; systematic comparison of behavior in this semi-natural habitat with behavior of chimpanzees in their fully natural habitat and with human adolescents in roughly similar situations. Hormone assays will be conducted on a regular basis over the next two years including measurement of several androgens, estrogen, progesterone, the gonadotropin LH, and biogenic amines. The research will be conducted by an inter-disciplinary group: ethology, psychiatry, biochemistry, statistics, and sociology. There will be a long-term, collaborative arrangement between the Stanford research and a longitudinal study of chimpanzee behavior in the natural habitat in Tanzania.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Mutations in parkin are largely associated with autosomal recessive juvenile parkinsonism (AR-JP). The underlying mechanism of pathogenesis in parkin-associated Parkinson's disease (PD) is thought to be due to the loss of parkin's E3 ubiquitin ligase activity leading to accumulation of parkin substrates due to failure of the ubiquitin proteasome system. A large number of possible parkin substrates have been identified, yet their role in the pathogenesis of PD due to parkin mutations have yet to be clarified. Moreover, the post-translational modifications that potentially regulate parkin's function are not known. We propose to generate and characterize parkin knockout mice to test the role of proteasome dysfunction in PD. Furthermore, we propose to characterize and identify parkin substrates, and identify potential post-translational modifications of parkin that regulate its function, thus providing important new information about the role of UPS dysfunction in PD. To accomplish these goals we propose the following specific aims. In Specific Aim #1 we will generate and characterize parkin knockout mice. In Specific Aim #2 we will evaluate the sensitivity of parkin knockouts to proteasome inhibitors. In Specific Aim #3 we will characterize the Role of Nitrosative Stress on Parkin Function. In Specific Aim #4 we will identify additional and potentially authentic parkin substrates in parkin knockout mice in response to proteasome inhibition. Identification and characterization of parkin substrates and regulatory mechanisms of parkin function may provide novel therapeutics and targets to prevent the toxic effects of this familial associated gene in the degenerative process of PD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Adolescents are at particular risk for acquiring HIV, and increasing numbers of young African American women are affected by the epidemic. We have developed a new theory-based model from the literature to investigate the impact of provider-patient interaction, mutual exchange of information and psychosocial development on the return for recommended visits to the Fulton STD clinic in Atlanta, Georgia. Previous studies indicated that health care providers could influence adherence to treatment and management of chronic diseases in adults. Most of these constructs have not been operationalized or tested with adolescents. Some questions concerning consumer opinions of providers have been adapted to the adolescent population especially with respect to patient satisfaction and access to care. In addition, a model of adolescent psychosocial maturity suggests that interpersonal skills may increase the ability of adolescents to communicate with providers. However, there is little information about the impact of providers on adolescents who are being treated for STDs to increase their likelihood of returning for future STD visits. For our last submission, we piloted the questionnaire with 60 African American adolescent girls and young women from the Fulton Clinic, obtained feedback about the draft questionnaire, and found that 33% of participants indicated high mutuality scores. After the second review, we have adjusted the plan in response to reviewers to refine our outcome variables, address potential confounders and developmental issues, and incorporate focus groups in the initial and final phases of the project. This project \"develops and tests new methods\" under the NIMH RO3 grants program, using a unique window of opportunity to complement the currently NIMH funded grant (R01 MH61210) designed to test an HIV prevention intervention (the HORIZONS Project) at a state supported STD clinic that serves a large population of African American adolescent girls aged 15-19 years old. This new initiative will develop and test culturally and developmentally sensitive measurement tools specifically for adolescent girls being treated for STDs concerning the association of provider-patient relationships and mutuality of exchange of information with return for subsequent health care visits. If these measures are useful, then an intervention can be designed to enhance provider-patient mutual exchange of information and reduce the future risk of STDs and HIV infection.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Natural products are extremely important sources of bioactive compounds for agricultural and pharmaceutical applications. Enzymes involved in secondary metabolism hold great potential as biocatalysts that may be used in the efficient synthesis of fine chemicals and high value pharmaceuticals. In this collaborative work between a metabolic engineering group and a structural biology group, we will harvest this potential towards the one-step synthesis of the blockbuster drug simvastatin (Zocor.). Simvastatin is currently synthesized from the natural product lovastatin via inefficient, multistep processes. Our proposed biosynthesis of simvastatin will result in a completely novel process that can be an attractive alternative over the current chemical routes. The central enzyme in this study is LovD, an acyltransferase from the lovastatin biosynthetic pathway. We have performed extensive, preliminary biochemical characterization of this enzyme to show that LovD is a simvastatin synthase, and can be potentially engineered into a powerful biocatalyst for simvastatin biosynthesis. This proposal will examine the following specific aims: AIM 1: Directed Evolution of LovD. We will use directed evolution methods to improve the catalytic efficiencies of LovD towards simvastatin synthesis. We have developed a high throughput screening assay based on the formation of simvastatin. AIM 2: Structure-Based Engineering of LovD. The X-ray crystal structure of LovD will be pursued in the Yeates Lab. Rational mutagenesis of key residues identified from structural analysis will be performed to probe LovD function and improve LovD catalytic properties towards simvastatin synthesis. AIM 3: Metabolic Engineering of E. coli as a whole cell biocatalyst. We will engineer the multidrug transporter system of E. coli to improve its efficiency in exporting simvastatin to the extracellular space. This will improve the conversion of the whole cell reactor at high product concentrations. AIM 4: Direct Biosynthesis of Simvastatin from A. terreus. We will metabolically engineer A. terreus to be blocked in lovastatin biosynthesis, but robust in simvastatin biosynthesis. Project Narrative We have proposed biochemical and structural studies to investigate a simvastatin synthase recently identified from our laboratories. We will use protein and metabolic engineering methods to develop a whole cell biocatalyst that can biosynthesize simvastatin. This work will represent an important milestone in biocatalysis, application of enzymes towards the synthesis of a compound as commercially important as simvastatin has not been reported. The successful outcome of engineering a natural product biosynthetic enzyme into a useful biocatalyst may lead to additional efforts to examine this class of fascinating enzymes from a biocatalysis prospective. At the same time, the outcome of the proposed work will provide important scientific insight into protein engineering, enzyme structure and function, E. coli membrane transport, and Aspergillus metabolism.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed MARC U*STAR Program at the Rio Piedras Campus seeks to improve an already exemplary program by increasing the number of trainees who enter prestigious graduate programs and with new strategies designed to ensure that more of its trainees will be in positions to enter into the mainstream of the nation's scientific community. To this end, the Program will continue to recruit highly talented students and provide them with a mentored intramural research experience that includes a well-defined project that leads to results which will be presented at scientific meetings or published and which will be described in a tesina. Emphasis will also be placed on an appropriate summer research at an external site, counseling and assistance in graduate school applications. In order to meet these objectives it will continue to seek positions for 34 trainees. It will establish direct communications with research supervisors who have recruited its ex-trainees in order to gain feedback on research training strategies. It will provide its trainees with workshops that improve their communications skills, particularly English writing, and will emphasize their participation in technical workshops. The Program also seeks funding to accelerate the development of the entry level Biology and Mathematics courses and to further improve those in Chemistry. These activities will not only increase the retention rate and reduce the time to graduation of all Biology and Chemistry majors, but in five years should increase the pool of MARC eligible rising Juniors by at least a 100 students. A series of science promotions will be included in order to direct more of these into careers into biomedical research. [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This Facility provides a variety of molecular biology techniques, genotoxicity tests and general tissue and bacterial culture techniques, as well as training in these, to AHFCC members. The Facility is both responsive and proactive to the community it serves. Room 201 in Unit 2 is used for tissue culture and molecular biology and room 324 for bacterial work. The Facility has equipment needed for basic molecular biology: small centrifuges, speed vacs, electrophoresis equipment for general analysis sequencing, and blotting. A tissue culture hood and incubator are in the large of the two laboratories. Artek counters are used to score the Ames and UDS assays. To tube-based thermocyclers are available for PCR experiments. Other major equipment includes an oligonucleotide synthesis unit and Pharmacia SMART HPLC system for micro-purification of proteins. The senior investigators requested the following new services: expression of human and mouse genes using Clontech's 588 Atlas arrays; 32/P-post- labeling the DNA adducts and the single cell gel electrophoresis (COMET) assay for their repair, and Telomerase assay. The requested deep freezer will enable us to offer controlled long-term storage of plasmids, bacterial and mammalian cells. We shall work with the Histopathology Facility to develop analytical routines for image analysis using the requested fluorescence/transmission microscope upgrade and software system. Services currently available include the requested fluorescence/transmission microscope upgrade and software system. Services currently available include: The Ames Salmonella mutagenicity assays, with or without metabolic activation. The 7000 strains, containing defined mutations, will be made available. The hepatocyte DNA repair assays (Williams' test) uses human or rodent hepatocytes in vitro and rats in the in vivo/in vitro version. Oligonucleotide syntheses include carcinogen- modified nucleotides. DNA analysis via Souther blot hybridization and sequencing of enriched PCR analysis. Individual RNA analysis via Norther blot hybridization will be supplemented by the Clontech array. Protein analyses include Western blotting and protein-DNA interactions via gel retardation assays and micro-purification. We supply S9 and bacterial and mammalian cultures. The two excellent technicians provide an experienced and stable resource of experimental techniques and train other AHFCC personnel in tissue culture methods and molecular biology techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this research is to define murine B cell subsets by surface markers, to determine the functional capacity of each subset with regard to stimulation requirements and effector molecules secreted, and to determine the interrelationships of the subsets. We will search for differences in mitogen receptors, and differentiation antigens and will relate these to known differences in membrane immunoglobulin and complement receptors. The functional capacity of each subset will be measured by stimulation with panels of mitogens and antigens known to elicit different types of responses in mixed cell systems. Neonatal suppression with anti-Lyb reagents should allow determinations of developmental relatedness of each subset. Finally, we will determine whether each subset has the ability to produce any immunoglobulin or whether limitations exist.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Glaucoma is the second most common cause of blindness worldwide. Our broad, long-term objectives are to investigate innovative technologies and methods that will accurately and reproducibly provide the earliest possible evidence of glaucoma and its progression so as to prevent blindness. We have improved on the program that we have pursued for the past ten years in a number of ways, but importantly we have assembled a unique collaborative group for this project. This team includes ophthalmologists, engineers, computer scientists and statisticians, as well as appropriate support personnel, from the University of Pittsburgh, Tufts University, Massachusetts Institute of Technology and Carnegie Mellon University. We have brought excellent investigators from disparate fields to bring new insights, knowledge and skills from outside of ophthalmology to bear on innovations in technology for glaucoma disease and progression detection. We will accomplish this via cross-sectional and longitudinal studies using cohorts of healthy, glaucoma suspect and glaucomatous subjects. Our Specific Aims are to (1) detect the earliest possible evidence of glaucomatous damage and progression. We will compare objective, quantitative ocular structural measurements obtained by ocular imaging and functional measurements, to test the prediction that changes structural functional change, and to characterize those changes, (2) advance optical coherence tomography (OCT) software innovations that assess the intra-retinal layers in the peripapillary and macular areas as well as the optic nerve head (ONH). This aim includes employing innovative image processing and image analysis techniques, as well as new techniques to improve scan quality post hoc, (3) identify the particular clusters of clinical characteristics distinct to specific glaucoma diagnostic technologies resulting in the earliest detection of glaucoma and its progression. We will use innovative automated machine classifiers and state-of-the-art statistical methods which use the best combination of parameters generated by the imaging devices in order to determine the optimal use of each device in assessing disease and progression, (4) advance micron-scale tomographic imaging using OCT for improved understanding of the anatomical and biomechanical properties of the ONH and intra-retinal substructure in the macular and peripapillary regions in health and in glaucoma. These are key areas involved in the glaucomatous process. This experiment is designed to improve our understanding of glaucoma and potentially create a new glaucoma diagnostic. Swept-source and ultra-high speed spectral- domain OCT will be used to obtain detailed information in the ONH, lamina cribrosa and retina. Rapid image acquisition by these devices will minimize OCT scanning artifact, and modulation of OCT light source wavelengths will allow optimization of imaging at various tissue depths. We expect that these studies will lead to our ability to detect glaucoma and its progression earlier than ever before with high sensitivity and specificity, enabling early intervention to prevent glaucoma blindness.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Hutchinson-Gilford Progeria Syndrome (HGPS) is a premature aging disorder that is lethal by age 16. It is caused by a defect in a nuclear envelope protein called Lamin A. Lamins are structural proteins that line the inner surface of the nuclear envelope. Humans have two types of lamins: A-types (Lamin A and C) and B-types (Lamin B1 and B2). All lamins, except Lamin C, are processed for membrane anchorage by the addition of a hydrophobic prenyl (lipid) group at the C-terminus. But for unknown reasons, Lamin A is further processed to remove the anchor. Thus, Lamin A is transiently rather than stably prenylated. Mutations that cause HGPS delete a recognition site in the Lamin A protein required for removal of the anchor. This defective Lamin A protein is called progerin, and is stably rather than transiently prenylated, resulting in organism-wide rapid aging by unknown mechanisms. The enzyme thought to be responsible for removing the prenyl anchor from Lamin A is called Zmpste24, which is a zinc-dependant prenyl protease. The experiments described in this proposal will establish the fruit fly Drosophila as a model to study human Lamin A processing. It is possible to incorporate human genes into the fly genome (so-called transgenic flies). When transgenically expressed in flies, normal human Lamin A is stably, rather than transiently, anchored in the fly nuclear envelope, with no apparent ill effects. This is in contrast to the human situation, in which stable membrane anchorage of Lamin A leads to disease, and suggests that no conserved prenyl proteases in flies are removing the anchor from human Lamin A. Thus the Drosophila nuclear envelope can serve as an ideal environment in which to confirm the interaction between human Lamin A and the protease Zmpste24, which is a critical step that goes awry in HGPS. In Specific Aim 1, normal or processing-defective human Lamin A proteins (including progerin) will be transgenically expressed in fruit flies together with human Zmpste24, and the localization patterns of Lamin A examined with immunofluorescence microscopy. Drosophila larvae possess huge, easily manipulated nuclei in their salivary glands, which permit detailed dissection of nuclear protein dynamics. Since human Lamin A is confined to the nuclear envelope in flies, co-expression with Zmpste24 should release membrane anchorage and cause re-localization into the nuclear interior, thus demonstrating a direct interaction between these two proteins. Specific Aim 2 comprises a genetic and molecular dissection of the Drosophila prenyl protease functions, firstly to confirm they do not interfere with the human processing assay from Aim 1, and secondly to establish the degree to which lamin processing is conserved in flies. Results from this work will support the use of Drosophila as a non-human in vivo environment in which to study human lamin biology, and will reveal underlying generalities about the nuclear envelope and its role in aging. PUBLIC HEALTH RELEVANCE: Hutchinson-Gilford Progeria Syndrome is a devastating and extremely rare premature aging disorder that kills children before the age of 16. The research described in this proposal will contribute to world-wide efforts to find a cure or at least a palliative. But it is also very probable that unraveling the biology of this unusual disease will go some way to explaining the root causes of aging in general, a genetic disease which affects us all.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Objectives. The goal of this project is to train a DVM/PhD anatomic pathologist in the field of translational biomedical research to facilitate transition to an independent established investigator. The research questions addressed in this fellowship application focus on the paucity of information regarding the pathogenesis of HIV- and FIV-induced oral disease in juveniles, and the corresponding lack of effective therapies to treat these syndromes. The aims described in this proposal will investigate the virologic impact, contributing factors, and pathogenic mechanisms of this juvenile condition, thereby providing baseline data to establish a novel approach and more efficacious treatment strategy in HIV-infected children. Specific Aims. 1) Characterize oral kinetics of in vivo FIV infection and the relationship to oral dysbiosis, immune dyscrasia, and oral lesion development. 2) Evaluate treatment response of retroviral-induced oral lesions to HAART and microbial-targeted therapy. Relevance to Public Health. Common therapies used to treat HIV-induced oral disease in adults are often unsuccessful in children, resulting in frequent disease recurrence and resistant microbial infections. There is a lack of information regarding virologic effects and HAART on the oral microbiota. It is of exceptionally high impact to define the pathogenesis of juvenile HIV associated oral disease, and test practical, attainable therapies for outcomes that result in reversal of underlying pathologies. Functional understanding of the pathogenic effects of virus-induced immunosuppression on oral disease in HIV-infected children is the first objective of this proposal. Implications of HAART and symptomatic therapy for gingival-stomatitis will be tested in Aim 2 to guide appropriate therapies for this complex disease syndrome. Collectively, this proposal will provide a focused analysis of both cause and cure, aimed to substantially improve management of this condition for more than 2.5 million HIV-infected children. Experimental design. Specific pathogen free (SPF) cats will be orally inoculated with a strain of FIV that reproducibly produces oral lesions. FIV viral and proviral loads in blood and saliva will be monitored over time and compared with hematologic data. Oral lesion development will be evaluated grossly and histologically, and changes in the oral microbiota and cytokine expression will be evaluated by 16S rRNA NGS and MIA, respectively. Following collection of baseline data, changes in viral/proviral status, oral microbiota, oral cytokine expression, and oral lesions will be measured in the presence and absence of highly active antiretroviral therapy (HAART). The synergistic effect of HAART on the resolution of oral lesions will be subsequently measured in the presence and absence of a novel microbial-targeted therapy comprised of dental hygiene, antimicrobial, and probiotic treatments.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research takes a social psychological perspective to gender differences in group interaction and performance. Previous research has documented consistent sex differences in group members' style of interactions, such that females (vs. males) display a greater amount of positive social - emotional activity and males (vs. females) display more task-oriented activity. Yet we know little about the origin of these differences or their consequences for task performance. The first two studies evaluated the extent to which gender differences in interaction (a) are manifestations of males' (vs. females') greater instrumental traits and females' (vs. males') greater expressive traits, (b) arise from the perception that males are generally more competent than females, and (c) arise from the perception that males have greater legitimate authority than females. A number of researchers have argued that group task performance is highly dependent on aspects of group interaction (e.g., Steiner, 1972), and thus gender differences in interaction would be expected to have implications for performance. Consistent with this idea, preliminary work by the PI suggests that all-male (vs. all-female) groups' higher task-oriented activities and lower positive social activities facilitate performance at brainstorming tasks. The third study will extend this work to assess whether all-female (vs. all-male) groups' interaction style facilitates performance at discussion tasks. The fourth study will then explore the link between the interaction style of mixed-sex groups and task performance. Finally, the fifth study will use meta-analytic techniques to estimate the magnitude of gender differences in group performance in prior research and to examine the relation between the size of this difference and attributes of the group task and number of group members.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Anticipated impact on Veterans' Healthcare: This project will strongly impact VHA patient care because: (a) VHA devotes significant resources to long term care, (b) the prevalence of dementia in long term care facilities is high, (c) the prevalence of dementia in the veteran population is high and is increasing, and (d) the VHA has identified pain control as a priority clinical and research area. Project Background: The goal of this study is to develop a pain intensity measurement strategy for persons with dementia and limited ability to self-report pain. Though many available instruments capture information about the pain of those with limited ability to self-report, there remain four major barriers in developing a practical and clinically relevant pain intensity measurement strategy for this vulnerable population: 1) uncertainty regarding the specificity of some behaviors typically counted as indicative of pain; 2) lack of a measure that goes beyond identification of pain presence to estimate pain intensity; 3) lack of an inviolable criterion or gold standard; and, 4) uncertainty regarding whether clinicians' globa ratings of patients' pain add incremental validity as compared to using observational pain behavior checklists alone. The research proposed here addresses these barriers. Objectives: Phase I: Building a Pain Behavior Measure for Persons with Dementia. Specific Aim 1: Develop an efficient and specific measure comprised of behavior observations that best discriminate pain from other constructs (e.g., agitation, boredom). Specific Aim 2: Evaluate the validity of item response theory-calibrated PIM-D scores in the cross-sectional sample of NH residents. Phase II: Responsiveness and Incremental Validity of Pain Measures. We will measure pain and relevant additional variables pre- and post-analgesic therapy in X nursing home residents who have dementia and limited ability to self-report. Specific Aim 3: Evaluate the responsiveness of the PIM-D to analgesic therapy. In a sample of 50 NH residents who receive analgesic therapy, a) evaluate the responsiveness of PIM-D scores, and b) compare responsiveness of the PIM-D to that of existing behavioral pain measures. Specific Aim 4: To evaluate the incremental validity of PIM-D scores compared to existing pain behavior measures and compared to nurse and NA gestalt pain intensity ratings in predicting changes in Expert Clinician Pain Evaluations. Project Methods: Item reduction will be achieved using a modified Delphi method with an expert panel. The total patient sample will be comprised of residents from 4 large VA Community Living Centers (CLCs; Philadelphia, Coatesville, Atlanta, Tuscaloosa). We will enroll veterans with moderate to severe dementia. Study measures will include evidence-based observer-recorded indicators of pain (pain-related behaviors, clinical gestalt pain ratings), and known correlates of pain (sleep, agitation, and depression). All participants will also be clinically assessed by a nurse practitioner.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A recent focus of development biology has been to isolate and study genes through to make critical decisions of cell fate and patterning in embryogenesis. Genetic studies in Drosophila indicate that genes containing the homeobox DNA-binding motif participate in such decisions.Other genes such as the vertebrate Myo-D gene, also have the compelling properties of developmental regulators. We propose to study regional specification and pattern formation in Xenopus embryos using as a focus the Xhox-1 homeobox genes and the Myo-D gene. In previous work we devised an overexpression assay to address the function of the Xhox-1A gene and demonstrated that it is likely to be involved in somite formation. Somitogenesis is a primary segmentation event which shapes the vertebrate body play. This finding is significant because it is the very first indication of a function for a vertebrate homeobox gene and it fulfills the broad expectation that homeobox genes will participate in critical developmental events. We propose to study further the role of Xhox-1A and a linked gene. Xhox-1B, in somite formation. We will use antibodies to investigate at cellular resolution the normal expression pattern of these genes. Furthermore, we will design dominant-acting mutants of the Xhox-1 genes which will be used to antagonize the action of the endogenous genes and thus perturb somitogenesis in new ways. We will also approach homeobox gene function at the biochemical level by studying the binding of Xhox-1A protein to DNA. We believe that in the long term this biochemical approach will reveal how stable developmental changes are affected. In Xenopus, most of the somite tissue becomes muscle. Nothing is known about how the patterning of muscle tissue (somitogenesis) meshes with commitment to the muscle lineage. Evidence suggests that the Myo-D gene is instructive in this commitment step. We will characterize the expression of the Xenopus Myo-D during early development and attempt to devise functional assays to test whether Myo-D gene expression can commit multipotent, embryonic ectoderm directly into the muscle lineage. These experiments are designed to investigate specific examples of cell commitment and patterning in vertebrate development. In addition, we will take a more general approach to the fundamental problem of how regional specification occurs in embryos. We will screen a pre-gastrulation cDNA library with probes for developmentally interesting genes such as other homeobox genes, believing that many of them will show restricted spatial expression and will indicate zones of cell commitment at early stages. The strength of this proposal lies in the fact that we already have a functional assay for the homeobox genes that we study and in the fact that we can relate these studies to the wealth of cellular and developmental information available for Xenopus embryos.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The radiation sensitizer Misonidzaole (MISO) can selectively potentiate the effectiveness of certain conventional chemotherapeutic agents in vitro and in vivo. In preliminary experiments we have demonstrated that this chemopotentiating effect of MISO can be significantly augmented by mild hyperthermia (41 degrees C, 1 hr) under oxygen-deficient, but not aerobic conditions, suggesting that the addition of hyperthermia to drug-sensitizer combinations might result in a significant therapeutic advantage. The overall objectives of the proposed research are 1) to evaluate this hypothesis in vivo comparing enhancement of tumor response (KHT sarcoma) and normal tissues damage when MISO-drug combinations are administered in conjunction with whole-body hyperthermia, 2) define parameters influencing the interactions among the three variables in vitro in controlled experimental environments and 3) determine the mechanism(s) of the interactions and the influence of oxygen concentration on their expression. Experiments indicate that hyperthermia, independent of its effect on the chemotherapeutic agent, increases the chemopotentiating efficiency of a dose of MISO, possibly by increasing nitroreductase (NR) activity, an oxygen sensitive reaction. This enhanced chemopotentiating efficiency might prove clinically significant as the success of chemopotentiation in patients may be limited by an inability to deliver adequate MISO doses. The proposed research will make extensive use of statistically- motivated experimental designs to determine the significance of temperature, oxygen-concentration, sensitizer dose, and sequencing on expression of chemopotentiation using cell survival, tumor-growth delay and nitroreductase activity as endpoints. Other experiments will utilize the multi-cell tumor spheroid model and selective trypsinization techniques to determine the effect of treatment on aerobic and hypoxic populations exposed simultaneously and to determine whether thermal enhancements are possible at MISO concentrations and pharmaco-kinetics achievable in the clinic. These later experiments will make use of a spheroid treatment device designed to permit pharmacokinetic modeling in vitro. Finally, the effect of heat on the chemopotentiation of nitrosourea-resistant, chemopotentiation- resistant Mer+ tumor cells (about 80% of human tumors express the Mer+ phenotype) will also be evaluated as preliminary experiments indicate that the addition of heat can overcome the resistance of this phenotype.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "OBJECTIVE: Obesity and chronic sleep deprivation have both become increasingly pervasive medical problems in recent years. Average sleep time has decreased over the last century by two hours. Chronically sleeping less has been associated with increased weight, various endocrine changes such as an increase in cortisol levels, a decrease in growth hormone levels and changes in the adypocite-derived hormone leptin and the gastric produced hormone ghrelin that, together may increase appetite and food intake. Immune alterations associated with obesity and sleep deprivation include subclinical inflammation, and a decreased resistance to infections, respectively. We have recently demonstrated that increased weight in subjects suffering from sleep disturbances and depression is associated with an increase in proinflammatory cytokines and with a decreased in anti-inflammatory cytokines. Such changes are detectable in the serum as well as in the sweat, as non invasively collected over 24h by the means of a sweat patch, specifically developed by us to study endocrine immune relationships in freely living subjects. The general goal of this proof-of-concept, controlled trial is to investigate the impact of increasing sleep time on endocrine and immune parameters in chronically sleep-deprived, obese subjects. STUDY POPULATION: 18 to 50 year old, obese (BMI 30-55) men and premenopausal women, who usually sleep less than 6 hours, mainly recruited from the Baltimore-Washington metropolitan area. Sleep duration will be assessed by the use of sleep logs and actigraphy. Secondary causes of sleep deprivation such as insomnia, psychological (depression), and medical conditions associated with poor sleep quality (including obstructive sleep apnea) will be exclusionary criteria. DESIGN: This is a randomized, 12-month duration, comparison-controlled clinical trial of a minimum additional 90 min of sleep per night (Intervention Group) or continuation of habitual short sleep schedule (Comparison Group). The proposed treatment is an educational and behavioral intervention aimed at increasing sleep in a non-pharmacological fashion. The main analysis of the study will be to determine if additional sleep will result in changes in the endocrine and immune profiles opposite to those associated with chronic sleep deprivation. Specifically, we hypothesize that 12 months of sleep extension will result in weight loss and subsequent corrections in the following hormonal levels: leptin, adiponectin, ghrelin, cortisol, and growth hormones, among others. The immune and inflammatory profile will be characterized by a decrease in C-reactive protein, TNF-alpha, IL-6 and other inflammatory cytokines, and a return of anti-inflammatory cytokines to a healthy range. At the end of the 12-month intervention study (Phase 1, Efficacy Study), all participants will be given information about the potential benefit of more sleep and encouraged to increase sleep time. Health teaching about proper nutrition and adequate exercise will also be provided at that time to the Comparison Group. All participants will be evaluated 6 months later to assess the effects of this intervention in a real-life situation (Phase 2, Effectiveness Study). As of August 31, 2008 70 subjects have been randomized. Two third of the participants are women and ore than 50% are minoritites. Subjects'retention in this prospective study has been excellent with less than 15% of subjects interrupting the study before completion. Most common reasons for interruption include move to a different geographical area and the clinical need for bariatric surgery or the need for drastic change sin life style (diet, physical exercise) above and beyond the standard of care. OUTCOME PARAMETERS: body weight, food intake, hormones, and cytokines.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The present R13 grant requests support for a joint Gordon Research Conference and Gordon Research Seminars (GRC/GRS) on Cannabinoid Function in the CNS to be held in 2013. We will exploit the innovative GRC/GRS format to accomplish three Specific Aims: 1. To promote and sustain a scientific forum that allows for in depth presentation and adequate discussion of cutting edge unpublished data in the area of Cannabinoid Function in the CNS. 2. To foster opportunities for cutting edge collaborative science that propels the next generation of scientific advances and unites basic researchers studying cannabinoid-related neuroplasticity with clinical researchers studying long term impact of altered neuoroplasticity in disease states. 3. To promote and mentor the next generation of cannabinoid scientists while encouraging diversity and inclusiveness in the cannabinoid field through the Gordon Research Seminar. The joint meeting format is warranted based upon the critical role of the endocannabinoid system in controlling neuronal excitability, the contribution f endocannabinoid dysregulation to disease states, the therapeutic potential of endocannabinoid modulators and the increasingly widespread use of both recreational (e.g. spice) and medicinal (e.g. Sativex, medical marijuana) cannabinoids in humans. The recent advent of cannabinoid- based medicines requires that continued research emphasis be placed on understanding cannabinoid effects in humans, especially long-term consequences (i.e. abuse liability, adverse side-effects, safety considerations). Clinical studies of cannabinoids have not been represented at any prior Gordon Research Conference on cannabinoids, mandating that program organizers specifically reach out to clinical researchers that would not typically attend a Gordon Research Conference focused on basic as opposed to clinical science. The format of the GRC thus provides an important opportunity to build bridges in interdisciplinary science between preclinical researchers studying neuroplasticity and clinical researchers studying human disease states marked by altered neuroplasticity (e.g. pathological pain, addiction, neurodegenerative diseases). The Gordon Research Seminar (GRS 2013: Endocannabinoids in Neurophysiology and Neuropathology) is organized by and for graduate students and post-doctoral fellows and has been highly ranked by trainees as a venue for providing mentoring and networking opportunities at a critical stage of trainee development. The GRS directly precedes the GRC (GRC 2013: Synapses, Circuits and the Human Brain). R13 support of the 2013 joint GRC-GRS is expected to promote mutually beneficial increases in diversity in the cannabinoid field. The present R13 grant is expected to foster opportunities for cutting edge collaborative science while ensuring the continued success and sustainability of a highly rated GRC devoted to cannabinoid research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application requests a further five years of support for an Institutional National Research Service Award to support multidisciplinary post-doctoral training in Genetic Epidemiology and Molecular Genetics and Neurobiology. We request support for six postdoctoral fellows (2 M.D. and 4 Ph.D.) for training in Genetic Methodology, Family Epidemiology and Behavior Genetics, Gene Mapping and Bioinformatics, Molecular Genetics, and Molecular Neurobiology. Recruitment will be staggered over two years to assure a high caliber of fellows. In addition to training in a primary area, fellows will be encouraged to obtain a broad understanding of the diverse skills in Psychiatric Genetics to facilitate their collaboration in (and leadership of) cross-disciplinary research teams. The fellowship will usually last three years, but one or two years may suffice for those with much pertinent experience. Fellows with a wide variety of backgrounds will be recruited including; Psychology, Psychiatry, Genetics, Statistics, Mathematics, Anthropology, Sociology, Biology, and Neuroscience. The training program uses an apprenticeship model, combining research under the mentorship of one or more experienced mentors with more formal training through seminars, didactic courses and individual reading. Major strengths of the program are; (i) the participation of a large multidisciplinary group of well-funded preceptors (n=l8) with expertise in statistical and computational genetics, molecular genetics and neuroscience; (ii) the study of quantitative and qualitative traits and the development of methods for the analysis of multivariate phenotypes and (iii) the availability of major epidemiological and genetic data sets (phenotypes and genotypes). Fellows may participate in ongoing linkage and candidate gene studies of Schizophrenia, Bipolar Disorder, Alzheimer's Disease, ADHD, Alcohol Dependence, Electrophysiological measures of CNS activity, Personality traits and Nicotine Dependence. Preceptors in this program have approximately 62 federally and non-federally funded grants providing many opportunities for training in all aspects of Psychiatric Genetics. The program is located in one of the nations leading Medical Schools with a rich array of basic and applied genetic research studies and educational opportunities. Thus, we expect the long tradition of successful mentoring and research training of scientists and physician-scientists from diverse intellectual backgrounds to continue.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Abstract - IICA With this current proposal, the Inter-American Institute for Cooperation on Agriculture (IICA) seeks to build on previous successes and implement high-impact interventions that directly reduce risks associated with importing food into the United States. Specifically, we propose to develop and test alternative technologies to greatly augment the availability of high-quality training opportunities using the existing Produce Safety Alliance (PSA) curriculum and other training materials, such as the On-Farm Readiness Review (OFRR), in support of the FSMA Produce Safety Rule (PSR). We will leverage the wide availability of mobile telephones throughout the Americas and develop web- and application-based training materials that not only incorporate requisite training per the curriculum but also provide easy links to relevant portions of the rule and other important documents so that the user can refresh their knowledge as needed. While a variety of training materials are available in English, we propose to strengthen the reach and impact of existing and new materials by making them available in Spanish as well. This approach will take advantage of the prevalence of access to mobile technology throughout the Americas to create a group of tools that address the needs of adult learners and non-traditional students. We will use well-tested approaches and learning techniques to ensure sustainable knowledge building and understanding around FSMA implementation. Working with partners in academia, we will design controlled experiments to test the efficacy of alternative training materials against in-person training techniques to better understand what is and what may be possible to achieve through alternative approaches. Using an iterative process, we will develop alternative training materials, package them into new delivery methods, and test and validated both the materials and delivery approaches through pilot projects. Based on the results of these pilot projects, we will modify and further test our approaches, with the goal of releasing final, validated products to a large, hemispheric audience. These ?final? products will then be evaluated over the life of the project to ensure their continued efficacy and provide inputs for corrective actions. IICA is the specialized agency for agriculture within the Inter-American System. Counting on over seventy-five years of experience and a network of over thirty offices across the Americas, IICA is an ideal partner to support FDA to address technical capacity needs around FSMA implementation by foreign governments and their industries in Latin America. IICA is and has been a strong U.S. government partner in Latin America and the Caribbean for many years and has the demonstrated capacity to implement FSMA training in the region.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Many cancer patients who are candidates for high dose chemotherapy do not have available a suitable related or unrelated donor of bone marrow or peripheral blood progenitor cells. Human umbilical cord blood (CB) is a unique source of transplantable hematopoietic cells that offers an alternative source of hematopoietic cells for these patients. The use of CB cells to date, has primarily been limited to small pediatric patients due to the low cell numbers in CB products. The focus of this proposal is to optimize ex vivo culture of CB to provide increased numbers of cells to enable the use of CB in adult patients. In addition, optimization of the expansion cultures will enable more rapid hematopoietic and immune recovery post transplant and potentially minimize other complications associated with transplantation of CB. The first aim of these studies is to evaluate improved expansion culture conditions in clinical trials. Preclinical studies will also be performed to evaluate other culture conditions that may provide enhanced platelet recovery. In addition, analysis will be performed of primitive cells in the expanded products as an indication of the effect of expansion on long term engrafting cells (stem cells). The second aim will optimize transduction cultures of expanded cells and subsequent clinical studies will incorporate these conditions to use marked expanded cells in patients. This will allow the evaluation of the contribution of expanded cells to long to engraftment. Correlative analysis will be performed comparing the contribution of the marked cells in patients with in vitro assays of primitive progenitor cells to determine the predictive value of the in vitro assays. The third aim will evaluate the immune recovery in patients receiving ex vivo expanded cells. Both phenotypical and functional analysis of T cells subsets will be performed The final goal of these studies is to determine culture conditions for the expansion of T cells in vitro for use a source of donor lympohocytes to induce an anti tumor effect in patients who relapse. In the absence of cultured T cells the patients who relapse have no alternate source of donor lymphocytes and would die from disease progression. The outcome of these studies will provide us with a better understanding of the potential clinical utility of CB. Optimal expansion cultures will enable the use of CB in adults and provide more rapid engraftment. Overall these studies will lead to improved outcome in these patients.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Leptin (L) and its receptor (L-R) are key players in the regulation of energy homeostasis and body weight. Complex formation between leptin and the extracellular portion of L-R results in the activation of Janus kinase 2 (JAK2) that is constitutively bound on the intracellular regions of the receptor. Leptin-instigated JAK2 signaling in hypothalamic nuclei reduces food intake and stimulates energy expenditure, while functional defects in either leptin or L-R result in morbid obesity, hyperglycemia, decreased insulin sensitivity, and hyperlipidemia. Despite the crucial impact of the leptin system on body weight and other physiological responses, little is known about the structure of the L/L-R complex and its association with JAK2. One of the reasons for this lack of insight is that both L-R and JAK2 have a relatively long and flexible multi-domain arrangement that has proved to be very challenging for both large-scale purification and implementation of X-ray crystallography. The present proposal aims to overcome these limitations in addressing the architectural prerequisites of L-R signaling by applying single- particle cryo-electron microscopy (cryo-EM) to characterize the holo-complex of full-length L/L-R and JAK2. Single-particle EM has emerged as a very powerful tool for the characterization of dynamic protein assemblies in relatively small concentrations and without the need for crystallization. We anticipate that the application of single-particle EM techniques on this system will reveal the architecture of the L/L-R assembly and JAK2 independently, and in complex. Given the underlying importance of this membrane-localized signaling complex in obesity, energy metabolism, and heart disease, the structural results obtained will be of very broad biomedical interest. Considering the current lack of structural information on any receptor/JAK complex, our studies will provide the general architectural framework for understanding how extracellular ligand binding on cytokine receptors results in intracellular JAK activation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Research Summary (For the funded R01 GM118553 proposal). Malaria, a disease caused by parasites of Plasmodium species, re- mains one of the most relevant infectious diseases; in 2015 over 200 millions of individuals had clinical malaria and over 500,000 individuals, mainly children, died from it. The infection starts when a Plasmodium-infected mosquito injects into the skin a small dose of sporozoites, a speci?c form of the parasite, which travel via the blood to the liver, infect hepatocytes, and form liver stages. Several vaccine candidates, including the most re- cent RTS,S vaccine, are aimed at eliminating sporozoites from the skin, blood, or hepatocytes. However, the low ef?cacy of such vaccines highlights the problem with lack of basic understanding of how Plasmodium sporozoites are eliminated by host immunity. CD8 T cells, a subset of lymphocytes, have been shown to play an important role in preventing clinical malaria by eliminating Plasmodium liver stages, speci?cally in radiation attenuated sporo- zoites (RAS)-based vaccines. Using intravital imaging, we have recently discovered that activated CD8 T cells form clusters around Plasmodium-infected hepatocytes in mice and that these clusters are important in parasite elimination. Mechanisms driving the formation of such clusters remain poorly de?ned and how activated CD8 T cells eliminate liver stages from the whole liver is not well understood. Another layer of complexity arises as the level of immunity needed for protection depends on a speci?c host-parasite combination. By combining math- ematical modeling and experiments we will provide quantitative insights into potential mechanisms that explain contribution of CD8 T cells to elimination of Plasmodium liver stages in mice. We will provide such insights via three complementary speci?c aims. In speci?c Aim 1, we will discriminate between alternative mechanisms of formation of CD8 T cell clusters around sporozoite-infected hepatocytes (T-cell intrinsic vs. T-cell extrinsic), de?ne the role of T cells, speci?c to irrelevant antigens, in the formation of clusters, and quantify the impact of T cell cluster size on the ef?ciency at which liver stages are eliminated. In speci?c Aim 2, we will determine the impact of structure of liver sinusoids on the ef?ciency of CD8 T search for rare sporozoite-infected hepatocytes and de- termine the speed at which moving CD8 T cells can localize the site of infection. Finally, in speci?c Aim 3 we will discriminate between alternative mechanisms for a larger number of memory CD8 T cells required for sterilizing protection against exposure to Plasmodium yoelii sporozoites as compared to Plasmodium berghei sporozoites. Completion of these aims will lead to a better understanding how CD8 T cells localize and eliminate Plasmodium liver stages from one of the major peripheral tissues, the liver. This understanding may help in designing more ef?cient immunization protocols of RAS-based malaria vaccines. In addition, deeper understanding of the mech- anisms by which CD8 T cells eliminate infections at peripheral sites may be also useful for the improvement of several others CD8 T cell-based vaccines such as those against HIV, HCV, and HSV.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Arterioles undergo major morphological changes during vasoconstriction. These changes consist of formation of an orderly pattern of periluminal ridges whose size and number vary systematically with the level of constriction. The cellular interactions which produce these changes have not been studied in detail, hence little is known about how the arteriolar wall coordinates rearrangement of its components as the vessel changes its diameter. The purpose of the present study is to explore the relationships between the degree of microvessel constriction, mural configuration, and the frequency and distribution of myoendothelial junctions (MEJ's). The working hypothesis is that points of attachment between the arteriolar endothelium and smooth muscle (i.e. MEJ's) provide focal areas of force transduction between these two cell types, and thereby determine deformations in the wall which occur by selectively exerting force along the luminal perimeter. The present study seeks to address this issue by undertaking an ultrastructural analysis of MEJ's in both dilated and constricted arterioles. Knowledge of the mechanical interactions between the cell of the arteriolar wall is important to our understanding of the pathology of hypertension and other cardiovascular lesions (e.g. cerebral vasospasm). To obtain arteriolar vessels for ultrastructural analysis, male golden hamsters are anesthetized with sodium pentobarbital (IP) and tracheostomized. Supplemental anesthetic is administered through a femoral vein cannula. The left cheek pouch is gently exposed, prepared for intravital observation as previously described (Duling, 1973) and suffused with a physiological salt solution at pH 7.4. Arterioles displaying appropriate responses to vasoactive substances are selected as experimental vessels. Vessels dilated with adenosine or constricted with norepinephrine are fixed for ultrastructural analysis in situ by immersing the entire pouch in fixative. Thin cross sections of arterioles are cut, stained and examined with a TEM. The frequency and distribution of MEJ's present in both dilated and constricted microvessels will be compared. Morphometric analysis of vessel cross sections will determine: 1) If MEJ's are labile entities, 2) any organized array of MEJ's associated with the periluminal ridges of constricted vessels, 3) the type of MEJ's extant between the endothelium and smooth muscle (e.g. tight junction, gap junction, etc.) and 4) the cellular origin of the process which ultimately produces the MEJ.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The approximately 1 ,000 G protein-coupled receptors (GPCRs) of humans mediate key signals - triggered by photons, odorants, hormones, and neurotransmitters - in brain, heart, blood vessels, white blood cells, and virtually every organ and endocrine gland. The fact that most of these signals represent potential targets for drug therapy justifies the broad-based strategy of this proposal, which aims to understand the conserved molecular mechanisms responsible for transmitting G protein-mediated signals between signaling molecules, in vitro and in the context of the cell. The first two aims test relations between structure and function of the GPCR and the G protein trimer at the level of individual molecules. Experiments with GPCRs aim to: a. use molecular probes to determine how the extracellular surface of a GPCR actually binds the activating ligand; b. engineer metal binding sites that activate a GPCR by inducing coordinated movement of its transmembrane helices, allowing us to infer how the natural ligand induces similar movements; c. identify sites on the GPCR's intracellular surface that interact specifically with peptides representing different parts of the trimeric target. To understand the conformational changes in a G protein trimer that mediate its activation by the GPCR, a second set of experiments will: a. test the hypothesis that the GPCR uses the beta-gamma subunit of the G protein trimer as a lever to open a route for bound GDP to exit from its binding pocket in the alpha subunit and thereby activate the trimer; b. determine how key structural elements of the G protein alpha subunit cooperate during the activation process, by constructing metal binding sites that restrict movements of these elements, relative to one another. The third set of experiments uses fluorescent probes and fluorescence energy transfer (FRET) to determine the locations of G protein alpha and beta-gamma subunits in intact cells and ask how hormonal activation affects their interaction. Biochemical experiments with pure G protein subunits indicate that activation in the test tube causes the alpha subunit to dissociate from the beta-gamma heterodimer; it is not known, however, whether such a dissociation accompanies activation in an intact cell responding to a hormone. Investigation of this question begins by constructing functioning G protein alpha and beta-gamma subunits attached to fluorescent tags; the fluorescent subunits are used to assess their subcellular distributions in intact cultured cells, and FRET between co-expressed tagged beta-gamma and alpha subunits will reveal whether hormones cause them to dissociate.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION (adapted from the abstract): Caring for critically ill patients suffering from multiple organ dysfunction syndrome (MODS) represents one of the most difficult challenges in critical care medicine. The high mortality of adults and pediatric patients with MODS warrants a greater understanding of pathophysiology at the molecular level. Elucidating the molecular mechanisms that cause MODS is a daunting task because critically ill patients are typically subjected to multiple, simultaneous, and/or sequential injurious stimuli. The \"multiple hits\" hypothesis of MODS states that the interactions of multiple stimuli, and the unexpected cellular responses that they may generate, can ultimately lead to organ failure. The investigators propose to undertake a reductionist approach whereby they will study the in vitro interactions of two fundamental, yet distinct, cellular responses that are ubiquitous during various forms of critical illness: the heat shock response, and activation of the transcription factor NF-kappaB. They have demonstrated that the heat shock response, a primitive cellular defense mechanisms, inhibits activation of NF-kB, which is involved in the regulation of many proinflammatory genes. Preliminary data indicate that inhibition occurs by two mechanisms. The proximal mechanism involves inhibiting phosphorylation and subsequent degradation of the NF-kappaB inhibitory protein, I-kappaBalpha. The distal mechanism involves heat shock response-mediated de novo expression of the I-kBa gene. Specific Aim I is designed to determine the mechanism by which the heat shock response inhibits phosphorylation of I-kBa. Specific Aim II is designed to determine the mechanism by which the the heat shock response induces expression of I-kBa, and proposes a fundamental reclassification of I-kB as not only an inhibitor of NF-kB, but also as a novel heat shock protein. Specific Aim III is designed to determine if heat shock response-mediated expression of I-kBa independently inhibits activation of NF-kB. By elucidating the mechanisms proposed in this application, they will provide novel insight regarding the interactions of these two fundamental cellular responses. Ultimately, it is hoped that this mechanistic insight, at the molecular level, will lead to a greater understanding of mechanism at the physiologic level, which in turn can form the basis of more rational and specific therapeutic strategies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY/ABSTRACT Incident epilepsy is more common in the elderly than at any other time of life. While some cases are due to stroke or known Alzheimer?s disease (AD), many cases have no known etiology. This proposal hypothesizes that late-onset epilepsy (LOE; epilepsy starting at age 60 or later, in the absence of stroke or other identified cause) is associated with cognitive decline and preexisting neuropathology such as amyloid deposition; i.e., LOE may be a marker for future cognitive impairment and dementia. The research will use data from the ongoing longitudinal Atherosclerosis Risk in Communities (ARIC) cohort study (Aims 1 and 2), as well as from a new cohort recruited as part of this grant (Aims 3 and 4). The specific aims are: 1) To test the hypothesis that late-onset epilepsy (LOE) is associated with cognitive decline, and the subsequent development of MCI or dementia. 2) To test whether A? 42/40 ratio is associated with LOE. 3) To determine whether individuals with clinically-identified LOE have lower cognitive scores than individuals without LOE. 4) (Exploratory) To compare AD biomarkers in patients with and without LOE, and correlate subclinical epileptiform activity with cognitive performance in patients with LOE. Defining whether there is an association of LOE with an elevated risk of future cognitive impairment could allow patients to have earlier interventions, which may help slow future cognitive decline. This is an application for a Mentored Patient-Oriented Research Career Development Award (K23). The overarching training goal of the proposed project is to provide the candidate with the skills necessary to become an independent researcher in epilepsy related to aging. Specific training goals are to obtain advanced training in epidemiology and biostatistics; to obtain training in recruiting and leading cohort studies; to acquire and apply knowledge of AD pathophysiology and the use of biomarkers; to gain didactic and experiential training in cognitive assessments in neurodegenerative diseases; and to advance professional development skills in preparing manuscripts and presenting at conferences. The career development plan includes classes in biostatistics and epidemiology at the Johns Hopkins School of Public Health; regular meetings with mentors; seminars in biostatistics and the epidemiology of aging; seminars in AD pathophysiology and research; and attending/presenting at national conferences. The candidate?s primary appointment is in the Johns Hopkins Department of Neurology, an extremely supportive environment with a strong history of junior investigators successfully transitioning to independent investigators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Genetic or mutational theories of cancer are supported by the mutagenic nature of most nonviral carcinogens, the age distribution of many cancers, the existence of genetic cancers such as bilateral retinoblastoma and the increased cancer incidence in some diseases as xeroderma pigmentosum. The development of techniques of gene isolation by DNA-mediated gene transfer in cultured animal cells provide a powerful tool for the analysis of the genetics of cancer. It has been shown that DNA from some human tumor-derived cell lines can transform normal cells in culture. We have also found that tumor cell lines derived from different human lung and colon carcinomas contain the same or very similar transforming gene. This gene has been found to be the human homologue to the Kirsten murine sarcoma virus oncogene (vi-Ki-ras). Our experiments describe the isolation of the transforming gene present in human colon and lung carcinomas. At present, we have molecularly cloned in 47.1 vectors over 40Kb of the c-Kirsten ras-2 oncogene from NIH3T3 tertiary transformants induced by DNA of two human lung tumors propagated in nude mice. The cloned sequences include the 5' end of the oncogenes (containing adjacent mouse host sequences) and four coding exons. Work is in progress to isolate the 3' end of the oncogenes including the fifth exon. We are currently sequencing the cloned exons of the genes in order to determine the mutation(s) which could be responsible for the activation of this oncogene in the human tumor cells. The availability of these genes will allow the comparative study of the types of human carcinomas containing the same transforming gene; the study of their structure and function, in an effort to understand the mutational events involved in these particular types of tumors; and the study of their role in the tumorigenesis in humans.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "(Adapted from investigator's abstract) The reliability of surgical and examination gloves has become a major concern among health care professionals as HIV and other viral diseases have spread. Current testing regulations and methods are not adequate to insure the level of protection which is now being demanded by the industry. The proposed research will test the feasibility of a production test using volatile halogenated compounds to detect and quantify leaks in surgical and examination gloves. Theoretical calculations show that these gases can detect holes that are one order of magnitude smaller that viruses. A halogen tracer gas test could provide a cost-effective and reliable way to test surgical and examination gloves. Research will focus on the development of the apparatus and methodology to test gloves, quantification of the technique parameters, and proposals for new standards.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Tat gene of human immunodeficiency virus (HIV) plays a central role in the activation and life cycle of HIV. Tat exerts its effects at the level of transcriptional initiation and elongation. We have previously reported the direct physical interaction between the HIV-1 Tat protein and the basal transcription factor, TBP/TFIID. Affinity chromatography demonstrated that wild type Tat, but not a transactivation mutant of Tat, was capable of depleting TBP/TFIID from cell extracts. These experiments represented the first demonstration of a basal transcription factor that binds, in an activation-dependent manner, to Tat. We now report that the Tat-TBP interaction can be detected in HIV-1-infected cells. The domain of TBP interacting with Tat has been mapped from amino acids 163 to 196 using deletion and site-specific mutants of TBP. This domain of TBP, which includes the H1 and S2 domain, is distinct from the H2 binding site for other activator proteins such as E1A. The interaction of Tat with TFIID regulates the binding of accessory proteins to TFIID. Tat stabilizes the interaction of TFIID with TFIIA in a gel shift assay. In addition, Tat competes for Dr1 interaction with TBP. Our results suggest that the basal transcription factor, TBP/TFIID, represents an important regulatory molecule in HIV transcription. The human immunodeficiency virus type 1 (HIV-1) transactivator Tat protein is essential for efficient viral gene expression and virus replication. Tat interacts with the basal transcription factor TFIID. This interaction is mediated through the amino acid 36-50 core domain of Tat. We now demonstrate that soluble peptide analogs of the Tat core domain are able to effectively block LTR transactivation. In cotransfection experiments, Tat peptide analogs inhibited Tat transactivation of an HIV-1 LTR-CAT reporter construct up to 80-fold. Inhibition of control promoters was approximately two-fold. Tat peptide analogs inhibited HIV virus replication by 85 percent in latently infected U1 cells induced with Tat. While both short and long peptide analogs (amino acids 36-50 vs 36-72) inhibited Tat transactivation in transient assays, the short peptides were more effective inhibitors of virus replication in U1 cells. Furthermore, U1 cells treated with the Tat peptide analog showed markedly delayed virus transmission when co-cultivated with parental U937 cells. The Tat peptide analog did not decrease expression of cellular genes including beta-actin, GAPDH and histone H2Bb.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Influenza, a vaccine-preventable disease, remains as an important cause of morbidity and mortality worldwide. All age groups are affected, but children experience the highest disease incidence while adults suffer the most serious disease complications and related mortality. The burden of seasonal influenza epidemics and the continuous threat of a potential pandemic influenza highlight the need for effective preventive strategies. During influenza epidemics, children are typically affected early and they serve as disease vectors, introducing influenza into the community. Influenza vaccines have proven to be efficacious in the prevention of disease in vaccinated individuals and it has been hypothesized that immunization of a significant proportion of children could have major beneficial effects in the community, through the interruption of influenza transmission. Therefore, school-based influenza immunization programs have been proposed as prevention models. Nevertheless, the effectiveness of these approaches has not been convincingly demonstrated in large scale interventions. If proven effective, school-based influenza immunizations could become one major strategy for prevention of both seasonal and pandemic influenza. In October 2005, a large school-based influenza immunization campaign was initiated in Knox County, TN. The campaign was successfully implemented, immunizing 24,281 (46%) school-aged children attending public schools in the county. The campaign achieved at least similar coverage in the subsequent 2006-2007 influenza season. Although this campaign was set up as a feasibility demonstration project, its impact on disease outcomes has not been examined. We hypothesize that this large 2-year intervention decreased the incidence of influenza related diseases in both vaccinated and unvaccinated Knox County residents. Vanderbilt investigators are currently performing active viral surveillance in Knox County (intervention) and Davidson County (control) to compare viral activity during the second year of the campaign (2006-2007 season). We propose to perform additional and complementary analyses using alternate data sources to measure the overall intervention effectiveness (direct and indirect effects) over both intervention years. We will utilize large electronic databases that systematically record healthcare encounters in the State of Tennessee, and the infrastructure and expertise of the New Vaccine Surveillance Network (NVSN), a prospective surveillance system for viral infections in children that is currently operating in Tennessee. The proposed research and career development plan complement Dr. Grijalva's training in Medicine, Public Health and Epidemiology. Furthermore, this proposal supports his efforts to develop the expertise needed to become an independent investigator focused on prevention of influenza-related morbidity and mortality and on vaccine program and policy evaluations. [unreadable] [unreadable] [unreadable]", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose a high risk/high impact study to develop an innovative circulatory support strategy using an implantable pump for the treatment of single functional ventricle. The most common form, hypoplastic left heart syndrome, in which the left ventricle fails to form in a way that is ever functional, is the leading cause of death from all birth defects in the first year of life. Currently, surgical repair is performed with three separate operations. In the first operation, blood flow to the lungs must be derived from a systemic-to-pulmonary arterial shunt to overcome the potentially elevated newborn pulmonary vascular resistance. The shunt, however, makes this procedure notorious for instability and mortality. In the second and third operations, the shunt is disconnected, and blood flow to the lungs is accomplished by connecting the vena cava to the pulmonary arteries. Stability and survival are greatly improved. Unfortunately, the second and third procedures cannot be safely performed in the newborn due to the unsolved obstacle of achieving passive blood flow through the reactive pulmonary circulation. We propose a unique application of a blood pump to temporarily augment flow from the great veins through the lungs until the potential for reactivity resolves. This process normally occurs most rapidly by 2 weeks and is nearly complete at 8 weeks of life. The problematic shunt is avoided, and the newborn pulmonary vessels are perfused by a low-pressure, high-volume flow from a systemic venous source similar to normal right ventricular hemodynamics. We hypothesize that under these conditions, the neonatal pulmonary arteries will mature normally. Once the transitional pulmonary vasculature has remodeled sufficiently to permit a systemic venous source to perfuse the lungs passively, the pump can be safely removed. We further hypothesize that the neonate will tolerate the systemic venous pressure elevation required to permit passive venous flow through the lungs. Using a newborn animal model of univentricular circulation, we will test these hypotheses through the following specific aim: To transition an assisted univentricular circulation (cavopulmonary pump + systemic ventricle) to an unassisted univentricular circulation in which the lungs are perfused by passive vena caval flow alone. We have generated pilot data which support feasibility of this innovative technique using currently available technology. We expect that successful completion of this approach will convert the current high risk three-stage surgical strategy to a much safer single-stage paradigm of newborn Fontan palliation of single ventricle cardiac disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Humans harbor a substantial Natural Killer (NK) cell compartment in their secondary lymphoid organs. These NK cells are enriched for immunoregulatory CD56brightCD16- cells, which are preferentially activated by dendritic cells (DCs). Our preliminary data demonstrate that NK cell activation by DCs restricts B cell transformation by the Epstein Barr virus (EBV) in vitro, especially when the NK cells are derived from tonsils, the secondary lymphoid organ of primary EBV infection. EBV is a human tumorvirus, which establishes latent infection in more than 90% of the human adult population. It causes tumors of epithelial and B cell origin in immune competent, and at increased frequencies in immune compromised individuals. The characterization of innate immunity to EBV is of particular interest, because failure of this initial immune control might result in increased viral titers and massive T cell expansion during primary immune responses, resulting in infectious mononucleosis. In order to characterize the role of DC/NK cell interactions during EBV infections, we plan to: 1. Characterize innate NK cell activation during EBV infection. We will dissect DC/NK cell synapse formation, DC activation by EBV derived dsRNA, and tonsillar DC subsets for their activation by EBV. 2. Analyze the protective effector functions of tonsillar NK cells. We will focus on cytokine production, cytotoxicity and assistance in protective T cell polarization by NK cells after DC activation via EBV infection. 3. Investigate EBV infection in vivo. In order to translate our in vitro findings into an in vivo model of primary EBV infection, we have reconstituted human immune systems in immune compromised mice, which could mount primary protective immune responses against EBV. This model will be explored to investigate NK cell expansion and activation during primary EBV infection, the contribution of NK cells to innate immune control of EBV, and their assistance in T cell polarization of adaptive immune control of EBV. These studies will characterize the mechanisms of NK cell activation by DCs, and protective NK cell functions against B cell transformation by oncogenic EBV in vitro and in vivo. Knowledge of the establishment of protective EBV specific immune control might suggest vaccination strategies against common EBV associated tumors like nasopharyngeal carcinoma and Hodgkin's lymphoma. PUBLIC HEALTH RELEVANCE: Epstein Barr virus (EBV) causes tumors of epithelial and B cell origin in the human population, like Hodgkin's lymphoma and nasopharyngeal carcinoma. This proposal plans to investigate how two cell types of the innate immune system, dendritic cells and Natural Killer cells, interact to initially control EBV and influence other immune cells to establish life-long protection from the development of EBV associated malignancies. An understanding of these initial events during the establishment of EBV specific immune control will help us explain why some individuals develop symptomatic acute infection with EBV, called infectious mononucleosis, and identify immune compartments that should be harnessed to efficiently vaccinate against EBV associated tumors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Approximately 28,000 individuals in North America have a subarachnoid hemorrhage from ruptured aneurysm each year. Most are young, healthy individuals with good neurological function after the hemorrhage who have the potential for returning to a vigorous and productive life. Many will develop devastating neurological deficits or perish, mostly from vasospasm and bleeding. Less than 60% of those who reach hospital will have a favorable outcome. There is considerable room for improvement. The long term objectives of this study are to decrease the mortality and morbidity of aneurysmal subarachnoid hemorrhage, principally by obtaining information which will lead to more effective management strategies for vasospasm and rebleeding. One approach entertained for decreasing these complications is early surgery which has many theoretically attractive features but remains one of the most controversial topics in neurosurgery. Accordingly, the principal aim of this study is to determine at which interval after subarachnoid hemorrhage surgery results in the most favorable outcome. Other specific aims include: determination of risk factors for rebleeding and the timing of these disastrous events; determination of the safety and effectiveness of antifibrinolytic agents and development of an administration schedule which optimizes the actions in preventing rebleeding and minimizes ischemic neurological complications; documentation of the diagnostic and prognostic value of CT scanning in subarachnoid hemorrhage; and development of a more reliable and universally acceptable neurological grading scale for patients with ruptured aneurysm.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although the fine structure of malarial parasites has been studied by many investigators, there remain many unanswered questions. Re-examination of the morphology of the parasites by the high voltage and scanning electron microscopes has been rewarding for understanding of the various subcellular organelles of malarial parasites. During the past year, we studied by electron microscopy the effect of immune serum on sporozoites of P. cynomolgi and P. berghei. When the sporozoites were incubated in immune serum, the surface coat became very thickened, while normal serum showed little effect on the sporozoite. Scanning electron microscopy indicated that the anterior end of the parasites was free of coat deposition and that the circum-sporozoite precipitate (CSP) reaction was resulted from coat deposition at the posterior end of the sporozoites. For the coming year, we will study microgametogenesis of P. gallinaceum and P. berghei and the effect of immune serum on the microgametes by electron microscopy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development of safer and more effective antipsychotic drugs requires the availability of sensitive probes of the dopamine (DA) receptor, which is a site of action of these agents. Research Biochemicals proposes a program to prepare novel substances with selective agonist or antagonist action at specific DA receptors for use by researchers in neuropharmacology. The objectives of Phase I research are: 1) the synthesis of novel aporphines and related compounds with irreversible binding activity at DA receptor sites. Structures will be formulated with particular emphasis on selective activity at presynaptic DA autoreceptors. Irreversible binding will be accomplished with alkylating and/or photoactivated functional groups. 2) the incorporation of highly selective DA receptor ligands into a practical affinity column. The prototype of this product will be a clebopride-functionalized support for use in isolating DA D2 receptors. 3) the preparation of hitherto unavailable enantiomerically pure pairs of known DA agonists and antagonists to permit stereochemical exploration of receptor surfaces. The new products generated in Phase I studies will be evaluated for commercial introduction to the expanding neuroscientific community.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recent studies in our laboratory indicate the presence of reactivity in normal and abnormal human sera by radioimmuno-precipitation proteins from type-C viruses of mice, cats and primates, analogous to reactivities that have been observed in normal sera of mice. We propose to further characterize these reactions to learn if there is any specificity for their presence or absence in malignancy and autoimmune disease. Reactivities of additional mammalian type-C viruses with human sera will be tested and family studies involving patients with diseases which might be related to type-C viruses based on animal models will be undertaken. Absorption with various leukemia and other tumor cells as well as normal cells will be performed, as well as further characterization of the reactant components in sera and viruses. A search for viral glycoproteins by surface radioiodination of leukemia cells, and viral studies of cultured leukemia cells are also planned. These studies will be conducted in parallel with a basic research program of regulation of type-C virus expression in the mouse. We feel there is considerable complementarity and synergy between the human and mouse programs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ViraCore is a biotechnology company that is creating a drug discovery tool that is capable of presenting complex receptors in a nanometer-sized particle format. Specific ally, ViraCore is using \"lipoparticles\", nanometer -sized particles surrounded by a lipid bilayer and embedded with membrane-bound receptors, to purify and solubilize complex receptors from the cell surface while maintaining their structuraI integrity. The purpose of this proposal is to use lipoparticles to develop monoclonal antibodies directed to conformationally complex epitopes of membrane- bound receptors. Many of the receptors and epitopes that we will target are difficult or, in some cases, impossible to target using traditional methods, including activated receptor conformations and intracellular structures. Lipoparticles present a unique technology for the development of important new classes of antibodies and vaccine candidates. This proposal focuses principally on antibodies to chemokine receptors, a class of membrane-bound receptors involved in HIV infection, AIDS pathogenesis, and immunity. However, the results of this proposal can have a broad impact on the development of antibodies, vaccines, and therapeutics against a wide variety of complex receptors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The principal objective of these studies is to investigate the potential role of astrocytes in the pathogenesis of hepatic encephalopathy (HE). Since the primary morphological change in HE is characterized by astrocytic degeneration, a derangement in astroglia may be a contributing factor to the development of encephalopathy. Given that astrocytes possess membrane receptors which influence glial function, as well as provide the chief mechanism for neuronal-glial communication, we propose that encephalopathy results from a toxin-induced breakdown in glial-neuronal interaction. Accordingly, we plan to investigate the potential involvement of astrocytic receptors and their second messenger' systems (cAMP, inositol phosphate) in primary astrocyte cultures. The effect of toxins implicated in HE (ammonia, short-chain fatty acids, mercaptans, phenol), their possible synergism and the effects of serum from patients with HE, on the affinity (Kd) and number (Bmax) of receptors will be determined. We have chosen to study the adrenergic, benzodiazepine, serotonin and histamine receptors, as there is evidence of their specific involvement in HE. These experiments will consist of saturation studies of specific ligands for each of the listed receptors. Where appropriate, the ability of the receptor to form high-agonist affinity complexes will also be examined. In addition, the effect of toxin exposure on basal and receptor agonist-stimulated levels of cAMP and inositol phosphate will be evaluated. Finally, the ability of agonists, antagonists and agents that act directly on the second messenger systems to modify toxin-induced changes, will be determined using light and electron microscopy. These experiments will, for the first time, delineate the effects of HE-related toxins on astrocyte receptor systems. It is anticipated that the characterization of specific receptor systems changes will elucidate possible mechanisms involved in HE and may lead to the formulation of new therapeutic strategies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Significance Borna disease virus (BDV) infects a broad range of warmblooded species (mammals and birds) to cause neurologic dysfunction. Evidence from several laboratories indicates that BDV may be associated with human neuro-psychiatric disorders. Preferential infection of limbic circuitry, disturbances in D2 and D3 neurotransmitter systems, and the behavioral and motor disturbances that accompany acute BDV infection in adult, immunocompetent rodents and ungulates suggest parallels with serious psychiatric disorders such as schizophrenia, bipolar affective disorder, and autism. Attempts to demonstrate BDV infections in humans through serologic and molecular analysis of peripheral blood samples have produced inconsistent results. Objectives We hypothesize that perinatal infections of rhesus macaques will manifest as subtle neurobehavioral and neuropathologic disturbances in a primate counter part to the neonatal rat model of persistent BDV infection. The objective of this pilot study is to develop a nonhuman primate model of persistent BDV infection, and determine if the clinical, behavioral and neuropathologic sequelae are consistent with human psychiatric disease. Results Intracranial inoculation of BDV in 2 juvenile rhesus macaques resulted in infection and fatal clinical disease at 5 and 8 weeks pi, respectively. Both animals developed the acute, encephalitic form of BDV infection, indicating that animals of this age are already immunologically mature enough to preclude immunologic tolerance and persistent infection. Both animals developed strong humeral immune responses to BDV-specific antigens, thus providing valuable primate reagents for refining and validating available serologic tests. In one animal, preliminary PCR assays have detected viral nucleic acids in samples of peripheral blood mononuclear cells. Two neonatal rhesus were inoculated intranasally. One developed encephalitis and was euthanatized 9 week p.i. The second neonate did not become infected following intranasal inoculation. To better approximate the immune competence of the neonatal rat, intraventricular inoculation of 2 fetuses (~90-100 day gestation) have been pe rformed in utero, using ultra-sound guided techniques. One pregnancy resulted in live-born infant with severe congenital abnormalities and neurologic deficits that was euthanatized at 5 days of age. BDV was detected in tissues and the infant had high titer of anti-BDV antibodies. The second pregnancy resulted in a near term fetal loss. Ultrasound guided inoculation (IP or IC) was performed on 2 additional fetuses of 50 days gestation. The IP inoculation resulted in fetal death ~60 days p.i. The second fetus (IC inoc) is viable and appears to be developing normally at ~43 days p.i. Future Directions The last pregnancy will be allowed to go to term, and the neonate will be monitored for persistent BDV infection. Other strategies for inducing immune tolerance to BDV in macaques will be investigated. KEY WORDS borna disease virus, neurologic dysfunction FUNDING NIH RR00169 Pilot Study - In Progress", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a proposal for the continuation of a Research Scientist Development Award, now termed an Independent Scientist Award (KO2). During the first term of this award 60 experimental and control monkeys were created that had either normal or atypical prenatal exposure to androgen. Androgen exposures were of 40 days or less duration and critically timed to the start of the second or third trimester of pregnancy. Prenatal androgen exposure was reduced by twice daily administration of flutamide, an androgen receptor blocker. Weekly injections of testosterone enanthate were used to increase androgen. The proposed K02 Award will allow the PI to devote the majority of his time to investigating the behavioral, neuroendocrine, and cognitive consequences of these alterations in prenatal androgen exposure in socially-living rhesus monkeys. These studies will provide fundamental information about how early hormone exposure affects the development of sexual and sex-typed behavior in a primate with a complex social organization and a long period of sexual development. One of the most striking human cognitive sex differences is in aspects of spatial cognition. The proposed support will allow the PI to develop computerized approaches to studying spatial cognition in group-living monkeys to take advantage of a large sample of rhesus monkeys with atypical androgen exposure created during the previous funding period. In addition, the increased time available for research will allow investigation of possible sex differences in brain structure using magnetic resonance imaging. If neuroanatomical sex differences are demonstrated they can then be pursued in the experimental subjects as they near adult development. During the term of this award the experimental and control subjects will go through puberty and enter adulthood. A goal of the proposed plan is to explore how alterations in the prenatal androgen environment affect the process of pubertal change and adult reproductive behavior. Because none of our hormonal treatments substantially altered the genitalia of experimental females there are no apparent physical barriers to pregnancy. It will be particularly interesting to determine whether substantially less drastic alterations in prenatal androgen exposure than those previously used produce marked changes in reproductive and maternal behavior. In males that have experienced reduced prenatal androgen exposure the PI will determine whether the timing of such hormonal alterations differentially affects their sexual behavior. These studies of nonhuman primates have particular relevance to issues of human gender development, particularly the occurrence of gender dysphoria. The last goal of the proposal is to allow the PI to gain a deeper understanding of human gender dysphoria by spending time at the Clarke Institute of Psychiatry in Toronto Canada, which has one of the world's most active research and treatment programs for childhood and adult gender dysphoria. The combination of these efforts will allow the PI to expand his research into new areas, while increasing his understanding of its relevance to important human mental health conditions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To optimize isolation, transduction and expansion of lymphocytes from HIV-1 infected individuals under the conditions which will be used for a phase 1 study, to examine kinetics and stochiometry of ribozyme protection of primary cells, to determine if HIV antigen driven proliferation can be used to expand transduced cells, to obtain populations enriched in effector lymphocytes for reinfusion and to explore alternative methods of controlling HIV replication in infected cells during transduction of vectors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this proposal is to localize the various types of DNA (A:T-rich, G:C-rich, and repetitive sequences of DNA) and the various types of chromosomal proteins on the chromosomes of Drosophila and mammals. Several methods for staining chromosomal DNA according to its base composition at the electron microscope level of resolution are suggested. An immunological approach to protein localization of chromosomes is outlined. The location of specific proteins will be correlated with the location of specific types of DNA. These findings will be related to the biochemical structure of chromosomes. The information derived from this study should give some insight into the mechanism of gene control in eukaryotic organisms, the mechanism of chromosomal condensation, and the function of chromosomal proteins and unusual sequences of DNA.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are numerous factors that determine the biological response of tissues that contain radioactivity such as radiosensitivity, distribution of radioactivity, type and number of radiations emitted by the radionuclide, biokinetics of the radionuclide, repair time, etc. Traditionally, the mean absorbed dose to the tissue is calculated to correlate the biological response with mean absorbed dose. However, nonuniform activity distributions in tissue at the multicellular and subcellular levels result in nonuniform doses and therefore have made it difficult to adequately correlate biological response with mean absorbed dose. This is an important problem in diagnostic and therapeutic nuclear medicine. In the case of diagnosis, the risk of the radiation insult can in principle be drastically underestimated and potentially lead to increased risk of inducing cancer. In contrast, patients can be over- or under-treated in radionuclide therapy of cancer. Over-treatment or under-treatment in radionuclide therapy of cancer can have very adverse consequences in the final outcome for the patient. While calculation of absorbed dose at the cellular level has been advocated as a means to address this problem, this has largely remained a theoretical exercise. The applicants hypothesize that the biological response of tissues containing incorporated radionuclides can be correlated with absorbed dose when calculated at the cellular level. To test the hypothesis, a novel in vitro multicellular cluster model will be used which allows tight control over variables. Multicellular clusters will be assembled with mammalian cells containing radioactivity and the cell survival fraction as a function of cluster activity will be determined for several different radiopharmaceuticals which emit alpha particles, beta particles, or Auger electrons. Different percentages of the cells will be labeled with the different radiochemicals to ascertain the impact of nonuniform distributions of radioactivity at the cellular and subcellular levels. By controlling the percentage of cells labeled, this model will also be used to ascertain the role of bystander effects in the biological effects of incorporated radioactivity. These data and cellular dosimetry calculations will be used to develop a theoretical model to predict response based on cellular absorbed dose and bystander effects. The outcome of this research is expected to have a major impact on understanding and predicting the biological response of tumor and normal tissue to nonuniform distributions of radioactivity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The TrialNet Natural History Study of the Development of Type 1 Diabetes is a part of the TrialNet research program, in which a group of National Institutes of Health-sponsored researchers from the United States and Canada are trying to learn more about the development and prevention of Type 1 Diabetes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Mouse Phenotyping Shared Resource (MPSR) was established in 2001 to provide expert, readily available, and affordable experimental pathology support to OSUCCC investigators utilizing animal models of human cancer. The MPSR specializes in the morphologic characterization of newly produced lines of genetically engineered mice, but also provides a variety of other customized pathology services, including necropsy, slide preparation, semi-quantitative histopathology for experimental studies, morphometry, hematology, clinical chemistry, consultation and referral. The MPSR is headed by Donna F. Kusewitt, D.V.M., Ph.D., a board-certified veterinary pathologist and NCI-funded Investigator and staffed by Deborah Devor Henneman, B.S., each with more than 20 years of experience in carcinogenesis and experimental pathology. The MPSR is strongly supported through the excellent veterinary resources (clinical pathology, histology, and immunohistochemistry service) and veterinary pathology expertise (10 board-certified veterinary pathologists) available in the Department of Veterinary Biosciences (VBS) at The Ohio State University (OSU). Since its inception in 2001, the MPSR has provided high quality experimental pathology services tailored to meet the needs of the growing number of OSUCCC investigators who employ animal models of human cancer. During this time, 23 funded CCC members consulted on 38 projects requiring 1,042 hours of MPSR service, representing 75% of its entire workload. The MPSR has also provided requisite information for grant applications and manuscripts for many of OSUCCC members. The volume of work performed in the MPSR has steadily increased since the service was initiated and, given the projected expansion of OSUCCC research programs, a continued increase in MPSR usage is anticipated. Indeed we provide documentation of 37 CCSG-funded investigators with over 40 projects requiring MPSR support in the near future. Finally, the MPSR also serves as an essential venue for training future mouse pathobiologists as part of an NCRRfunded T-32 training grant, \"Mouse Pathobiology: Models of Human Disease\".", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary/Abstract Reversible protein lysine acetylation is a fundamental posttranslational modification observed in histone and non-histone proteins. Lysine acetylation can alter protein-protein and protein-DNA interactions, protein stability, and enzyme activation/deactivation. Among the major regulators of lysine acetylation is the histone deacetylase (HDAC) family. Of the 18 known human HDACs, 11 are metal-dependent hydrolases related to the acetylpolyamine amidohydrolases (APAHs). The HDACs contribute to the regulation of gene expression and many other critical cellular processes. Notably, abnormal lysine acetylation is observed in multiple human disorders, including cancer; thus HDACs are a validated drug target. Despite their critical biological functions and clinical roles as drug targets, little is known about the molecular basis for HDAC substrate specificity and inhibition. This is particularly the case for HDACs 10 and 11, which are the least well characterized of the metal-dependent HDACs. Our preliminary studies coupled with phylogenetic comparisons suggest that HDACs 10 and 11 may function as dual acetyllysine and acetylpolyamine deacetylases with unique substrate binding site architectures. However, how HDAC10 and 11 could accommodate small polyamine substrates as well as large protein substrates containing sissile acetyllysine moieties is unclear. In addition, while classic HDAC inhibitors such as SAHA are known to inhibit HDACs 10 and 11, the molecular basis for this inhibition is unknown as no HDAC10-inhibitor or HDAC11-inibitor complex structures are available. In fact, no structure of HDAC11 is available, despite the fact that HDAC11 represents a unique class of HDAC due to its limited sequence identity with other HDACs. We propose to study structure-function relationships for HDACs 10 and 11 to establish a molecular foundation for understanding substrate recognition, catalysis, and inhibition. Due to a lack of structural and mechanistic studies focusing on HDACs 10 and 11, we are currently unequipped to design HDAC isozyme-specific inhibitors. I propose to study the molecular mechanisms of HDAC substrate recognition and inhibition by (1) exploring the structural basis of HDAC10 substrate specificity; (2) defining the structural basis of HDAC10 inhibition; and (3) determining structure-function relationships for HDAC11. As mentioned above, aberrant lysine acetylation is a hallmark of certain cancers and other human disorders; therefore HDACs are critical drug targets. Currently, four broad-specificity HDAC inhibitors are FDA-approved for cancer chemotherapy, but isozyme-specific HDAC inhibitors are mostly unavailable. Our studies aim to better understand the structure and function of poorly characterized HDACs with the goal of facilitating the design of specific HDAC inhibitors for use in human disorders, particularly cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There are many diseases for which there are no vaccines and others for which the vaccines are not optimal or have significant side effects. The objectives of this project are to characterize viral antigens, determine targets of humoral and cell mediated immunity, and use this information to develop candidate vaccines. Live recombinant viral vaccines, DNA vaccines and recombinant protein vaccines are being developed. The vaccines are first tested in small animals and then in non-human primates. Presently we are working on vaccinia virus-vectored vaccines for HIV.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project is designed to test the hypothesis that specific areas of the brain such as substantia nigra and basal ganglia are especially vulnerable to oxidative stress and that their response to biochemical oxidative reactions (or oxidative stress) is defective in Parkinsons disease (PD) as well as in normal aging. Mitochondrial and synaptic membranes may lose their function due to inadequate response to oxidation an-or from direct damage induced by cytotoxic products of oxidation (e.g., cholesterol oxidation products). Levels of oxidation products and loss of function could be small under steady state conditions. Therefore, we will perturb the system by inducing oxidation of membranes in vitro (using primarily endogenous oxidants) in order to amplify existing differences. Our preliminary studies show that mitochondrial cholesterol is more oxidizable in PD than in controls. Mitochondrial and synaptosomal fractions will be oxidized by incubation with oxidizing agents (e.g., arachidonic and linoleic acid hydroperoxide plus metal ions like iron, mixtures of iron and ascorbic acid, nitric oxide with and without superoxide etc.). Different levels of oxidation of membranes can be defined by the extent of oxidation of two membrane lipids - tocopherol and cholesterol. Since the profile of cholesterol and tocopherol oxidation products would be characteristic for various types of oxidations, information on the mechanism of oxidation will be obtained. Synaptosomes and mitochondria from young and old rats, and PD and control human autopsy brain samples will be compared in all studies. Similar studies on oxidation of platelets (as an accessible model of neuronal membranes in humans) from PD and control subjects will be conducted. The extent of oxidation of membranes will be monitored and quantitated by determining the concentrations of tocopherol and cholesterol and their oxidation products as well as by estimating thiobarbituric acid reactive substances and the decline in total suIfhydryl concentrations. Oxidizing conditions which yield maximal differences between the control and experimental groups will be established first and then the cholesterol and tocopherol oxidation products will be isolated, identified, and quantitated by GC and GC-MS. A study of the effect of monoamine oxidase(MAO) on mitochondrial oxidation will be done by incubating mitochondria with dopamine, norepinephrine or serotonin (substrates for MAO) which releases in situ hydrogen peroxide and/or other oxidants derived from it. The oxidation will be followed and oxidation products characterized as with brain fraction& The effect of MAO inhibition by deprenyl upon the oxidation will also be studied. Number of similarities exist between the uptake of serotonin by platelets and brain. Serotonin uptake by platelets and synaptosomes at various levels of oxidation will be studied to examine the effect of oxidation upon membrane function. The data will be used to determine whether membrane damage induced by oxidative stress exists in PD and/or in normal aging. The data from the platelet studies may also provide a laboratory tool for the preclinical diagnosis of PD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Transformation in retroviral systems is characterized by abnormal cell growth and differentiation. Expression of the v-abl protein tyrosine kinase encoded by Abelson murine leukemia virus (Ab-MLV) induces transformation in pre-B lymphocytes. These transformed cells are arrested at the pre-B cell stage based upon their immunoglobulin (Ig) gene rearrangements and cell surface antigens. Cells transformed by a temperature sensitive (ts) mutant of Ab-MLV undergo a high frequency of Ig light chain gene rearrangement soon after inactivation of the v-abl kinase at nonpermissive temperature. In addition, up-regulated expression of at least three genes important to Ig gene rearrangement (RAG-1, RAG-2 and germline kappa gene) occurs at the nonpermissive temperature. This system provides an opportunity to study the pathways mediating differentiation arrest as a component of retroviral transformation. The goals of this study are: (1) To identify functional domains of v-abl protein which mediate differentiation arrest and the pathways used to transmit these signals. A series of abl mutants and protein tyrosine kinases will be used in genetic complementation studies to determine which functional domains mediate this process. To identify pathways by which differentiation arrest is mediated, over-expression of c-ras, c-raf and c-myc will be tested, as will dominant negative forms of these genes. Proteins to be tested will be co-expressed with ts v-abl in a single pre-B cell; the differentiation phenotype of these cells will then be assessed after inactivation of the ts v-abl kinase. (2) To determine what genes are up-regulated upon release from differentiation arrest. The v-abl protein appears to block light chain rearrangement by suppressing expression of genes important for this process. The increased expression of RAG-1, RAG-2, and germline kappa gene in ts transformants at nonpermissive temperature supports this idea. These events are necessary, but probably not sufficient, to initiate rearrangement. The differential display technique will be used to identify other genes with altered expression prior to Ig rearrangement. Combining efforts to investigate both mechanisms of regulation and their targets may contribute to a better understanding of normal pre-B cell differentiation and how disruption of this process contributes to the transformed phenotype. This may provide clues to therapeutic strategies for human diseases of abnormal B lymphocyte maturation, including immunodeficiencies and both abl and non-abl related lymphoid malignancies.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This three-year project tests certain strategies for inducing program changes and innovations in community mental health centers (CMHCs). The first is the improvement of evaluation technology through development of specialized program evaluation techniques specifically aimed to be more relevant and compelling in program decision-making and planning. Termed \"option-evaluation techniques,\" these methods involve the seeking out or clarification of program alteration opportunities, which are then evaluated with respect to expected output (\"yield\") and associated costs or resource factors. Shaping of specific evaluation techniques for needs-assessment, process studies and outcome studies to fit this approach are proposed. The second strategy is the development of program-change potential in other CMHCs through the use of technical assistance aimed at increasing their evaluation capability and use of evaluation findings. The technical assistance program involves intensive workshops, on-site visits, and computerized data analysis via remote terminal for a number of the nation's CMHCs. This effort also involves developing measures of \"evaluation achievement\" to use in testing for effects of the assistance program over time. A second feature is the use of measures of organizational \"readiness for evaluation\" as variables mediating the effectiveness of assistance in helping generate evaluation capability and utilization. A variety of non-experimental and quasi-experimental methods are used to determine relationships between evaluation achievement and readiness measures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal requests funds to continue studies of the reinforcing or euphoric property of opiates. The studies are designed to test the hypothesis that this property is mediated by the pathways activated during self-stimulation behavior and that these pathways constitute the \"normal\" channels for mediating motivational effects in the mammalian central nervous system. The studies fall into two categories, neurophysiological-behavioral and strictly behavioral. In the first group, single and multiple unit recordings are made in the awake, behaving rat during and after intraventricular or intracerebral application of opiates, opiate-like natural substances, and opiate antagonists. In these neurophysiological studies the neuron populations studied include those at rewarding sites, those whose spontaneous rate of discharge is depressed or enhanced during self-stimulation behavior, those that respond to naturally rewarding stimuli, and those containing monoamines. The goal is to sample for opiate action that correlates with behavioral stimulant effects and thus might be indicative of hedonic effect, in nerve cell populations defined on anatomical and functional grounds as possibly implicated in organizing the drive-reward underprinnings of motivated behavior. In the second group, rats are tested for self-stimulation after single or repeated opiate treatment at reward sites selected on the basis of a variety of behavioral parameters, for escape behavior from brain stimulation after opiate administration, for self-administration of opiates at reward sites, and for gross activity. Doses of opiates, schedules of drug administration are designed to produce a stimulant effect; so far as the activity test is concerned, the aim is to determine whether a stimulant action or dependence can be established in rats depleted neonatally of brain monoamines. The ultimate aim is to contribute to an understanding of opiate action on the one hand and to a central motivational mechanism on the other.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: This application is for the renewal of an NEI Core Grant in support of the research programs of 16 principal investigators holding 23 active NEI individual research grants and an NEI Institutional Training grant. The previous funding cycle generated 100 publications in the major areas of aging and diabetes, extraocular muscle biology and diseases, ocular immunology and inflammation, and retinal biology and diseases. The current investigators form the nucleus of the Case Visual Sciences Research Center (VSRC), a broader group of 40 vision researchers in 19 different basic and clinical science departments at the Case Medical Center (Case Western Reserve University School of Medicine and University Hospitals of Cleveland). Since our initial Core Grant award in 1997 and its renewal in 2002 with five Modules, these modules have continued to evolve over the current funding cycle due to a changing investigator group and scientific direction, expanded services and new technologies. The five modules that are proposed are: 1) Tissue Culture and Hybridoma, 2) Histology, Microscopy and Imaging, 3) Molecular Biology, 4) Specialized Animal Resource, and 5) Proteomics. Major new functions since our previous submission include hybridoma production, live cell imaging, laser capture microdissection, and a full service proteomics facility to acquire and analyze protein expression in ocular tissues. With the recent renewal of the T32 Training grant, these modules will also continue to serve as an integral resource for pre- and postdoctoral trainees in the Case Visual Sciences Training Program. The Visual Sciences at Case has receives very strong institutional support, including, construction of a centralized facility for Core Grant modules and functions, recruitment funds and supplemental support for technical salaries and supplies in the core facilities. The change in P.I. from Dr. Lass to Dr. Pearlman will ensure continuity in leadership and direction of the Core Grant, which will enable collaborative goals and objectives while enhancing individual programs, and facilitate Case VSRC investigators'continuing efforts toward analysis of fundamental mechanisms in ocular and visual system development, function, and pathogenesis of diseases of the eye and visual system.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: DNA damage in the form of apurinic/apyrimidinic (AP) sites is induced by cytotoxic agents resulting in base substitution mutations and blocks to DNA replication. The DNA base excision repair (BER) enzyme, AP endonuclease (APE), is a multifunctional protein involved in DNA base excision repair, oxidative signaling, transcription factor regulation, cell cycle control, and apoptosis. APE is essential for the repair of AP sites thus maintaining cellular and genetic integrity. Deficient expression of APE results in a heightened sensitivity to radiation and alkylating agents with resultant tissue damage. Logically it follows that APE may also play a role in the sensitivity to malignant cells to DNA damaging therapeutic agents. Many cancer therapeutic agents will induce apoptosis or programmed cell death, however little is known about the relationship of DNA repair enzymes (particularly BER) and apoptosis. Preliminary observations indicating that APE expression can be suppressed in myeloid leukemia cells by retinoic acid or DMSO while inducing apoptosis, has led to the hypothesis that: Decreased expression of APE is functionally related to apoptosis in myeloid leukemia cells. The specific aims are: Specific Aim #1 What is the relationship of APE expression to myeloid leukemia cell differentiation and apoptosis? Specific Aim 2: What is the role of APE protein phosphorylation in APE expression, function and apoptosis. Specific Aim #3: Determination of the molecular mechanisms that are responsible for down-regulation of APE expression upon induction of apoptosis. We propose to initiate studies on the relationship of APE expression and phosphorylation to cell differentiation, apoptosis, and sensitivity to cytotoxic agents, with the long-term goal of developing ways to enhance the chemo/radio sensitivity of leukemia cells by manipulating expression of APE.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Previous studies supported by NIH grant HL46213 (\"Molecular Interactions of Factor XI\"), for which continued support is sought, have focused attention on FXI and its interactions with other plasma proteins, i.e., thrombin, FXIa, FXIIa, prothrombin, high molecular weight kininogen (HK), and FIX, with the platelet plasma membrane and with various inhibitory molecules, e.g., protease nexin II (PN2), in the initiation and regulation of blood coagulation. These studies, utilizing NMR and x-ray crystallography, provide three separate structures, one of the catalytic domain of FXIa in complex with the KPI domain of PN2, one of the Apple 4 (A4) domain dimer and one of the zymogen FXI dimer that provide the basis for the future proposed investigations. The long-term goals of the present proposal are to elucidate the molecular mechanisms involved in the interaction of FXI/FXIa with protein and cell surface ligands involved in its activation and with plasma protein and cell surface ligands involved in the expression and regulation of FXIa enzymatic activity. Specifically these studies propose to determine the mechanism and physiological importance of FXI homodimer formation by a mutational analysis of the A4 dimer interface to prepare a monomeric FXI molecule;to study the role of the A4 domain in FXI dimerization in equilibrium unfolding studies;and to determine whether the dimeric structure of FXI is required for FXI activation to FXIa by thrombin or by FXIIa in the presence or absence of activated platelets or glycocalicin. They will examine the hypothesis that the activation of FXI to FXIa is accompanied by a major conformational change that promotes the efficient activation of FIX by exposing a substrate-binding site in the heavy chain (A2 and/or A3 domains) of FXIa for efficient binding and activation of FIX. Finally they will determine whether the dimeric structure of FXIa is required for efficient FIX-activation and to differentiate between alternative mechanisms to account for the failure of monomeric FXIa to activate FIX on the activated platelet surface. Recent observations resulting in the recognition of the essential role of FXI in hemostasis and thrombosis concern the relationship of its domain structure to its biological function, its molecular and cellular interactions, the elucidation of pathways for activation of FXI, and the expression and regulation of its enzymatic activity. It is hoped that information gleaned from the proposed studies will result in novel approaches to targeting FXI/XIa in the development of new anticoagulants that prevent thrombosis (heart attacks, strokes and pulmonary emboli) without bleeding complications.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There continues to be substantial controversy concerning neural control of the cerebral circulation. Over the past several years, our laboratory has demonstrated that there are several factors which modulate the response of cerebral vessels to stimulation of peripheral sympathetic nerves. Of these factors, species variation and temporal course of the response are of the utmost importance. The cerebral vessels of cynomolgus monkeys are far more sensitive to sympathetic nerve stimulation than cerebral vessels of cats or dogs. Furthermore, in the cynomolgus monkey, cerebral vasoconstriction induced by continuous maximal electrical stimulation of the sympathetic nerves persists in most areas of the brain for less than 90 seconds. Thus, the escape phenomenon is very important in cerebral vessels. In addition to these new observations concerning factors which modulate the response of cerebral vessels to sympathetic nerve stimulation, we have recently applied a new method--the pulsed Doppler flow meter--to measure cerebral blood velocity in small cerebral vessels on the surface of the brain. The major objective of this proposal is to further elucidate the role of peripheral sympathetic nerves in the control of cerebral vascular resistance. The proposed studies will be performed in both awake and anesthetized cynomolgus monkeys and the temporal course of the response of cerebral vessels to sympathetic stimulation will be explored in detail under a variety of experimental conditions. Responses of cerebral vessels to sympathetic nerve stimulation will be assessed with the pulsed Doppler method and with carefully timed injections of labeled microspheres. In the proposed experiments I will study possible mechanisms that permit cerebral vessels to escape from the vasoconstrictor effects of sympathetic nerve stimulation. I will also study effects of sympathetic nerve stimulation on cerebral vascular resistance in awake and anesthetized monkeys during hemorrhagic hypotension, severe hypertension, and chemoreceptor stimulation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Acquired immunodeficiency syndrome (AIDS) is an immunoregulatory disorder characterized by the presence of Kaposi's sarcoma or opportunistic infections. CD4+ T cells and mononuclear phagocytes are the target cells for the human immuno- deficiency virus (HIV), the etiologic agent of AIDS. Monocyte/macrophages (M-phi) are the major reservoir of HIV and, therefore, a critical concern for the host is the functional capacity of M# from HIV+ individuals to destroy opportunistic pathogens. Disseminated histoplasmosis is seen with increasing frequency as a complication of AIDS, particularly in areas where Histoplasma capsulatum (Hc) is endemic. AIDS patients with disseminated histoplasmosis have a high rate of relapse even after apparently successful therapy with amphotericin B. Therefore, it is imperative that we gain a better understanding of the interaction of Hc with M-phi from HIV+ patients. The goal of the proposed study is to characterize the intrinsic defects in the function of monocyte/M-phi from patients with HIV infection with respect to their ability to defend against Hc. The specific aims of this proposal are: l) To quantify the capacity of M-phi from HIV-infected patients to bind and to phagocytose Hc yeasts and microconidia compared to M-phi from HIV- individuals; 2) To determine if M-phi from HIV-infected patients provide a more than normal permissive environment for the conversion of Hc microconidia into the pathogenic yeast phase, or permit more rapid multiplication of yeasts than normal M-phi; and 3) To quantify the capacity of M-phi from HIV-infected patients to be activated by colony stimulating factors (CSFs) to inhibit the intracellular multiplication of Hc yeasts compared to M-phi from HIV- individuals. As changes in M-phi function may correlate with disease progression and not merely HIV infection, monocyte/M-phi will be studied from different patient groups, from those who are at risk for HIV infection but who are not infected to patients with AIDS. In addition, studies will be performed on normal M-phi infected with HIV in vitro to determine if HIV infection alone can cause M-phi dysfunction with respect to its interaction with Hc.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have developed a high pressure liquid chromatographic technique for determination in human urine concentrations of antipyrine and its metabolites, norantipyrine, 4-hydroxyantipyrine, and 3-hydroxyantipyrine. This method involved studies to determine optimal buffer pH and molarity, ratio of solvents in the mobile phase of the system, and effects of centrifugation temperature during assay. Experiments performed disclosed interesting variations from one subject to another both in the amount of each metabolite excreted with unit time, as well as in the time of appearance of each metabolite. We are now in a position to perform family studies to determine which of these factors that exhibited interindividual variations are heritable and what the mechanism of heritable transmission is.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The proposed research has two objectives: 1) to investigate the genetic and physiological control mechanisms of tetrahydrofolate metabolism; 2) to determine the role of proteases in the metabolism and differentiation of yeast. Mutants altered in THFA metabolism will be analyzed genetically and biochemically. The THFA interconversion enzymes will be purified to see whether they occur in an enzyme aggregate. The intracellular localization of these enzymes will be determined. Mutants with alterations in the function or regulation of proteases will be isolated and characterized both with respect to genetics and biochemistry. The consequences to the cell of such alterations in protease function will be examined. This research should contribute to our understanding of regulatory mechanisms underlying transport of proteins through membranes, and may contribute to our understanding of the mechanisms governing transitions between cell states (meiosis and mitosis).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "As a resubmission, the proposed five-year plan will train me and foster my launch into an independent interdisciplinary research career. My multidisciplinary background in Biomedical Engineering, Electrical Engineering and Mechanical Engineering fields and my outstanding experience in advanced biomechanical modeling and electrophysiological modeling research have fostered my interest and given me unique advantages in conducting interdisciplinary biomedical research in Urinary Incontinence (UI). The immediate goal is to prepare for a career as an independent scientist and to develop a novel subject-specific electromechanical pelvic model based urethrovaginal support and urethral function assessment (UUFA) technique to minimally invasively and quantitatively assess these two etiologic factors and to investigate the specific changes associated with aging effects that cause the increase in prevalence of SUI in women. The central hypothesis behind this research is that urethrovaginal support and urethral function in women and aging effects on female SUI can be minimally invasively and quantitatively assessed and characterized using a subject-specific pelvic modeling approach. Primary Hypothesis #1 is that urethral function in women can be minimally invasively and quantitatively assessed using a subject-specific electrophysiological pelvic modeling approach. Primary Hypothesis #2 is that urethrovaginal support function in women can be non-invasively and quantitatively assessed using a subject-specific biomechanical pelvic modeling approach. Primary Hypothesis #3 is that aging will cause poor urethrovaginal support and/or urethral dysfunction which will lead to SUI. Primary Hypothesis #4 is that a female continence profile can be created to predict the status of continence in women in women clinically and to suggest optimal therapeutic modalities for specific patients. We propose four Specific Aims (SA) to test these Hypotheses: SA #1: To develop a subject- specific electrophysiological pelvic model based periurethral muscle fatigue assessment (UMFA) technique to minimally invasively and quantitatively assess urethral function in women. Primary Hypothesis #1 will be tested. SA #2: To develop a subject-specific biomechanical pelvic model based urethrovaginal support function assessment (USFA) technique to non-invasively and quantitatively assess urethrovaginal support function in women. Primary Hypothesis #2 will be tested. SA #3: To investigate aging effects on urethral function and urethrovaginal support in young/elderly female subjects with/without SUI using the UUFA technique which consists of the UMFA and USFA techniques. Primary Hypothesis #3 will be tested. SA #4: To develop a female SUI profile as a clinical standard of female SUI by utilizing the UUFA technique to quantitatively characterize SUI etiologic factors in urethrovaginal support and urethral function in women. Primary Hypothesis #4 will be tested. At the completion of this project, it is our expectation that we will have established and validated the UUFA technique and the feasibility of its application in clinical studies, and have achieved a better understanding of the contributions of specific changes associated with aging and menopause to these etiologic factors in women.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Utilizing a 4 pound fully automatic portable blood pressure recorder with a conventional sphygmomanometer cuff, a group of hypertensives (essential, labile, renal types) will have their blood pressures and heart rates recorded at 15 minute intervals during an 8 hour period while engaged in their usual work activities. A voice channel allows the subject to indicate with each pressure recording, the time, whether standing, sitting or lying, his physical activity, his emotional state and any physical symptoms. Such recordings will be made prior to and during treatment with single or multiple antihypertensive agents. Similar recordings at work will be made for apparently healthy controls, not on medications, of comparable age and sex. Where possible and feasible, some patients and controls will have 24 hour recordings including sleep at home. Such data would be unique and of great potential value in understanding blood pressure behavior in normals and hypertensives and the benefits and side effects of chemical treatment in the latter group. Patients referred for unexplained dizziness or for atypical headaches or angina will wear the device to determine if unusual blood pressures correlate in time with such symptoms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although there is significant evidence that 2,3,7,8-tetrachlorodibenzodioxin (TCDD) alters lymphocyte function in mice, there have been very few studies which examine the effects of TCDD on human lymphocytes. While there have been a number of inadvertent exposures to TCDD, via industrial accidents or environmental contamination, there have been no consistent immunological defects observed in exposed individuals. Studies in German chemical workers have demonstrated alterations in lymphocytes subpopulations and functional parameters 20 years after exposure to TCDD. As part of a continued effort to assess the health effects of environmental exposures to TCDD it is imperative that we examine endpoints which have the potential to be sensitive predictors of human exposure and to correlate these biomarkers with blood TCDD levels, determine influence of potential confounding variables on biomarker expression, and evaluate inter- and intra-individual variation. Identification of additional biomarkers may aid in identifying individuals or populations at greater risk of developing disease or neoplasia after exposure to dioxins and related compounds. Samples of peripheral blood lymphocytes were obtained from several cohorts of control or TCDD-exposed individuals. These samples were mitogen-stimulated in culture, alone or in the presence of TCDD, and the cells and culture supernatants harvested after 72 hr. To date we have examined supernatants from a group of workers who were employed at a chemical plant in Hamburg-Moorfleet, Germany operated by Boehringer-Ingelheim from 1951-1984 that produced organochlorine herbicides and pesticides (including 2,4,5-trichlorophenoxyacetic acid) and opioids. TCDD contamination at this plant was confirmed in 1984 and significant increases in mortality have been observed in a cohort of workers employed at this plant for at least 3 months from 1952-1984. Peripheral blood for lymphocyte isolation and serum dioxin analysis was obtained from a cohort of 110 individuals in 1992, in collaboration with clinicians at the University of Mainz (Detlev Jung and Johannes Konietzko) and a biostatistician at the German cancer Research center. In vitro treatment with TCDD resulted in a significant decreases in the mitogen-stimulated production of the T Helper 1 type (TH1) cytokines Interleukin 2 (IL2) and Interferon gamma (IFNgamma) in ymphocyte cultures from this cohort. We believe the alterations TH1 cytokines are a result of the in vitro exposure as there is limited correlation with standard biomarkers of TCDD exposure such as total TCDD plasma levels or total plasma TEQ. In contrast, there appears to be a dose-related correlation between individual TCDD body burden and decreased secretion of the T Helper 2 type cytokine Interleukin 4. Significant correlations were also seen between in vitro TCDD-inducible ethoxyresorufin-O-deethylase activity and secretion of tumor necrosis factor alpha. While it appears that there may be some significant effects on immunoglobulin (Ig) secretion, particularly for IgA and IgG2, interindividual variability is quite high, and sophisticated statistical analyses will be required to tease out exposure-specific effects. In collaboration with Dr. Joseph Haseman, the dataset is undergoing multivariate analysis to examine correlations between immunologic effects and in vivo and in vitro biomarkers of exposure, and to determine the effects of confounding variables such as gender, smoking status, etc.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Parathyroid hormone (PTH) is a major regulator of mineral homeostasis and bone metabolism. Intermittent PTH therapy is approved for treatment of osteoporosis, yet the cellular mechanisms underlying the biologic effects of PTH are incompletely understood, including the molecular basis for the observation that low-dose, intermittent administration of PTH elicits net bone formation, whereas continuous administration of high-dose PTH causes predominantly bone resorption. The actions of PTH on bone and kidney are mediated by the Type 1 PTH receptor (PTH1R), a G protein-coupled receptor (GPCR).beta-arrestins (1 and 2) are cytoplasmic molecules that regulate GPCR activity. We previously reported that arrestins inhibit PTH-stimulated cAMP signaling and trigger cellular internalization of PTH and its receptor in vitro. Thus, our primary hypothesis is that beta-arrestin2 is a key modulator of the activity of PTH in bone. In support of this hypothesis, our preliminary data indicate that compared to wild-type (WT) mice, beta-arrestin2 null (beta-arr2-/-) mice have reduced bone mass and architecture, and importantly, an altered skeletal response to intermittent PTH that is characterized by increased bone resorption leading to decreased net anabolic effects on trabecular bone. Moreover, compared to WT, osteoblasts from beta-arr2-/- mice exhibit increased and sustained cAMP signaling in response to PTH. Thus, to further explore the role of beta-arr2 in mediating the skeletal response to PTH we will pursue three specific aims: 1) Determine the skeletal response of ovariectomized beta-arr2-/- and WT mice to intermittent PTH, thereby allowing examination of the role of beta-arr2 in the context of high bone remodeling induced by estrogen deficiency; 2) Investigate the arrestin-mediated regluation of PTH-stimulated intracellular signaling and gene expression by comparing primary osteoblastic cells from beta-arr2-/- and WT mice; and 3) Determine the skeletal response to intermittent and continuous PTH administration in transgenic mice with targeted overexpression of beta-arrestin2 in well-differentiated osteoblasts (OC-beta-arr2-GFP-Tg+). In summary, the overall goal of this project is to improve our understanding of the mechanisms regulating the activity of PTH in bone. By conducting complementary in vivo and in vitro experiments in mice deficient for beta-arrestin2 and in mice overexpressing beta-arrestin-2 in mature osteoblasts, we will provide novel insights into the mechanisms that underlie the distinct skeletal response to intermittent versus continuous PTH administration. Information gained from the proposed studies will be instrumental for developing new PTH1R ligands with improved signaling and biologic activity profiles for treatment of osteoporosis and other metabolic bone disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Cocaine abuse is a major public health problem in the United States with no accepted pharmacologic treatment. We propose a placebo-controlled, randomized, double-blind outpatient study evaluating imipramine as a treatment for cocaine abuse in 80 chronic users. The study is designed to: 1) determine the efficacy of imipramine as an outpatient medication treatment for cocaine abuse 2) determine the efficacy of imipramine in blocking the cocaine euphoria and/or diminishing the craving for cocaine in abusers 3) examine the physiologic response to cocaine via a controlled laboratory challenge with cocaine before and after imipramine 4) determine the prevalence and types of psychiatric disorders in chronic cocaine abusers and contribute to the determination of whether \"cocaine dependence\" should be a part of the official nosology of mental disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Growing evidence indicates a reciprocal relationship between the HPA axis and the immune system which may be relevant to disease expression. Since patients with AIDS exhibit abnormalities in the HPA axis, a thorough understanding of the basic physiology of neuroendocrine-immune interactions is an important prerequisite for determining the potential role of neuroendocrine function in the development of this disease. Nevertheless, adrenal steroid hormones, the final product of HPA axis activation, have complicated and inconsistent effects on in vivo immune function. Recent studies in our laboratories suggest that further understanding of these complex yet important immunologic effects of adrenal steroids will be greatly enhanced by focusing on adrenal steroid receptors in immune cells and tissues. Our results indicate that Type I and II adrenal steroid receptor binding is differentially expressed in various immune tissues and there is considerable heterogeneity between immune tissues in receptor subtype activation following in vivo hormone exposure. Moreover, we have found a highly significant relationship between adrenal steroid receptor activation in vivo and immune function. These findings in conjunction with the proposed studies will advance our knowledge of the mechanisms involved in the interaction between adrenal steroids and the immune response. The specific aims of the proposed project are as follows: Except where indicated, all experiments will be conducted on Sprague Dawley rats. For Specific Aim 1, the distribution of Type I and II receptor positive cell types in sections of spleen, thymus and mesenteric lymph nodes will be antibodies. In addition, these antibodies will be used in conjunction with antibodies to specific cell surface markers to determine receptor subtype expression in defined immune cell subpopulations using flow cytometry. For Specific Aim 2, selective Type I and II receptor agonists and antagonists will be administered in vitro and in vivo and the immunologic effects of these agents on the following immune functions will be determined: T cell proliferation, II-2 secretion, T cell independent B cell proliferation, NK cell activity, monocyte II-1 secretion. Specific Aim 3 will examine the relationship between receptor activation and the noted immune variables in a range of in vivo physiological conditions known to differ in steroid bioavailability. Effects of diurnal variation, sex, strain, and duration of endogenous corticosterone exposure will be evaluated. Finally, Specific Aim 4 will examine immune function in animals with downregulated adrenal steroid receptors secondary to chronic steroid exposure or aging. Taken together, these studies, which are a major component of the PI's RSDA, are designed to provide a foundation for understanding the role of neuroendocrine function in immune-related diseases such as AIDS.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Umbilical cord blood (CB) is used increasingly to restore hematopoiesis in transplant patients lacking sibling, or unrelated donors. A major disadvantage of CB transplantation (CBT) is the low cell dose with resultant delays in neutrophil and platelet engraftment, as well as a high rate of engraftment failure. A major hypothesis of this grant is that ex vivo-expanded CB progenitors will provide more rapid hematopoietic reconstitution and less engraftment failure than unmanipulated CB. In the two clinical CB expansion trials conducted during the last RO1 funding period, CD133+ progenitors were isolated and cultured ex vivo in a liquid culture system targeting committed, or more primitive hematopoietic progenitors. In those sequential trials, trends in faster time to engraftment were seen, but significant losses of CD34+ progenitors occurred. To avoid the need for positive selection, which results in a substantial loss of CD34+ cells, we have developed an alternative approach that involves the ex vivo co-culture of CB mononuclear cells (MNC) with bone marrow-derived mesenchymal stem cells (MSC). We hypothesize that MSC, by functioning as a surrogate hematopoietic 'niche', will provide a more suitable microenvironment for the expansion of lineage-committed CB hematopoietic progenitors than afforded by current liquid suspension culture systems. As accrual to the current CB expansion trials nears completion, this competitive renewal application will focus on the clinical evaluation of CB expansion in MSC co-cultures, with the long-term goal of improving neutrophil and platelet engraftment in CBT patients. Although a low cell dose is clearly the chief limitation of CBT, a number of investigators have also reported a deficit in the homing of CB cells to the marrow. Thus, it is conceivable that even with optimal ex vivo expansion, inadequate homing may limit the rapidity of engraftment which is the goal of this proposal. The homing defect has been attributed to low levels of fucosylation of cell surface molecules responsible for binding to P- and/or E-selectins expressed by the marrow microvasculature. This interaction is a key component of the recruitment of hematopoietic progenitors to the marrow. We hypothesize that increasing the level of CB cell surface fucosylation will improve interactions with selectins, thereby improving homing and engraftment. Aim 3 will evaluate the modification of unmanipulated and expanded CB progenitor cells with a fucosyltransferase, as a means to facilitate their recruitment to the marrow. PUBLIC HEALTH RELEVANCE: Umbilical cord blood is being used increasingly for many patients with high-risk hematological malignancies and genetic disorders who do not have a bone marrow donor, but is associated with a longer time to engraftment compared to marrow. The expansion of cord blood in the laboratory prior to infusion appears to shorten time to engraftment; additionally the treatment of the cord blood with a sugar molecule prior to infusion appears to improve its ability to home to the bone marrow quickly. Both these strategies will be explored in the grant thereby making the transplant safer for patients in terms of infectious and bleeding complications and will thus improve survival.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This proposal deals with the development of a radiation detector system that will make possible the direct measurement of the structure of charged particle tracks in a gas. Such a detector will permit the measurement of energy e.g., the diameter of a DNA molecule. This information, which is critical to development and evaluation of mechanistic biophysical models for understanding radiation effects, has hereto fore not been measurable. When a charged particle penetrates a gas, electrons produced during ionization events quickly become thermalized. The proposed detector system will determine the positions and number of these electrons by imposing a suitable RF field, causing the electrons to agitate and excite molecules of the gas which will then fluoresce producing photons that will be detected by the external optical detector system. Thus, each point of light will represent the position of a single electron. In Phase I experimental proof of the concept was made that a single charged particle track can be detected optically. In this proposal the operation of the system would be greatly enhanced so that CCD detectors can be used to obtain quantitative information from images of all of the electrons produced in the track of a single particle.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The purpose of this investigation is to identify the effect of single-dose disulfiram on human cytochrome P4502A6 activity in vivo, using coumarin 7-hydroxylation as the in vivo probe for human P4502A6. Our hypothesis is that disulfiram will have no effect on in vivo P4502A6 activity and we expect to find that coumarin clearance and 7-OH-coumarin formation clearance will not be significantly diminished by disulfiram.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Work from our laboratory has focused on the pathophysiology of Epstein-Barr virus (EBV), a gamma herpesvirus which is associated with several human malignancies including B-cell lymphoma and nasopharyngeal carcinoma. We have also explored the role that psychological stress plays in the modulation of the steady state expression of EBV early proteins and how stress could be a factor in the risk for EBV associated disease. This proposal will focus on how EBV can induce inflammation in vitro and in mice. We have explored the hypothesis that EBV-encoded early proteins can induce immune changes observed in patients infected with EBV independent of their role in viral replication. We discovered that EBV-encoded deoxyuridine triphosphate nucleotidohydrolase (dUTPase) up-regulates the production of several proinflammatory cytokines including TNF-a, IL-1, IL-6, IL-8, as well as IL-10 in macrophages. When purified EBV-encoded dUTPase was inoculated into mice the protein was capable of producing symptoms compatible with cytokine-induced sickness behavior. The data demonstrate that EBV-encoded dUTPase can induce sickness behavior in mice resulting in increased body temperature and decreased body mass and physical activity. Furthermore, there is new and extensive literature linking chronic inflammation with an increased risk for cancer. Our data provide a new perspective on how a latent herpes virus, such as EBV, when reactivated by stress or other factors, could cause immune dysregulation, the activation in NF?B in macrophages and the upregulation of proinflammatory cytokines with possible implications for EBV associated clinical symptoms and disease, including EBV associated tumors. PUBLIC HEALTH RELEVANCE: This multidisciplinary research grant will provide new information on how stress can have an impact on the pathophysiology on herpesvirus associated diseases.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We propose to study the synthesis, physical properties and utility in synthetic organic chemistry of several classes of compounds which share the distinction of having multi-center electron-rich bonds of the type usually called hypervalent. Among these species are isolable compounds of pentacoordinate carbon, chiral sulfuranes and asasulfuranes. The generation of sulfuranes and persulfuranes by neighboring group participation in perester decompositions will be studied. The generation of radical pairs, and larger arrays of radicals, within the solvent cage by such reactions will receive attention. The study of planar triarylmethyl radicals and cations is to be continued with particular emphasis on the complexes formed in the solid state between such radicals and cations and their possible electrical conductivity. Analogues of compounds from earlier studies in these areas which have shown activity in the NCI Cancer Chemotherapy Screening Program will be prepared and submitted for testing.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This application requests funding to identify risk factors for chronic back pain disability. The proposed study will draw on our existing data base of physical, psychological, and work-related premorbid data. This data base is unequalled in population size and scope of independent variables. Furthermore, this study will provide the longest follow-up of any such study to date. A better understanding of risk factors would provide a solid foundation for establishing appropriate programs to prevent chronic back pain disability, to enhance return to work, and to reduce the impact back pain has on the industrialized nations of the world. Our goal is to continue monitoring our subject population of 3,020 individuals so that risk factors for the development of chronic back pain disability can be identified. Our efforts to establish this data base have been supported to date by NIOSH; however, NIOSH has stated that the evaluation of chronic disability is beyond their scope of interest and has limited our funding to analysis for the prediction of acute industrial back injuries. To stop without evaluating chronic back pain disability would ignore the 10% of back injuries that cause the most suffering and account for approximately 80% of the total cost for back problems. We have already found that the first 26 subjects who developed disabling back problems of at least a three-month duration have a significantly different fitness level than their age-matched controls. With two more years of follow-up we estimate another 15 - 21 subjects will develop chronic back pain disability. An increased number of subjects in the chronically disabled category would add to the statistical power for the evaluation of other variables. Our results to date indicate that it is highly probable that analysis of premorbid data can predict chronic back pain disability and greatly increase our understanding of this expensive healthcare problem.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The liver is an organ with strong innate immunity, which plays an important role in host defense against microbial infection and tumor transformation. Emerging evidence suggests that innate immunity as well as a variety of cytokines produced by innate immune cells also contribute to the pathogenesis of acute and chronic liver diseases. Our laboratory has been actively studying the role of innate immunity and its associated cytokines in liver injury and repair. During the fiscal year, we have generated floxed STOP-CD59 knockin mice (ihCD59), in which expression of human CD59 only occurs after Cre-mediated recombination. By using this model, we studied the effect of cell death on activation of innate immunity and liver regeneration. Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration. Cell ablation is a powerful tool for studying cell lineage and/or function; however, current cell-ablation models have limitations. Intermedilysin (ILY), a cytolytic pore-forming toxin that is secreted by Streptococcus intermedius, lyses human cells exclusively by binding to the human complement regulator CD59 (hCD59), but does not react with CD59 from nonprimates. Here, we took advantage of this feature of ILY and developed a model of conditional and targeted cell ablation by generating floxed STOP-CD59 knockin mice (ihCD59), in which expression of human CD59 only occurs after Cre-mediated recombination. The administration of ILY to ihCD59+ mice crossed with various Cre-driver lines resulted in the rapid and specific ablation of immune, epithelial, or neural cells without off-target effects. ILY had a large pharmacological window, which allowed us to perform dose-dependent studies. Finally, the ILY/ihCD59-mediated cell-ablation method was tested in several disease models to study immune cell functionalities, hepatocyte and/or biliary epithelial damage and regeneration, and neural cell damage. Together, the results of this study demonstrate the utility of the ihCD59 mouse model for studying the effects of cell ablation in specific organ systems in a variety of developmental and disease states.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Numerous neurological diseases are characterized by a sex difference. The neuropathology often includes infiltration of immune cells, with this immune infiltration potentially contributing to disease pathogenesis. Since it is known that sex differences exist in the immune system, this confounds investigations into sex differences in the CNS. Thus, we will use bone marrow chimeras to investigate sex differences in the CNS. By varying sex chromosomes or sex hormones in hosts reconstituted with a common immune system, one can ascertain the role of sex chromosomes and sex hormones on the brain response to injury. We will use one of the most inflammatory of all CNS disease models, the multiple sclerosis model, experimental autoimmune encephalomyelitis (EAE), to show applicability of this approach to a variety of neurological diseases. We will employ mice which differ in the complement of sex chromosomes (XX vs. XY), while having the same gonadal type, to determine the effect of sex chromosomes in the absence of confounding effects of exposure to different types of sex hormones. Specifically, in aim #1 we will determine whether the greater severity of EAE in XX, as compared to XY-, mice is due to sex chromosome effects in the CNS. In aim #2, we will determine if the sex chromosome effect in the CNS during EAE is due to the dose of X or Y genes. Finally in aim #3, we will use mice which differ in gonadal type, female vs. male, while having the same sex chromosome complement (XX vs. XX Sry) to determine whether the greater severity of EAE in female, as compared to male, mice is due to sex hormone effects in the CNS. PUBLIC HEALTH RELEVANCE: This is an exploratory (R21) grant to determine the effect of sex chromosomes and sex hormones on the central nervous system's response to an immune attack using the multiple sclerosis model, experimental autoimmune encephalomyelitis. This proposal will establish a model system to determine the effect of sex chromosomes and sex hormones on a variety of neurological diseases characterized by a sex difference.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION(Adapted from applicant's abstract): The conversion of adenosine to inosine by RNA editing represents an increasingly important mechanism for generating diversity in neurotransmitter receptor expression. Such RNA modifications have been shown to alter both the ion permeation and electrophysiological properties of ligand-gated ion channels and to modulate the efficacy of receptor G-protein interactions. Recent studies in our laboratory have demonstrated that ADAR2, a double-stranded RNA-specific adenosine deaminase involved in the editing of glutamate and serotonin receptor transcripts, can selectively modify its own pre-mRNA, suggesting a negative autoregulatory mechanism by which ADAR2 can modulate its own expression level to prevent editing at aberrant sites. The long-term objectives of these studies are to define the cellular processes by which such RNA editing events can modulate neurotransmitter receptor expression and function in the central nervous system. To identify the cis-active regulatory sequences necessary for ADAR2 editing, a tissue culture model system will be utilized that exhibits RNA processing patterns analogous to those observed in vivo. Analyses of RNA from cells transfected with a variety of mutant ADAR2 minigene constructs will serve as the primary methodology for these mapping studies. An in vitro RNA editing system, utilizing purified, recombinant ADAR proteins, will also be used as a mapping technique and to further define the ADAR proteins responsible for site-specific modifications of ADAR2 pre-mRNA. The effect of ADAR2 protein levels on the specificity of adenosine modification will be examined using an inducible tissue culture system and an in vitro editing system to test whether over expression of ADAR2 can lead to the modification of known ADAR2 substrates(GluR-Band 5-HT2CR) at aberrant sites competition studiesbetweenADAR2 pre-mRNA and otherADAR2 substrates will be performed to determine if such competition represents a cellular mechanism for maintaining appropriate ADAR2 protein levels. To examine the molecular and phenotypic consequences of ADAR2 misregulation, genetically modified strains of mice will be generated in which ADAR2 levels have been altered by over expression or by selective ablation of ADAR2 auto editing. Multiple, independent mouse lines will be assessed for gross alterations in animal phenotype, changes in the level of ADAR2 expression/activity, alterations in editing patterns for previously identified ADAR2 substrates and increases in mRNA-derived inosine content. It is anticipated that these studies will provide new insights concerning the cellular processes modulatingADAR2 expression as well as the effects that such modulation confer upon the expression of neurotransmitter receptor isoforms.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of this research effort are to determine the natural history of diabetic retinopathy, and to evaluate the effect of photocoagulation therapy on specific forms of diabetic retinopathy. The Contractor will perform the function of a participating clinical center under the Diabetic Retinopathy Study.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The project documented the frequency of primary intracranial neoplasms in the pediatric populations of Rochester, Minnesota, and the state of Connecticut. In addition, using the records-linkage system available for residents of Rochester, Minnesota, we investigated the magnitude and risk factors for cerebrovascular disease in infants and children. The same Rochester, Minnesota records-linkage system was used to determine temporal trends in the incidence rates of cerebral palsy as well as the distribution of clinical subtypes and survival by clinical subtype, for the year 1950-1976. For the state of Minnesota, sex-specific neonatal mortality rates (NMR) in gestational age/birthweight risk subgroups were delineated for the years 1970-1976, and sex- and birthweight-specific NMR trends were determined for the years 1967-1976. The same record linkage system has been used to identify all possible cases of complex partial seizures occurring in the years 1960-1980. A case-control study is being designed to identify risk factors associated with the occurrence of such seizures.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY/ABSTRACT Electronic nicotine delivery systems (ENDS) have emerged in the US market, with use and awareness rapidly increasing in recent years, particularly in young adults. While ENDS may facilitate harm reduction in smokers, ENDS represent significant health risks, including addiction in the nicotine-nave (e.g., young adults). From a socioecologic perspective, the literature regarding tobacco retail indicates that place characteristics such as neighborhood demography and policy context influence retailer location and marketing, and these factors impact individual tobacco use. However, this literature is in its infancy in regard to ENDS and particularly to vape shops, which are stores exclusively devoted to ENDS sales. Vape shops have proliferated in the US and are unique in their product offerings, marketing, and overall retail environment (e.g., tasting bars). Vape shops, as well as the 2nd and 3rd generation ENDS they sell, have particular appeal to young adults. A particularly important and timely macro-level factor that may impact ENDS use and distribution channels is the impending FDA Deeming Regulation on ENDS and other tobacco products. The Deeming Regulation involves a range of policies implemented in the next 3 years (e.g., mandatory age verification and prohibiting free samples beginning in Aug 2016, mandatory health warning labels effective Aug 2018, manufacturers required to submit a new tobacco product application by Aug 2018, etc.). These regulations are likely to impact vape shop survival as well as their marketing and the overall vape shop experience, given young adults are the biggest segment of vape shop clientele (who will require ID), the social experience of tasting bars (which will no longer be allowed), and the history of ENDS being promoted as safe and for cessation or harm reduction (with products and ads requiring health warnings). A well-integrated program of research is needed to examine the multilevel impact of regulation on ENDS retailers, as well as on ENDS marketing, specifically among vape shops, given that they are a unique retail settings that have a particular impact on young adult ENDS use. Leveraging a Socioecological Framework, we will draw data from 6 metropolitan statistical areas (MSAs) representing the CDC-defined regions and the gradient of tobacco control in order to address 3 inter-related aims: 1) examine density and survival of vape shops over time and across contexts in relation to FDA regulation, local policies, and other sociocontextual factors (e.g., neighborhood context, density/proximity of convenience stores); 2) examine vape shop marketing and POS practices (e.g., age verification, free sampling, health warnings) over time and across contexts in relation to FDA regulation, local policies, and other sociocontextual factors (e.g., neighborhood context, density/proximity of convenience stores); and 3) examine young adult ENDS use over time and across contexts in relation to spatial access to vape shops and convenience stores, ENDS advertising exposure, local policies, and sociocontextual factors. This research will document the impact of regulation on this industry and provide an evidence base for legislation regarding zoning and vape shop marketing/POS practices to protect high-risk populations. We will disseminate findings with an explicit focus on informing public health policy and practice regarding ENDS, as well as future research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The JAK-STAT (Janus Kinase - Signal Transducer and Activator of Transcription) signaling pathway activated by more than 35 different cytokines or growth factors has been implicated in a variety of cellular functions in the hematopoietic, immune, neuronal and hepatic systems. Ethanol inhibition of JAK-STAT activation may be a mechanism by which ethanol causes a wide variety of disorders in many organs. Acute exposure of rat hepatocytes to 25-100 mM ethanol significantly inhibits interleukin 6-, interferon gamma- or growth hormone-induced STAT activation. Chronic ethanol consumption significantly attenuates hepatic STAT3 activation induced by partial hepatectomy or interleukin 6. Acute ethanol (25-100 mM) exposure also significantly attenuates thrombopoietin- or interleukin 6-induced STAT activation in hematopoietic cells (BAF3/mp1 cells, platelets, U937, and AF10 cells), and ciliary neurotrophic factor-induced STAT activation in neuronal cells (NBTF neuroblastoma). The target sites of ethanol in the JAK-STAT signaling pathway will be explored by analyzing the effects of ethanol on cytokine-induced STAT phosphorylation, dimerization, translocation, JAK phosphorylation, receptor phosphorylation and ligand-receptor binding. The effects of chronic ethanol on the JAK-STAT pathway will be explored by examining JAK-STAT activation in vivo in rats fed with an ethanol containing diet. Identification of ethanol inhibition of the JAK-STAT signaling pathway in various tissues will enhance our understanding of the pathogenesis and progression of a wide variety of disorders caused by ethanol, such as antiregenerative effects, thrombocytopenia and fetal alcohol syndrome, and may also prove helpful in the design of novel drugs with therapeutic potential in alcohol-related disorders.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary: Aim #1. The function of Cx43 in the ciliary epithelium will be studied in three mouse models. In the nestin-cre /Cx43flox/flox line, Cx43 is selectively deleted from the pigmented (PE) layer of the ciliary epithelium resulting in a decrease in aqueous humor secretion and backflow of plasma proteins into the aqueous compartment. In the GjaUrt line, Cx43 immunostaining in the ciliary epithelium is globally reduced and also shows backflow of plasma proteins. The pax6alpha-cre/Cx43flox/flox line shows a loss of Cx43 from the non-pigmented epithelial (NPE) cells and a decrease in intraocular pressure. The three models will be studied in parallel using light and electron microscopy, immunohistochemistry, dye transfer methods and measurement of mouse intraocular pressure. Aim #2. Epithelial cells in CxSOnull lenses exhibit a striking reduction in mitotic index during the first postnatal week. Since replacement of Cx50 with Cx46 does not restore the normal growth rate, Cx50 must provide a unique functionality relating to epithelial cell proliferation. We will test two hypotheses regarding that function. The first is that Cx50 channels exhibit unique properties of selectivity, permitting the exchange of regulatory signals between cells. The second hypothesis is that Cx50, but not Cx46, regulates mitotic progression by C-terminal domain interactions with cytoplasmic proteins. To distinguish between channel properties and signaling involving connexin cytoplasmic domains, we will employ \"channel-dead\" Cx50 mutants that traffic normally to the plasma membrane and assemble into gap junctional plaques. One mutant with these properties is already characterized and others will be identified. The mutants will be introduced into CxSOnull mice as transgenes using a novel lentivirus approach. This strategy insures an ability to test multiple mutant genes very quickly. If these mutants rescue lens growth, this will strongly implicate the cytoplasmic domains of Cx50 as critical for the regulation of mitotic rate. If not, the properties of the intercellular channel are key and a rescue of mitotic rate will be tested by the introduction of tailless Cx50 mutants. Relevance: These studies will investigate the functional roles of gap junctions in the formation of aqueous humor and lens growth in the eye. They will permit a better understanding of the pathologies involved in glaucoma and cataract.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Differentiating cultures of embryonic skeletal muscle are being studied from two points of view. (1) A protein factor in embryo extract that is required for differentiation is being purified and characterized by standard biochemical methodology. (2) It has been found that the permeability of myoblasts to some fluorescent dyes changes abruptly as the cell differentiates. The molecular basis of this change is under investigation.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The adenylate cyclase and guanylate cyclase of cultured cells have been further characterized. Prostaglandin El activates a.c. in a highly cooperative manner. The calcium transport activity of the internal membranes of fibroblasts has been found to increase as cells crowd together and fall when they are dispersed. Two phosphoproteins, filamin and microtubule-associated proteins have been purified and characterized. The former binds to actin, the latter to tubulin. BIBLIOGRAPHIC REFERENCES: Davies, P., Shizuta, Y., Olden, K., Gallo, M., and Pastan, I.: Phosphorylation of filamin and other proteins in cultured fibroblasts. Biochem. Biophys. Res. Commun., 74: 300-307, 1977. Thomopoulos, P., Kosmakos, F.C., Pastan, I., and Lovelace, E.: Cyclic AMP increases the concentration of insulin receptors in cultured fibroblasts and lymphocytes, (in press) 1977.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The transcription factor NF-kappaB plays key roles in regulating gene expression in both normal physiological processes and in diseases. It is required for normal embryogenesis as well as immune responses to infections. Disregulation of NF-kappaB has also been implicated in a large number of human diseases from inflammation to cancer. Although much progress has been made in the past decade on understanding the early signaling mechanisms leading to NF-kappaB activation, how NF-kappaB transmits its signal after its binding to DNA in the nucleus has only begun to attraction attention in recent years. A marine natural product named pateamine A was found to be a potent immunosuppressive agent, inhibiting the T cell receptor-stimulated transcription of interleukin-2 gene. Preliminary studies revealed that pateamine A selectively inhibits NF-kappaB signaling and to a lesser extent, that of AP-1. A systematic examination of the known steps in the NF-kappaB signaling pathway revealed that pateamine A does not affect early signaling steps up to the DNA binding by NF-kappaB. Instead, pateamine A was found to block the transactivation activity of NF-kappaB, making pateamine A a valuable probe to study the nuclear signaling mechanism of NF-kappaB. To identify the molecular targets of pateamine A, a biotin-pateamine A conjugate was synthesized, which enabled the detection, isolation and identification of two pateamine A-binding proteins. These pateamine A-binding proteins have not been previously shown to be involved in NF-kappaB regulation. The main objective of this application is to confirm the molecular interaction between these proteins and pateamine A both in vitro and in vivo and to verify the physiological relevance of these proteins as mediators of the inhibition of NF-kappaB by pateamine A. The physiological functions of these proteins in regulating nuclear signaling by NF-kappaB will be investigated. As these proteins have been implicated in several cellular processes, whether pateamine A affects the other known cellular activities of the putative pateamine A-binding proteins will be investigated. Lastly, gene chip analyses will be performed to determine the specificity of pateamine A for the IkappaB-NF-kappaB signaling pathway by comparing the gene expression profiles of mouse embryo fibroblasts treated with pateamine A and those derived from NF-kappaB and IkappaB kinase knockout animals. In addition, crystal and solution structures of the complexes between pateamine A and its binding proteins will be obtained to facilitate the future design and synthesis of novel and simplified pateamine A analogs as candidates for development of anticancer and immunosuppressive drugs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Many medicinal reagents for the treatment of cancer originate from natural resources. To obtain sufficient quantities of these molecules and to improve their biological profile it is often necessary to produce these compounds synthetically. Monanchocidin, a member of the crambescidin family, is an attractive target for synthesis due to its complex structure and promising inhibition of cancer cell growth. An innovative strategy is proposed to address the synthesis of this molecule employing palladium catalyzed asymmetric allylic alkylation (AAA) and a novel hydroguanidination/cyclization sequence. The objective of this proposal is to develop a concise enantioselective synthesis of monanchocidin using newly developed reactions. We hypothesize that the pentacyclic guanidine can be rapidly assembled stereoselectively by developing a transition metal catalyzed hydroguanidination reaction. The specific aims of this project are; (1) to utilize palladium-catalyzed asymmetric allylic alkylation to construct the pyrolidine core of monanchocidin and other crambescidin family members, (2) develop and implement a novel transition-metal catalyzed hydroguanidination-spirocyclization method to assemble the pentacyclic guanidine, (3) develop an expedient synthetic route to access the fatty acid side chain in monanchocidin, and (4) develop a fragment coupling strategy that provides an efficient and modular route toward the first total synthesis of monanchocidin. The proposed research will be executed in an efficient manner by synthesizing key fragments enantioselectively and coupling them to the pyrolidine core of monanchocidin; assembled via palladium catalyzed asymmetric allylic alkylation (AAA). A novel method for construction of the pentacyclic guanidine of monanchocidin will be developed using a transition metal catalyzed hydroguanidination. An examination of transition metals typically used in hydroamination reactions will be conducted and the most efficient of these will be further optimized and employed in the proposed synthesis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Recently, a family of five muscarinic acetylcholine receptor genes was identified. The encoded receptors are novel targets for more specific and effective cholinergic therapies for individuals with Alzheimer's disease (AD), but, because of their pharmacological similarities, functions of the subtypes are not well defined. The abundance and localization of individual receptor proteins in the brain, their regulation by cholinergic input, and their involvement in AD have never been studied. In the proposed investigations, subtype-specific polyclonal antisera will be used to evaluate the five native muscarinic receptor proteins in control rat and human brain as well as alterations in receptors in a model of experimental cholinergic deafferentation and in cases of AD. By immunoprecipitation, the relative abundance of proteins will be determined in basal forebrain, neocortex, and hippocampus in rats. By immunocytochemistry, the proteins will be precisely localized and colocalized with cholinergic forebrain neurons. The assessment of receptor number and localization after lesioning of the fimbria-fornix and basal forebrain in rats will provide infomation about the pre- and postsynaptic localization of the subtypes and the cellular and molecular bases for changes induced by deafferentation. Lastly, muscarinic receptor proteins in human brain will be identified, and changes that occur in cases of AD compared to age- and postmortem-matched controls will be measured. These studies will identify the best targets for subtype-specific cholinergic drugs and will provide new neurochemical markers for selective populations of neurons that are potentially at risk in AD.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aim of this study was to determine the immunogenicity of recombinant hepatitis B vaccine when administered intramuscularly to premature infants. Ninety percent of preterm infants responded to the hepatitis B vaccine when the first dose of vaccine was given just before hospital discharge. This study is now closed, and the data are being analyzed.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Particulate matter (PM) and its adsorbed contaminants are associated with adverse health effects, including an increased risk of cancer. The level of PM10, particulates 10 um or less, in the Paso del Norte (PdN) air basin regularly exceeds federal guidelines, potentially exposing people to harmful toxicants. To date, research has not been undertaken to study biological effects of PM locally. In the proposed study, PM filters will be collected from air monitoring stations in El Paso, TX and Juarez, Chihuahua; organic material will be extracted and evaluated for biological effects. Cultured hepatoma cells will be exposed to extracts to determine Ah receptor mediated biological activity. Ah receptor ligands are known to cause numerous adverse health effects. Further, extracts will be used in the Ames test for mutagenicity, with and without metabolic activation. Finally, cells will be incubated with extracts and DNA damage will be measured with the Comet assay. The results of the study will also compare the use of a number of cell types with the goal of identifying the most sensitive and cost effective model system. This study will provide valuable baseline data on the potential for adverse health effects of PM in an airshed that has received almost no scientific attention.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project focuses on the kinetics, metabolism and human toxicology of dichloroacetate-DCA and chloral hydrate-CH, potentially harmful metabolites of trichloroethylene-TCE. Humans exhibit polymorphisms of MAAI that may possess different kinetic properties toward DCA. Human haplotype variability may influence DCA's kinetics and toxicology. These hypotheses will be tested by accomplishing the following Specific Aims: Specific Aim 1: Quantify the influence of DCA, at exposure levels ranging from environmental, [unreadable]g/kg/d to clinical, mg/kg/d, on human liver MAAI and tyrosine catabolism. This aim tests the hypotheses that: there is a dose-dependent effect of DCA on human hepatocellular tyrosine metabolism in general and on the accumulation of potentially hepatotoxic tyrosine intermediates in particular; DCA inhibits hepatic MAAI expression in vivo in humans in a dose-dependent manner;and DCA, tyrosine and/or their metabolites form adducts with MAAI. Specific Aim 2: Establish the relationship between human MAAI haplotype and DCA and tyrosine metabolism. This aim tests the postulates that MAAI haplotype determines, and thus can predict: dose-dependent DCA kinetics and biotransformation and DCA's effects on tyrosine metabolism. Specific Aim 3: Determine the in vivo kinetics and biotransformation of CH in healthy adults and the influence of CH and DCA on each other's metabolism. This specific aim will examine three postulates: CH is metabolized in adults to DCA; CH, via DCA formation, inhibits its own metabolism and that of tyrosine;and these effects are dependent upon exposure level but not upon gender.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Uterine leiomyomas (fibroids) are the leading indication for hysterectomy in the United States. Despite the morbidity and high medical costs associated with fibroids, there has been little epidemiologic study of this condition in the United States. Uterine leiomyomas are histologically identifiable as benign smooth muscle tumors with varying amounts of associated fibrous tissue. Many women have more than one uterine leiomyoma, but each appears to be clonally distinct. Several specific cytogenetic changes have been identified in tumor tissue, but most show no chromosomal abnormalities. These benign tumors are hormone-dependent. They develop after puberty and regress after menopause. Both estrogen and progesterone are considered important stimulants, or at least permissive factors for tumor growth. To address the research needs in this field we have designed four studies. The first is a large epidemiologic study, the NIEHS Uterine Fibroid Study, designed to 1) estimate the age-specific cumulative incidence of leiomyomas in black and white women, aged 35-49, 2) identify risk factors for the condition, 3) compare growth mediating factors in tumor and matching myometrial tissues collected at time of hysterectomy, and 4) to identify factors associated with development of fibroid symptoms including pelvic pain and uterine bleeding. The second study (Fibroid Growth Study, Shyamal Peddada, PI) is a clinical study of fibroids designed to describe fibroid growth and compare the growth-mediating factors in growing vs nongrowing tumors. The third study, Postpartum Uterine Regression, monitors fibroid change with pregnancy and postpartum uterine regression. The fourth study, a prospective study of fibroid incidence, is currently under development. In this study we will enroll women before they have fibroids and follow them over 5 years for fibroid incidence. After estimating the age-specific incidence of uterine fibroids for black and white women, we began to examine risk factors for uterine fibroids. Pregnancy is protective, though not those that occur before the mid twenties. Alcohol appears to increase risk. In two cases we have replicated findings from animal models of fibroids. We find that the location of fibroids is somewhat different for parous and nonpauous women, and that prenatal exposure to DES is associated with increased development of fibroids. Increasing LH is associated with increased prevalence of the tumors, though LH may not be having direct proliferative effects, as we had hypothesized. We find no evidence for increased risk of fibroids with oral contraceptive use or with variability in menstrual-cycle length. We also explored our data on body fat and exercise. We find a small increase in risk with increased BMI (similar to other studies), and we also find that exercise is protective. As in the recent cohort analyses, smoking was not associated with risk in our data. We also collected questionnaire data for exploratory analyses on early-life exposures and several environmental/occupational exposures. While few factors showed associations with fibroid development, we did find an association of childhood use of insect repellent with fibroids. This may merit further investigation given the possible link between insect repellents and breast cancer. We measured fasting insulin and IGF-I in blood specimens collected from participants, hypothesizing both would be risk factors for fibroids. Surprisingly both tended to be protective, and diabetics were actually significantly less likely to have fibroids. We examined vitamin D status in relation to prevalence of fibroids both with the biomarker of hydroxylated vitamin D and with questionnaire data on time outside. With both methods, women with low vitamin D status had higher fibroid prevalence, and the findings were consistent for blacks and whites. We are beginning to examine dietary factors that may be related to fibroids. Regardiing fibroid symptoms, we found that urinary incontinence was significantly associated with fibroid size. We are currently examining the relationship between fibroid size and/or location and menstrual bleeding. The Fibroid Growth Study data have been analyzed and we conclude that: 1) spontaneous regression of fibroids occurs, 2) fibroids from the same woman grow at different rates, despite a uniform hormonal milieu, 3) fibroid size does not predict growth rate, and 4) age-related differences in fibroid growth between blacks and whites may contribute to the higher symptom burden for black women. We are currently examining short-term changes in growth of fibroids. In our study that monitored fibroid change during pregnancy and/or postpartum uterine regression we found that 36% of solitary fibroids were lost during the pregnancy/postpartum. Tumors that remained tended to have lost volume. We are now analyzing data to identify factors affecting the extent of fibroids reduction. We starting enrollment for the prospective study of fibroid incidence in October of 2010.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This project will address methodology development needs that are important in chronic disease population research. These include non- parametric estimator of multivariate survivor functions as relevant to disease prevention trials with multiple clinical outcomes and to family studies in genetic epidemiology. The closely related auxiliary data problem under which one uses data on multiple short-term response variables to strengthen the analysis of a primary clinical outcome will also be considered. Both estimators based on Peano series representations of the survivor function and likelihood-based estimators will be considered. Measurement error in key exposure and confounding variables can be a critical factor in determining observational study reliability. A more flexible than usual measurement model will be studied for application to such difficult to measurement exposures as nutritional and physical activity factors. Regression calibration estimators and empirical score function estimators of relative risk parameters will be studied based on data from this new measurement model in conjunction with an objective measures (biomarker) subsample. A third aim combines aspects of multi-variate failure time analysis and co-variate measurement modeling to develop new estimators of pairwise dependency in ages at disease onset of family members in the context of genetic epidemiology studies with measurement error in environmental factors. This project will also continue our studies of the role of various population study designs in the chronic disease research agenda as a function of the biases, measurement issues, and other limitations of ecologic studies, analytic epidemiologic studies, small intervention trials and full-scale intervention trials using the Women's Health Initiative database as a principal motivating setting.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our ability to translate new scientific knowledge rapidly into improvements in patient and population health continues to be limited by numerous challenges. The Northwestern University Clinical and Translational Sciences Institute (NUCATS) was launched in 2007 to create a home supporting clinical and translational (C&T) science at Northwestern and beyond. Northwestern University (NU) is a leading national research university affiliated with nationally-renowned clinical partners. NUCATS is nationally recognized as an innovation leader in several key areas of C&T science, pioneering novel and scalable approaches in the areas of preclinical therapeutic development, health and biomedical informatics systems, research networking platforms and training programs to promote multi-disciplinary team science; and creation and leadership of a regional consortium of the three Chicago-area CTSAs to promote community engagement activities and practice-based research. In the present proposal, we describe our second generation institute, NUCATS 2.0. The Mission of NUCATS 2.0 is Speeding transformative research discoveries to patients and the population. Our Vision is to transform NUCATS into a national model of a highly integrated academic nexus that continually increases the quality, safety, efficiency and speed of innovative C&T research, by pursuing three Global Aims: 1) To speed translation and improve efficiency by integrating processes and programs that connect researchers with a continuum of resources, training, funding opportunities, and strategic partners; 2) To develop and implement innovative systems to identify, evaluate, facilitate and disseminate scientific breakthroughs, and improve the quality, safety and cost-effectiveness of C&T research; and 3) To promote Northwestern's culture of collaboration, innovation and translation through team-building, education, and training to empower the multi-disciplinary translational research teams of tomorrow. NUCATS 2.0 will be an innovative leader driving the rapid translation of life-saving and health promoting scientific discoveries to improv clinical practice and population health.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The two target populations have in common their potential for exposures to chemicals with resultant negative health outcomes. Both populations must have hazardous materials emergency response training as required under OSHA's Hazardous Waste Operations and Emergency Response standard (29 CFR 1910.120 or 40 CFR 311). In recent years. National Incident Management System (NIMS) mandates and development of the National Response Framework (NRF) have resulted in a nationwide approach to preparedness for all types of emergencies that reaffirms the critical role of first responders in dealing with these emergencies. Unfortunately, federal funds for preparedness assistance have not reached first responders in amounts sufficient to meet their needs for training. The populations targeted by this proposal typically have training budgets that are insufficient to meet the training needs, as indicated by the attached letters of support, particularly when travel is required to receive training. The current economic downturn has significantly eroded each population's ability to provide this much needed training and therefore makes the proposed cooperative agreement even more important.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Environmental toxicants with the ability to directly or indirectly modify the cellular thiols of lymphocytes can initiate immune dysfunctions resulting in increased incidence of infectious diseases, cancer and/or autoimmune diseases. The majority of our analyses will employ T cells, the regulators of the immune system; however, we will also compare the thiol characteristics and sensitivities of T and B cells; furthermore, we will evaluate the thiol-related biochemical differences between mouse and human lymphocytes. Preliminary data suggests that these species have differential thiol characteristics. The ability of exogenous and endogenous thiols, thiol-blockers, oxidants and anti-oxidants, as well as irradiation, to alter the subcellular mechanisms necessary for the activation and growth of lymphocytes will be assessed. In order to accomplish this objective we first must evaluate some thiol characteristics key to the biochemical activation of lymphocytes. The lymphoid subsets have different thiol sensitivities which we believe are due, in part, to variances in their exofacial thiols. Biochemical and cellular/molecular biology technics will be utilized to assess the influences of thiol modulation at various levels within the cell (exofacial, transmembranal, cytosolic, and nuclear). Novel thiol reactive chemicals and antibodies will be used to probe these cellular domains. Various cDNA probes, monoclonal antibodies, and anti-sense oligonucleotides will be used to assess the regulation of the mRNA expression, protein expression (and distribution) and need for thiol modulation of cellular cysteinyl proteins believed to be involved in lymphocyte activation. Some specific proteins to be assayed include protein kinase C, phospholipase C, calmodulin, and ornithine decarboxylase as well as some proto-oncogenes (c-myc; c-fos). The homogenous nature of cloned T cells, T cell hybridomas, and T cell tumor lines will aid in the analysis of the thiol-related sensitivities and activities of the subsets. Subcellular differences between the T cell subsets and cell lines will be compared. Evaluation of some biochemical changes involved in the stimulation of the different types of T cells will aid in the assessment of differences between non-proliferating and rapidly proliferating T cells and the processes involved in modulating their activation. The mechanism(s) of cell injury and modulation of activation and growth by toxicants with thiol reactivities will be further defined by our analyses. In addition, information may be obtained on numerous diseases which concomitantly have aberrant immune functions and redox states such as AIDS, Bloom's Syndrome, Ataxia Telangiectasia and Down's Syndrome.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The role of poxviruses, subviral particles isolated from virions or purified poxviral DNA in causing abnormal proliferation or neoplastic transformation of human and murine cells will be investigated. The structure and replication of ortho and paravaccinia DNA's will be studied in order to define: (a) the nature of the viral chromosomal structure and (b) the viral \"replisome\", in order to gain insight into the structure and replication of intrinsically more complex eukaryotic cell chromosomes. Alterations in cellular membranes following infection with various vaccinia strains will be compared and defined to: (a) determine how such alterations affect cell growth and cell-cell recognition; (b) function in providing \"targets\" recognized by the host's immune system. Restriction enzyme analysis and molecular hybridization techniques will be used to probe the genetic relatedness of poxviruses and their diversification in nature. This will provide the basis for future studies of the genetic information needed for determining host range specificity, pathogenecity and cell proliferation or killing after infection of cells with poxviruses.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The assembly and release of HIV-1 particles is quite complex, in that a number of viral and cellular RNAs and proteins participate, and while much has been learned about this process, much remains to be discovered. The Gag protein is central to the assembly and release of HIV-1 and all other retrovirus particles. A key function of the Gag protein that will be one focus of tis proposal is the recruitment and packaging of viral genomic RNA (gRNA). Reciprocally, gRNA may help to drive the recruitment of additional Gag molecules into a virion as assembly progresses. However, the full extent to which RNA participates in particle assembly in cells, and how Gag:RNA interactions change as assembly is initiated and completed remain to be defined. The experiments proposed in specific aim 1 will be the first to determine precisely how HIV-1 Gag and gRNA interact in cells. These studies could identify the very first as well as subsequent interactions between Gag protein and gRNA as well as between gRNA and antiviral cytidine deaminases. Following assembly, HIV-1 particles depart from the surface of the infected cell, to colonize neighboring or distal uninfected cells. During the previous funding period, we identified an IFN-induced membrane protein, termed tetherin, as a host defense molecule that inhibits HIV-1 particle release. Our work has demonstrated that tetherin acts directly, to retain nascent virions on the cell surface, but key mechanistic details of how tetherin functions, and what role t plays in preventing the dissemination of viral infection in cultured cells and in vivo remain to b determined. In specific aim 2, we will perform a series of in vitro and in vivo experiments to address these key questions.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract The signals and processes by which diverse stimuli lead to keratinocyte cytokine production and cytotoxicity are presently not well understood. Moreover, chronic stimulation by agents such as ultraviolet radiation leads to malignancy. Recent studies have demonstrated that ultraviolet B radiation (UVB) damages human keratinocytes (HK) in part by inducing oxidative stress and cytokine production. Severe UVB damage to the keratinocyte can also result in apoptosis, or programmed cell death. Previous studies by our group and others have indicated that keratinocyte damage, especially via agents that result in oxidative stress results in the production of glycerophosphocholines (GPC) that can act as agonists for the Platelet-activating Factor (PAF) or the peroxisome proliferator activated receptor gamma (PPAR) systems. The long-term objective of this ongoing research proposal is to elucidate the role of these oxidized GPCs in the acute UVB-induced photoresponse. Three specific aims are designed to test the hypothesis that UVB-mediated oxidative damage to the keratinocyte results in the production of novel ox-GPCs that activate the PAF-R and PPAR systems. These novel lipids augment cytokine production, modulate apoptosis, and induce keratinocyte proliferation: 1). The effect of these ox-GPCs on acute UVB-mediated inflammation, cytokine production, DNA damage/apoptosis and proliferation will be assessed in mice deficient in PAF-R and epidermal PPAR; 2) Mass spectrometric methodologies will be used to structurally characterize ox-GPCs produced in response to UVB; and 3) The role of these ox-GPCs in the acute UVB photoresponse in humans will be determined. We anticipate that these studies using novel in vitro and in vivo model systems and novel methodologies will result in the characterization of ox-GPCs with PAF-R- and PPAR- agonistic effects in acute UVB-induced keratinocyte cytokine production, cytotoxicity, and proliferation. This information will aid in the understanding of the mechanism(s) by which oxidative stress can be involved in cutaneous pathophysiology. Since these mediators would be produced in response to oxidative stress in other organ systems, these studies should provide important insights into and tools to study the impact of ox-GPCs in human pathophysiology.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Aging and diabetes are associated with increased generation and deposition of Advanced Glycation Endproducts, or AGEs, the products of nonenzymatic glycation/oxidation of proteins/lipids. As human subjects age, the incidence of insulin resistance and, often, frank hyperglycemia rises. Long-lived proteins of the peripheral nervous system (PNS) are highly susceptible to AGE modification. AGEs, an heterogeneous class of molecules, may modify cellular function by an array of distinct mechanisms. One such mechanism is by their ligation of the signal transduction receptor, RAGE. RAGE also interacts with S100/calgranulins and amphoterin (or high mobility box group 1, HGMB1), which display diverse functions, such as neurite outgrowth and inflammation. Among the complications of diabetes, symmetrical neuropathy of the sensory and autonomic nervous systems affects significant numbers of diabetic subjects. Evidence accrued from the first five years of this Project has suggested an innate, survival role for RAGE in response to acute peripheral nerve injury. In euglycemia, transient and sharply-limited upregulation of RAGE (particularly in mononuclear phagocytes [MP] and neuronal/axonal structures) and its ligands is linked to RAGE-dependent inflammation and neurite outgrowth that contribute beneficially to regeneration. In diabetes, regeneration consequent to acute injury, such as crush, is significantly impaired. We hypothesize that in diabetes, chronic and sustained accumulation of AGEs and RAGE ligands, and upregulation of RAGE, perturbs homeostatic mechanisms in the peripheral nerve, leading to chronic dysfunction. Upon superimposed acute nerve injury, RAGE-dependent mechanisms sustain inflammation and neuronal dysfunction, and thwart regeneration. This proposal will address the fundamental question of whether RAGE/RAGE signaling globally, or specifically in cells of MP lineage or neurons, is protective or destructive to peripheral neurons in diabetes. We propose that in euglycemia, acute engagement of RAGE/RAGE signaling in both cell types is protective for peripheral neurons, but that chronic activation in diabetes promotes long-term neuronal dysfunction in the absence or presence of superimposed acute injury. This Project is critically linked to Project 1, as studies in chronic AB-enrichment in the CMSstrongly support deleterious and maladaptive roles for RAGE in neuronal stress. As antagonism of RAGE nears clinical trials in Alzheimer's Disease and diabetes, it is imperative to dissect the beneficial vs maladaptive impact of RAGE in both the CMS and PNS, and in chronic stress vs acute injury. Projects 1 and 2 are uniquely positioned to address these questions due to the long-time collaboration of the leaders of these Projects. Project 2 will employ Cores A and B through all five years of this Program.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "PROJECT SUMMARY The International Consortium on Brain and Behavior Copy Number Variants (IBBC-CNVs) is a collaborative effort of 9 institutions with complementary experience and expertise in phenomics and genomics. The 22q11.2 and 16p11.2 loci are associated with significant risk for neuropsychiatric disorders across the lifespan. The clinical presentations are heterogeneous, manifesting in a range of developmental neuropsychiatric disorders, including Attention Deficit Hyperactivity, Anxiety, Autism Spectrum, and Psychosis Spectrum Disorders. Taking a `genetics first' approach of ascertaining patients based on known, homogeneous genetic etiologies will allow us to overcome barriers posed by the genetic and phenotypic complexity of idiopathic developmental neuropsychiatric disorders. We postulate that CNVs exert a large main effect on psychopathology, but the nature and degree of psychopathology observed in CNV carriers is multifactorial, with contributions from additional rare and common genetic variants, as well as environmental factors. Therefore, dissecting the effects of major CNV hits as well as additional rare and common variants on dimensional measures of psychopathology can elucidate the combined contribution of genetic mechanisms to psychiatric conditions and build models of risk prediction. Notably, the presentation and course of psychopathology in the CNVs resemble these features in idiopathic disorders. Therefore, beyond the specific genetic syndromes investigated, such a cross-CNV effort will identify convergent risk mechanisms for developmental neuropsychiatric disorders that are of relevance to the broader population. We propose to dissect dimensional measures of psychosis, social-emotional processing and neurocognition, and their genetic and environmental modifiers, to elucidate the architecture of risk for neuropsychiatric disorders in CNV carriers. Prospective evaluation with dimensional measures relevant to neuropsychiatric disorders will be applied to a cohort of 2000 individuals with 22q11.2 and 16p11.2 deletions and duplications (500 per group) and their relatives as feasible. In addition, categorical psychiatric diagnoses will be assessed in CNV carriers. Recruitment for prospective phenotyping will leverage existing large cohorts that carry these reciprocal CNVs, many of whom have already been ascertained and characterized with a range of phenotypic measures. New whole genome sequencing (WGS) will be performed in CNV carriers that have not yet been sequenced. We will also utilize existing genetic data from the largest available case-control samples diagnosed with SZ, ASD, and ADHD in the PGC to generate polygenic risk scores. Finally, we will examine family and environmental factors that contribute to the heterogeneity of presentation and developmental course in CNV carriers. This project will establish common phenomic and genomic resources. Our ability to conceive such a large-scale study capitalizes on our existing successful collaborations, complementary expertise, and institutional commitments to achieve these goals.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aims of this competitive renewal application are to maintain and expand the present program for students participating in cancer research. The ultimate goal is to increase the number of health professionals, especially those from under-represented minorities, that will pursue clinical and/or research careers in cancer. The current grant award, Nutrition-Cancer Education Program (R25 CA47877) began September 1, 1988 and will end August 31, 1992. The grant award included two program components: Nutrition-Cancer Curriculum (Program I) and Short Research Experiences (Program II). This renewal application addresses the Short Research Experiences only, since Program I is not renewable. During years 01 through 04 of this award, the number of applications to the program have increased each year as has the quality of the students receiving the awards. Additional, there has been a higher percentage of minority student recipients when compared to the percentage of minority students enrolled in the Louisiana State University School of Medicine. In this application, the following activities are proposed: * continued funding for students from the health professions to participate in the Short Research Experiences in cancer/oncology; * expansion of the assistantships from 12 to 18 positions, including two positions for outstanding high school students; * new approaches for the recruitment of minority participation; * modification of the educational component of the Short Research Experiences; * use of newly-developed tools for evaluation of the Short Research Experiences; The above activities will be implemented in order to expand the educational and research opportunities of students in healthcare in Louisiana. It is anticipated that these opportunities will eventually contribute to cancer control in a state with excessive cancer rates, especially among the minorities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Comprehensive examination of RNA structure and function can yield insights Impossible to giean from analysis of individual molecules. To aid understanding of the large and rapidly growing corpus of RNA structural data, we propose to create a structural classification of RNA (scaR). This database will organize features at four levels of complexity: three-dimensional motifs, tertiary interactions, structural domains, and whole molecules. Each of these may have multiple means of organization; for example, both by loop size and by base stacking. To initially define significant recurrent features, the classification will begin with manual inspection of all RNA structures. As the project progresses, observed characteristics will be formalized into algorithms that will be applied to perform rapid, reproducible, and accurate analysis and classification of RNA structures. [unreadable] [unreadable] The structural features will be mapped into a functional classification, and the associations between the two will be used to study general relationships between aspects of RNA structure and their function. To explore the evolutionary basis of RNA structure and the dependence of structure upon sequence, detailed links will be forged between classified RNAs of known structure and their evolutionarily related families. [unreadable] [unreadable] This classification will immediately elucidate the prevalence and variability of features in RNA structure. In addition, it is expected to have applications in RNA molecular modeling, molecular design and engineering, and RNA ligand and drug design. It will greatly enhance our understanding of RNA structure-function relations, the correlation between RNA structure and physical and biological properties, and evolutionary effects on RNA structure. The database will also provide a convenient and logical means for experts and neophytes alike to access and interpret the growing wealth of RNA sequence and structural data.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have shown in our laboratory that rhesus monkeys with multiple inoculations of Brugia malayi L3 develop lymphatic pathology and immune responses to B.malayi adult worm antigen (BmA) that resemble, in many respects, those of human lymphatic filariasis patients. One similarity of the immune response between human and rhesus with brugian filariasis is that both exhibit T-cell unresponsiveness to BmA in vitro. In an attempt to overcome the cellular unresponsiveness to BmA, eight multiple-infected rhesus monkeys whose cells exhibited unresponsiveness to BmA and two uninfected controls were studied. PBMC were co-cultured with antigen and one of the following cytokines that have been reported to be effective in reconstituting T-cell proliferation interleukin (IL)-2, IL- 12, anti-IL- 10, anti-IL-4 and anti-IL- 13. We were able to overcome antigen-specific unresponsiveness in only a minority of the rhesus monkeys studied. Co-culture with either IL-2 or IL-12 were the only conditions which resulted in a slightly enhanced proliferation to BmA in these animals. Even high concentration of every cytokine was unable to render a significant antigen-specific proliferation. The results indicate that more than one distinct immunological mechanism may account for the antigen-unresponsiveness in rhesus monkeys infected with brugian filariasis.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This longitudinal study, now in its sixth year, assesses the effects of increased calcium intake on bone mass formation in females during puberty and early adolescence using double-blind, placebo-controlled trials of calcium supplements.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objective of this program is to determine by a study of the nucleotide sequences in oncornavirus RNA whether the large 35S RNA subunits comprising the genome RNA are identical or unique polynucleotide chains. Regions of the RNA at the 5'-termini and adjacent to the poly(A) segments will be examined. In addition, the nature of the linkage of the 35S RNA subunits will be examined as well as the binding site(s) for the 4S RNA molecule which primes DNA synthesis by the viral RNA-dependent DNA polymerase.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This is a multi-center, randomized, open-label, parallel group trial to compare the efficacy of 9 ug interferon alfacon-1 administered sub-q three times per week for 72 weeks or 1 X day for 48 weeks to the same dose administered TiW for 48 wks in subjects with chronic HCV infection, previously untreated with interferon. Efficacy is defined by undetectable serum HCV RNa levels 24 wks following cessation of therapy.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long term objectives of the proposed studies is to determine the regulation of UDP-glucoronosyl transferase (UDP-GT) inducer-mediated changes in thyroid follicular cell proliferation and apoptosis. This proposal describes a series of experiments that address how these chemicals alter rodent thyroid physiology, both in vivo and in vitro. Preliminary studies with other goitrogens suggest that UDP-GT inducers will effect thyroid follicular cell proliferation and apoptosis. Previous studies have emphasized feedback production of thyroid stimulating hormone (TSH) as the sole contributor to thyroid follicular cell proliferation, and ultimately, thyroid neoplasia. Recent findings such as, 1) while TSH inhibits apoptosis, propylthiouracil (PTU) treatment leads to an induction of apoptosis, and 2) insulin-like growth factor-1 (IGF-1) potently enhances TSH-mediated proliferation, strongly suggest that in addition to TSH that bother\" factors are also important in regulating thyroid follicular cell proliferation and death. The findings from these studies are of importance to human health because there are several therapeutic and environmental agents that induce UDP- GT activity, decrease serum thyroid hormone, and potentially increase serum TSH concentrations. Yet, the relative importance of other factors is unknown. The studies outlined in this proposal will increase the knowledge base in the regulation of thyroid-follicular-cell proliferation and apoptosis, thus, enabling future studies with UDP-GT inducers to accurately assess which changes in thyroid physiology are impor nt.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DCCPS supports web-based, online, electronic and other communication and education efforts as part of its mission. These activities enable the organization to be more interactive with stakeholders by sharing important information about research portfolios and the resulting tools, products and scientific advances.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-range goals of this project are: (1) to evaluate the effects of prenatal exposure to environmental chemicals on the subsequent reproductive capacity of the offspring; (2) to investigate the mechanisms involved in the production of subfertility in mammals as a result of their in utero exposure to foreign chemicals; (3) to assess the transplacental carcinogenic potential of these compounds; (4) to study the physiologic disposition and metabolism of these compounds in the pregnant animal and fetus; (5) to study chemico-biological interactions of transplacental toxicants, with special emphasis on structure-activity relationships; (6) to determine if prenatal exposure to environmental agents can alter the cytoplasmic receptors for steroid hormones in reproductive tract tissues; (7) to develop and utilize organ culture systems to study the effects of environmental chemicals on the fetal ovary and reproductive tract in vitro; and (8) to evaluate the above animal models as predictors of human response. Special attention is given to diethylstilbestrol (DES).", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Virtual product design and assessment has become a valuable tool in the development and evaluation of new buildings (and consumer products) as well as the interior design of automobiles. While these processes are beginning to work well for whole body designs targeted at accommodating population anthropometries, particularly in an effort to reduce injuries associated with awkward postures (Corlett et al. 1980, Chang et al. 2003, Chaffin 2005, Perez 2005), they have not been applied to hand tool or device design. It has been shown that repeated use of hand tools by individuals with hand dimensions significantly different than for which the tool was intended has resulted in an increased potential for hand-related injuries (Cobb et al. 1996, Meagher 1987, Kar et al. 2007, Markison 2007), particularly when significant force is required to operate the tool (Sancho-Bru et al. 2003, Molteni et al. 2008). Virtual hand models have been ineffective in addressing such anthropometric tool-fit issues. Current technological impediments related to available virtual hand models include (1) a lack of a solid geometry hand with accurate surface representation and accurate joint kinematics defined by centers of rotation, (2) a limited capacity in accounting for anthropometric scaling (e.g. the relationships between individual finger segment dimensions) (3) missing information on predicting how the hand grips objects and the forces applied during gripping, and (4) an inability to account for tissue compliance (particularly at the finger tips and palmar surface). The proposed project intends to address these impediments by developing a 3D geometric hand model, to be integrated into the current CAD design software products used by design engineers (e.g., SolidWorks, ProE) to evaluate the interaction between the hand and a new product. The proposed Phase 1 research will result in a virtual hand model that specifically targets the first two impediments (listed above) and will establish a draft framework (to be used to scope future research) for the necessary parameterization to address (3) and (4). At the conclusion of phase 1, a stand-alone software program with a scalable geometric hand representation with kinematically realistic articulating digits will be produced. The virtual hand model will be developed using the open-source SimTK core libraries (Sherman et al. 2005, Delp et al. 2007, Schmidt et al. 2008). The bones and joints of the hand will be modeled using rigid body structures to mathematically replicate laboratory recorded hand anthropometry, joint centers, and kinematics. The surface representation of the hand will be modeled using a similar method used by rigid body spring models (RBSM) (Kawai 1980). An integrated hand model, incorporating both the proposed surface-deformation/skeletal model and existing muscle models, would provide a powerful analysis tool for 1) understanding hand related injury mechanisms associated with grip posture and force and 2) optimizing tool design in-silico, prior to workplace deployment. PUBLIC HEALTH RELEVANCE: There currently exist no methods for design engineers to assess prospective design changes in a virtual environment as related to the overall tool-hand fit with respect to different population hand sizes. This has led to hand tool designs that are inappropriate for a large number of users and whose repetitive use will result in an increased potential for injury (Meagher 1987, Markison 2007). The overall goal of this project is to develop a scalable, virtual hand model that can be used to evaluate and determine appropriate hand-tool coupling interfaces, information that can be used to design hand tools to accommodate the hand sizes and hand shapes of end users.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We have found that the receptor FPRL1 recognizes Alzheimer's disease-associated amyloid beta peptides and promotes proinflammatory responses in the brain. We used mouse FPRL1 counterpart mFPR2 as a model for further understanding of the receptor in the progression of the disease. We found that mFPR2 in microglial cells, the macrophages in the brain, is upregulated by diverse proinflammatory stimulants, including agonists for the pattern recognition receptors and cytokines. Increased expression of mFPR2 on microglial cells enhances the capacity of the cells to endocytose amyloid beta peptide, therefore the receptor may profoundly affect the progression of the disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Modern biomedical imaging technologies allow for generation of high-resolution digital 3D images of many microscopic biological objects. However, effective strategies remain to be developed for automatic quantitative and statistical analysis of such microscopic structures' 3D morphologies. This proposal is aimed at developing algorithms for automatic morphometric analysis of neurons in intact brains. Quantitative and statistical characterization of individual neurons' spatial locations and their 3D projection patterns is not only essential for understanding brains' complexity, diversity, and plasticity with single-cell resolution, but also critical for elucidating subtle cellular pathological mechanisms underlying various neurological/mental/psychological disorders. New expertise will be explored to advance technologies in multiple areas of biomedical imaging, such as image computation and simulations of complex tissues. A GAL4-independent binary transcriptional system has been developed to label specifically the entire morphologies of the Drosophila olfactory learning and memory center, the mushroom bodies (MBs). In conjunction with MARCM (Mosaic Analysis with a Repressible Cell Marker) technologies, one can independently label various single MB neurons and the whole MBs in the same brains. Meanwhile, new algorithms have been developing to conduct automatic morphing (morphological deformation & matching) of irregular-shaped 3D objects. A virtual average MB will be constructed via statistical characterization of pair-wise morphing among multiple \"standard\" MBs. Morphometric analysis of distinct MBs and spatial mapping of individual MB neurons will then involve establishing point-to-point correspondence between the MBs of interest or the MBs, in which specific single MB neurons are differentially labeled, and the statistical model MB. Thus, one may be able to detect automatically and describe quantitatively any given MB's structural deviations and to identify individual MB neurons based on their 3D neuronal location/projection patterns.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The nicotinic acetylcholine receptor (AChR) is a pentarneric integral membrane protein with the subunit composition alpha2beta-gamma-delta . The five subunits form an ion channel at their center whose opening and closing is controlled by the binding of acetylcholine. The nicotinic AChR is muscle is the best-studied of a large family of neurotransmitter receptors that include the receptor for GABA glycine and the neural AChR. Glutamate receptors are formed according to a similar pattern, but are more distantly related. Our laboratory studies the structure, function, and assembly of the nicotinic AChR n muscle. We investigate both the properties of the receptors in muscle cells and in heterologous cells in which the AChR is expressed after transfection of receptor subunit cDNA. Because of the similarity of members of the family, findings that related to the AChR serve as a model for other receptors, which are targets for drugs that are relevant to the treatment of mental illness.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The goal of this research is to develop novel, biologically functional DNA nanostructures that dramatically enhance the reproducibility, sensitivity, and spatial density of chip-based DNA assays. These nanostructures will improve applications ranging from point-of-care diagnosis to genomic arrays used in basic research by enabling the development of next generation screening technologies that are faster, more sensitive, more reliable, and possibly more cost effective than those presently available in the life sciences market. To accomplish the stated goals, Nanolnk will develop a DNA patterning methodology based on Dip Pen Nanolithography (DPN) to generate sub-micron sized features of DNA on solid surfaces. This multidisciplinary effort will involve life and physical scientists at Nanolnk, MEMs and instrumentation engineers at our fabrication facility, in addition to support from outside experts in the fields of DNA microarrays and microfabrication. DPN, built upon the technique of Atomic Force Microscopy (AFM), allows one to deposit materials uniformly in a direct-write fashion on surfaces with nanoscale spatial precision. This strategy offers significant advantages over current microarray printing technologies that suffer from poor spot to spot reproducibility in terms of size, shape, and oligonucleotide density, as well as reproducibility across microarray slides. Preliminary work has demonstrated that the DPN technique can be used to deposit 12mer synthetic oligonucleotides on surfaces with extremely uniform sub-100 nm to several micron scale features. The DNA nanostructures formed robust films and exhibited selectivity in binding to complementary oligonucleotides. Thus, DPN can be used to generate uniform features of synthetic DNA far smaller than can be obtained with other spotting or photolithography techniques. In Phase I, Nanolnk will demonstrate feasibility of the DPN-based approach for generating sub-micron scale DNA nanostructures on glass surfaces. The resulting nanostructures will be analyzed using existing fluorescence probe technology to provide benchmarking standards for comparison to conventional microarray assays. In addition, for applications in life sciences and biomedicine, it is desirable and advantageous in terms of speed and throughput to extend the serial patterning capability of DPN to a parallel methodology. Thus, concurrent with ink development and patterning optimization, microfabricated parallel multipen arrays will be explored as a means for faster, simultaneous writing of multiple DNA inks.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Background: START, a Study of Trauma and Reduction of HIV Transmission, is a cross-sectional study that will examine whether traumatic stress contributes to potentially amplified transmission (PAT) risk behavior (the co-occurrence of detectable HIV viral load and HIV transmission risk) among stimulant-using men who have sex with men (MSM). Stimulant-using MSM are most heavily affected by HIV, despite the widespread availability of antiretroviral therapy (ART).1 Treatment as Prevention (TasP), defined as expanded ART access to achieve sustained viral suppression, is a promising biomedical prevention approach that will be utilized. While literature supports that early ART initiation decreases HIV transmission rates, stimulant-using MSM were excluded from these landmark trials. Stimulant-using MSM are known for low utilization and low engagement in HIV-related care thereby resulting in higher viral loads and high HIV transmission rates.2 The overarching objective of this proposed study is to identify modifiable risk factors associated with PAT to inform the development of innovative, theory-based interventions to optimize the effectiveness of TasP in stimulant-using MSM. Resulting information will inform the development of HIV/AIDS prevention approaches enhancing the effectiveness of TasP in stimulant-using MSM, thereby decreasing HIV transmission rates. Relation to Training: This proposal is designed to provide in-depth research training. Pathway analysis will be conducted to understand the role of psychosocial factors in the relationship between traumatic stress and PAT. The results will provide preliminary data for the preparation of a K99/R00 grant, which will be used to develop and pilot test a behavioral intervention to reduce traumatic stress symptoms and HIV transmission in stimulant- using MSM. This F31 proposal is a logical step in becoming an independent nurse investigator. Innovation: The study is innovative in three ways. First, PAT, a co-occurring outcome variable, will be utilized to model an outcome most closely linked with high HIV-viral transmission efficiency; second, the potential mediational role of psychosocial factors in the relationship between traumatic stress and PAT will be explored; and third, novel techniques will be utilized, including (a) the Balloon Analogue Risk Task (BART) measure of impulsivity while eliminating the limitation of a self-report measure requiring self-insight and awareness, and (b) Audio Computer Assisted Self Interviewing (ACASI), which is used in behavioral research to minimize recall and social desirability bias and enhance the veracity of self-report, thereby increasing internal and external validity.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "There is an increase in the risk of bone fracture with aging and adult diabetes, and this increase cannot be solely explained by changes in bone mineral density (BMD). One barrier to new diagnostic tools and treatments is that the underlying cause for the disproportionate increase in fracture risk among diabetics and the elderly is currently unknown. Consequently, there is a need to identify the biophysical basis of the age- and diabetes-related changes in bone that decrease fracture resistance, not just bone strength. Addressing this, the proposal aims to determine whether increases in advanced glycation end-products (AGEs) explain the effect of diabetes and aging on the fracture resistance of bone (as characterized by fracture properties) and to determine whether an AGE inhibitor and/or an antioxidant can both improve the quality of bone structure and increase the fracture properties of bone. Aim 1 will determine the role of bone structure, BMD, and AGEs in the effect of Type 2 Diabetes (T2D) and aging on the fracture properties of bone. In the first experiment, bones will be collected from T2D rats and non-diabetic rats at 24 weeks and 32 weeks of age. In the second experiment, bones will be harvested from aging rats at 4 months (young), 12 months (adult), and 24 months (old) of age. All the bones will undergo extensive analysis in order to identify the relative contribution of compositional properties such as BMD and AGEs and structural properties such as moment of inertia to a set of biomechanical properties including traditional measurements of strength and new measurements of fracture toughness and fatigue life. Aim 2 will evaluate how exogenous glycation of collagen affects the fracture properties of bone. This aim investigates whether the direct accumulation of AGEs within the extracellular matrix of bone decreases the fracture resistance of bone and whether the AGE inhibitor pyridoxamine, a B6 vitamin, protects against such a change. With appropriate controls, both human cortical bone and rodent bone will be incubated in diabetic concentrations of glucose with and without the inclusion of pyridoxamine. After quantifying the concentration of pentosidine, a biomarker for AGEs, and BMD, each specimen will be subjected to a mechanical test. In the case of the human bone, tests will determine the effects of increasing AGE on bone strength, post-yield energy dissipation, fatigue life, and crack-initiation & crack-growth toughness. For the rodent bones, tests will determine the effect of increasing AGE on fatigue life and fracture toughness, the ability to resist crack propagation. Aim 3 will assess the efficacy of pyridoxamine and N-acetylcysteine to increase fracture resistance of bone in an aging rat model of T2D through changes in bone structure, BMD, and AGEs. In this translation aim, the role of oxidative stress and AGE accumulation in the aging and diabetic effects on bone will be investigated using these two compounds. Starting at 4 months of age, non-diabetic and T2D rats will drink water, water with pyridoxamine, or water with N-acetylcysteine, an antioxidant. After 4 months, 8 months, and 14 months (aging) of treatment, bones will be harvested for extensive analysis to determine the effects of treatment on the structural, compositional, and biomechanical properties of bone. Additionally, histological and cell culture assays will assess the biological effects of the compounds on oxidative stress and osteoblast differentiation. To achieve these aims, Micro-Computed Tomography will quantify volumetric BMD; high performance liquid chromatography will quantify the concentration of crosslinks and collagen content; and thermal gravimetric analysis will quantify the collagen and mineral fractions as well as water content. Statistical models will determine the relative contribution of the structural and the compositional properties to the fracture properties of bone. This will address the relevance of targeting oxidative stress and AGEs to improve the bone health of Veterans and prevent bone fractures. The long-term goal is to identify the factors affecting the important determinants of fracture resistance and developing accurate diagnostic assessments of fracture risk.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Although antiestrogens are known to be effective against many metastatic breast cancers and are presently undergoing extensive clinical trials, little is known of their mechanism of action beyond the fact that they mimic estradiol in binding cytoplasmic estrogen receptor and translocating it to the cell nucleus. I therefore propose to compare selected antiestrogens with active estrogens at four critical steps of estrogen action: 1) temperature-dependent estrogen-induced receptor transformation; 2) receptor translocation rate into cell nuclei; 3) receptor binding to specific nuclear acceptor sites; and 4) antiestrogen retention by nuclear receptor and its relation to clearance from the general circulation. Three model systems will be compared: the normal rat and mouse uterus, the dimethylbenzanthracene-induced rat mammary tumor system, and the MCF-7 human breast cancer cell line. The goal will be discovery of the critical differences between estrogen and antiestrogen action, particularly in mammary tumor cells and particularly as correlated with tumor responsiveness to antiestrogen therapy. The results should help improve the design of atiestrogen therapy alone or in combination with chemotherapy, should aid in the search for more effective antiestrogens, and should also contribute substantially to knowledge of normal estrogen action and the nature of hormone dependence in breast cancer.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Radiation-induced gastrointestinal syndrome (RIGS) results from a combination of direct cytocidal effects on intestinal crypt and endothelial cells and subsequent loss of the mucosal barrier, resulting in microbial infection, septic shock and systemic infiammatory response syndrome. Currentiy, there is no therapy for RIGS. Irradiation induces apoptosis of crypt endothelial cells, intestinal stem cells (ISC) and enterocytes within hours. We rationalized that the acute loss of cells in situ requires rapid compensation of their functions and this was best achieved with cell replacement therapies, e.g., blood transfusion for hemorrhage. The stroma of solid organs contains a variety of supporting cells, such as, mesenchymal and microvascular endothelial cells, macrophages and lymphocytes. These stromal cells provide the niche and could supply critical growth factor/signals for ISC regeneration. For example, upon intestinal mucosal disruption, resident macrophages in the intestinal submocosal layers are activated by pathogen-derived ligands for Toll-like receptors (TLR) and transmit regenerative signals to ISCs. We thereby propose intestinal regenerative therapy with a combination of systemic administration of growth factors and cell replacement therapy to salvage Gl function post-radiation exposure. In order to develop an stem cell-based therapeutic strategy for RIGS, we hypothesized that combinations of: a) intestinal stem cell growth factor, R-spondinl (R-spol), b) TLR ligands, and c) transplantation of bone marrow-derived endothelial progenitor cells (EPC) and mesenchymal stem cells (MSC) would restore the IR-damaged ISC niche, protect against IR-induced cell death and provide growth signals for host ISC regeneration, thus providing protection and mitigation from RIGS. Aim I Pathophysiologic Mechanisms, Discovery and Validation of Molecular Targets in RIGS Aim II will investigate whether acceleration of ISC regeneration could mitigate/protect RIGS by administration of a Wnt agonist, R-spondinl and a BMP antagonist. Aim III will examine whether repair of the ISC niche by TLR activation and/or bone marrow-derived adherent stromal cell-based therapies could mitigate RIGS in mice. The final goal is to identify radio-mitigating factors secreted by stromal cells.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This grant application requests five years of renewed support for the Arizona Alzheimer's Disease Core Center (ADCC). Scientifically, the ADCC is intended to optimize the development and use of its Cores, so as to capitalize on Arizona's scientific and organizational resources in the understanding, very early detection, and tracking of Alzheimer's disease (AD) and in the discovery of disease-slowing and prevention therapies. Organizationally, the ADCC is intended to establish a leading model of statewide collaboration in AD research. The Administrative Core provides the leadership and support needed to optimize the development, interaction, and use of its Cores. Working closely with researchers inside and outside Arizona, the National Alzheimer's Coordinating Center (NACC), and other AD Centers, it promotes the development and progress of AD-related studies and collaborations. It administers a program for the statewide solicitation, competitive review, and support of pilot studies. It helps solve the challenges and fulfill the opportunities associated with the ADCCs statewide collaborative model, and ensures the ADCC's accountability to the NIA. The Clinical Core maintains a large pool of clinically well characterized and annually assessed research subjects for the scientific study of AD and aging. The subjects include patients with AD and other dementias, patients with mild cognitive impairment (MCI), and normal controls, almost all of whom are enrolled in a brain donation program, and a growing pool of Latino and American Indian research subjects. This Core ensures the comparability and productive and appropriate use of subjects and data from its five clinical sites. It also promotes the productive and appropriate scientific use of longitudinally followed subjects, DNA, and data from three independently funded ancillary programs, yielding new information about the transition from cognitively normal aging to cognitive decline in persons at differential risk for AD. The Data Management and Statistics Core maintains the ADCC's database, helps ensure the quality of data and the protection of subject confidentiality, and provides statistical services in a manner that best serves the needs of the statewide ADCC. It works closely with researchers, NACC, and other AD Centers, sharing data in the most productive, timely, and appropriate way. The Neuropathology Core provides neuropathological diagnoses and extremely high-quality brain tissue from expired Clinical Core subjects to support research studies in Arizona and around the world. It also promotes the productive and appropriate use of biological materials from a large additional number of clinically and neuropathologically well characterized non-demented elderly subjects from its independently funded ancillary brain donation program, helping to address a critical need in the AD research community. The Education and Information Core provides training, innovative educational and outreach programs, and strategic partnerships to promote the development of AD-related researchers, address needs of professional and family caregivers, provide information about the ADCC, and address unmet needs of Arizona's American Indian and rapidly growing Latino communities.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Over the past five years, this Purdue University team has been collaborating with colleagues at Rutgers University to investigate the computational and experimental infrastructure for continuous manufacturing (CM) of solid oral dosage products. This work has built on a decade of research under the NSF supported Center for Structured Organic Particulate Systems. In parallel, the PI and Purdue co-workers have investigated CM approaches to small molecule API manufacture at micro and intermediate scales. This work has demonstrated the essential roles of process modeling, process analytical technology, active process control, intelligent process monitoring, material tracking and real time risk assessment. Moreover, current work is showing how these components must be linked through an integrated data management and informatics infrastructure in order to achieve the desired Industry 4.0 functionalities. While CM is an important development for the pharmaceutical industry, it is not a universal solution that meets all manufacturing needs, either technically or economically: often hybrid batch-continuous or fully batch modes can be advantageous. Moreover, given the major investment in existing batch facilities in the generic \\manufacturing sector, there is an unmet need to bring Industry 4.0 functionalities to those manufacturers and thus significantly improve quality and reduce cost of medicines. The goal of this proposal is to expand the research on CM to develop and demonstrate a framework for the design and operation of batch and hybrid small molecule manufacturing systems. This will be achieved through five aims: Aim 1: Development of Pharmaceutical Model Library for small molecule and oral drug product manufacture Aim 2: General framework for the optimal synthesis of processes for small molecule-based API and product manufacture, encompassing the spectrum from batch to fully continuous processes Aim 3: Development of an Industry 4.0 real-time process management framework (RTPM). Aim 4: Demonstration of these technologies using several case-studies including high cost/low volume and high volume/low cost generic drugs at three different scales (lab, pilot and industrial) Aim 5: Development of instructional modules for conducting training programs for both FDA staff and industry as well as web-based access to tools and cases studies via pharmaHUB. This work will result in the development of the tool set necessary to implement Industry 4.0 across the pharmaceutical sector as well as the demonstration of the framework for systematic design and operation of processes for several specific drugs, including the effects of scale. The case studies will serve to inform and promote innovative manufacturing practices across the numerous batch and hybrid batch-continuous facilities existing worldwide. Moreover, by complementing the progress made in CM, it will enable the FDA to develop effective guidelines on the application of Industry 4.0 functionalities across the industry.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Glucose uptake by muscle and fat tissue is a primary mechanism for energy storage and represents a main control mechanism for glucose concentration in the circulation. A detailed understanding of the intracellular signaling system that mediates insulin-triggered translocation of the glucose transporter (GLUT4) will likely provide new findings for attacking diseases such as diabetes and obesity. The main goal of the proposal is to develop a kinetic screen to identify adipocyte regulatory proteins that control dynamic parameters for PIP3 signaling and for GLUT4 translocation and endocytosis. We will establish a wide-field TIRF system to measure the translocation and endocytosis rates of GLUT4 transporters, as well as the generation and degradation rate of PI3P lipids, in individual cells under 12 different perturbation conditions in parallel. We will create an RNAi library to target the approximately 800 adipocyte signaling/secretory proteins and a library of 800 expressed dominant negative and constitutively active expression constructs for complementary pertubation studies. In contrast to earlier approaches in which screens were based on end-point assays in fixed cells, the live-cell strategy proposed here focuses instead on the identification of adipocyte signaling and secretory proteins that control important kinetic parameters in insulin triggered PIP3 signaling as well as in GLUT4 translocation. In the second part, we will execute, analyze and interpret a screen of the insulin-PIP3-GLUT4 signaling network with this new approach and will also perform a screen of cross-talk between insulin and TNF-alpha signaling. These screens will be used to identify new players in the insulin-GLUT4 signaling network and to gain a better understanding of kinetic control mechanisms. The project will also provide a test case for a kinetic screening strategy and a test case for a combined screen using RNAi, as well as DN and CA expression constructs.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Complimentary and alternative medicines (CAM) are amongst the most commonly used therapies worldwide and in the US for chronic liver diseases. Silymarin, the active ingredient of the milk thistle, Silybum marianum, is one of the commonest used herbal medications but its efficacy in chronic liver disease is unproven. Liver disease secondary to both hepatitis C (HCV) and non-alcoholic steatohepatitis (NASH) represent 2 of the commonest liver diseases in the US and there is a need for the study of silymarin in these diseases in a controlled clinical trial. We are proposing three collaborative clinical trials to evaluate silymarin versus placebo in HCV patients who have never received treatment, in HCV patients who have failed treatment and in patients with NASH. These will be double blind, placebo controlled phase l/ll trials focusing on dose finding, safety and efficacy of silymarin. The endpoints of the trials will include histological improvement in liver disease and multiple secondary surrogate markers to understand both how silymarin works and to get a preliminary idea of the endpoints to be utilized in larger Phase 3 trials. We also intend to evaluate important secondary factors such as expectations of patients on CAM and effect of silymarin on quality of life. The goal of these studies is to determine whether there is a rationale for larger Phase 3 studies of silymarin in patients with chronic liver disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dopamine (DA) signaling has received considerable attention for its role in reward-related processes, including the motivation to seek drugs and relapse in response to drug-associated cues (1-4). Destruction of DA cells in the ventral tegmental area (VTA), or DA terminals in regions such as the nucleus accumbens, disrupts drug self-administration (5-6) and administration of DA receptor antagonists attenuates the ability of drug-associated stimuli to promote reinstatement (7). In human addicts, striatal DA release in response to drug-associated stimuli is associated with increased drug craving and future relapse (8,9). Though DA has been generally implicated in drug taking and relapse, DA systems have thus far not been manipulated with the temporal precision and cell-type specificity required to isolate their role in specific aspects of those behaviors. Optogenetic tools have been applied to target DA neurons in transgenic mice, demonstrating that DA signaling supports behavioral conditioning and facilitates instrumental responding for food (20-21). More recently, a Th:Cre transgenic ratline was developed that allows for the selective targeting of DA neurons with optogenetic methods (22-23) in more complex behavioral paradigms optimized for use in rats. In this proposal, I will utilize Th:Cre rats, incorporating in vivo optogenetics and electrophysiology in combination with sophisticated behavioral analyses, to probe the causal contribution of DA signaling to different aspects of instrumental cocaine intake and relapse in response to Pavlovian cocaine stimuli. First, in Aim 1 I propose to test the sufficiency of VTA DA neuron activation to modulate cocaine self-administration and cocaine cue-induced reinstatement. Second, in Aim 2 I propose to test the necessity of DA signaling, via inhibition of VTA DA neurons, for cocaine intake and reinstatement. Additionally, the VTA contains a heterogeneous mixture of not only DA neurons, but also a substantial fraction of non-DA neurons that contribute to motivational processing (11-19), but little is known about how different populations of neurons in the VTA encode drug-related behaviors. Thus, in Aim 3 I propose to characterize the firing patterns optogenetically-identified VTA DA neurons (16) during self-administration and reinstatement.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The development of regulatory behaviors necessary for state, motor, and emotion control may have certain underlying physiological processes in the parasympathetic nervous system that play a role in their functioning. One specific psychophysiological correlate of regulatory behavior that has been extensively studied in the existing literature is that of heart rate variability, or vagal tone. Low heart rate variability in children has been associated with maladaptive outcomes (i.e., conduct disorder;Pine et al., 1996) and high heart rate variability has been associated with better social competence (Eisenberg et al., 1995). Thus, it is critical to understand the trajectory of vagal tone from infancy to childhood and how it both influences and is influenced by development. The objectives of the current application are 1) to examine the stability of baseline vagal tone and vagal regulation longitudinally in infants from 3 months to 3 years of age, 2) to examine the concordance between infants'behavioral and vagal responses to challenge tasks, 3) to examine the influence of factors endogenous (i.e., temperament) and exogenous (i.e., maternal sensitivity) to the infant on vagal trajectories over time, 4) to examine factors related to ethnicity and family income as predictors of vagal trajectories and 5) to examine baseline vagal tone and vagal withdrawal trajectories as predictors of socio-emotional functioning assessed by maternal report at 36, 48, and 60 months of age. The Durham Child Health and Development Study has demographic, maternal report, behavioral, and physiological data on 199 infants and their families at seven time points (3, 6, 12, 18, 24, 30, and 36 months). At each visit, infant vagal tone and behavioral response was measured before, during, and immediately following various challenge tasks meant to elicit negative reactivity and regulation from infants. Further, maternal behavior toward her infant during a free play and puzzle task was video recorded for behavioral coding. Our current understanding of vagal tone as a physiological index of regulatory functioning is limited. It is important to move beyond concurrent assessments of vagal response and identify patterns and trajectories that develop over time. The proposed analyses will allow us to examine the development of vagal tone over the first three years as a predictor of typical and atypical socio-emotional outcomes. Relevance to Public Health: The proposed study is designed to provide longitudinal analyses of the development of infant heart rate variability (i.e., vagal tone), and the regulation of vagal tone, across the first three years of life. This physiological system may underlie the processes of emotion regulation and executive functioning and has been associated with both positive and maladaptive behavioral outcomes in children. Understanding the various child and family characteristics that are related to its development will provide new insights into individual differences in such areas as self-regulation of emotion, responses to challenge such as the transition to school, and the acquisition of social competency in peer groups.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronic drug administration can produce allostasis, a maladaptive state related to drug tolerance. This proposal investigates nitrous oxide (N2O)-induced allostatic changes and the motivational consequences of being in an allostatic state. Allostasis refers to a disordered form of homeostatic regulation wherein a regulated variable, or one or more of its controlling determinants, persistently functions at levels significantly different from control values, potentially compromising an individual's health or viability. An allostatic model of drug addiction posits that biobehavioral control systems regulate variables relevant to drug taking behavior and that these control systems are vulnerable to drug-induced allostatic changes which promote the development of addiction. The proposed studies use a sophisticated experimental model that combines direct and indirect calorimetry so that core temperature and its determinants (metabolic heat production and heat release) can be simultaneously measured, enabling rigorous determination of allostatic dynamics during repeated N2O administrations. This thermoregulatory model system also provides a sensitive method for determining the motivational consequences of allostasis. Preliminary data indicate that adolescent rats are especially prone to develop drug-induced allostatic changes, suggesting that increased susceptibility to allostasis development may be a critical etiological factor for the heightened vulnerability to drug addiction during adolescence. Specific Aim (SA) 1 compares allostasis development in adolescent versus mature rats over a range of N2O concentrations, determines whether these allostatic processes stabilize, and explores how they can be extinguished. SA 2 compares the thermally motivated effects of a range of N2O concentrations in adolescent versus mature rats and assesses the motivational effects of an allostatic state during adolescence. In addition, the relationship between initial sensitivity, allostasis development and N2O self-administration behavior will be investigated. SA 3 examines whether factors measured in the allostatic state (N2O concentration, core temperature, heat loss, heat production) can be used as predictors of motivated behavior. This work has practical and theoretical importance for understanding the mechanisms underlying drug addiction. The proposed research has the added relevance of investigating an abusable inhalant during the adolescent period which NIH has identified as an important, yet understudied, research area. PUBLIC HEALTH RELEVANCE: A form of homeostatic dysregulation known as allostasis is suspected to play an etiologic role in the development of drug addiction. The proposed research uses an understudied inhalant to investigate drug-induced allostasis during a developmental period (adolescence) that is known for its heightened susceptibility to drug addiction. The findings of this research will contribute to our understanding of the pathogenesis and treatment of drug addiction.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Incontinence and urinary symptoms of frequency and urgency are prevalent in post-menopausal women. This project is designed to evaluate the effectiveness of topical estrogen in the behavioral treatment of urinary symptoms and incontinence.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The aim of this project is to clone and characterize the gene for a form of syndromic retinitis pigmentosa (RP), called. Hallervorden-Spatz syndrome (HSS) and characterized by abnormal electroretinogram, lipofuscin accumulation in the retinal pigment epithelium, and pigmentary retinopathy. Other features include dystonia, due to massive iron accumulation in the basal ganglia, and progressive deterioration leading to early death. Though lipid peroxidation is an hypothesized mechanism leading to the HSS phenotype, no knowledge exists of the molecular or biochemical defect in HSS. We have a unique opportunity to map the HSS gene using linkage analysis (homozygosity mapping) in a consanguineous Amish family with multiple affected members. Once the HSS gene has been mapped, we will search for mutations in candidate genes, as well as identify novel transcribed sequences that may contain the HSS gene. We will then characterize the gene and its protein product through homology studies to known sequences. Knowledge of the molecular basis of this disease will lead to a better understanding of the pathophysiologic process causing its pleiotropic effects. Understanding the etiology of a rare disease will often illuminate the mechanism at work in common, related diseases. Furthermore, by studying syndromic RP, we can use information about all of the syndrome manifestations (e.g. patterns of tissue expression, common metabolic or developmental pathways) to theorize a disease mechanism. Inference of a pathophysiologic process from a defective gene has proved frustrating for the forms of RP that are due to mutation in retina-specific genes. The HSS gene is not retina-specific, and a defect in it must account for rod photoreceptor degeneration as well as regional brain iron accumulation. Once the HSS gene is cloned and characterized, the other pathologic changes may provide a context for understanding the mechanism of pigmentary retinopathy. Since defects in this non-retina-specific process may cause other forms of syndromic and isolated RP and may be integral in disorders of lipofuscin accumulation, including aging macular degeneration, identification of the HSS gene may lead to greater understanding of RP as well as the macular dystrophies associated with sene ence. The HSS project forms the research core of Dr. Hayflick's training to become an independent biomedical investigator. Dr. Michael Litt, internationally recognized in the field of genetics, will he her primary sponsor with Dr. Richard Weleber, accomplished in the study of hereditary retinal diseases, as secondary sponsor. Unique strengths of her training program include a period of intense study in the Visiting Investigator Program through the National Center for Human Genome Research, which will provide her with expertise that will complement but not duplicate existing University research strengths, and the Oregon Health Sciences University and Department of Molecular and Medical Genetics research environments, which effectively foster collaboration with a diverse group of outstanding investigators.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Project Summary Antimicrobial resistance has evolved due to decades of inappropriate antibiotic prescribing by clinicians. In the emergency department (ED), nearly 50% of the 10 million antibiotic prescriptions written each year are inappropriate or unnecessary. However, antimicrobial stewardship programs (ASPs) for the ED have yet to be created. Failure to develop ED-based ASPs is a result of barriers unique to the ED, including lack of ED formatted guidelines, erratic workflow, rapid decision making, and diagnostic uncertainty. Electronic health record-based clinical decision support (EHR-CDS) can be customized to present ED- specific antibiotic recommendations at the point of care, in the usual clinical workflow. Therefore, EHR-CDS has great potential to overcome ED barriers and serve as platform for ED-based ASP. In this proposal, we build upon our preliminary single-center research to develop a multicenter EHR-CDS for antibiotic prescribing for two common pediatric ED infections, community acquired pneumonia (CAP) and urinary tract infection (UTI). National CAP and UTI antibiotic prescribing guidelines exist, but have not been adapted into ED context. We will use rigorous dissemination and implementation methods that this team has successfully used previously to create multicenter EHR-CDS. We also build upon our preliminary work to create a novel, multi- center EHR-CDS within the Pediatric Emergency Care Applied Research Network (PECARN). We will conduct workflow analyses and identify EHR triggers to determine the optimal timing for EHR-CDS activation, necessary to minimize alert fatigue. At each of the 3 participating sites, we will also develop an EHR-based mechanism to determine guideline adherent antibiotic prescribing for CAP and UTI. Through integration of the formatted ED antibiotic treatment guidelines for CAP and UTI, workflow analyses, and EHR activation triggers analyses, we will configure a prototype EHR-CDS for antibiotic prescribing. We will conduct heuristic review and subsequent scenario-based user testing at each site to produce a functional, scalable prototype EHR- CDS. Though we will initially build the EHR-CDS using a single her (with the largest market share), we will simultaneously develop a web-service version of the CDS for further scalability. Finally, as we have for prior studies, we will create automated clinician feedback reports using EHR data to provide antibiotic prescribing practices; these will complement the EHR-CDS as part of the overall ED-based ASP. The adaptable scalable EHR-CDS for CAP and UTI, and the provider feedback reports, will serve as the centerpieces for generalizable ED-based ASP. Upon completion of the work in this proposal, we will be poised to conduct a multicenter trial to test the effect of our EHR-CDS in pediatric and general ED settings.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The Quality Assurance Laboratory (QAL) in the Comparative Medicine Branch at the National Institute of Environmental Health Science (NIEHS)performs testing on the microbiological and/or chemical contamination of critical aspects of the overall NIEHS research program. Our primary task is monitoring the animal research program for microbiological and/or chemical contamination that may affect animal health and welfare, as well as, the physiological responses of animals used in research studies. Our research focuses on the physiological effects of natural or contaminating compounds in the micro-environment (e.g., animal feed, caging, bedding, water, etc.)of the research animals. Our goal is to assure that we minimize exposure to environmental compounds that may effect animal health and welfare or alter physiological responses of the animals resulting in unacceptable variability. Historically, QAL studies have focused on the potential for endocrine-disrupting compounds (EDCs) being present in the micro-environment of the research animals and the impact these compounds may have on study outcomes, especially reproductive development. For these studies, pre-pubertal CD-1 mice are weaned at post-natal day (PND) 15 and given test compounds orally (via diet, water or gavage) from PND 15 to PND 35 or until vaginal opening (VO) occurs which is a developmental milestone affected by hormones. We have shown that natural phytoestrogens (e.g., daidzein and genistein) present in commercially available rodent diets or added to diets free of these compounds can significantly (P less than 0.01) accelerate the time of VO in CD-1 mice. We have also shown that the total metabolizable energy (ME) in the diet can significantly affect these hormone-sensitive endpoints, although the predictability of this variable was less powerful than the phytoestrogen content. We have shown that the estrogenic mycotoxin zearalenone is ubiquitious in commercially available corn-cob bedding. 154 of 189 (84%) of the samples were naturally contaminated with zearalenone at levels ranging from 100 to 7,000 ppb (mean 500 ppb). We have shown that levels of 5-10 ppm can significantly advance the time of VO in immature CD-1 haired and SKH-1 hairless mice. We are currently performing studies looking at the effects of autoclave sterilization of rodent feed on the physical and chemical properties of the feed. Sterilization of rodent feed is important to prevent the introduction of microorganisms that may alter animal health or physiological response, and autoclave sterilization is a widely used method. Previous studies found that autoclaving rodent feed resulted in the production of acrylamide, as does any high starch containing food (e.g., potatoes, breads, etc.). These studies also determined that the levels of acrylamide produced by autoclaving the feed were high enough to cause measurable genotoxic effects (DNA adducts). Our study determined that increasing the sterilization temperature used resulted in an increase in the harness of the pelleted feed, as well as, the concentration of acrylamide. We are currently measuring the in-vivo effects of these levels of acrylamide.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of our laboratory is to identify the neural circuitry and neurochemical and neurophysiological mechanisms that underlie the expression and control of rage and aggressive behavior. The primary focus of the present grant application, which represents a major new direction in our research program, is to identify and characterize the roles of serotonin and cytokines in the medial hypothalamus in regulating these forms of aggression. This rationale for this application is based upon our most recent preliminary studies. They provide evidence that, in the medial hypothalamus, 5-HT1A and 5-HT2 receptors and cytokines, IL-lbeta and IL-2, powerfully modulate defensive rage behavior in the cat. The overarching hypothesis is that differential cytokine effects upon defensive rage and predatory attack are mediated principally through distinct neurotransmitter receptors of which serotonin and possibly GABA are primary candidates. Five experiments are proposed to test this hypothesis. The first will utilize immunocytochemical and neuroanatomical methods to characterize the pathway from the PAG to the medial hypothalamus mediating defensive rage and its relationship to serotonin axons and pre-terminals in this region. The second experiment will determine the effects of 5-HT1A and 5-HT2 receptors in the medial hypothalamus upon defensive rage. The third experiment will determine: the role of IL-1beta in the medial hypothalamus upon defensive rage, its relationship to 5-HT2 receptors, the distribution of IL-1R in this region and its relationship to serotonin axons and pre-terminals as well as to c-Fos labeled neurons following the expression of defensive rage. The fourth experiment will seek to determine the role of IL-2 in the medial hypothalamus upon defensive rage and their underlying neurotransmitter-receptor mechanism. The fifth experiment will identify the effects of activation of 5-HT and the above cytokines upon predatory attack behavior. The discovery that cytokines in the brain play a significant role in defensive rage represents a most significant observation. It has provided an entirely new direction of research - a direction in which the focus will address how cytokines and related substances in the brain may play critical roles in the expression and control of aggression and rage. By identifying the mechanisms underlying these effects, the proposed studies are of further significance because the strategies utilized here can now be applied for the study of how cytokines in the brain may regulate other behavioral processes.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "To evaluate strategies that test the safety and effectiveness of therapeutic approaches/regimens to reduce the probability of the emergence of antibiotic drug resistance by minimizing unnecessary drug exposure. Trial title: A Multi-Center, Randomized, Open-Label, Comparative Study to Assess the Safety and Efficacy of a Treatment Algorithm to Reduce the Use of Vancomycin in Adult Patients With Blood Stream Infections Due to Staphylococci.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our goal is to scintigraphically detect, stage and ultimately treat ovarian carcinomas, using radiolabeled monoclonal antibodies preferentially reactive with epithelial ovarian carcinomas. Monoclonal antibodies raised against ovarian cancer, already shown capable of radiolocalizing human ovarian carcinoma xenografts in nude mice, will be further evaluated in that model. The effects of antibody dose, route of administration (intravenous as compared to intralymphatic or intraperitoneal), fragmentation, and labeling, will be evaluated to optimize antibody localization to tumor. Since ovarian carcinoma is generally a regional disease within the peritoneum and draining lymphatics, we expect this regional antibody delivery to significantly enhance tumor localization. Antibody localization will be evaluated by gamma camera scanning, tumor excision with gamma counting, immunoperoxidase staining, and microautoradiography. In parallel experiments, the specificity of existing and new monoclonals raised against ovarian cancer will be further evaluated by immunohistochemical techniques. The radiotherapeutic potential of the best monoclonals for imaging will then be evaluated in vitro using clonogenic assay techniques. The dose of radiolabeled antibody required to produce tumor kill of ovarian carcinoma lines in vitro will be determined. Based on these studies, the best antibodies will then be evaluated for radiotherapeutic potential in our nude mouse model of disseminated intraperitoneal human ovarian carcinomatosis. A variety of dosing schemes will be tested therapeutically with evaluation for the number of clonogenic ascites cells, mean survival, and cure as indices of the efficacy of radiotherapy. The monoclonal antibodies with the best localization to ovarian carcinoma xenografts in vivo, as well as showing the greatest in vitro specificity for ovarian cancer, will be evaluated in phase I imaging trials in patients with advanced ovarian cancer who are scheduled for a second debulking procedure. This will assure histologic, autoradiographic and gamma scan assessment of antibody localizaiton to tumor. This study will provide unique information regarding the optimal means of delivering radioantibody to ovarian tumors and about these tumor's sensitivity to radioantibody. This should result in a means of scintigraphically following residual ovarian carcinoma post-diagnosis and may eliminate the need for second-look diagnostic procedures. If adequate specific localization of radioantibody to ovarian cancer is achieved, then this approach may prove useful in the radioimmunotherapy of this common and lethal disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "ABSTRACT Autism spectrum disorder (ASD) is a devastating neurodevelopmental disorder of undetermined etiology and without effective therapeutics. Recent advances in genomic approaches have led to the identification of over 65 ASD risk genes. Despite the genetic heterogeneity of ASD, several lines of evidence suggest that these genes share common molecular underpinnings. Therefore, we hypothesize that these genes will converge upon shared phenotypes when inhibited in model organisms and that these phenotypes will be the most central to the neuropathology of ASD. Identifying such convergent phenotypes requires a high-throughput system for modifying many ASD genes in parallel and assaying their effect(s) on embryonic brain development. Here we propose to leverage the diploid vertebrate tetrapod, Xenopus tropicalis, and the CRISPR/Cas9 system to identify convergent phenotypes among ASD genes during brain development. By injecting Cas9 protein and a single guide RNA (sgRNA) against an ASD gene at the two-cell stage, animals will be generated in which exactly half the body (separated by the midline) is mutant, allowing for direct comparison of mutant and control cells in the same animal. This will be performed for approximately 65 ASD risk genes, and the effects of ASD gene loss will be assayed by imaging neurons throughout embryogenesis using fluorescent reporters and by in situ RNA hybridization for neuronal cell fate specification markers. By employing fluorescence activated cell sorting (FACS), next generation RNA sequencing, and weighted gene co-expression network analysis (WGCNA), convergent transcriptional signatures of ASD gene loss will be characterized. The biological pathways indicated by these convergent signatures will be validated through targeted manipulation of key genes. Importantly, the validated biological pathways may provide clues about the observed convergent phenotypes. By combining the high-throughput capability of the CRISPR/Cas9 system and a tractable vertebrate model organism with a reliably-associated set of ASD genes, this study aims to understand the neuropathology of ASD at a critical developmental period, which should provide critical insights into the etiology of this disorder.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronobiology is the study of biological rhythms. There are a number of chronobiological effects which may have important clinical implications for antitumor therapies. For example, chronobiological effects on the replication of some types of tumor cells, the expression of certain drug toxicities, and the number of cytotoxic effector cells have been described. Interferon therapies are currently being evaluated in clinical trials for their potential as antitumor agents. This proposal focuses on possible chronobiological effects which may effect the potencies of various interferon therapies. We propose to evaluate the chronobiological effects on tumor growth of interferons administered (1) as single agent therapy; (2) as multiple interferon therapies (IFN-gamma plus IFN-alpha or IFN- beta); (3) as multiple agent therapies with hyperthermia; and, as multiple agent therapies with phenytoin. These studies will be performed both in vitro, using B-16 melanoma cells in cloning studies, and in vivo, using the B-16 melanoma/C57/B1/6 tumor system. Simultaneous treatments and various sequential treatments will be compared to identify the most potent treatment protocol. We also propose to evaluate the chronobiological effects on IFN-mediated bone marrow suppression of colony stimulating factor administration. These studies will be performed both in vitro using bone marrow colony growth assays and in vivo using total white blood cell counts as an indicator of bone marrow suppression.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The long-term goal of the laboratory is to gain a structural understanding of the molecular organization and function of biological macromolecules at the biological membrane interface. Our focus of this study is to understand physical mechanisms of ion selectivity, conductance regulation, channel gating of ion channel proteins, and the structural basis of molecular specificity for oligomerization by X-ray crystallography. The first specific goal is to determine the three-dimensional structure of the tetramerization domain of Shaker potassium channel. We have obtained three-dimensional crystals of the soluble domain of this channel that diffract X-ray beyond 2.0 Angstroms. Crystals belong to the space group I4 with one subunit molecule per asymmetric unit. We have also obtained three-dimensional crystals of the soluble, domain of a channel from a different subfamily of potassium channel Shaw that diffract X-ray to about 3.0 Angstroms. The second specific goal is to determine the atomic structure of an inwardly rectifying potassium channel ROMK1 from rat kidney. Towards this goal, the underlying hypothesis of our novel strategy is that the lipid- facing exterior of a channel protein can be mutated systematically without perturbing its channel function. Site-directed mutagenesis was used as a perturbation method in order to identify the lipid-facing residues. Based on this study, simultaneous changes have been introduced at sites in its transmembrane lipid-facing exterior of the channel, which do not alter the channel function. The surface of this exterior-modified channel has a sequence motif known to form a specific helix-to-helix interaction, through which the exterior-modified channel itself is solubilized by binding of synthetic amphipathic peptides to the channel exterior in the aqueous phase without the use of detergents. Understanding the structural basis for tetramerization will provide an essential knowledge on the physical nature of channel diversity. From the atomic model of the ROMK1 channel, we seek to establish the exterior- modification strategy that will be generally applicable to structural studies of a variety of integral membrane proteins such as ion channels and transmembrane receptors.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Neutrophil priming following trauma and sepsis is a key event implicated in causing Adult Respiratory Distress Syndrome (ARDS) and Multi-Organ Failure Syndrome (MOSF). Priming of the respiratory burst by cytokines following injury and sepsis results in excessive superoxide production by NADPH oxidase leading to auto-inflammatory tissue damage. Numerous studies have focused on details of how NADPH oxidase produces superoxide. A similar investigation into the mechanisms of priming has been relatively lacking. Our long-term goal is to develop a detailed molecular understanding of this priming phenomenon. Our previous studies and preliminary observations show that different signaling pathways activated by trauma and sepsis converge to phosphorylate two serine residues in a key protein of the NADPH oxidase, p47phox. We propose that through this phosphorylation, p47phox integrates upstream signaling events to control the level of NADPH oxidase activity by regulating protein-protein interactions within neutrophils. In the studies outlined in this proposal the consequences of priming-induced phosphorylation and the identity of proteins associating with NADPH oxidase components following priming will be investigated using neutrophil cell-biological and protein chemistry approaches. The results from these studies may assist in the development of novel therapies aimed at limiting the auto-inflammatory tissue damage patients suffer as a result of sepsis and trauma.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "During this period, the collaborative team is working to validate and characterize previously identified hits from primary SENP screening. As a center, the NCGC has fostered and maintained over 130 active collaborations with both NIH and extramural investigators, facilitating drug discovery efforts across the entire spectrum of human disease. These efforts have led to dozens of high-throughput screens and a number of medicinal chemistry campaigns to further improve on screening hits, providing our collaborators and the general research community with publications and a variety of promising small molecule probes and leads. In addition, the NCGC has worked to advance a number of informatic initiatives to make better use of existing drug and disease target information and provide the general public with easily accessible resources, further catalyzing the development of new therapies for human disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "OBJECTIVES: 1. To disseminate and coordinate sickle hemoglobin education in the community; 2. To counsel patients with sickle cell disease and their families and establish ongoing relationships with hospitals, clinics and other health agencies; 3. To develop educational materials on sickle hemoglobin to be used locally, nationally and internationally; 4. To make available to the medical community and general public a resource for the laboratory diagnosis of sickle and other hemoglobin variants; 5. To determine the molecular structure and intermolecular contact sites of hemoglobin S in sickle cell fibers; 6. To understand the mechanisms of translational and transcriptional control of the synthesis of specific hemoglobins; 7. To study the renal adaptation of patients with sickle cell anemia to overproduction of uric acid and the association of hyperuricemia and gout; 8. To determine monocyte bactericidal capacity in patients with sickle cell anemia and restore same in vitro by incubation with sickle serum cleared of red cell fragments; 9. To ascertain whether variations in severity of anemia among patients with sickle cell anemia are due primarily to differences in rate of red cell production or in red cell destruction and whether regulation of erythropoiesis is defective in such patients; 10. To study the role of prophylactic transfusion therapy in care of pregnant patients with sickle cell anemia.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Alveolar epithelial cells (AEC) cover the alveolar surface of the lung, which is the major acid-excreting organ in mammals. Because these cells are subject to an inconstant extracellular milieu with respect to pCO2, the regulation of intracellular pH (pHi) in alveolar epithelial cells is of particular significance. In previous work it has been established that alveolar pneumocyte pHi is regulated by several ion transport mechanisms that are capable of effecting acid/base entry or extrusion from the cell. These ion transport mechanisms are plasma membrane proteins asymmetrically distributed across polarized epithelial cell surfaces, thereby making transepithelial transport of acid/base equivalents possible. Their distribution with respect to membrane polarity across the alveolar epithelium, whose apical lining fluid pH is acidic (6.9), has not previously been established. We will investigate the mechanisms whereby acid/base transport by AEC is regulated by studying three hypotheses: (1) Acid/base transport by AEC occurs via specific ion transport mechanisms having a characteristic distribution with respect to membrane polarity across the cells, and which represent the activity of specific transport protein isoforms. (2) These ion transport mechanisms are both acutely and chronically regulable in AEC with respect to activity, expression, and polarity by specific extracellular stimuli that can thereby modulate acid/base transport under normal conditions and in disease states. (3) Regulation and localization of acid/base transport mechanisms in AEC influence alveolar fluid pH. The objectives of the studies proposed herein are therefore to further characterize the acid/base transport mechanisms in AEC, and in particular, to study regulation of their activity, expression, and polarity by specific exogenous factors and lung injury. We will use rat alveolar epithelial cell monolayers cultured on tissue culture-treated polycarbonate filters for many of these studies. This model is especially well suited for studies on alveolar epithelial polarity and acid/base transport. We will validate our findings using in situ studies on rat alveolar epithelium. The long term goals of this project are to define the importance of acid/base transport in intracellular homeostasis of alveolar epithelial cells, and to demonstrate the role of acid/base transport by alveolar epithelial cells in modulation of the extracellular alveolar milieu.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Abstract - Project 4: Cure rates for pediatric patients with relapsed or metastatic solid tumors remain unacceptably low. Cancer immunotherapies hold great promise, but scores of disappointing studies highlight our relative ignorance in understanding the immunosuppressive microenvironment within solid tumors. Because of their central role in mediating immunosuppression, tumor associated macrophages (TAMs), typically ?polarized? to a so-called M2- like immunosuppressive phenotype, and myeloid-derived suppressor cells (MDSC), are thought to be important therapeutic targets. We have found a clinically viable strategy that simultaneously reduces TAMs/MDSC (we dub ?myelolytic?) and polarizes the microenvironment (via oncolytic virus infection), resulting in significant antitumor efficacy. We hypothesize that targeting TAM and MDSC by combining ?myelolytic? therapies with pro- inflammatory therapies activates innate antitumor mechanisms that cause cancer regressions and reshapes the solid tumor microenvironment to be more permissive to cellular immunotherapies. In aim 1, we will determine the mechanism(s) by which combined myelolytic-virotherapy drives tumor regressions. We will use novel technologies such as fluorescent nanodiamonds to determine effects on innate immune cell phagocytosis of tumor cells. We will utilize the Genomics & Immune Monitoring Shared Resource Core B directed by Dr. Elaine Mardis to conduct flow cytometry with time-of-flight mass spectrometry and single cell transcriptomics to determine the effects on immune cell composition and polarization. We will also utilize gain- and loss-of-function approaches to determine if loss of MDSC are critical for enabling tumor regressions with myelolytic-virotherapy. We will also test combination therapies in xenograft and immunocompetent models of other cancer types to confirm its generalizability (osteosarcoma, Ewing sarcoma, rhabdomyosarcoma, neuroblastoma). In aim 2, we will determine the effects of myelolytic-virotherapy on T cell-mediated immunotherapies. We will examine the effect of myelolysis alone and combined with virotherapy on the efficacy of antitumor T cells in a T cell exhaustion setting and with CAR-T cells (with Project 1 Leader Dean Lee and co- investigator Ruoning Wang, PI-DDN U01 member). We will work with Core B to examine the effects on T cell clonality using TCR sequencing. In aim 3, we will determine whether combined myelolytic-virotherapy enhances the efficacy of NK-based cellular therapies. We will work with Project 1 Leader Dean Lee and Project 2 Leader Mitch Cairo to study the effects on adoptive NK and CAR-NK cell therapy. Overall, with this project we will further elucidate, test and develop strategies to modulate the tumor microenvironment to facilitate innate immune cells as cancer therapy. Our findings may be applicable across a broad panel of pediatric cancer types and thus fits well into the aims of the Pediatric Immunotherapy Discovery and Development Network.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The pulmonary involvement in disorders of immune or inflammatory regulation can range from a primary manifestation such as with sarcoidosis or a relatively minor manifestations such as seen with the Mendelian Susceptibility to Mycobacterial Disease immune deficiencies. While both of these can be associated with granulomatous inflammation in the lung, the clinical pulmonary manifestations are quite different as are the management strategies. The Pulmonary Clinical Medicine Section (PCMS) has sought to capitalize on the close collaboration with the Laboratory of Clinical Infectious Diseases and other branches within the NIAID and NIAMS focused on these disorder to describe the pulmonary manifestations of known and emerging immune and inflammatory diseases. This has included characterizing the lung manifestations of newly described immune dysregulatory syndromes associated with mutations in such as cytotoxic T-lymphocyte-associated protein 4 (CTLA4) and phosphoinositide 3-kinase (PI3K)and well known and characterized disorders such as chronic granulomatous disease. The PCMS has been actively involved in characterizing the protean lung manifestations of GATA Binding Protein 2 (GATA2) deficiency have provided unique insight into the role of alveolar macrophage dysfunction in development of pulmonary alveolar proteinosis and intracellular control of mycobacteria and fungi in the lung. Ongoing work in this area include characterization of lung manifestations of Autoimmune Lymphoproliferative Syndrome (ALPS) and Autoimmune Polyendocrinopathy, Candidiasis, and Ectodermal Dystrophy (APECED) Syndrome.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "RESEARCH EDUCATION CORE SUMMARY The goal of the Research Education Core is to attract and catalyze a new generation of diverse research professionals and biomedical scientists who are committed to elucidating and addressing the cancer disparities outcomes faced by so many in South Carolina and the US. During the five-year U 5 4 project period, undergraduates at South Carolina State University (SCSU) and junior faculty at SCSU and the Medical University of South Carolina (MUSC-HCC) will be designated as South Carolina Cancer Disparities Research Center (SC CADRE) Scholars. Building from multiple ongoing efforts, the leaders of the SC CADRE are proposing a dual-level, integrated Research Education Core with an explicit focus on cancer health disparities. Expanded research education and training opportunities will be directed to both levels of Scholars. To realize the goal of the SC CADRE Research Education Core, the SCSU and MUSC-HCC Core Co-Leads plan to establish a new undergraduate SC CADRE Honors Program in Applied Oncology Sciences Research within the Department of Biological and Physical Sciences at SCSU to increase the pipeline of highly skilled, diverse undergraduates who pursue careers in biomedical research and cancer disparities. A new cancer research career development program at SCSU and MUSC-HCC will also be implemented for junior faculty who are interested in establishing independent careers in cancer disparities research. Within the Research Education Core, the SC CADRE undergraduate and junior faculty Scholars will interact as they participate in coursework and/or career development programming, weekly SC CADRE Research Club meetings, monthly seminar series, and an annual SC CADRE Cancer Health Equity Research Symposium. Through the Research Education Core, SCSU will hire two new junior faculty Scholars and MUSC-HCC will hire one new junior faculty Scholar, all of whom will be early-stage investigators (ESIs). Both levels of Scholars will actively engage in numerous SC CADRE enrichment activities and activities planned by the Community Outreach Core, which will work directly with Research Education Core leadership to s t r e s s the importance of community engagement in developing, conducting, and disseminating cancer disparities research.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Understanding how memory develops in the immune system is an important issue in basic science and in clinical medicine, both in terms of promoting an immune response and, conversely, in blocking the pathway that leads to an overactive state (allergy, autoimmunity). However, the mechanisms underlying the generation of memory lymphocytes are poorly understood. This proposal is aimed at answering the important question of whether the generation of memory T cells is associated with a stable alteration of cytokine expression. We focus here on the generation of mouse CD4+ memory T cells that help B cell antibody responses (memory helper T cells) in relation to attainment of the capacity to secrete IL- 4. We wish to distinguish whether there are pre-existing CD4+ subsets committed to this IL-4 potential, or instead whether such subsets are generated de novo by a maturation process following antigenic stimulation. We will examine TCR-V gene usage of CD4+ subsets by establishing T hybrids from a 2B4 TCR Vbeta (or Valpha) transgenic mouse to determine whether the cell subsets capable of rapid IL-4 secretion (\"IL-4+\") show restricted TCR V-gene usage, evidence for antigenic selection. Then we will attempt a direct demonstration of IL-4- naive cell transition to IL-4+ cells at the single cell level. Finally, we will examine whether memory T helper cells are already committed to a specific cytokine potential or not. If we can demonstrate that IL-4+ cells are produced during the process of memory T cell generation, then this provides an important concept of lymphocyte maturation; that is, specialization of effector function similar to memory B cell generation characterized by immunoglobulin isotype switch. These studies will provide greater understanding of the generation and function of memory cells in the immune system, both in normal and diseased states.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "We established that neurons present in dorsal root ganglia (DRG), similar to leukocytes, express a wide variety of receptors for cytokines, chemokines, opioids, anandamide and other neuropeptides. We previously showed that prior exposure to chemokines such as MIP1&amp;#945;results in PKC mediated desensitization of the chemotactic response to opioids by opioid receptors, and thus potentially enhances pain. This decrease in the analgesic effect of opioids was evident from the enhanced tail flick assay of rats administered MIP1&amp;#945;or RANTES prior to an analgesic opioid into the PAG of the CNS. We then extended these earlier studies by showing that prior administration of chemokines Asensitized and primed the calcium flux of capsaicin or anandamide stimulated vanilloid (TRPV1) algesic receptor on DRG neurons. This response also increased pain as shown by the enhancement of paw withdrawal in response to the intrathecal administration of the chemokine prior to capsaicin in vivo. This sensitization of the vanilloid receptor was also PKC dependent. Consequently, proinflammatory chemokines can increase pain both by suppressing opioid and enhancing vanilloid receptor responses. Based on these studies, we predicted that the anti-inflammatory effects of adenosine, which also interacts with GiPCR, might have effects on chemokine receptors. Indeed our studies showed that prior addition of adenosine results in suppressing the in vitro chemotactic response of leukocytes to a variety of chemokines. Furthermore, prior in vivo injection of adenosine inhibited the in vivo influx of leukocytes into a murine air pouch by about 90%. This cross-desensitization of chemokine receptors by adenosine A2a receptors was PKA dependent. The role of adenosine as an immunosuppressive effector molecule also has been reported to mediate the cell contact dependent effects of Tregs and to interfere with host resistance to tumors. Thus, studies of adenosine effects are relevant to tumor biology and immunology. These studies therefore reveal novel pathways of receptor mediated intercommunication of inflammatory as well as painful stimuli. Means of interfering with these PKC and PKA dependent signals and the pathophysiological relevance of this receptor cross-talk to inflammation and pain need to be further evaluated. Our current project focuses on neuroimmune interactions contributing to pain sensation in cancer patients funded by an INIP postdoctoral IRA financial grant from NIAID and NCI. As previously shown, chemokine receptor cross-talk suppresses analgesic opioid receptors, but enhances algesic transient receptor potential channel (TRP) receptors, thus resulting in painful inflammation. a) In collaboration with Dr. Jeffrey Cohen, NIAID we have investigated this in a cotton rat herpes virus infection model for chemotherapy induced Herpes Zoster. Herpes infection of dorsal root ganglia results in extremely painful inflammatory responses along nerve tracts. It has been reported that VZV infection produces TLR ligands and we have found that peripheral neurons present in dorsal root ganglia express TLR3, 7 and 9 which when stimulated express mRNA for many cytokines and chemokines. In addition, TLR ligand stimulation of neurons upregulate the expression of TLRs and TRPV1. Furthermore, preincubation of neurons for 16 hours with the TLR ligands, enhances the calcium flux induced by capsaicin stimulation of TRPV1. Consequently, products of the herpes virus interacting with these TLR's can either directly or indirectly, by inducing chemokines, enhance the response of TRPV1 pain receptors, providing one possible basis for herpes Zoster neuralgesia in immunosuppressed cancer patients. These studies in a pain model indicate that the peripheral pain reported by many cancer patients may be addressed by effective regulation of neuroimmune molecules.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "DESCRIPTION: It is hypothesized that estrogen deficiency induces a decrease in the rate of fat oxidation which leads to accelerated gain of visceral fat mass and its associated insulin resistance. It is further hypothesized that viscera fat accumulation will be prevented by hormone replacement therapy through restoration of fat oxidation. To test this hypothesis, 100 women in early menopause will be metabolically characterized for substrate oxidation, fat distribution, and insulin sensitivity prior to being randomly assigned to receive either hormone replacement therapy or placebo for 2 years. Metabolic testing will be repeated at 2, 6, 12, 18 and 24 months.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Our knowledge about the organisation of cells, biological specimen structures, composition and properties in submicroscopic detail of biological samples is associated to a large extent to transmission electron microscopy (TEM). The Electron Microscopy Core (EM Core) Facility provides advice, technical services, training, equipment and facilities to NHLBI intramural scientists if their research requires electron microscopy (EM) to clarify questions. Using an electron microscope offers the advantage of increasing both the magnification of an object and the resolution over other imaging tools. These could be issues involving subcellular, supramolecular or macromolecular structure at a level of resolution below that obtained by a light microscope. The NHLBI EM Core Facility has supported projects using the following techniques in the past year: 1. Chemical fixation, embedding, ultra-thin sectioning and transmission EM digital imaging of tissues and cell culture. 2. EM immunocytochemistry, including immunogold, nanogold with silver enhancement and immunoperoxidase localisation of proteins and other antigens within and on the surface of tissues and cells by pre-embedding techniques. 3. Negative staining of large proteins, polymers and supramolecular structures as well as lipid and membrane vesicles for transmission EM digital imaging. 4. Rotary shadowing of large protein molecules, DNA and other macromolecules. 5. Preparation of platinum replicas of cytoskeletons, partially lysed cells and freeze-fractured/freeze- dried tissues and cells. 6. Chemical fixation, critical point drying, sputter-coating and scanning EM digital imaging of small organisms, organs, tissues and cells, as well as other materials such as artificial matrices. 7. Thawed cryosection immuno labelling technique (also called Tokuyasu method). 8. Immuno correlative light and electron microscopy (CLEM) on Tokuyasu cryosections. 9. Cryo-electron microscopy of vitreous sections (CEMOVIS) to observe biological samples in their most native, fully hydrated state. 10. High-pressure freezing (HPF) and freeze substitution (FS) allows improved morphological preservation compared to the conventional preparation. These techniques are available as a service to be performed by the EM Core Staff for the customer or they are available to be learned by users who will be trained to be independent users of the EM Core. In the past year, the NHLBI EM Core supported a total of 89 recorded projects for 37 investigators within the NIH. 62% of the services were requested by NHLBI research groups. The remaining 38% are divided as follows: NHGRI (8%), NCI (8%), NIA (1%), NIAID (2%), NIDCR (10%), NINDS (7%), NINR (1%) and OD (1%). We have also trained several postdoctoral fellows, students, and contractors in the use of the scanning and transmission electron microscopes and sample preparation techniques.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Dr. Philippe Szapary plans a career as a clinician investigator at the University of Pennsylvania School of Medicine, focusing his research on the critical evaluation of complementary and alternative medicine (CAM). His training will include formal course work towards a Master's of Science in Clinical Epidemiology with a specific focus on patient-oriented research (POR). To supplement his formal research training, he also plans to take courses in pharmacognosy and ethnobotany at the University of the Sciences in Philadelphia (USP). Using his formal training in POR and botanical pharmacology, Dr. Szapary plans to establish himself as an independent investigator focusing on new and existing cardiovascular CAM therapies. Environment: The University of Pennsylvania and nearby USP are uniquely suited to provide complementary training and resources for this award. At Penn, the Center for Clinical Epidemiology and Biostatistics will provide the formal POR research training. The Cardiovascular Risk Intervention Program and the Division of General Internal Medicine will provide study subjects. The General Clinical Research Center will provide ancillary support including nurses, a dietitian, a biostatistical programmer and laboratory services. At USP, the Department of Pharmacognosy will provide technical support in the analysis of study drugs. Research: In the last 10 years, patients have markedly increased their use of dietary supplements to treat and prevent chronic medical conditions like atherosclerotic cardiovascular disease. This widespread use continues despite little scientific evidence of benefit from randomized controlled trials (RCTs). Current estimates suggest that 30 percent of Americans are hypercholesterolemic. Serum cholesterol remains one of the strongest predictors of risk for coronary artery disease, the leading cause of death in Americans. Herbal therapies have multiple mechanisms of action making them attractive in the prevention of a multifactorial disease process like atherosclerosis. Two Ayurvedic herbals, gugulipid and curcuminoids, appear to have hypolipidemic, antioxidant and anti- inflammatory effects in humans. In this protocol, Dr. Szapary will primarily study the hypolipidemic effects of standardized extracts of curcuminoids and gugulipid in two RCTs. These trials will assess the safety and efficacy of these two agents when used alone and in combination, as done in Ayurvedic medicine, over a 3 to 6 month period. In addition, Dr. Szapary will examine their effects on state of the art biomarkers of oxidant stress and vascular micro-inflammation, both of which are important processes in the pathogenesis of atherosclerosis. These studies will help define the role of these herbal therapies in the prevention of atherosclerosis, and serve as a model for the critical evaluation of dietary supplements in cardiovascular disease.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Objectives: Sensorineural hearing loss (SNHL) is strongly associated with many aspects of military service including blast injury. The objectives of this proposal are to improve the prevention and treatment of SNHL in veterans. Research Design: To improve our understanding of cellular processes underlying hair cell (HC) damage and loss, the contributions of molecular isoforms within known HC damage and survival pathways will be evaluated by determining their expression, and by evaluating HC loss in gene knockout mice. To improve SNHL prevention, pharmaceutical agents that can protect cochlear HCs from damage will also be identified. To improve treatment for SNHL, transcription factors that enhance the ability of Atoh1 to induce the conversion of mammalian cochlear non-sensory cells into HCs will be identified. In addition, modifications to mechanisms of gene silencing will be studied to identify epigenetic limitations on the ability of nonsensory inner ear cells to adopt features of the HC phenotype. Methodology: Studies will be performed using in vitro cultures of inner ear sensory epithelia, and in vivo studies of noise damage to the cochlea. For identification of HC protectants and transcription factors, innovative high-throughput methods have been developed to permit rapid, but targeted, screening of pharmaceuticals and induced gene expression in mammalian cochlear epithelia. Progress over the past period of funding: Mechanisms of ototoxin entry into HCs have been identified; cellular patterns of reactive oxygen species accumulation into HCs have been determined and antioxidant treatments evaluated; two novel intracellular pathways of HC protection have been defined; three gene mutations that lead to deafness have been identified; a critical period of HC development in which transplantation is possible has been determined; two novel transcription factors that enhance the conversion of nonsensory cochlear cells into HCs have been identified, and epigenetic manipulation of the cochlear sensory epithelium to enhance gene expression induced by a viral vector has been performed. Clinical Relationship: The prevention and treatment of SNHL is of great importance to veterans and to the VA. The effects of SNHL on veterans' quality of life are substantial. SNHL and tinnitus also account for more disability compensation in the VA than any other disorder, and rehabilitation costs are high. The proposed research is targeted at developing new and improved therapies for prevention and treatment of this important health problem.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This research is directed toward synthesis and derivatization of novel DNA-binding peptides and detailed characterization of their DNA-binding and -condensing properties. A series of low molecular weight branched oligolysines and oligoarginines with a defined coupling site will be synthesized. We will determine the effect of number and positioning of positive charges on DNA condensation. Selected compounds will be derivatized with effectors such as receptor ligands, membrane-destabilizing peptides, carbohydrates and PEG. DNA binding properties of the various derivatives and the physical characteristics of resulting DNA complexes will be examined. Considering parameters like hydrophobicity and propensity of amino acids for particular secondary structures, we will introduce a limited number of rational mutations into a known membrane-destabilizing hexapeptide sequence and select peptides with highest membrane-destabilizing activities. The dependence of transfection efficiency in cell culture on DNA complex composition will be examined. Using the GCG package, membrane destabilizing peptides have been selected and compared for their primary, secondary and tertiary structure. This information and things like hydrophobicity indices etc. are essential in finding and designing new peptide sequences for the above-described project. The facilities provided by the Computer Graphics Laboratory are absolutely necessary for the continuation of this work. GL", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Chronic cigarette smoking is very common in alcohol use disorders (AUD). Nevertheless, the effects of chronic smoking on in-vivo measures of brain injury in alcohol-dependent individuals have not been studied. Thus, it is unknown if the full extent of brain atrophy, cell membrane and microstructural injury, derangement of neurometabolism, low cerebral blood flow, and neurocognitive dysfunction described in alcohol dependence are solely attributable to chronic alcohol use, or if chronic smoking influences these measures. Preliminary results from our ongoing magnetic resonance (MR) and cognitive studies indicate that chronic smoking appears to exacerbate alcohol-induced abnormalities in brain morphology, neurochemistry, and blood flow, and may adversely affect recovery of surrogate markers of neuronal and cell membrane/myelin integrity as well as aspects of neurocognition during short-term abstinence from alcohol. The main goal of this competing continuation is therefore to test in treated alcohol dependent individuals (i) if chronic smoking compounds alcohol-induced neurobiological brain injury, (ii) if chronic smoking effects are functionally significant, and (iii) if chronic smoking influences recovery of alcohol-induced neurobiological and neurocognitive dysfunction during abstinence from alcohol. We will continue to apply an integrative approach of combining information from different MR modalities and neurocognitive testing in the same individual longitudinally to assess the neurobiological and functional consequences of chronic smoking in treated alcohol-dependent individuals. Specifically, we will continue our longitudinal 1.5 Tesla MR studies to quantitate regional brain structure, brain metabolites (reflecting neuronal and myelin viability), and regional cerebral blood flow as well as repeat comprehensive neurocognitive testing. In addition, new MR studies at high magnetic field (4T) will improve the quality of cerebral blood flow and diffusion data, and will measure cortical levels of glutamate and Y-aminobutyric acid, amino acids critically involved in the initiation and maintenance of substance dependence. Relationships of MR-derived and neurocognitive measures will determine the functional relevance of neurobiological measures and their changes over time and test the effects of chronic smoking on functional neurocircuitry in alcoholism. This application is responsive to PA-05-074 in that the proposed research will improve our understanding of the mechanisms underlying alcohol- and smoking induced brain injury and its potential reversibility with abstinence from alcohol. Chronic smoking may have hitherto unrecognized but significant contributions to these neurobiological processes and their cognitive and clinical consequences. The knowledge to be gained from this translational research can be used directly for public education, in new approaches to pharmacologic and behavioral interventions for AUD, and in monitoring treatment outcome.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "A method for attaching fluorescent receptor groups at specific sites in large RNA molecules will be developed. The sensitivity of the labels to the solvent environment and to energy transfer from nucleic acid bases, proteins, and metal ions will be used as a way of investigating RNA secondary and tertiary structure. A small, functionally interesting region of the E. coli 16S ribosomal RNA (the S8/S15 binding region) will be isolated as a well-defined fragment and used to test the fluorescence labeling method. The fluorescent labels and other physical chemical methods will be used to confirm the predicted secondary structure of the region (seven potential helices) and to determine sites for tight Mg ion binding which associate with tertiary structure. The binding of two ribosomal proteins known to recognize this region, S8 and S15, will be examined in detail to determine what RNA recognition features are used by each and whether the proteins influence the structure of the RNA fragment.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "The objectives of these projects are to: 1) develop equipment for use by investigators in Viral Oncology, 2) develop narrated, slide presentations for use by intra- and extra-mural members of oncogenic virus research on the hazards of laboratory procedures and equipment, 3) develop a training workshop for \"Certification of Biological Safety Cabinets\", 4) prepare monographs and safety notes on current hazards that occur within the laboratory, 5) assist in the development of standards for production and procurement of Class II biological safety cabinets, 6) provide consultation services on biohazards to Viral Oncology intra-and extra-mural personnel.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "Puresyn, Inc. has developed purification processes for recombinant adeno-associated virus gene therapy vectors utilizing its proprietary chromatographic resin, PolyFlo. In Phase I, two tandem chromatography processes were developed: heparin affinity and PolyFlo and tandem PolyFlo in two different modes. Each process resulted in product of high purity and recovery that exceeds gradient centrifugation processes. We demonstrated each process removes significant amounts of host and viral contaminants. We are seeking Phase II funding to concentrate on AAV production to include large-scale purification up to 5 x 10e14 particles per run without compromising purity, yield or biological activity. We will refine and expand our current processes to assure they are applicable to AAV serotypes 1, 2 and 5. We will also address the significant problem of AAV aggregation. Finally, we will conduct an in vivo mouse study to determine the correlation between purity, biological activity and safety through the evaluation of immune and histopathological responses to the vector. Results of this research will have immediate impact for those engaged in the use of recombinant AAV vectors for gene delivery because it will allow the large-scale development of AAV vectors, It will have positive commercialization implications for PoIyFlo and Puresyn, Inc. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The common mold, Aspergillus fumigatus, is an opportunistic pathogen and the most frequently isolated causative agent of invasive aspergillosis. The disease is often contracted by immunocompromised hosts, including cancer patients, with reported mortality rates >80%. Our long-term goals are to develop a vaccine and to provide better means for the early diagnosis and monitoring of aspergillosis. Our specific hypotheses are: 1) certain aspergillus antigens have an immunoprotective effect, and 2) some antigens may serve as indicators of early stages of aspergillus infections. We base our hypotheses on the following observations. First, mice vaccinated with crude fungal protein extracts or viable conidia survive infection following immunosuppression (e.g., with corticosteroid treatment). Second, the fungus actively secretes a variety of specific proteins and peptides at different stages of its lifecycle. More than 60 antigens and allergens of Aspergillus have been described (1-4). However, only one of them, the polysaccharide galactomannan, is routinely assayed to diagnose Aspergillus infections. Based on these observations, our experimental focus is on the identification and structural and immunological characterization of A. fumigatus antigens. We plan to utilize and improve novel mass spectrometric techniques to detect and identify proteins and peptides of fungal origin from complex mixtures such as culture supernatants, bronchoalveolar lavage (BAL) fluid, blood and urine.", "meta": {"pile_set_name": "NIH ExPorter"}}
{"text": "As a result of Phase I researches, a perfected design of a simple, reliable and economical phase modulation system has been achieved. Desig parameter perturbation and component selection has lead to this simplified and reliable design of a dual wavelength time sharing phase modulation spectrometer (dual wavelength PMS). The frequency has been optimized at 200 MHz; time-sharing of the oscillator frequency between two diode lasers at different wavelengths, laser diode driver circuits, fiber optic coupling, standardized detector heterodyne njection, amplitude and sensitive phase detection circuit, together with time sharing demodulation are engineered. This instrument had undergone rigorous electromechanical tests and has a remarkable stability corresponding to a draft of less than 0.04degree/hr of phase drift and a noise level of 0.9 mm or approximately 0.2degree of phase. Tissue signals are of the order of 4 cm in amplitude and thus signal to noise ratios of 5:1 with a 5 sec time constant at 3 cm input/output distance are feasible. The remarkable features of time and frequency domain spectroscopy in providing the optical path length for photon migration in tissues is well recognized. Phase I set forth to exploit design parameters of a simplified system suitable for biomedical research and for clinical studies. Phase II will focus on construction of dual wavelength phase modulation systems that will bring to the clinic the advantages of TRs in an economical and feasible device. We propose to pursue product development and testing under Phase II support leading to commercial exploitation under Phase III; that is, production prototype, FDA approval, marketing and sales.", "meta": {"pile_set_name": "NIH ExPorter"}}