Kawasaki syndrome (KS) is the most relevant cause of heart disease in children living in developed countries. Intravenous immunoglobulin (IVIG) has a preventive function in the formation of coronary artery abnormalities and a poor strictly-curative action in established coronary damage. More than two decades ago, the Harada score was set to assess which children with KS should be subject to administration of IVIG, evaluating retrospectively a large cohort of patients with regard to age, sex and laboratory data. Nowadays, high dose IVIG is administered to all children with a confirmed diagnosis of KS, but a tool for predicting non-responsiveness to the initial infusion of IVIG has not been found. The prediction of IVIG resistance is a crucial issue, as recognising these high-risk patients should consent the administration of an intensified initial treatment in combination with IVIG in order to prevent coronary injuries. Few reports have focused on factors, referring to both clinical parameters and laboratory data at the onset of KS, in order to predict which patients might be IVIG non-responsive. We have analysed three different risk scores which were formulated to predict IVIG resistance in Japanese children with typical KS, but their application in non-Japanese patients or in those with incomplete and atypical patterns of the disease has been studied in a fragmentary way. Overall, our analysis showed that early and definite ascertainment of likely IVIG non-responders who require additional therapies reducing the development of coronary artery involvement in children with KS is still a challenge. Almost half a century has passed since Kawasaki syndrome (KS) was first reported by Tomisaku Kawasaki as an enigmatic disease affecting children, and, today, KS still remains uncannily dangerous due to the intrinsic risk of damaging the vascular system, mostly the coronary arteries, in 25% of untreated patients. It is no coincidence that KS stands as the most relevant cause of heart disease in children of developed countries, not only in Japan where the disease is largely observed and recognized ,2. Many i.e., within nine days of onset, with the eventual development of CAA. The study was set in two parts: the first part aimed to evaluate doses and types of IVIG, the second aimed to assess the elective indications of IVIG for children with KS. In particular, Harada analysed data from 865 naïve patients who had not been treated with IVIG, corticosteroids or nonsteroidal anti-inflammatory drugs, and the scoring criteria included seven items: white blood cell and platelet counts, C-reactive protein (CRP), hematocrit, albumin, age at diagnosis, and male sex. Children with KS who satisfied at least four of these seven criteria were thought to be candidates for IVIG treatment (List 1) [Methods to predict which children are at greatest risk of developing CAA have been extensively sought to determine the general prognosis of KS. The first scoring model was formulated in 1991 by Harada, but it was not intentionally created to assess a general risk of cardiovascular involvement: it merely tried to identify KS children at higher risk for potential complications, as different issues related to cost and availability of IVIG made it problematic at that time to treat all children with KS in Japan. The “Harada score” was developed by retrospectively correlating routine laboratory and clinical findings in the early stages of KS, List 1. The Harada score (1991) was extrapolated by a multicentre study carried out to evaluate the effectiveness of different doses and kinds of intravenous globulin (IVIG) in 865 naïve patients with Kawasaki syndrome, analysed retrospectively with regard to laboratory data, age, and sex: the score was an attempt to establish a set of criteria for starting treatment with IVIG.3/mm3White blood cell count > 12 × 104/mm3Platelet count < 35 × 10C-reactive protein > 3+Hematocrit < 35%Albumin < 3.5 g/dLAge < 12 monthsSex MaleThe Presence of at Least 4 of these 7 Criteria prompted the Administration of Intravenous Immunoglobulin to the Child with Kawasaki Syndrome:et al. [Although this score should not be used to triage children who do not need IVIG, its influence was dispelled over time because, nowadays, IVIG is necessarily given to “all” KS patients. In 2014 Tewelde et al.  performeet al. [Different researchers have focused on better recognising early predictors of IVIG resistance with the hope of developing risk scoring algorithms and estimating the probability of a successful response to IVIG. In 2009, Son et al.  indicateet al. .et al. [Another scoring model was created by Nakano et al.  in 1986,et al. [In 1998, Beiser et al.  construcet al. [In 2000, after a multivariate analysis of 82 KS patients, Fukunishi et al.  noted thThe prediction of IVIG resistance has been also evaluated by three more recent risk scores, formulated in the last decade and based on children with typical KS: these three scoring systems have been considered and schematized in et al. [In 2006, Egami et al.  defined et al. [However, in the same year, Tremoulet et al.  . The authors concluded that resistance to IVIG therapy might be heralded by elevated levels of IL-6, and that IVIG-resistance should contemplate IL-6 in the scoring system.Sato et al.  determini.e. the ratio of siblings to population incidence of KS, is only slightly lower than that of type 1 diabetes and about four times higher than that of allergic asthma [The contribution of genetics to KS susceptibility and even to responsiveness to IVIG is a matter of great debate. It has been reported that siblings of KS patients have a 6 to 10-fold greater incidence of the disease than the general population . In addic asthma . These ac asthma ,35. TherChildren suspected to have KS should receive prompt treatment with IVIG (2 g/kg of body weight as a single dose) within 10 days of illness onset . Many re


P < 0.05); the odds of being aware of bTB were 9.4, 7.3, and 2.1, respectively. In conclusion, these demonstrate the urgent need for public health authorities to intervene in bTB control. The risk of bTB transmission as indicated by the personnel's practices and awareness levels in Bauchi State could be prevented through the use of protective clothing (PPEs).Bovine tuberculosis (bTB) is a chronic infectious and contagious zoonotic disease of domestic animals, wild animals, and humans. It poses a public health threat and economic losses due to abattoir condemnation of infected carcasses during meat inspection of slaughtered animals. Bovine tuberculosis is widespread in Africa including Nigeria affecting both cattle and humans, particularly Northern Nigeria. A prospective survey was conducted from June to August 2013 in the three Zonal abattoirs of Bauchi State, Nigeria. A total of 150 structured close-ended questionnaires were administered to abattoir personnel to assess their level of awareness of bTB. This study was aimed at determining the level of public health awareness, attitude, and practices of abattoir workers of bTB in Bauchi State, Nigeria. There was a statistically significant association between respondents' awareness of bTB and their occupational status, age, and duration of exposure to cattle carcasses ( Bovine tuberculosis (bTB) is a chronic infectious and contagious zoonotic disease of domestic animals, wild animals, and humans . It also Mycobacterium bovis which is a member of Mycobacterium tuberculosis complex [ Mycobacterium tuberculosis complex (MTBC), include Mycobacterium tuberculosis, M. bovis, M. microti, M. caprae, M. africanum, M. canettii, and M. pinnipedii. Mycobacterium africanum consists of a rather heterogeneous group of strains isolated from humans in Africa [ Mycobacterium bovis, otherwise known as the bovine tubercle bacillus, is the cause of bovine tuberculosis and the organism may be transmitted by aerosol or droplets of exudates containing the bacilli. It can be transmitted by ingestion of feed and water contaminated with urine, faecal material, or exudates from diseased animals that contain the tubercle bacilli [ M. bovis via direct inoculation [ M. tuberculosis and is an occupational risk associated with performing autopsies), this skin lesion can occur in persons handling infected meat. It is very rare and generally self-limiting. Because M. bovis is either enzootic or found sporadically in much of the developing world, there is clearly a risk of cow to human transmission by either ingestion or inhalation [ M. bovis disease in humans is more common.Bovine tuberculosis is caused by complex , 10. Then Africa . Mycobac bacilli . In rareculation . Referrehalation . As a re M. bovis of the tubercle bacillus irrespective of the host. Bovine tubercle bacillus has one of the broadest host ranges of all known pathogens. The species has been reported in domesticated and feral Bovidae. Other species in which the disease has been reported include goat, sheep, pig, horse, cat, dog, fennec fox, bison, buffalo, badger, wild and feral pig, antelope, camel, man, and nonhuman primates [The bovine tubercle bacilli is usually assigned to bTB in cattle and sometimes could be used to denoteprimates . Cattle primates .2 (250 g/m2), and per capita income of $983 [Bauchi State occupies a total land area of 49,119 km² representing about 5.3% of Nigeria's total land mass and is located between latitudes 9° 3′ and 12° 3′ north of the equator and longitudes 8° 50′ and 11° east of the Greenwich Meridian . The staBauchi State has a total of 55 tribal groups in which Hausa, Fulani, Gerawa, Sayawa, Jarawa, Bolewa, Karekare, Kanuri, Fa'awa, Butawa, Warjawa, Zulawa, and Badawa are the main tribes. This study was carried out in Bauchi, Katagum, and Misau Local Government Areas (out of the 20 LGAs), each representing the three senatorial zones as Bauchi south, Bauchi north, and Bauchi central with populations of 493,810, 295,970, and 263,487, respectively. vice versa. Assessment of their level of awareness was based on biodata (demographic features), knowledge (zoonotic nature and symptoms of TB in humans), attitude (vaccination status of abattoir staff), practice (use of protection while handling carcasses), and other risk factors and scoring recall format of “0–8” was adopted as criteria for a respondent to be regarded for analysis as described by Waller et al. [A total of 113 structured close-ended questionnaires were retrieved from and analysed after the distribution to abattoir staff focused on assessing the level of knowledge, attitude, and practice of bTB transmission from slaughtered cattle to man andr et al. . Personnχ2) was used to determine possible association between variables and M. bovis. Odds ratio (OR) and 95% confidence interval were calculated to measure the strengths of association between variables and bTB (M. bovis). Tables and bar charts were constructed using Microsoft Excel 2010. Values of P < 0.05 were considered significant.Data were analysed using statistical package for social sciences (SPSS) version 20.0. Chi- square (P < 0.05) association between awareness of the respondents of bTB and their occupational status, age, and duration of exposure to cattle carcasses, and the odds of being aware of bTB by their level of awareness were 9.36, 7.29, and 2.06, respectively (Majority of the respondents had contact with cattle for over 3 years (81.40%) . A signiectively . Howeverectively .P < 0.05) association between awareness of the respondents (abattoir staff) of bTB and their occupational status, age, and duration of exposure to cattle carcasses. However, most of the abattoir staff believed in the importance of use of protective clothing while working, but very few of them did not know the importance. Likewise, on its zoonotic nature, most of them knew that bTB can be contracted from cattle. Some of the respondents had even received prophylactic BCG vaccine while others had not received it and few did not know their vaccination status. Therefore, this clearly indicated their awareness of bTB and its zoonotic risk, probably due to awareness campaign and researchers that visit the abattoir for sampling and data collection.In the public health risk analysis of bTB found in this study, there were more respondents in Bauchi abattoir (53.10%) compared to Misau and Katagum abattoirs. Butchers made up of most of the respondents were then followed by other abattoir staff and veterinarians. Many of them had not used protective clothing, despite the protection it gives against zoonotic transmission, clearly indicating their high risk of contracting bTB by occupation. This study showed significant  risk associated with the respondents' awareness and practices of bTB found in this study had agreed with the report of Cadmus and Adesokan  who repoP = 0.001) between reactor cattle and human TB cases in households, indicating the high zoonotic risk of bTB among exposed individuals. Despite their awareness of the zoonotic risk, these figures had clearly indicated their high risk of contracting the disease due to exposure and negligence, as only few of the respondents (32.38%) used protective clothing in practical sense. Moreover, the exception was that a significant number (59.81%) of abattoir personnel of Bauchi State reported consuming boiled milk in recognition of the risk of contracting bTB and Brucellosis in fresh milk (unpasteurised milk). These findings were contrary to the report of Tigre et al. [The statistically significant association between awareness of the respondents of bTB and occupational status found in this study had agreed with the report of Tigre et al.  who carre et al.  in a croMycobacterium bovis, the cause of tuberculosis in cattle, sometimes causes disease in humans: the finding that majority of the respondents have had contact with cattle for over 3 years (81.40%) could be a risk factor that promotes transmission from cattle to humans. Transmission from cattle to humans is mainly by ingestion of raw cattle products from infected animals, and transmission by inhalation is possible when there is prolonged contact. This agreed with the report of Byarugaba et al. [ Mycobacterium responsible for the disease, whether M. bovis causes disease in humans. Conclusions from this study showed that large proportions of the respondents (90.27%) were aware of zoonotic bTB. However, some (32.38%) used protection while working with cattle carcasses. This showed their high risk of contracting bTB and other zoonoses endemic in the study area. These findings also demonstrated the urgent need for the state public health authorities to intervene. The risk of bTB transmission as indicated by the practices and awareness levels of the abattoir personnel in Bauchi State could be prevented through the use of protective clothing.a et al. , a risk


After publication of the original article  the auth1, Denise St-Cyr-Tribble2†, Marianne Xhignesse3†, Andrew Grant4†, Christine Brown1† and Marie-France Langlois1*Jean-Patrice BaillargeonThe new order, however, should be as listed below:1, Denise St-Cyr-Tribble2†, Marianne Xhignesse3†, Andrew Grant4†, Christine Brown1†, Thomas G. Poder5 and Marie-France Langlois1*Jean-Patrice Baillargeon


Mobile health technologies to detect physiological and simple-analyte biomarkers have been explored for the improvement and cost-reduction of healthcare services, some of which have been endorsed by the US FDA. Advancements in the investigations of non-invasive and minimally-invasive molecular biomarkers and biomarker candidates and the development of portable biomarker detection technologies have fuelled great interests in these new technologies for mhealth applications. But apart from the development of more portable biomarker detection technologies, key questions need to be answered and resolved regarding to the relevance, coverage, and performance of these technologies and the big data management issues arising from their wide spread applications. In this work, we analyzed the newly emerging portable biomarker detection technologies, the 664 non-invasive molecular biomarkers and the 592 potential minimally-invasive blood molecular biomarkers, focusing on their detection capability, affordability, relevance, and coverage. Our analysis suggests that a substantial percentage of these biomarkers together with the new technologies can be potentially used for a variety of disease conditions in mhealth applications. We further propose a new strategy for reducing the workload in the processing and analysis of the big data arising from widespread use of mhealth products, and discuss potential issues of implementing this strategy. There have been intensifying efforts to explore mobile health  technologies for delivering healthcare at reduced costs and for facilitating more precise and personalized medicine1244Although these physiological and simple-analyte biomarkers cover many disease conditions, their coverage is substantially limited for such prevalent diseases as cancers, infectious, respiratory, digestive, endocrine and nervous system diseases, as indicated by the disease-coverage profiles of the 73 FDA endorsed, and 94 physiological and simple-analyte biomarker candidates described in the literatures  and S2. 12Some genetic, proteomic and metabolomic molecular biomarkers have been clinically used and many more such molecular biomarker candidates  have been discovered for diagnosing and monitoring diseases, directing treatments and predicting patient responses679109121314From the investigations and opinions described in the literatures listed in Here, we address some of these questions by analysing (1) biomarker detection capability of the literature-reported new technologies with specific focus on their detection sensitivity, required sample volume, test time, and costs with respect to experimentally-determined biomarker levels in patients and the detection limits, and (2) the disease coverage, patient populations, and the diagnostic, prognostic, and theragnostic sensitivity and specificity of the literature-reported non-invasive and minimally-invasive finger-prick molecular biomarkers for mhealth applications with respect to the detection limits of the new detection technologies. We also discuss the feasibility and practical issues of adopting a new strategy for reducing the heavy workload of mhealth data processing by automated electronic pre-screening of the big biomarker screening data.The detailed information of 73 mhealth apps endorsed by the US FDA was obtained by manually checking the descriptions of the apps listed in FDA 510(k) medical device databaseThe performance of the biomarkers in diagnosing, prognosing or theragnosing specific conditions has been statistically measured by sensitivity (the proportion of the condition-positive samples that are correctly identified as positive) and specificity (the proportion of the condition-negative samples that are correctly identified as negative)The new biomarker-detection technologies combined with mobile phone or the equivalent imaging devices have been explored for detecting at least 23 molecular biomarkers including 11 non-invasive ones . These n912The second group of technologies primarily employ innovative P-ELISA in combination with a scanner, printer or digital camera based image-processing facility to achieve a detection sensitivity as high as 33.7 fg/mL2931Overall, 12 or 52.2% of the 23 tested molecular biomarkers are detectable by these new technologies at low concentrations (0.3–810 pg/mL and 4–50 ng/mL for 8 and 4 biomarkers respectively). Although the detectable concentrations of these 23 biomarkers are roughly 10-fold higher than those of the conventional technologies2413Analysis of the 664 literature-reported non-invasive molecular biomarkers (examples in inThe diagnostic performance of 88 (or 29.7%) of the 296 diagnostic biomarkers for 43 diseases and the prognostic performance of 24 25.5%) of the 94 prognostic biomarkers for 14 conditions have been reported in the literature  codes for defining, studying and managing diseases and treatmentsMolecular biomarker-based mobile health technologies have the potential to significantly improve the efficiency and quality of healthcare for a variety disease conditions particularly those with large patient populations that cannot be solely covered by physiological and simple-analyte biomarkers. Some of these biomarkers combined with the new detection technologies are readily applicable for mhealth applications. The increased workload in processing and analyzing high volumes of mhealth data may be efficiently managed by an electronic system that facilitate automatic pre-screening and analysis of the biomarker data for filtering potential patients likely in need of further attention and evaluation.How to cite this article: Qin, C. et al. The Assessment of the Readiness of Molecular Biomarker-Based Mobile Health Technologies for Healthcare Applications. Sci. Rep.5, 17854; doi: 10.1038/srep17854 (2015).


Protein microarray technology has gone through numerous innovative developments in recent decades. In this review, we focus on the development of protein detection methods embedded in the technology. Early microarrays utilized useful chromophores and versatile biochemical techniques dominated by high-throughput illumination. Recently, the realization of label-free techniques has been greatly advanced by the combination of knowledge in material sciences, computational design and nanofabrication. These rapidly advancing techniques aim to provide data without the intervention of label molecules. Here, we present a brief overview of this remarkable innovation from the perspectives of label and label-free techniques in transducing nano-biological events. Microarrays are now recognized as powerful tools for predictive and early disease diagnosis, as well as research in many biological fields. In each microarray, also known as ‘biochips’, thousands of reaction spots (capturing agent spots) are arrayed onto a few square centimeters. It is a remarkably versatile and useful platform, particularly when dealing with very limited sample volumes in genomics , proteomProtein microarrays are mainly constructed to achieve two primary objectives ,13: (i) Protein microarrays are supported by three key technologies: (i) production of functional capture agents; (ii) surface chemistry; and (iii) development of high-throughput detection methods. These technologies must be tied with one another in order to provide valuable protein detection. To date, dozens of exciting reviews on protein microarrays have been reported  high signal-to-noise ratio; (ii) high spatial resolution; and (iii) good reproducibility. While offering broad practicality, methods need to have low instrumentation costs, rapid determination and robustness, while producing good qualitative results. Although there is not yet a general sensing method that can be ubiquitously applied to detect the cluster of biomolecules in various samples, there are several candidates that can be considered.In this review we focus on five candidate label detection methods, namely, (1) fluorescent labeling, (2) isotopic labeling, (3) chemiluminescent labeling, (4) electrochemically active probe labeling, and (5) nanoparticle labeling, and five candidate label-free detection methods, namely, (6) mass spectrometry (MS), (7) microcantilevers, (8) quartz-crystal microbalance (QCM), (9) surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR), and (10) anomalous reflections of the gold surface (AR). The features of the different methods with respect to their availability for protein microarrays are summarized and discussed below. A label is defined as any foreign molecule that is chemically or temporarily attached to the molecule of interest to detect molecular presence or activity, which can potentially alter its intrinsic properties. It requires a labeling process as a preparation step that is usually low yield, combining synthesis and purification. Fluorescent, chemiluminescent, and nanoparticle labeling usually involves covalent bonding through coupling chemistries, while some of electrochemically active probe labeling requires only temporary attachment of intermolecular bonding. Isotopic labeling implicates “light” and “heavy” elements being incorporated into target molecules to result a detectable difference.Meanwhile, label-free detection methods utilize molecular biophysical properties such as molecular weight , refractive index  and molecular charge to monitor molecular presence or activity. Furthermore, these methods can be used to track molecular events in a real-time manner. In a typical biosensing process, molecular interactions are transduced as mechanical, electrical, or optical signals, and are thus detectable without any label probes. The main advantage for label-free detection is that more direct information can be acquired, as the methods use only native proteins and ligands.Fluorescent label detection methods are the most common and convenient techniques to transmit information from molecular events. Fluorescence probes are stable, easily manipulated, and provide good sensitivity and resolution . These aFluorescence probes are currently available in many forms, from quantum dots and small organic molecules to fluorescent proteins with a range of brightness and specificity that can be selected based on need ,31. The We also developed new fluorescent labeling detection methods utilizing the excluded volume effect of the target molecule or fluorometric/colorimetric changes in the detection moiety . In the One of the earliest methods for molecular detection involved the use of radioisotopes that enables accurate quantification of protein abundance . Althouget al. introduced stable isotope labeling using amino acids in cell cultures (SILAC) . W. Wet al.ions (2) . HoweverIn gold surface modification approach, in order to improve the sensitivity of detection of biomolecular interactions in the AR method, three-dimensional (3D) nanostructures on gold surfaces with a series of well-defined structures of poly(amidoamine) dendrimers (PAMAMs) from generation 2 to 4  b 104]. . 104]. SεAu = −1.567 + 4.764i would be moved towards ε = −1 + i. Subsequently, gold-containing alloys (Au-Ag) or composites (Au-Ag2O) were prepared. Measurement of monomolecular layers with different thicknesses, and biotin-antibiotin IgG assay confirmed the predicted higher sensitivity as a result of dielectric constant optimization. The dielectric constants of these newly produced substrates, as measured by ellipsometer, correspond with theoretically enhanced levels of sensitivity.We also developed gold-containing alloy or composite approach. The use of an AR substrate with an optimized dielectric constant would improve the sensitivity when compared to that obtained with conventional gold substrates due to an optimal complex reflection coefficient at the surface . IdeallyIn Metal-insulator-metal (MIM) approach, a new AR platform based on a metal-insulator-metal (MIM) structure c has beeObserving molecular events requires methods that are highly sensitive to minute changes, subsequently producing a high signal-to-noise ratio. To date, there are no general sensing platforms that can be ubiquitously applied to detect the constellation of biomolecules of various samples  with high sensitivity, large linear dynamic range, good speed, low cost and simple design. Major limitations include problems such as: (i) time-consuming preparation steps; (ii) non-native signal interference; (iii) unsuitability for in-situ observation; (iv) necessity of sophisticated, high cost equipment; and (v) need for specialized technicians and motivated researchers to keep improving or combining principles in detection methods.The development of label-free methods in recent years has revealed unprecedented information on molecular interactions, largely because observations are taken based on the characteristics of the biomolecules themselves. Furthermore, the issues of false positives and conjugated labels can be avoided. This could provide more reliable dynamic constant parameters that involve molecular interactions. From another perspective, detection methods that require no labeling are easily reproducible. Thus, these methods hold great potential to be further developed into a unique class of microarray analytical techniques. Nevertheless, stepping up these methods to match the commercial viability of current labeling methods will remain a challenge.All examples given here have shown that microarray using label or label-free methods require assistance from advanced chemistry and material sciences, nano-fabrication and computational design. In the near future, it is hoped that more practical detection methods will be developed.


The BTB domain is a conserved protein-protein interaction motif. In this study, we identified 56 BTB domain-containing protein genes in the silkworm, in addition to 46 in the honey bee, 55 in the red flour beetle, and 53 in the monarch butterfly. Silkworm BTB protein genes were classified into nine subfamilies according to their domain architecture, and most of them could be mapped on the different chromosomes. Phylogenetic analysis suggests that silkworm BTB protein genes may have undergone a duplication event in three subfamilies: BTB-BACK-Kelch, BTB-BACK-PHR, and BTB-FLYWCH. Comparative analysis demonstrated that the orthologs of each of 13 BTB protein genes present a rigorous orthologous relationship in the silkworm and other surveyed insects, indicating conserved functions of these genes during insect evolution. Furthermore, several silkworm BTB protein genes exhibited sex-specific expression in larval tissues or at different stages during metamorphosis. These findings not only contribute to a better understanding of the evolution of insect BTB protein gene families but also provide a basis for further investigation of the functions of BTB protein genes in the silkworm. Drosophila melanogaster) [The BTB  domain, also known as the POZ domain, is an evolutionarily conserved protein-protein interaction motif consisting of approximately 100 amino acid residues. This domain was originally identified in the bric-a-brac, tramtrack, and broad complex proteins in the fruit fly (ogaster) . Severalogaster) , 5, 6.The members of the BTB protein gene family are functionally involved in a variety of biological events in eukaryotes, including developmental timing , oogenesHomo sapiens), 178 in the nematode (Caenorhabditis elegans), 195 in the mouse (Mus musculus), 77 in the mouse-ear cress , 85 in the fruit fly, and 85 in the malaria mosquito (Anopheles gambiae). The silkworm (Bombyx mori) is considered an economically important insect and has been used widely for determining the genetic basis of biological events in the order Lepidoptera. To date, only one BTB protein, broad complex (BR-C) protein of the BTB-ZF subfamily, has been identified in the silkworm, where it has been shown to mediate ecdysone signaling [Genome-wide characterization of the BTB protein gene family has been performed in several species. The numbers of genes belonging to this family vary greatly among species ; 183 areignaling , 30. In ignaling , comparehttp://www.silkdb.org/silkdb/) and KAIKObase (http://sgp.dna.affrc.go.jp/KAIKObase/), in a local BLASTp search. The predicted BTB protein genes were further annotated through an online BLAST search in NCBI. The same approach was used to identify BTB protein genes in four other insects: the fruit fly, the honey bee , the red flour beetle , and the monarch butterfly . Two online programs, SMART (http://smart.embl-heidelberg.de/) [http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi) [BTB protein genes were identified in thesilkworm genome using the amino acid sequence of the conserved BTB domain as a query against the gene collections downloaded from two silkworm genome databases, SilkDB (erg.de/)  and a CDpsb.cgi) , were ushttp://wolfpsort.org) [http://www.cbs.dtu.dk/services/TargetP).Silkworm BTB protein genes were mapped on the chromosomes based on the single- nucleotide polymorphism (SNP) linkage map for the silkworm . The subort.org)  and TargThe full amino acid sequences and BTB domain sequences of the identified BTB proteins of the silkworm and other surveyed insects were used to perform a multiple sequence alignment by the ClustalX program . The aliBased on microarray data for gene expression in various tissues of silkwormlarvae on day 3 of the fifth instar (L5D3) , we dete Dazao strain were reared on fresh mulberry leaves at 25°C under 12-hour light/12-hour dark cycle. Silkworm larval tissues, including silk gland, midgut, fat body, hemocyte, head, integument, Malpighian tubule, ovary, and testis, were isolated from female and male larval individuals on L5D3. The procedures for RNA extraction, cDNA synthesis, and RT-PCR reaction were the same as previously described [http://dx.doi.org/10.1155/2014/865065).RT-PCR experiments were further used to examine the expression patterns of silkworm BTB protein genes in multiple tissues of silkworm larvae on L5D3. Silkworms of theescribed . The priTo identify BTB protein genes in the silkworm genome, we used the amino acid sequence of a common BTB domain (SMART accession number: SM00225) and the BTB domain sequence of BR-C protein as queries to search against the predicted silkworm gene collection. As a result, 56 BTB protein genes were retrieved from the silkworm genome . SMART aAccording to previous reports  and on t BTBD20, BTBD21, and ZBTB13) were assigned to 22 chromosomes  of the BTB zinc finger subfamily, were grouped more closely than other BTB protein genes. Curiously, the members of each of the remaining subfamilies, except for BTB-Kelch and MATH-BTB, each of which contained single gene, were grouped irregularly with the members of other subfamilies.Given that BTB fold of the BTB proteins has been shown to be structurally well conserved , we used BTBD13 and BTBD14, the clade of BTBD3 and ZBTB3, the clade of BTBD18 and ZBTB13, and the clade including BTBD5, BBTB1, and the BTB-BACK-PHR subfamily. The remaining BTB protein genes were grouped into different clades in an irregular manner.We also constructed a phylogenetic tree of silkworm BTB protein genes using the complete amino acid sequences predicted by these genes. As shown in To comprehensively characterize the evolution of insect BTB protein genes, we identified BTB protein gene families in four insect species for which the whole-genome sequence is available. As listed in  BmZBTB6 and BmZBTB5, two silkworm BTB-ZF protein genes that encode BR-C and fruitless, respectively, were grouped together into a clade with their homologs from other surveyed insects and they showed an orthologous 1 : 1 : 1 : 1 : 1 relationship among all five insect species. This was the same as grouping clades containing the orthologs of the following BTB protein genes from the silkworm: BmZBTB3, BmZBTB7, BmZBTB9, BmZBTB10, BmBTBD1, BmBTBD15, BmABTB1, and BmBBTB4.We constructed a phylogenetic tree including all of the BTB protein genes identified in the silkworm and other surveyed insects using the amino acid sequences of their BTB domains . In this ABTB2, BBK3, and BBK4 were specifically expressed in the testis. ZBTB6 expression was present in the Malpighian tubule, integument, fat body, and midgut. ZBTB4 was mainly expressed in the hemocytes, head, and integument. MBTB showed ubiquitous expression in the ovary, testis, midgut, and Malpighian tubule. In addition, RT-PCR examination confirmed the testis-specific expression of ABTB2, BBK3, and BBK4 in silkworm larvae  . Based om larvae . BTBD6, ABTB2, BBTB3, BBK3, BBK4, BBK5, ZBTB5, and ZBTB7, displayed sexual dimorphism; the expression of BTBD6, ABTB2, BBK3, and BBK4 was male-specific, whereas that of BBTB3, BBK5, ZBTB5, and ZBTB7 was female-specific.Using the microarray data on gene expression during silkworm metamorphosis from the larval to adult stages, we surveyed the developmental expression profile of the silkworm BTB protein genes. The results showed that 27 of the identified BTB protein genes were expressed during silkworm metamorphosis . RemarkaMembers of the BTB protein gene family are characterized by the presence of a BTB domain that mediates protein-protein interactions , 41, 42. BR-C (ZBTB6) or fruitless (ZBTB5), two well-studied insect BTB protein genes, were grouped into a clade on the phylogenetic tree , which has been shown to be involved in male genital development and survival in the fruit fly [ ABTB2 gene suggests that this gene may be involved in the regulation of male genital development in the silkworm. In addition, we noted that ZBTB5 gene showed no sex-dependent expression on L5D3 but was specifically expressed in the female silkworm during the pupa-adult transition. Given that during oogenesis the growth and meiosis of the oocytes and egg formation occur during the pupa-adult transition of the silkworm [ ZBTB5 may play key roles in the regulation of silkworm oogenesis. Future studies are necessary to determine how sex-specific expression of these BTB protein genes functions during silkworm development.Another important finding of our study is that several silkworm BTB protein genes exhibited spatiotemporally specific expression profiles. Interestingly, silkwormruit fly , 50. Thesilkworm , we specTable S1: List of the primers used in this study.Table S2: Inventory of BTB protein genes in other four insect species, including the fruit fly, honey bee, red flour beetle, and monarch butterfly.Click here for additional data file.Click here for additional data file.


However, among non-HAART users-ever, the oldest age group had the highest incidence rate ratio compared to younger individuals ). While the incidence of KS decreased with age, older HIV-infected persons who do not receive HAART are still at increased risk of KS. As KS remains an important malignancy among HIV-infected persons, earlier HIV diagnoses and HAART initiation, particularly in older HIV-infected persons is warranted.The life expectancy for HIV-positive individuals has improved over time due to increasing access to highly active antiretroviral therapy (HAART). Yet, as the HIV-positive population ages, their risk of developing cancers also increases. Studies of Kaposi's sarcoma (KS) among elderly HIV-infected persons are quite limited. We examined the age patterns of KS incidence and an association between age and KS risk in a US cohort of 3458 HIV-infected men, the Multicenter AIDS Cohort Study (MACS). Poisson distribution was used to calculate incidence rates and respective 95% confidence intervals (95% CIs). Cox proportional hazards regression was performed to examine the association between age and KS risk. There were 534 incident KS cases with a total follow-up time of 25,134 person-years. The overall KS incidence rate was 2.13 per 100 person-years (95% CI: 1.95–2.32) . Overall, KS frequency and incidence declined with age, even in the oldest age group ( Kaposi's sarcoma (KS) is caused by human herpesvirus-8 (HHV-8), which was first discovered in 1984 by Chang et al. ).AIDS patients have an increased risk of certain types of cancer due to their immunocompromised status –6. As HIIn HIV-infected persons, KS is still one of the most common cancers. KS can have profound effects on quality of life and may become a debilitating condition, especially in the context of the other comorbidities associated with AIDS. An HIV-infected person is 3640 times more likely to have KS than an HIV-uninfected person . AlthougStudies of KS among older HIV-infected persons and AIDS patients are limited. Recent reports indicate that cancer incidence has risen rapidly among this aging population , 15. It Data used for the current study were obtained from the public dataset (release 18) of the Multicenter AIDS Cohort Study (MACS), an ongoing cohort study of HIV-infected and uninfected men who have sex with men (MSM) in the US. MACS is described in detail elsewhere . BrieflyThe MACS study protocol includes a baseline visit and follow-up visits at 6-month intervals. During both baseline and follow-up visits, study participants were administered a standardized questionnaire by trained interviewers. Information obtained through the interview included: sociodemographic and behavioral characteristics , health service utilization, medical history, medication use , and HIV-related symptoms , 18. Alsn = 3334). Thus, the remaining sample of 3458 men who were either HIV-seropositive at baseline or seroconverted during the cohort was used for the current analysis.MACS has recruited 6792 HIV-infected and HIV-uninfected MSM. For the current analyses, participants were excluded if they remained HIV-uninfected for the duration of follow-up . Race/ethnicity was categorized into three groups . Employment status was dichotomized into current employment and no current employment. Education level was grouped into three groups . Individual gross income was categorized into four groups . Current smoking status was defined as “Yes” or “No”.HAART use was defined as a binary variable , according to the DHHS/Kaiser Panel . AccordiIncident cases of KS among HIV-seropositive men were defined as persons who did not have KS at the baseline visit but developed KS during the follow-up period. Incident cases of KS among HIV-seroconverters were defined as persons who did not have KS at the seroconversion visit but developed KS during the follow-up period. The overall time to follow-up was from 1984 (starting at first enrollment) to 2005 (the cutoff period for the provided public dataset). Among HIV-seropositive men, follow-up time was defined for each subject as the time from the date of enrollment into the cohort to the date of diagnosis for KS cases, or to the date of last recorded visit for subjects who did not develop KS. For HIV-seroconverters, follow-up time was from the date of seroconversion to the date of KS diagnosis for KS cases, or from the date of seroconversion to the date of last recorded visit for KS-free participants. Age at diagnosis or last visit was calculated for KS cases or KS-free participants, respectively. It was then, categorized into three groups as mentioned above.P < 0.0001), we included only CD4 + cell count into the adjusted models. Covariates included in the models were current employment, education level, individual gross income, current smoking status, CD4 + cell count (as a time-dependent variable), and enrollment period. Enrollment period was included in the adjusted models because a previous analysis using MACS data showed that there was a difference in HAART adherence between Caucasians and African Americans between the two enrollment periods  Table Ptrend < 0.0001). Those who did not receive HAART were significantly more likely to develop KS than those who did receive HAART in each of the age groups <40, 40–49, and ≥50 . The protective effect of HAART was also observed in all three race/ethnicity groups. A similar observation was found when stratifying by HIV-seropositive only versus HIV-seroconverters only  is similar to that found in the HIV/AIDS Cancer Match Study (HACM) that examined Caucasian MSM from six states and five metropolitan areas during the 1981–1996 period (incidence rate: 2.4 per 100 person-years) . Given tThe declining trend of KS incidence with age found in our analysis is also consistent with findings from other studies , 28, 29.Ptrend < 0.0001). Also, the IRR of KS was the highest in the oldest age groups for both HIV-infected men and HIV-seroconverters. Although the incidence of KS is decreasing, it is important to note that KS and non-Hodgkin lymphoma are still the most common cancers in the US AIDS population [Another important point from our analysis is that although there is a declining trend of KS incidence with age, the risk of KS development is higher in every age group among those without HAART compared to those with HAART (pulation  despite pulation . Furtherpulation . Specifipulation  reportedpulation , 38 repopulation –41, partWe also found that the incidence of KS decreased in both older age groups; however, in multivariable analyses, the risk of KS in the oldest group was higher than that in 40- to 49-year-olds. Further studies are, therefore, warranted to confirm the KS incidence patterns among the older population with or without HAART usage.The main strength of our analysis was the use of the largest cohort study among HIV-infected men in the US to examine the age patterns of KS incidence. This well-designed cohort allowed us to calculate accurate estimates of KS incidence with age. The design of this cohort also makes it possible to identify the date of seroconversion and to calculate follow-up time accurately. Also, too our knowledge, current analysis is one of the only studies to examine KS in HIV-seroconverters. One limitation of this analysis is that because the cohort only included MSM and most of them were Caucasian American, our results may not be generalizable to HIV-infected women or to mixed race/ethnic populations. Another limitation is that since the data provided were followed to 2005, the definition of HAART used in our study might not be comparable to the new definition in 2011, particularly because several new HIV medications have been approved since 2005. The other limitation is that information on KSHV serostatus was not available in the current provided dataset.In summary, we found declining incidence rates of KS with age among HIV-infected men in the largest US cohort. Furthermore, we reported that HAART has great impact in decreasing the overall KS incidence in this cohort. Among the HIV-infected older population, the KS risk is higher in those without HAART compared to those with HAART. Our findings are consistent with results from previous studies and provide information on the directions of age and period effects on KS risk. Our results have substantial clinical and public health implications related to the low usage of HAART in HIV/AIDS-affected populations, especially in older populations. As KS remains an important malignancy among HIV-infected persons, earlier HIV diagnosis and HAART initiation, particularly in older HIV-infected persons should remain a public health priority.


Early microbiological documentation may reduce attributable mortality and excessive use of broad-spectrum antibiotics in ventilator-associated pneumonia (VAP). Using bronchoalveolar lavage (BAL) and endotracheal aspirates (ETA), we studied a new molecular biology-based approach to detect and quantify bacteria in less than 3 hours. This prospective multicenter trial aimed at comparing the microbiological results obtained using this molecular protocol (easyMAG® system) and semiquantitative culture in suspected VAP.Staphylococcus aureus, Pseudomonas aeruginosa and Enterobacteriaceae. Quantification was performed using qPCR standard curves, by converting the cycle threshold to CFU/ ml.ETA and BAL samples were consecutively collected during 10 months in adult patients in four ICUs of France. The molecular method includes a preprocessing liquefaction for ETA before DNA extraction. DNAs were extracted using the easyMAG® system. Real-time PCR (qPCR) was run using the ABI7500FastDx PCR instrument. The results presented here concern: A total of 125 suspected VAP were included from 122 patients. In total, 125 BAL and 107 ETA were collected. Sex ratio (M/F) was 76%, and CPIS ≥6 was calculated in 74.6% of the suspected VAP patients. Mean ventilation duration before sampling was 6 days. Seventy-eight percent and 65% of the BAL and ETA culture were positive respectively. Correlations between molecular method and culture on BAL and ETA are reported in Table Sensitivity and specificity of the new molecular approach for these main bacteria found in VAP could enable targeted first-line antibiotic therapy. In the future, the development of this approach will aim at obtaining a bedside diagnostic in only a few hours.


BRS) would be reduced with hypoxia and this effect would be blunted when carotid chemoreceptor activity was reduced with low‐dose dopamine. Fifteen healthy adults (11 M/4 F) completed two visits randomized to intravenous dopamine or placebo . On each visit, subjects were exposed to 5‐min normoxia (~99% SpO2), followed by 5‐min hypoxia (~84% SpO2). Blood pressure  and heart rate (ECG) were measured continuously and scBRS was assessed by spectrum and sequence methodologies. scBRS was reduced with hypoxia (P < 0.01). Using the spectrum analysis approach, the fall in scBRS with hypoxia was attenuated with infusion of low‐dose dopamine (P < 0.01). The decrease in baroreflex sensitivity to rising pressures (scBRS “up‐up”) was also attenuated with low‐dose dopamine (P < 0.05). However, dopamine did not attenuate the decrease in baroreflex sensitivity to falling pressures . Present findings are consistent with a reduction in scBRS with systemic hypoxia. Furthermore, we show this effect is partially mediated by the carotid body chemoreceptors, given the fall in scBRS is attenuated when activity of the chemoreceptors is reduced with low‐dose dopamine. However, the improvement in scBRS with dopamine appears to be specific to rising blood pressures. These results may have important implications for impairments in baroreflex function common in disease states of acute and/or chronic hypoxemia, as well as the experimental use of dopamine to assess such changes.The purpose of the present investigation was to examine the contribution of the carotid body chemoreceptors to changes in baroreflex control of heart rate with exposure to hypoxia. We hypothesized spontaneous cardiac baroreflex sensitivity (sc The carotid body chemoreceptors are located bilaterally at the bifurcation of the common carotid artery and are responsible for monitoring the partial pressure of oxygen in arterial blood , nonsmokers without chronic diseases, and taking no medications known to affect cardiovascular, respiratory, or autonomic function. Women were not pregnant (confirmed by negative pregnancy test prior to participation) and were studied in the placebo phase of oral contraceptive use . Subjects refrained from alcohol, caffeine, and exercise for 24 h and fasted for 12 h prior to the dose–response visit and the study visits.Fifteen participants  completed a dopamine dose–response visit and two study visits randomized to intravenous infusion of dopamine or placebo . All participants (22–42 years of age) were healthy, nonobese . Subjects breathed through a two‐way non‐rebreathing valve connected to a switching valve. Each trial consisted of 15 min of room air breathing followed by hypoxic response testing  2 μg · kg−1·min−1, (4) 3 μg·kg−1·min−1, (5) 4 μg·kg−1·min−1. The hypoxic ventilatory response was calculated as follows: (1) for each administration of 100% nitrogen, the three largest consecutive breaths were averaged; (2) simultaneous oxygen saturation (%SpO2) was collected and the nadir was recorded for each nitrogen administration, (3) chemosensitivity to hypoxia was assessed as the slope of the linear regression line for average ventilation (L·min−1) and nadir oxygen saturation (%SpO2) from each nitrogen breathing trial  that resulted in a maximum reduction in the hypoxic ventilatory response with minimal effect on cardiovascular measures was used for the subsequent study visit (average dose: 2.3 ± 0.3 μg·kg−1·min−1). See Limberg et al.  or placebo . Subjects and research personnel were blinded to condition. Only pharmacy and nursing staff were aware of study condition until each subject completed both visits, at which time, all other research personnel were unblinded.An intravenous catheter was inserted in the dominant arm for dopamine/placebo infusion. A brachial arterial catheter  was inserted in the nondominant arm using aseptic techniques under local anesthesia (2% lidocaine) to measure beat‐to‐beat blood pressure . Heart rate was monitored with a three‐lead electrocardiogram  and arterial oxygen saturation by pulse oximetry . Subjects breathed on a mouthpiece/mask connected to a two‐way, non‐rebreathing valve. Breath‐by‐breath tidal volumes , respiratory rate, and inspired/expired gasses  were assessed and inspiratory time was calculated.P > 0.05) followed by ~5‐min hypoxia . Hypoxic conditions were poikilocapnic and hypoxemia was achieved by titrating inspired oxygen levels using a gas blender  to achieve an oxygen saturation of ~85% . Arterial blood gasses were confirmed in a subset of subjects . The average transition phase from room air to the desired SpO2 was ~4 min  and the 5‐min time period for hypoxia began when ~85% SpO2 was achieved.Subjects rested supine. Following a 10‐min infusion  period, subjects were exposed to 5‐min normoxia  (Mulder and Mulder n = 2) were excluded from the analysis. Values belonging to the identified sequences were formed into xy‐pairs and a regression curve was fitted, with the slope of the curve equaling baroreflex sensitivity (ms·mmHg−1). The absolute  and relative (% [Hypoxia‐Normoxia]/Normoxia × 100]) changes in scBRS with hypoxia were compared between saline and dopamine conditions.All hemodynamic measurements were collected digitally using a PowerLab data acquisition system  with a sampling rate of 1000 Hz. To determine baroreflex sensitivity, a standard spectrum analysis method was used . Normal oscillations in heart rate and blood pressure were manipulated via fast Fourier transform to divide variability into frequency components. Cross‐spectrum analysis was used to compare variability in frequency between the R‐R interval and systolic blood pressure spectra. Spontaneous cardiac baroreflex sensitivity (scBRS) was defined as the mean gain of the low‐frequency (0.04–0.15 Hz) transfer function of the resulting cross spectrum after phase correction with a strong spectra‐to‐spectra correlation . The primary analysis was to examine the effect of hypoxia on baroreflex sensitivity under saline and dopamine conditions. A one‐way repeated measures analysis of variance was used to examine the effect of hypoxia on main outcome variables. In all cases, distributional assumptions were assessed and nonparametric tests  were used when necessary. One‐way repeated measures analysis of variance was used to examine the differences in the effect of saline/dopamine on change  variables. Further comparisons using Pearson correlations were conducted post hoc to determine the potential contribution of secondary outcome variables  on changes in baroreflex sensitivity. An alpha of −2), and not taking medications. When assessed during the dopamine dose–response visit, the ventilatory response to acute hypoxia (a measure of carotid body chemoreflex sensitivity) was significantly reduced with low‐dose dopamine .Subject demographics are provided in Table P < 0.01) that were not different between saline and dopamine conditions . Changes in blood pressure  were neither observed with hypoxia (P > 0.05) nor low‐dose dopamine infusion (P > 0.05).Steady‐state hemodynamic responses are reported in Table P < 0.01) which resulted in significant reductions in oxygen saturation (P < 0.01). The inspired oxygen level necessary to achieve the same reduction in oxygen saturation was higher under dopamine conditions (15.1 ± 0.4%) when compared to saline conditions . Steady‐state hypoxia resulted in significant increases in tidal volume (P < 0.01) and minute ventilation (P < 0.01), with no change in respiratory rate (P > 0.05). These responses were not significantly different between saline and dopamine conditions (range: P = 0.11–0.85). The rise in minute ventilation contributed to a slight fall (−0.5 ± 0.1%) in end‐tidal CO2 (P < 0.01) that was not different between saline and dopamine conditions . Inspiratory time was significantly increased with hypoxia, but only under saline conditions .Steady‐state ventilatory responses are reported in Table P < 0.01), and this was observed using both spectrum and sequence methodologies. Using the spectrum analysis approach, the decrease in scBRS with hypoxia during saline conditions was attenuated with low‐dose dopamine infusion . The decrease in baroreflex sensitivity to rising blood pressures (Up‐Up Sequences) with hypoxia during saline was also attenuated with low‐dose dopamine . However, when the hypoxia‐mediated decrease in scBRS was assessed from Down‐Down Sequences, dopamine did not attenuate the fall .Baroreflex sensitivity is reported in Table R = −0.51, P < 0.01), tidal volume , and respiratory rate . The relative (%) fall in scBRS (Up‐Up Sequences) with hypoxia was also correlated with the change (%) in heart rate  and minute ventilation . The relative (%) fall in scBRS (Down‐Down Sequences) with hypoxia was correlated with the change (%) in heart rate , respiratory rate , and minute ventilation .When examining the potential contribution of secondary outcome variables on changes in baroreflex sensitivity, we found the relative (%) fall in scBRS (Low‐Frequency Transfer Function) with hypoxia correlated with the change (%) in heart rate , spontaneous cardiac baroreflex sensitivity is significantly reduced. Results from this study strengthen these findings and those of others  and adrenergic (α and β) receptors. D2 receptors play a main role in the depressant effect of dopamine, including reduced neurotransmitter release from type 1 glomus cells of the carotid body, followed by decreased neural output and carotid sinus drive  of dopamine – similar to that used in this study – are thought to primarily target dopaminergic (D1 and D2) receptors  and sympathetic (Down‐Down) arms of the baroreflex, desensitization of the carotid chemoreceptors results in a significant improvement in the parasympathetic  component of the autonomic nervous system only. These data suggest that the activation of the carotid chemoreceptors during hypoxia results in a primary attenuation of vagally mediated responses. However, it is important to note that although a rising sequence (Up‐Up) could be due to increased parasympathetic drive, parasympathetic withdrawal could also be the primary mechanism of a falling sequence (Down‐Down). Therefore, although our data provide important insight, our analysis does not allow for the strict identification of underlying mechanisms at this time.The carotid chemoreceptors and arterial baroreceptors interact at a variety of levels. Although the location of this interaction was not directly examined in this study, it is reasonable to propose the change in baroreflex sensitivity may be attributed to an interaction between afferent activity of the carotid chemoreceptors and arterial baroreceptors at the level of the nucleus tractus solitarius  was less under dopamine conditions when compared to saline  from a subset of subjects (n = 6) suggests this is an unlikely explanation. Third, this study focused on cardiac baroreflex sensitivity around the operating point of resting blood pressure and therefore results cannot be extrapolated out to sympathetic baroreflex sensitivity and/or more extreme swings in blood pressure. Further, we are unable to comment on the possibility of baroreflex resetting  were not different between experimental conditions , thereby limiting any effect of changes in CO2 on main findings.Although our data show that the carotid body chemoreceptors contribute to the reduction in scBRS during hypoxia, there are some important experimental considerations. First, a reduction in baroreflex sensitivity with hypoxia is not a universal finding (Cunningham et al. ne Table . Althougne Table ], and alns Table . We specns Table . Importans Table . Lastly,The carotid body chemoreceptors have recently been identified as a potential therapeutic target for sympathetically mediated conditions, such as heart failure and hypertension (Paton et al. There are no competing interests and no relevant conflicts of interest.


In the solid-state photodimerization of cinnamic acid polymorphs, can mechanical properties tell us whether Schmidt ‘minimal molecular movement’ or Kaupp ‘molecular migration’ is more important? It appears to propagate in a two-dimensional manner from internal nuclei, rather than in a one-dimensional manner from the surface. Careful 13C-CPMAS NMR studies of the photodimerization of the two DCMA polymorphs and conversely nanoindentation measurements on systems such as the α-forms of o-methoxy- and o-ethoxycinnamic acids would provide a useful cross-correlation study of the two methods and indicate whether mechanical properties can indeed dominate in the progress of these solid-state reactions.Bringing ancillary techniques to bear on a problem can give surprises, as well. For example through  al. 2009 have sho


Introduction. The PiCCO transpulmonary thermodilution technique provides two indices of cardiac systolic function, the cardiac function index (CFI) and the global ejection fraction (GEF). Both appear to be correlated with left ventricular ejection fraction (LVEF) measured by echocardiography in patients with circulatory failure, especially in septic shock. The aim of the present study was to test the reliability of CFI as an indicator of LVEF in patients with cardiogenic shock. Methods. In thirty-five patients with cardiogenic shock, we performed (i) simultaneous measurements of echocardiography LVEF and cardiac function index assessed by transpulmonary thermodilution (n = 72) and (ii) transpulmonary thermodilution before/after increasing inotropic agents (n = 18). Results. Mean LVEF was 31% (+/−11.7), CFI 3/min (+/−1), and GEF 14.2% (+/−6). CFI and GEF were both positively correlated with LVEF . CFI and GEF were significantly increased with inotropic infusion . A cardiac function index <3.47/min predicted a left ventricular ejection fraction ≤35% (sensitivity 81.1% and specificity 63%). In patients with right ventricular dysfunction, CFI was not correlated with LVEF. Conclusion. CFI is correlated with LVEF provided that patient does not present severe right ventricular dysfunction. Thus, the PiCCO transpulmonary thermodilution technique is useful for the monitoring of inotropic therapy during cardiogenic shock. Hemodynamic monitoring is essential for the diagnosis and therapeutic management of critically ill patients . There aThe aim of the present study was thus to evaluate the reliability of CFI as a marker of left ventricular ejection fraction in patients with cardiogenic shock. We hypothesized that CFI is correlated with LVEF, increases with inotropic infusion and is not altered with fluid expansion in patients with cardiogenic shock.This prospective observational study was conducted in a 13-bed ICU in a university hospital.Patients were included if they met the following criteria: presence of a cardiogenic shock and monitoring by a transpulmonary thermodilution device. Inclusion was possible during the initial evolution of cardiogenic shock .persistent hypotension ,cardiac index (CI) less than 2.2 L/min per m² without dobutamine infusion and/or patient already receiving inotropic agent because of a low CI. CI was assessed either by echocardiography or by transpulmonary thermodilution , 18.Cardiogenic shock was defined as a persistent hypotension resulting from heart failure in the presence of adequate intravascular volume . CirculaExclusion criteria included the following: age <18 years; no available echocardiography (absence of sufficient echogenicity); septic shock and/or septic cardiomyopathy; and patient treated with intra-aortic balloon pumping . Nonsinus rhythm , right ventricular failure, and therapeutic hypothermia for cardiac arrest were not considered as exclusion criteria.The following data were recorded: age, sex, past medical history (such as chronic cardiac insufficiency and nonsinus rhythm), simplified acute physiology score (SAPS II), cause of cardiogenic shock, need for mechanical support, renal replacement therapy, and use of vasopressor and/or inotropic agent during measurement and during the ICU stay.A 5-French thermistor-tipped catheter was placed into the femoral artery and a central venous catheter was inserted into a central vein (jugular or subclavian vein); both were connected to the PiCCO system. Cardiac output (CO) and volumetric parameters were measured with the thermodilution technique and obtained after injection of 15 mL of cold isotonic saline 0.9% (<8°C) via the central venous catheter.The CO was calculated from the thermodilution curves, according to the Stewart-Hamilton algorithm; the mean of three consecutive injections was recorded.Volumetric parameters were calculated from the mean transit time (MTt) and the exponential downslope time (DSt) of the thermodilution curve: (i) intrathoracic thermal volume (ITTV) was obtained by the product of CO × MTt; (ii) pulmonary thermal volume (PTV) was obtained by the product of CO × DSt; (iii) the global end-diastolic volume (GEDV) represented the difference between ITTV and PTV: GEDV = ITTV − PTV = CO * MTt – CO * DSt.−1.CFI is the ratio between CO and GEDV: CFI = CO/GEDV, expressed in minGEF is defined as the ratio of the stroke volume (SV) to the quarter of the GEDV: GEF = SV/(GEDV/4), expressed as a percentage.The PiCCO monitor automatically calculated the two cardiac systolic function indices, namely, CFI and GEF.each time an echocardiography was performed,before and 30 minutes after the initiation or increase in inotropic agent,before and immediately after a volume loading: the reason for administering fluid infusion was systematically recorded .The transpulmonary thermodilution parameters were recorded in 3 different situations:A transthoracic echocardiography was performed with a Vivid 3 (Philips) by a specially trained cardiologist. LVEF was obtained by the biplane Simpson's method or was visually estimated , 20.visually right ventricular dilatation and/or right ventricular wall motion abnormalities,tricuspid annular plane systolic excursion (TAPSE) ≤15 mm,systolic pulmonary artery pressure (PAPs) ≥35 mmHg.The operator systematically looked for evidence of a right ventricular dysfunction; a severe right heart failure was defined in this study by the association of three abnormalities , 22:visuStatistical analyses were performed using the GraphPad software, version 4.0, and STATA software, version 9.0.Continuous variables are expressed as mean (+/− standard derivation). Categorical variables are expressed as percentages. The correlations were tested using a Pearson test.The comparison of variables between before and after therapeutic intervention was performed with a nonparametric test (Wilcoxon matched pairs signed-rank test).The receiver operating characteristic (ROC) curve was constructed to study the ability of CFI to predict a LVEF ≤35%.P < 0.05 was considered statistically significant.A n = 15) and end-stage cardiomyopathy (n = 6). Overall ICU mortality rate was 54%.Thirty-five patients were studied between January 2009 and November 2012. μg/kg/min), and 63 were under dobutamine infusion (mean dosage of 9.4 μg/kg/min). For the 72 pairs of measurements, mean LVEF was 31% (+/−11.7), whereas mean CI, CFI, and GEF were, respectively, 2.6 L/min/m2 (+/−0.8), 3/min (+/−1), and 14.2 (+/−6). As shown in P < 0.0001, r = 0.52 , r2 = 0.27) in the 72 pairs of measurements. A significant correlation was also established between GEF and LVEF , r2 = 0.27).Seventy-two pairs of CFI/LVEF measurements were obtained, for which thermodilution and echocardiography variables are described in Eighteen thermodilution measurements were obtained before and after the onset or increase in dobutamine infusion . CI, CFIOnly 4 measurements before and after volume expansion were performed: 3 fluid loadings were decided because the patient was deemed to respond positively to fluid administration . CI, CFI, GEF, and GEDV were not altered by volume expansion .In the 72 pairs of measurements, a CFI value <3.47 allowed diagnosing a LVEF ≤35% with a sensitivity of 81.1%  and a specificity of 63% . The area under the ROC curve was 0.8  .P = 0.28; P = 0.34)  .The present study shows that CFI, obtained by the transpulmonary thermodilution function, is a reliable indicator of left ventricular ejection fraction in cardiogenic shock. Indeed, CFI was found to be statistically correlated with LVEF and to increase with inotropic infusion while not altered by fluid expansion. Moreover, a CFI <3.47 allowed predicting a LVEF ≤35% with good sensibility (81.1%) and specificity (63%).r = 0.87; P < 0.0001). A similar correlation was also described in 2009 by Jabot et al. [r = 0.67; P < 0.0001) involving 39 patients in which LVEF was obtained by transparietal echocardiography. In these studies, however, few patients presented severe cardiac dysfunction, as only 15% of the population was admitted for cardiogenic shock. Our study is the only study to assess CFI as a marker of LVEF in patients with severe cardiac impairment, as witnessed by a mean LVEF of 31%. Interestingly, the correlation between CFI and LVEF was slightly lower in our study than in the two previous studies . Because CFI represents the ratio between CO and GEDV, a severe dilatation of cardiac cavity such as that observed in chronic atrial fibrillations may underestimate CFI (and thus LVEF estimated with CFI). Combes et al. [Assessing LVEF in critically ill patients admitted to the ICU is a key aspect of their hemodynamic management, since detecting a low LVEF may lead to specific therapy such as inotropic infusion. Echocardiography is the gold standard for LVEF estimation: it is a reliable, safe, and noninvasive technique; however, echocardiography requires specific equipment and a competent operator 24/24 h, which is currently not possible in all ICUs. Thus, by providing two cardiac function indices, the PiCCO system could provide an interesting alternative to echocardiography in the assessment of LVEF. Our work confirmed the results of the two previous studies regarding the validity of CFI as an indicator of LVEF in critically ill ICU patients: Combes et al.  first det et al.  from 96 s et al.  excludedIn addition, CFI may also track the changes in LVEF under inotropic agent. Indeed, as previously demonstrated by Jabot et al. , we showr = 0.46); while the observed correlation coefficient was lower than the correlation for the whole population (r = 0.69), the difference between the two values was not statistically significant. In the present study, when considering the patients without a severe RV dysfunction (n = 58), CFI and LVEF are positively correlated (r = 0.55); this correlation coefficient is closed to the rate for the whole studied population (r = 0.52).A serious weakness of CFI is probably the existence of a right ventricular (RV) impairment. Since CFI represents the ratio between CO and GEDV, it reflects global myocardial contraction, in other words, left and right ventricular function , 23. HenConsidering that RV failure may underestimate LVEF or lead to a false negative, the PiCCO system cannot surrogate echocardiography for the assessment of LVEF. In clinical practice, a low CFI should alert the clinician to an impairment in systolic function (left and/or right): an echocardiography must therefore be performed to discriminate between right or left ventricular dysfunction; echocardiography can also provide critical information regarding diagnosis such as segmental wall-motion or valve abnormalities.In conclusion, this study demonstrates that CFI is significantly correlated to LVEF in cardiogenic shock provided that patient does not present severe isolated right ventricular dysfunction. The PiCCO system is a simple and easily reproducible technique, which provides a consistent estimation of LVEF. However, it does not replace echocardiography: a low CFI should alert the physician to a possible impairment of LV systolic function, and an echocardiography must be performed to exclude right ventricular impairment. Once the LV dysfunction is confirmed, CFI allows a consistent monitoring of LV function under inotropic treatment.


Substantial evidence suggests that the phasic activity of dopamine neurons represents reinforcement learning’s temporal difference prediction error. However, recent reports of ramp-like increases in dopamine concentration in the striatum when animals are about to act, or are about to reach rewards, appear to pose a challenge to established thinking. This is because the implied activity is persistently predictable by preceding stimuli, and so cannot arise as this sort of prediction error. Here, we explore three possible accounts of such ramping signals: (a) the resolution of uncertainty about the timing of action; (b) the direct influence of dopamine over mechanisms associated with making choices; and (c) a new model of discounted vigour. Collectively, these suggest that dopamine ramps may be explained, with only minor disturbance, by standard theoretical ideas, though urgent questions remain regarding their proximal cause. We suggest experimental approaches to disentangling which of the proposed mechanisms are responsible for dopamine ramps. Dopamine has long been implicated in reward-motivated behaviour. Theory and experiments suggest that activity of dopamine-containing neurons resembles a temporally-sophisticated prediction error used to learn expectations of future reward. This account would appear to be inconsistent with recent observations of ‘ramps’, i.e., gradual increases in extracellular dopamine concentration prior to the execution of actions or the acquisition of rewards. We explore three different possible explanations of such ramping signals as arising: (a) when subjects experience uncertainty about when actions will be executed; (b) when dopamine itself influences the timecourse of choice; and (c) under a new model in which ‘quasi-tonic’ dopamine signals arise through a form of temporal discounting. We thereby show that dopamine ramps can be integrated with current theories, and also suggest experiments to clarify which mechanisms are involved. Ideas from the field of reinforcement learning (RL) have played an important role in neuroscientific theories of how animals choose actions to gain rewards and avoid punishments. Prominently, it has been suggested , 2 that However, recent reports of ramp-like increases in dopamine concentration preceding self-initiated instrumental responses –13 and dOne possibility is that these signals have no functional importance, for instance being the result of a process of gated release , 15. In We start by sketching what might be considered the ‘standard’ computational account of dopamine and examining the confounding experimental phenomena.s the expected discounted sum V(s) of the rewards that will be encountered during a trialrt is the reward delivered at time t, and 0 ≤ γ ≤ 1 is a discount factor that controls how much weight is given to future relative to immediate rewards. Crucially, the definition of this state value function satisfies a (Bellman) consistency condition with respect to each possible next state s′:δt is defined asV(s). It is exactly this TD error that phasic dopaminergic activity has been hypothesized to represent.In the main class of TD models of the phasic dopamine response , 2, 16, learning , 17, 18.ξ = 0.7s set to match experimental observations , are acquired by convolving time-varying activity pause in burst firing ) rather than the phasic activity of dopaminergic neurons. These quantities are known to be related ; we assurvations . In othe the DRF , albeit  the DRF , other s the DRF ). Indeedt firing , 24, tho gradually ramps up as the goal is approached and peaks at the same value regardless of the time taken to reach the goal or the distance travelled to reach it, assuming a fixed reward size (γ)Vγ(st + 1) with the DRF that we have assumed leads to a signal which looks comparatively over-smoothed  resolution of uncertainty about the timing of action within an actor-critic, leading to a prediction error shortly preceding the action itself; (b) positive correlations between the time of action and dopamine levels generated by dopaminergic gain control of the decision-making process; and (c) a quasi-tonic signal replacing the average reward in the exponentially discounted setting. These explanations, along with the possibility mentioned earlier that release from dopamine axons might be directly occasioned by a form of spillover from cortico-striatal activity, are by no means mutually exclusive. This prompts a need for experimental test.γ < 1), such as in the temporally-extended tasks considered here, since it is in this case that the quantity (1 − γ)Vγ(st + 1) should be visible as a ramping signal.Note that the various cases of ramps may be caused by different, or combined, mechanisms. Indeed, the possible explanations that we considered mainly in the context of pre-response transients, in which ramp-like signals are observed leading up to completion of an instrumental action, were somewhat distinct from the explanation offered for ramping in the spatial reward task, in which the subjects are already engaged in acting. Nevertheless, the account of discounted vigour suggested in the latter case should be relevant in all contexts where some degree of discounting is probable  and long-latency (17%) trials, respectively. Closer examination of the empirical response distribution in such studies would be of interest for future work.Two assumptions in the proposed TD account merit further comment. Firstly, while we assumed that the actor’s choice of when to press the lever immediately follows cue presentation, one can imagine variability in when the actor makes decisions about when to act. For example, it might be that the animal initially fails to notice the cue, or is otherwise engaged  when thA more general problem for a TD account of pre-response signals is that while there is abundant evidence of a systematic temporal relationship between the time at which a cue indicating reward availability is presented and subsequent phasic activity in dopaminergic neurons, there is little or no evidence of such a relationship between the time of the phasic response and when a subsequent instrumental action—necessary to obtain the reward—is emitted. For example, Ljungberg et al.  found thSuggestively, striatal  neurons in monkeys show various patterns, including ramp-like increases in activity, before self-initiated movements , 92, 93.A range of previous work has considered the vagaries of the representation and processing of time. We noted that possible sources of uncertainty included partial observability of the actor’s choices in the case where the critic does not have direct access to this information, and possible timing uncertainty in the case where it does. Implications of partial observability for TD models of dopamine have been explored in previous work, notably by Daw and colleagues , though We showed that ramping dopamine signals can be generated by a mechanistic decision-making model in which dopamine sets the gain of value-based accumulation. Furthermore, we saw that this direct coupling of dopamine to decision-making could generate a negative correlation between the size of TD error and decision time, consistent with the experimental observation that a larger phasic response of dopaminergic cells to a start cue is associated with a shorter latency of behavioural response .This route to ramping signals is primarily statistical, arising from trial-averaging. On any individual trial, dopamine ramping towards the time of decision may or may not occur, though it is certainly more typical when dopamine fluctuations incorporate a strong, TD-related phasic component. To the best of our knowledge, whether pre-response transients in NAc reliably precede the animal’s response on individual trials, or may reflect trial-averaging, is unknown.Mathematical analysis of the time-varying gain DDM that we described is given by Moehlis et al.  and, indAlso of relevance is the biologically-detailed neural network model of the basal ganglia proposed by Frank  in whichγ)〈Vγ(st + 1)〉 in the discounted reward model plays an equivalent role to the average reward rate ρ in the average reward model. Since values often  change modestly as a result of the passage of time, this signal is quasi-tonic, and thus a candidate for what would be recorded using a technique such as FSCV. This signal can explain the ramping phenomena observed by Howe et al. [We reconciled an apparent inconsistency between the definitions of TD errors in the cases of average and discounted reward via an analysis in which ramp-like signals would be expected to emerge. In particular, we suggested that the quantity 〈Vγ(st + 1)〉 is appropriate for controlling vigour, changes in the running speed of rats in Howe et al.’s study do not show a close match to the temporal profile of dopamine concentrations. However, one would not necessarily expect a straightforward relationship between these variables, given that the subjects must negotiate environments without crashing into walls. Howe et al. used T-, M-, and S-shaped mazes, whose turns, unsurprisingly, led to decreases in velocity  cannot be captured in an error-free manner if the state  is represented in particular, over-generalizing manners, for instance by units whose activity is governed by the square, rather than linear, distance to the goal. In this case, a ramping prediction error turns out to arise via persistent representational error. Place cells [in silico, rather than in vivo.Gershman  considerce cells –131 provce cells . Howeverce cells , and thuMorito and Kato  have alsThe most pressing consideration is a set of experiments that can test and refine or reject these various mechanisms, and understand how they might work together. Perhaps the most straightforward to test is the last suggestion, since it is unique in its dependence on discounting. Given that the rate of this should be sensitive to things like the reliability of the environment , it woulTesting the second suggestion could be accomplished using photo-uncaging of dopamine in the accumbens , since Testing the relationship between actor and critic is particularly tricky, since we know so little about the implementation (or indeed existence) of either and, in particular, the micro- or nano-scopic nature of choice over time . NeverthMore generally, key issues surround the relationships between the number of dopamine cells that are active, the phasic and tonic activity of those neurons, the spatiotemporal profile of the concentration of dopamine at receptor targets in the accumbens, and the action of this dopamine on those receptors . This information is key for making qualitative and ultimately quantitative progress.