title: Cell-penetrating peptide-conjugated antisense oligonucleotides restore systemic muscle and cardiac dystrophin expression and function

text: Antisense oligonucleotides (AOs) have the potential to induce functional dystrophin protein expression via exon skipping by restoring in-frame transcripts in the majority of patients suffering from Duchenne muscular dystrophy (DMD). AOs of morpholino phosphoroamidate (PMO) and 2′-O-methyl phosphorothioate RNA (2′Ome RNA) chemistry have been shown to restore dystrophin expression in skeletal muscle but not in heart, following high-dose systemic delivery in murine models of muscular dystrophy (mdx). Exploiting the cell transduction properties of two basic arginine-rich cell penetrating peptides, we demonstrate widespread systemic correction of dystrophin expression in body-wide muscles and cardiac tissue in adult dystrophic mdx mice, with a single low-dose injection of peptide-conjugated PMO AO. This approach was sufficient to restore uniform, high-level dystrophin protein expression in peripheral muscle and cardiac tissue, with robust sarcolemmal relocalization of the dystrophin-associated protein complex and functional improvement in muscle. Peptide-conjugated AOs therefore have significant potential for systemic correction of the DMD phenotype.


title: From Bench to Cageside: Risk Assessment for Rodent Pathogen Contamination of Cells and Biologics

text: Many newly developed animal models involve the transfer of cells, serum, or other tissue-derived products into live rodents. These biologics can serve as repositories for adventitious rodent pathogens that, when used in animal studies, can alter research outcomes and result in endemic outbreaks. This review includes a description of some of the biologics that have inadvertently introduced infectious agents into in vivo studies and/or resulted in endemic outbreaks. I also discuss the points of potential exposure of specific biologics to adventitious rodent pathogens as well as the importance of acquiring a complete developmental and testing history of each biologic introduced into a barrier facility. There are descriptions of specific cases of mycoplasma and lactate dehydrogenase–elevating virus (LDHV), two of the most common organisms that contaminate cells and cell byproducts. The information in this article should help investigators and animal resource program personnel to perform an appropriate risk assessment of biologics before their use in in vivo studies that involve rodents.


title: Whooping Cough and Parkinson's Disease

text: de Pedro-Cuesta J (Instituto de Salud Carlos III, Centro Nacional de Epidemiología, Departamento de Epidemiología Aplicada, c/ Sinesio Delgado, 6, 28029 Madrid, Spain), Gudmundsson G, Abraira V, Gudmundsson G, Lōve A, Tulinius H, Veiga J, Almazán J and Petersen I J for the EUROPARKINSON Preparatory Activity Research Group. Whooping Cough and Parkinson' Disease. International Journal of Epidemiology 1996; 25: 1301–1311. BACKGROUND: We reported high levodopa use and prevalences of Parkinson's Disease (PD) in periodically, time-clustered, Icelandic cohorts born after major whooping cough epidemics (MWCE). METHODS: In order to quantify a possible relationship between age at first post-birth MWCE and risk of PD we: 1) calculated cumulative incidences of PD during the period 1954–1963 in one-year Icelandic cohorts born between 1869 and 1927, using raw material from a reported survey; 2) identified MWCE from 1869 onwards in Iceland; 3) estimated cohort ages at onset of incidence period and at first MWCE; and 4) combined the above-mentioned information using log-linear models. In addition, we studied the prevalence of levodopa users in Icelandic birth cohorts during a recent period. RESULTS: The curves of the above-mentioned incidences and prevalences in one-year birth-cohorts showed: 1) a similar, age-related, inverted V profile; and 2) a systematic notchy pattern, with peak values for one or both measurements for cohorts born during or after each of nine MWCE identified during the period 1869–1927. When 13 cohorts bom in years with MWCE were excluded from the analysis, the risk of PD rose with age at first defined MWCE, with the linear increase being 8.4% per year (95% Cl: −0.1–18.3%). CONCLUSIONS: These results are consistent with reported effects of age at exposure in animal models of toxic parkinsonism, age-related changes in the dopamine receptor-GPT-binding protein-adenylatecyclase system observed in rats treated with pertussis toxin, and some PD epidemiological features. They suggest that pertussis neurotoxicity could be causally related to PD worldwide.


title: Human Microbiota-Associated Swine: Current Progress and Future Opportunities

text: Gnotobiotic (GN) rodent models have provided insight into the contributions of the gut microbiota to host health and preventing disease. However, rodent models are limited by several important physiological and metabolic differences from humans, and many rodent models do not dependably replicate the clinical manifestations of human diseases. Due to the high degree of similarity in anatomy, physiology, immunology and brain growth, the domestic pig (Sus scrofa) is considered a clinically relevant model to study factors influencing human gastrointestinal, immune, and brain development. Gnotobiotic piglet models have been developed and shown to recapitulate key aspects of GN rodent models. Human microbiota-associated (HMA) piglets have been established using inocula from infants, children, and adults. The gut microbiota of recipient HMA piglets was more similar to that of the human donor than that of conventionally reared piglets harboring a pig microbiota. Moreover, Bifidobacterium and Bacteroides, two predominant bacterial groups of infant gut, were successfully established in the HMA piglets. Thus, the HMA pig model has the potential to be a valuable model for investigating how the gut microbiota composition changes in response to environmental factors, such as age, diet, vaccination, antibiotic use and infection. The HMA also represents a robust model for screening the efficacy of pre- and probiotic interventions. Lastly, HMA piglets can be an ideal model with which to elucidate microbe–host interactions in human health and disease due to the similarities to humans in anatomy, physiology, developmental maturity at birth, and the pathophysiology of many human diseases.


title: Highly efficient in vivo delivery of PMO into regenerating myotubes and rescue in laminin-α2 chain-null congenital muscular dystrophy mice

text: Phosphorodiamidate morpholino oligomer (PMO)-mediated exon skipping is among the more promising approaches to the treatment of several neuromuscular disorders including Duchenne muscular dystrophy. The main weakness of this approach arises from the low efficiency and sporadic nature of the delivery of charge-neutral PMO into muscle fibers, the mechanism of which is unknown. In this study, to test our hypothesis that muscle fibers take up PMO more efficiently during myotube formation, we induced synchronous muscle regeneration by injection of cardiotoxin into the tibialis anterior muscle of Dmd exon 52-deficient mdx52 and wild-type mice. Interestingly, by in situ hybridization, we detected PMO mainly in embryonic myosin heavy chain-positive regenerating fibers. In addition, we showed that PMO or 2′-O-methyl phosphorothioate is taken up efficiently into C2C12 myotubes when transfected 24–72 h after the induction of differentiation but is poorly taken up into undifferentiated C2C12 myoblasts suggesting efficient uptake of PMO in the early stages of C2C12 myotube formation. Next, we tested the therapeutic potential of PMO for laminin-α2 chain-null dy(3K)/dy(3K) mice: a model of merosin-deficient congenital muscular dystrophy (MDC1A) with active muscle regeneration. We confirmed the recovery of laminin-α2 chain and slightly prolonged life span following skipping of the mutated exon 4 in dy(3K)/dy(3K) mice. These findings support the idea that PMO entry into fibers is dependent on a developmental stage in myogenesis rather than on dystrophinless muscle membranes and provide a platform for developing PMO-mediated therapies for a variety of muscular disorders, such as MDC1A, that involve active muscle regeneration.


title: Molecular dynamics-derived conformation and intramolecular interaction analysis of the N-acetyl-9-O-acetylneuraminic acid-containing ganglioside G(D1a) and NMR-based analysis of its binding to a human polyclonal inununoglobulin G fraction with selectivity for O-acetylated sialic acids

text: The influence of 9-O-acetylation of G(D1a), yielding G(D1a) ((e)Neu5,9Ac(2) with a 9-O-acetylated sialic acid moiety linked to the outer galactose residue, on the spatial extension and mobility of the carbohydrate chain and on recognition by a natural human antibody is analysed. To study a potential impact of the O-acetyl group on the overall conformation of the carbohydrate chain, molecular dynamics (MD) simulations of oligosaccharide chain fragments of increasing length starting from the non-reducing end have been carried out for the first time in this study. They revealed a considerable loss in chain flexibility after addition of the internal N-acetylneuraminic acid onto the chain. Besides MD calculations with different dielectric constants, the conformational behaviour of the complete oligosaccharide chain of the 9-O-acetylated G(D1a) ganglioside was simulated in the solvents water and dimethyl sulfoxide. These solvents were also used in NMR measurements. The results of this study indicate that 9-O-acetylation at the terminal sialic acid does not influence the overall conformation of the ganglioside. An extended interaction analysis of energetically minimized conformations of G(D1a) ((e)Neu5,9Ac(2)) and G(D1a), obtained during molecular dynamics simulations, allowed assessment of the influence of the different parts of the saccharide chains on spatial flexibility. Noteworthy energetic interactions, most interestingly between the 9-O-acetyl group and the pyranose ring of N-acetylgalactosamine, were ascertained by the calculations. However, the strength of this interaction does not force the ganglioside into a conformation, where the 9-O-acetyl group is no longer accessible. Binding of G(D1a) ((e)Neu5,9Ac(2)) to proteins, which are specific for 9-O-acetylated sialic acids, should thus at least partially be mediated by the presence of this group. To experimentally prove this assumption, a NMR study of 9-O-acetylated G(D1a) in the presence of an affinitypurified polyclonal IgG fraction from human serum with preferential binding to 9-O-acetylated sialic acid was performed. The almost complete disappearance of the intensity of the 9-O-acetyl methyl signal of the G(D1a) (clearly indicates that the assumed interaction of the 9-O-acetyl group with the human protein takes place.


title: Altered lipid metabolism in a Drosophila model of Friedreich's ataxia

text: Friedreich's ataxia (FRDA) is the most common form of autosomal recessive ataxia caused by a deficit in the mitochondrial protein frataxin. Although demyelination is a common symptom in FRDA patients, no multicellular model has yet been developed to study the involvement of glial cells in FRDA. Using the recently established RNAi lines for targeted suppression of frataxin in Drosophila, we were able to study the effects of general versus glial-specific frataxin downregulation. In particular, we wanted to study the interplay between lowered frataxin content, lipid accumulation and peroxidation and the consequences of these effects on the sensitivity to oxidative stress and fly fitness. Interestingly, ubiquitous frataxin reduction leads to an increase in fatty acids catalyzing an enhancement of lipid peroxidation levels, elevating the intracellular toxic potential. Specific loss of frataxin in glial cells triggers a similar phenotype which can be visualized by accumulating lipid droplets in glial cells. This phenotype is associated with a reduced lifespan, an increased sensitivity to oxidative insult, neurodegenerative effects and a serious impairment of locomotor activity. These symptoms fit very well with our observation of an increase in intracellular toxicity by lipid peroxides. Interestingly, co-expression of a Drosophila apolipoprotein D ortholog (glial lazarillo) has a strong protective effect in our frataxin models, mainly by controlling the level of lipid peroxidation. Our results clearly support a strong involvement of glial cells and lipid peroxidation in the generation of FRDA-like symptoms.


title: Loss of mitochondrial peptidase Clpp leads to infertility, hearing loss plus growth retardation via accumulation of CLPX, mtDNA and inflammatory factors

text: The caseinolytic peptidase P (CLPP) is conserved from bacteria to humans. In the mitochondrial matrix, it multimerizes and forms a macromolecular proteasome-like cylinder together with the chaperone CLPX. In spite of a known relevance for the mitochondrial unfolded protein response, its substrates and tissue-specific roles are unclear in mammals. Recessive CLPP mutations were recently observed in the human Perrault variant of ovarian failure and sensorineural hearing loss. Here, a first characterization of CLPP null mice demonstrated complete female and male infertility and auditory deficits. Disrupted spermatogenesis already at the spermatid stage and ovarian follicular differentiation failure were evident. Reduced pre-/post-natal survival and marked ubiquitous growth retardation contrasted with only light impairment of movement and respiratory activities. Interestingly, the mice showed resistance to ulcerative dermatitis. Systematic expression studies detected up-regulation of other mitochondrial chaperones, accumulation of CLPX and mtDNA as well as inflammatory factors throughout tissues. T-lymphocytes in the spleen were activated. Thus, murine Clpp deletion represents a faithful Perrault model. The disease mechanism probably involves deficient clearance of mitochondrial components and inflammatory tissue destruction.


title: Reevaluating the effect of Brefeldin A (BFA) on ganglioside synthesis: the location of GM2 synthase cannot be deduced from the inhibition of GM2 synthesis by BFA

text: Brefeldin A reversibly disassembles the Golgi complex, causing mixing of the Golgi cisternae with the ER while the trans Golgi network persists as part of a separate endosomal membrane system. Because of this compartmental separation, Brefeldin A treatment has been used to map the sub-Golgi locations of several Golgi enzymes including GM2 synthase. We previously proposed that GM2 synthase might be located in a distal portion of the Golgi complex which in the presence of Brefeldin A would be separated from the substrate ganglioside GM3 present in the mixed ER-Golgi membrane system. In the present study we show using GM2 synthase chimeras that GM2 synthesis was blocked by Brefeldin A when GM2 synthase was distributed throughout all Golgi subcompartments or even when it was restricted to the medial Golgi. Because these findings opposed our speculation regarding a distal location of this enzyme, we sought an alternative explanation for the inhibition of ganglioside synthesis by Brefeldin A. However, Brefeldin A did not degrade GM2 synthase, prevent its homodimerization, or inhibit its in vitro activity. Brefeldin A did result in the conversion of a portion of membrane bound GM2 synthase into a soluble form which has minimal capability to produce GM2 in whole cells. However, this conversion was not sufficient to explain the nearly total loss of GM2 production in intact cells in the presence of Brefeldin A. Nevertheless, the results of this study indicate that Brefeldin A-induced inhibition of ganglioside synthesis cannot be used to deduce the location of GM2 synthase.


title: Characterization of Stool Virome in Children Newly Diagnosed With Moderate to Severe Ulcerative Colitis

text: BACKGROUND: Viral infections have been suggested as possible triggers for the onset of ulcerative colitis (UC). METHODS: We employed VirCapSeq-Vert, a high-throughput sequencing virus capture platform, to examine the stool virome of children with newly diagnosed moderate to severe UC. We surveyed fecal samples collected at presentation, after symptom remission, and from a control group diagnosed with irritable bowel syndrome. RESULTS: Seventy subjects with UC (mean age 13 years, 45 had moderate symptoms, 25 had severe, 69 of 70 had a Mayo endoscopy subscore 2/3) were studied. We detected a wide range of animal viruses that were taxonomically classified into 12 viral families. A virus was present in 50% of fecal samples collected at presentation, 41% of samples collected after remission, and 40% of samples in our control group. The most frequently identified viruses were diet-based gyroviruses. The UC cohort had a significantly higher prevalence of anelloviruses compared with the control cohort. However, we did not identify a single virus that can be implicated in the onset of UC and did not find an association between UC disease severity and viral presence. CONCLUSION: Presence of virus in stool was not associated with the onset of pediatric UC.


title: Microbial Considerations in Genetically Engineered Mouse Research

text: Microbial infections have long been of concern to scientists using laboratory rodents because of their potential to confound and invalidate research. With the explosion of genetically engineered mice (GEM), new concerns over the impact of microbial agents have emerged because these rodents in many cases are more susceptible to disease than their inbred or outbred counterparts. Moreover, interaction between microbe and host and the resulting manifestation of disease conceivably differ between GEM and their inbred and outbred counterparts. As a result, infections may alter the GEM phenotype and confound interpretation of results and conclusions about mutated gene function. In addition, because GEM are expensive to produce and maintain, contamination by pathogens or opportunists has severe economic consequences. This review addresses how microbial infections may influence phenotype, how immunomodulation of the host as the result of induced mutations may modify host susceptibility to microbial infections, how novel host:microbe interactions have led to the development of new animal models for disease, how phenotype changes have led to the discovery of new pathogens, and new challenges associated with prevention and control of microbial infections in GEM. Although the focus is on naturally occurring infections, extensive literature on the use of GEM in studies of microbial pathogenesis also exists, and the reader is referred to this literature if microbial infection is a suspected culprit in phenotype alteration.


title: Neutralization of chemokines RANTES and MIG increases virus antigen expression and spinal cord pathology during Theiler's virus infection

text: The role of chemokines during some viral infections is unpredictable because the inflammatory response regulated by these molecules can have two, contrasting effects—viral immunity and immunopathologic injury to host tissues. Using Theiler's virus infection of SJL mice as a model of this type of disease, we have investigated the roles of two chemokines—regulated on activation, normal T cell-expressed and secreted (RANTES) chemokine and monokine induced by IFN-γ (MIG)—by treating mice with antisera that block lymphocyte migration. Control, infected mice showed virus persistence, mild inflammation and a small degree of demyelination in the white matter of the spinal cord at 6 weeks post-infection. Treatment of mice with RANTES antiserum starting at 2 weeks post-infection increased both viral antigen expression and the severity of inflammatory demyelination at 6 weeks post-infection. MIG antiserum increased the spread of virus and the proportion of spinal cord white matter with demyelination. Overall, viral antigen levels correlated strongly with the extent of pathology. At the RNA level, high virus expression was associated with low IL-2 and high IL-10 levels, and RANTES antiserum decreased the IL-2/IL-10 ratio. Our results suggest that RANTES and MIG participate in an immune response that attempts to restrict viral expression while limiting immunopathology and that anti-chemokine treatment poses the risk of exacerbating both conditions in the long term.


Diversity in the sialic acids


title: Protective role of mouse MBL-C on intestinal mucosa during Shigella flexneri invasion

text: Mannan-binding lectin (MBL) is a C-type serum lectin, which is believed to play an important role in the innate immunity against a variety of pathogens. MBL can bind to sugar determinants of a wide variety of microorganisms, neutralize them and inhibit infection by complement activation through the lectin pathway and opsonization by collectin receptors. Given that small intestine is a predominant site of extrahepatic expression of MBL, here we addressed the question whether MBL is involved in mucosal innate immunity. The carbohydrate recognition domain (CRD) genes of mouse MBL-C (mMBL-C) were cloned and expressed in Escherichia coli. Recombinant mMBL-C-CRD binds to Shigella flexneri 2a in a calcium-dependent manner and that interaction could be blocked by the anti-mMBL-C-CRD antibody. mMBL-C-CRD protein could inhibit the adhesion of S. flexneri 2a to intestinal mucosa, while administration of anti-mMBL-C-CRD antibody caused an increased level of bacteria adhesion in vitro. Administration of recombinant mMBL-C-CRD protein reduced the secretion of IL-6 and monocyte chemoattractant protein 1 from primary intestinal epithelial cells stimulated with S. flexneri 2a. Furthermore, neutralization of MBL activity by anti-MBL-C-CRD resulted in a significant increase in the number of S. flexneri 2a that colonized the intestines of BALB/c mice and attenuated the severity of inflammation seen in the areas of bacterial invasion. These findings suggest that mMBL-C may protect host intestinal mucosa by directly binding to the bacteria.


title: Characterization of human vascular endothelial cadherin glycans

text: The glycosylation pattern of human vascular endothelial cad-herin (VE-cadherin), purified from cultured human umbilical cord vein endothelial cells, was analyzed. VE-cadherin was metabolically radiolabeled with D-[6-(3)H]glucosamine, isolated by immunoprecipitation, purified by SDS-PAGE and in-gel digested with endoproteinase Asp N. Oligosaccharides were sequentially released from resulting glycopeptides and analyzed by chromatographic profiling. The results revealed that VE-cadherin carries predominantly sialylated diantennary and hybrid-type glycans in addition to some triantennary and high mannose-type species. Highly branched, tetra-antennary oligosaccharides were found in trace amounts only. Immunohistochemical labeling of VE-cadherin and sialic acids displayed a codistribution along the intercellular junctions in endothelial cells of human umbilical arteries, veins, and cultured endothelial monolayers. Ca(2+)-depletion, performed on cultured endothelial cells, resulted in a reversible complete disappearance of VE-cadherin and of almost all sialic acid staining from the junctions. Sialidase treatment of whole cells caused a change of VE-cadherin immunofluorescence from a continuous and netlike superstructural organization to a scattered inconsistent one. Hence, cell surface sialic acids might play a role in VE-cadherin organization.


title: New Building, Old Parasite: Mesostigmatid Mites—An Ever-Present Threat to Barrier Rodent Facilities

text: Mesostigmatid mites are blood-sucking parasitic mites found in wild rodent populations. Periodically they can also become a problem for laboratory rodent colonies, particularly when building construction or renovations disturb colonies of commensal (building) rodents that had been acting as hosts. Mesostigmatid mites infest both rats and mice and, unlike the more common rodent fur mites (Myobia, Myocoptes, and Radfordia sp.), can survive for long periods in the environment and travel considerable distances in search of new hosts. They easily penetrate barrier caging systems, including individually ventilated cages, thus circumventing the usual precautions to protect rodents from infection. The two mites reported in laboratory rodent colonies, Ornithonyssus bacoti and Laelaps echidnina, also bite humans and have the potential to transmit zoonotic diseases. Once the mites gain access to a colony, eradication requires elimination of commensal rodent reservoirs in addition to insecticide treatment of both the laboratory rodents and the environment. In view of the undesirability of insecticide use in the animal facility, it is advisable to investigate the effectiveness of preventive treatments, such as environmental application of insect growth regulators or silica-based products. This article summarizes available information on mesostigmatid mites and their laboratory incursions, and provides suggestions for diagnosis, treatment, and control based on the author’s experience with several outbreaks at a large academic institution.


title: Respiratory Symptoms or Signs on the Day of the Study Alter Pulmonary Function in Teenagers†

text: Pulmonary function variables (forced expiratory flows and vital capacity, static lung volume, alveolar N(2) slopes closing volume and closing capacity) were compared in a group of 10–16 year old children with (n=65) and without (n=440) symptoms or signs of mild acute respiratory infection (‘common cold’). Symptomatic children had a significant impairment of forced expiratory vital capacity and flows, with no change In static volumes, alveolar N(2) slopes and closing volumes. The effect was present only In boys (in whom RV/TLC and phase III slope were borderline abnormal), was more evident in older children (13 to 16 years) and was independent of the smoking habits or the presence of chronic respiratory symptoms. The results are in favour of a predominant involvement of upper airways, but signs of lower airways dysfunction are present in boys and in children 13 to 16 years old.


title: Biological roles of oligosaccharides: all of the theories are correct

text: Many different theories have been advanced concerning the biological roles of the oligosaccharide units of individual classes of glycoconjugates. Analysis of the evidence indicates that while all of these theories are correct, exceptions to each can also be found. The biological roles of oligosaccharides appear to span the spectrum from those that are trivial, to those that are crucial for the development, growth, function or survival of an organism. Some general principles emerge. First, it is difficult to predict a priori the functions a given oligosaccharide on a given glycoconjugate might be mediating, or their relative importance to the organism. Second, the same oligosaccharide sequence may mediate different functions at different locations within the same organism, or at different times in its ontogeny or life cycle. Third, the more specific and crucial biological roles of oligosaccharides are often mediated by unusual oligosaccharide sequences, unusual presentations of common terminal sequences, or by further modifications of the sugars themselves. However, such oligosaccharide sequences are also more likely to be targets for recognition by pathogenic toxins and microorganisms. As such, they are subject to more intra- and inter-species variation because of ongoing host—pathogen interactions during evolution. In the final analysis, the only common features of the varied functions of oligosaccharides are that they either mediate ‘specific recognition’ events or that they provide ‘modulation’ of biological processes. In so doing, they generate much of the functional diversity required for the development and differentiation of complex organisms, and for their interactions with other organisms in the environment.


title: Golgi localization of glycosyltransferases: more questions than answers

text: The structures of cellular oligosaccharides are determined by a series of processing reactions catalyzed by Golgi glycosidases and glycosyltransferases. While there are subtle cell type differences in Golgi enzyme subcompartmentation, in general, glycosylation enzymes are localized within the Golgi cisternae in the same sequence in which they act to modify oligosaccharide substrates. The possibility that this enzyme subcompartmentation may control the types of oligosaccharides expressed by a cell has led to an interest in the signals and mechanisms directing enzyme localization in the Golgi cisternae. All glycosidases and glycosyltransferases characterized thus far have very little sequence homology that might suggest a common Golgi retention signal, but they do share a similar domain structure. They are all type II transmembrane proteins consisting of an amino terminal cytoplasmic tail, a signal anchor transmembrane domain, a stem region, and a large luminal catalytic domain. Their lack of sequence homology suggests that these proteins' Golgi retention signals are not linear amino acid sequences, but most likely involve general characteristics or conformations of larger protein domains. The peptide sequences required for Golgi retention of the N-acetylglucosaminyltransferase I (GlcNAcTI), (β1,4-galactosyltrans- ferase (GalT) and α2,6-sialyltransferase (ST) have been extensively studied. To do this, researchers created mutant and chimeric proteins, expressed these in tissue culture cells, and localized these proteins using immunofluorescence microscopy or immunoelectron microscopy. The cell surface expression of deletion mutants suggested that the deleted sequences were necessary for Golgi retention. Then, if these sequences were fused to a non-Golgi reporter protein and this chimeric or hybrid protein was retained in the Golgi, then these sequences were also sufficient for Golgi retention. Due to differences in reporter proteins used to construct these chimeric proteins, different cell types used for protein expression, different levels of protein expression, and different methods of cell surface protein detection, these experiments have led to somewhat confusing results. However, in general, it appears that the GalT relies primarily on its transmembrane domain for Golgi retention, while the GlcNAcTI and ST have requirements for their transmembrane regions, sequences flanking these regions, and luminal stem sequences. Based on these results, two potential Golgi retention mechanisms have been proposed and are now being tested. The observation that glycosyltransferase transmembrane domains are frequently sufficient for Golgi retention has led to the first of these models, the bilayer thickness model. This model proposes that the shorter transmembrane domains of Golgi proteins prevent them from entering cholesterol-rich transport vesicles destined for the plasma membrane,.and that this leads to Golgi retention. The second of these models is supported by the role of multiple protein domains in the Golgi retention of some proteins. This model, the oligomerization/kin recognition model of Golgi retention, proposes that the formation of insoluble protein homo-oligomers or very large hetero-oligomers prevents protein movement into transport vesicles destined for later compartments. Initial work suggests that the bilayer thickness mechanism may play a role in the retention of some Golgi retained proteins; however, it is not the sole retention mechanism. Other evidence suggests that an oligomerization/kin recognition mechanism may be more common, but definitive proof for its general use in Golgi protein retention is lacking. More research is required to further elucidate the sequences and particularly the mechanisms of Golgi retention. In the future, we hope to be able to explain the cell type differences in glycosylation enzyme Golgi subcompartmentation, the different sequence requirements for the Golgi retention of the same enzyme in various cell types, and whether differences in glycosylation enzyme Golgi subcompartmentation change the types of oligosaccharides made by a cell.


title: The role of α4 integrin in Theiler’s murine encephalomyelitis virus (TMEV)-induced demyelinating disease: an infectious animal model for multiple sclerosis (MS)

text: Natalizumab, which is an antibody against α4 integrin, has been used for the treatment of multiple sclerosis (MS). In the present study, we investigated both the role of α4 integrin and the therapeutic effect of HCA3551, a newly synthesized orally active small-molecule α4 integrin antagonist, in the development of TMEV-induced demyelinating disease (TMEV-IDD). The mRNA levels of α4 integrins were significantly up-regulated in the CNS of mice with TMEV-IDD as compared with naïve mice (*p<0.05). HCA3551 treatment in the effector phase significantly suppressed both the clinical and histological development of TMEV-IDD. The number of infiltrating mononuclear inflammatory cells in the CNS was significantly decreased in the mice treated with HCA3551 (**p<0.01). The labeling indices for CD68 antigen and the absolute cell numbers of TNF-α-producing CD4(+) T cells and IFN-γ-producing CD8(+) T cells were significantly decreased in the CNS of mice treated with HCA3551 (*p<0.05). HCA3551 treatment in the effector phase might inhibit the binding of α4 integrin to VCAM-1, thereby decreasing the number of MNCs in the CNS.


title: Development of Saudi e-health literacy scale for chronic diseases in Saudi Arabia: using integrated health literacy dimensions

text: OBJECTIVE: Health literacy has become a global issue, and it is important that patients and individuals are able to use information technology to access health information and educational services. The research objective is to develop a Saudi e-health literacy scale (SeHL) for measuring e-health literacy among Saudis suffering from non-communicable diseases (NCD). METHODS: Overall, 14 relevant papers in related interdisciplinary fields were reviewed to select the most useful literacy dimensions. From these articles, we extracted the most common dimensions used to measure e-health literacy across the disciplines. Multiple workshops with multidisciplinary team members reviewed and evaluated items for SeHL. RESULTS: Four key aspects of e-health literacy—use of technology/media, information-seeking, usefulness and confidence—were identified and integrated as e-health literacy dimensions. These will be used to measure e-health literacy among Saudi patients with NCDs. A translation from Arabic to English was performed in order to ensure that translation process was accurate. A SeHL scale was developed to measure e-health literacy among Saudi patients. By understanding e-health literacy levels, we will be able to create a patient-education system to be used by patients in Saudi Arabia. CONCLUSIONS: As information technology is increasingly used by people of all ages all over the world, e-health literacy has been identified as a key factor in determining health outcomes. To date, no comprehensive scale exists to assess e-health literacy levels among speakers of Arabic, particularly among people with NCD such as diabetes, cardiovascular diseases and hypertension.


title: Post-translational modifications distinguish cell surface from Golgi-retained β1,4 galactosyltransferase molecules. Golgi localization involves active retention

text: β1,4 Galactosyltransferase (GalT) is a membrane-bound enzyme localized predominantly to the trans-Golgi cisternae. Our previous studies have shown that the transmembrane domain of bovine GalT plays a critical role in Golgi localization (Teasdale,R.D., D'Agostaro,G. and Gleeson,P.A., J. Biol. Chem., 267, 4084–4096, 1992). Here we have compared the localization and post-translational modifications of fulllength bovine GalT with a GalT/hybrid molecule where the transmembrane domain of GalT was replaced with that of the transferrin receptor. GalT/hybrid molecules were expressed on the surface of transfected cells; however, differences were observed in the distribution of the hybrid molecules between transfected COS and murine L cells. In transfected COS cells, the GalT/hybrid protein was expressed efficiently at the cell surface, with little Golgilocalized material, whereas in stable murine L cells, which expressed lower levels of the construct, hybrid molecules were detected both at the cell surface and within the Golgi apparatus. Expression of the GalT constructs in either COS or L cells produced two glycoprotein products which differed in molecular mass by 7 kDa. The difference in size between the two products is due to post-translational modiications which are inhibited by brefeldin A and are therefore likely to occur in the trans-Golgi network (TGN). Very little of the high-molecular-weight species was detected for full-length GalT, whereas it was a major product for the GalT/hybrid protein. Only the higher molecular weight species was expressed at the cell surface. Thus, this additional 7 kDa post-translational modification distinguishes molecules retained within the Golgi apparatus (lower M(r) species) from those transported through the TGN to the cell surface. These studies indicate that (i) the level of expression influences the intracellular distribution of GalT/hybrid molecules and (ii) the localization of full-length GalT involves active retention within the Golgi stack, and not retrieval from later compartments. After treatment of membrane preparations from stable L cell clones with a heterobifunctional cross-linking agent, full-length bovine GalT molecules were found almost exclusively as high-molecular-weight aggregates, suggesting that GalT exists as an oligomer or aggregate. This ability to oligomerize may be a requirement for Golgi retention.


title: The deubiquitinase USP15 antagonizes Parkin-mediated mitochondrial ubiquitination and mitophagy

text: Loss-of-function mutations in PARK2, the gene encoding the E3 ubiquitin ligase Parkin, are the most frequent cause of recessive Parkinson's disease (PD). Parkin translocates from the cytosol to depolarized mitochondria, ubiquitinates outer mitochondrial membrane proteins and induces selective autophagy of the damaged mitochondria (mitophagy). Here, we show that ubiquitin-specific protease 15 (USP15), a deubiquitinating enzyme (DUB) widely expressed in brain and other organs, opposes Parkin-mediated mitophagy, while a panel of other DUBs and a catalytically inactive version of USP15 do not. Moreover, knockdown of USP15 rescues the mitophagy defect of PD patient fibroblasts with PARK2 mutations and decreased Parkin levels. USP15 does not affect the ubiquitination status of Parkin or Parkin translocation to mitochondria, but counteracts Parkin-mediated mitochondrial ubiquitination. Knockdown of the DUB CG8334, the closest homolog of USP15 in Drosophila, largely rescues the mitochondrial and behavioral defects of parkin RNAi flies. These data identify USP15 as an antagonist of Parkin and suggest that USP15 inhibition could be a therapeutic strategy for PD cases caused by reduced Parkin levels.


title: Distinct requirements for IL-6 in polyclonal and specific Ig production induced by microorganisms

text: The role of IL-6 in Ig production induced in the mouse by lactate dehydrogenase-elevating virus (LDV), Toxoplasma gondii or lipopolysaccharide (LPS) was assessed. Following infection with LDV, a strong activator of B cells, an early and transient IL-6 production was observed, that originated predominantly from macrophages. Whereas LDV-induced B lymphocyte proliferation appeared independent of IL-6, mice deficient for this cytokine showed a marked reduction in their total T-dependent IgG2a production when compared to their normal counterparts. By contrast, specific responses directed against either LDV or non-viral antigens administered at the time of infection were not decreased in the absence of IL-6. Similarly, polyclonal, but not anti-parasite IgG2a production triggered by T. gondii infection was strongly dependent on the presence of IL-6. Finally, T-independent total IgG3 secretion triggered by LPS was also markedly reduced in IL-6-deficient mice. These results suggest that IL-6 plays a major role in T-dependent and T-independent polyclonal Ig production following B lymphocyte activation by viruses, and parasites, but not in specific antibody responses induced by the same microorganisms.


title: Characterization of epithelial IL-8 response to inflammatory bowel disease mucosal E. coli and its inhibition by mesalamine

text: BACKGROUND: Mucosally adherent E. coli are found in inflammatory bowel disease (IBD) and colon cancer. They promote release of the proinflammatory cytokine interleukin-8 (IL-8). We explored mechanisms for this release and its inhibition by drugs. METHODS: IL-8 release from colon epithelial cells in response to mucosal E. coli isolates from IBD, colon cancer, and controls was characterized at the cellular and molecular level. RESULTS: IL-8 response of HT29 cells was greater with Crohn's disease (689 ± 298 [mean ± SD] pg IL-8/mL at 4 hours, n = 7) and colon cancer isolates (532 ± 415 pg/mL, n = 14) than with ulcerative colitis (236 ± 58 pg/mL, n = 6) or control isolates (236 ± 100 pg/mL, n = 6, P < 0.0001). Bacterial supernatants contained shed flagellin that triggered IL-8 release. For whole bacteria the IL-8 response to E. coli that agglutinate red blood cells (548 ± 428 pg IL-8/mL, n = 16), a function that correlates with epithelial invasion, was greater than for nonhemagglutinators (281 ± 253 pg/mL, n = 17; P < 0.0001). This was particularly marked among E. coli that, although flagellate, could not release IL-8 from TLR5-transfected HEK293 cells. IL-8 release was mediated by extracellular-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) and inhibited by mesalamine, but not hydrocortisone, at therapeutic concentrations. CONCLUSIONS: Mucosa-associated E. coli shed flagellin that elicits epithelial IL-8 release but this may only become relevant when the mucosal barrier is weakened to expose basolateral TLR5. Adherent and invasive IBD and colon cancer E. coli isolates also elicit a flagellin-independent IL-8 response that may be relevant when the mucosal barrier is intact. The IL-8 release is MAPK-dependent and inhibited by mesalamine.


title: Viral Aetiology and Epidemiology of Acute Respiratory Infections in Hospitalized Saudi Children

text: A total of 1429 Saudi children of either sex and under 5 years of age who were admitted to King Khalid University Hospital, Riyadh during a three year period (April 1993–March 1996) with complaints suggestive of acute respiratory tract infections (ARTI) were investigated for viral aetiology of the infection. Viruses could be detected in 522 (37 per cent) cases with respiratory syncytial virus (RSV) the most commonly detected (79 per cent) followed by parainfluenza type 3 (8 per cent). Detection of influenza A, B and adenoviruses accounted for 6 per cent, 3 per cent and 2 per cent respectively. Except for parainfluenza virus type 3 the peak of activity of the respiratory viruses was during the winter months (October–February). Parainfluenza virus type 3 could be detected all year round but epidemics can occur in the hottest months of the year (June–August) when the temperature can exceed 40°C. Association between clinical manifestation, type of causative agent, and age was evaluated in 137 infected Saudi children in the first year of the study (April 1993–March 1994). The majority of our cases presented with bronchiolltis (58 per cent) while only 26 per cent had bronchopneumonla. There was a significant association between bronchiolitis and lower age groups (0–6 months), with RSV as the major causative agent of bronchiolitis cases (88 per cent).


title: Bronichiloitis obliterms-current concepts

text: We review current concepts abut the clinical manifestations,diagnosis and treatment of patients with bronchiolitis obliterans (BO) with emphasis on clinical/pathological correlations and recent developments. BO is a relatively rare disease, but its incidence is probably higher than generally believed and is continuously rising, partly because of better recognition, but also because of increased exposure to industrial fumes, and its occurrence in lung transplantation. BO is characterized histologically by varying degrees of obliteration of the lumen of the respiratory bronchioles by organizing connective tissue often extending into the alveoli (‘proliferative’ BO with organizing pneumoni+BOOP) or by more extensive fibrosis and scarring of the more proximal, conductive bronchioles (‘constrictive’ BO). Diverse clinical conditions have been associated with the development of BO, notably viral and mycoplasma infection, toxic fume exposure and immune reactions in the setting of a collagen vascular disease, drug reaction or organ transplantation. The clinical course and features of BO may vary considerably according to the aetiology, histological pattern and stage of the disease. The most common presentation is that of a progressive dry cough and dyspnea, associated with diffuse patchy interstitial lung infiltrates on chest X-ray. In the more advanced cases, lung function tests show either restrictive or obstructive defects, depending on the extent of alveolar involvement, and hypoxemia without CO(2) retention. The diagnosis is often possible on clinical grounds, however, in a seriously ill patient uncertainty should be resolved by tissue diagnosis, preferably by open lung biopsy. Treatment is based on symptomatic therapy. The use of corticosteroids is controversial, but common. Patients with BOOP are exceptional, in that there may be no underlying condition (‘idiopathic’ BOOP or cryptogenic organizing pneumonia—COP), a restrictive ventilatory defect is usual and the response to corticosteroids often remarkable.


title: Comparison of Fresh and Room-Aged Cigarette Sidestream Smoke in a Subchronic Inhalation Study on Rats

text: Two experimental types of cigarette sidestream smoke (SS) were compared in a subchronic inhalation study on rats. Fresh SS (FSS) was generated continuously from the reference cigarette 2R1. Room-aged SS (RASS) was generated by aging FSS for 1.5 h in a room with noninert surfaces with materials typically found in residences or offices. Male Sprague-Dawley rats were head-only exposed to three dose levels of each SS type and to filtered, conditioned fresh air (sham-exposure) for 6 h/day, 7 days/week, for 90 days. Room-aging resulted in decreased concentrations of various SS components, e.g., total participate matter (TPM) and nicotine, while other components, such as carbon monoxide (CO), were not affected. The CO concentrations were 6, 13, and 28 ppm for both SS types. TPM concentrations were between 0.6 and 8.7 (μg/liter and thus up to 100-fold above the maximum of average concentrations of respiratory suspended particles reported for environmental tobacco smoke. Slight reserve cell hyperplasia in the anterior part of the nose as well as hyperplastic and metaplastic epithelial changes in the larynx were the only observed dose-dependent findings. The metabolism of benzo(a)-pyrene-as a proxy for polycyclic aromatic hydrocarbon metabolism-was induced in the nasal respiratory epithelium and in the lungs while no effect was seen in the nasal olfactory epithelium. The lowest-observed effect level was 6 ppm CO or 0.6 μg TPM/liter. Most of the effects seen were less expressed in RASS- than in FSS-exposed rats when compared on the basis of the CO concentrations. When compared on the basis of TPM, these effects were equally pronounced for both SS types, suggesting a major role of particulate matter-associated compounds. All findings reverted to sham control levels following a 42-day postinhalation period.


title: Computational Epidemiology

text: Born from a desire to predict the future, epidemiology has largely been limited to studying the past. Now, computational epidemiology researchers are harnessing computing power to crack the complicated mystery of how diseases spread. Born from a desire to predict the future, epidemiology has largely been limited to studying the past. Now, computational epidemiology researchers are harnessing computing power to begin to crack the complicated mystery of how diseases spread.


More Models of Infection: It's Epidemic


title: Modeling Epidemics with Dynamic Small‐World Networks

text: In this presentation a minimal model for describing the spreading of an infectious disease, such as influenza, is discussed. Here it is assumed that spreading takes place on a dynamic small‐world network comprising short‐ and long‐range infection events. Approximate equations for the epidemic threshold as well as the spreading dynamics are derived and they agree well with numerical discrete time‐step simulations. Also the dependence of the epidemic saturation time on the initial conditions is analysed and a comparison with real‐world data is made.


title: Spatial Epidemic Modelling in Social Networks

text: The spread of infectious diseases is highly influenced by the structure of the underlying social network. The target of this study is not the network of acquaintances, but the social mobility network: the daily movement of people between locations, in regions. It was already shown that this kind of network exhibits small world characteristics. The model developed is agent based (ABM) and comprehends a movement model and a infection model. In the movement model, some assumptions are made about its structure and the daily movement is decomposed into four types: neighborhood, intra region, inter region and random. The model is Geographical Information Systems (GIS) based, and uses real data to define its geometry. Because it is a vector model, some optimization techniques were used to increase its efficiency.


title: Self‐organized criticality in human epidemiology

text: As opposed to most sociological fields, data are available in good quality for human epidemiology, describing the interaction between individuals being susceptible to or infected by a disease. Mathematically, the modelling of such systems is done on the level of stochastic master equations, giving likelihood functions for real live data. We show in a case study of meningococcal disease, that the observed large fluctuations of outbreaks of disease among the human population can be explained by the theory of accidental pathogens, leading the system towards a critical state, characterized by power laws in outbreak distributions. In order to make the extremely difficult parameter estimation close to a critical state with absorbing boundary possible, we investigate new algorithms for simulation of the disease dynamics on the basis of winner takes all strategies, and combine them with previously developed parameter estimation schemes.


title: Three‐dimensional Numerical Simulation of Gas‐particulate Flow around Breathing Human and Particulate Inhalation

text: It is important to predict the environment around the breathing human because inhalation of virus (avian influenza, SARS) is recently severe worldwide problem, and air pollution caused by diesel emission particle (DEP) and asbestos attract a great deal of attention. In the present study, three‐dimensional numerical simulation was carried out to predict unsteady flows around a breathing human and how suspended particulate matter (SPM, diameter∼1 μm) reaches the human nose in inhalation and exhalation. In the calculation, we find out smaller breathing angle and the closer distance between the human nose and pollutant region are effective in the inhalation of SPM.


title: Epidemiologic Insights into Stone Disease as a Systemic Disorder

text: Examining the epidemiology of stone disease can provide insight into etiology. There is a growing body of evidence that stone disease is not simply a disorder of the kidney. In fact, nephrolithiasis is clearly a systemic disorder. Conditions associated with stone disease include the classic ones such as inflammatory bowel disease and primary hyperparathyroidism. More recent studies have demonstrated strong associations with obesity, gout, diabetes and hypertension. Future studies will help uncover the underlying common pathophysiologic abnormalities.


title: Computational Evolutionary Methodology for Knowledge Discovery and Forecasting in Epidemiology and Medicine

text: Humanity is facing an increasing number of highly virulent and communicable diseases such as avian influenza. Researchers believe that avian influenza has potential to evolve into one of the deadliest pandemics. Combating these diseases requires in‐depth knowledge of their epidemiology. An effective methodology for discovering epidemiological knowledge is to utilize a descriptive, evolutionary, ecological model and use bio‐simulations to study and analyze it. These types of bio‐simulations fall under the category of computational evolutionary methods because the individual entities participating in the simulation are permitted to evolve in a natural manner by reacting to changes in the simulated ecosystem. This work describes the application of the aforementioned methodology to discover epidemiological knowledge about avian influenza using a novel eco‐modeling and bio‐simulation environment called SEARUMS. The mathematical principles underlying SEARUMS, its design, and the procedure for using SEARUMS are discussed. The bio‐simulations and multi‐faceted case studies conducted using SEARUMS elucidate its ability to pinpoint timelines, epicenters, and socio‐economic impacts of avian influenza. This knowledge is invaluable for proactive deployment of countermeasures in order to minimize negative socioeconomic impacts, combat the disease, and avert a pandemic.


title: Reaction‐Superdiffusion Systems in Epidemiology, an Application of Fractional Calculus

text: Spatially extended stochastic processes in epidemiology lead to classical reaction‐diffusion process, when infection spreads only locally. This notion can be generalized using fractional derivatives, especially fractional Laplacian operators, leading to Lévy flights and sub‐ or super‐diffusion. Especially super‐diffusion is a more realistic mechanism of spreading epidemics than ordinary diffusion.


title: Optimal Conditions for the Control Problem Associated to a Biomedical Process

text: This paper considers a mathematical model of infectious disease of SIS type. We will analyze the problem of minimizing the cost of diseases trough medical treatment. Mathematical modeling of this process leads to an optimal control problem with a finite horizon. The necessary conditions for optimality are given. Using the optimality conditions we prove the existence, uniqueness and stability of the steady state for a differential equations system.


title: Epidemics Modelings: Some New Challenges

text: Epidemics modeling has been particularly growing in the past years. In epidemics studies, mathematical modeling is used in particular to reach a better understanding of some neglected diseases (dengue, malaria, …) and of new emerging ones (SARS, influenza A,….) of big agglomerates. Such studies offer new challenges both from the modeling point of view (searching for simple models which capture the main characteristics of the disease spreading), data analysis and mathematical complexity. We are facing often with complex networks especially when modeling the city dynamics. Such networks can be static (in first approximation) and homogeneous, static and not homogeneous and/or not static (when taking into account the city structure, micro‐climates, people circulation, etc.). The objective being studying epidemics dynamics and being able to predict its spreading.


title: Describing dengue epidemics: Insights from simple mechanistic models

text: We present a set of nested models to be applied to dengue fever epidemiology. We perform a qualitative study in order to show how much complexity we really need to add into epidemiological models to be able to describe the fluctuations observed in empirical dengue hemorrhagic fever incidence data offering a promising perspective on inference of parameter values from dengue case notifications.


title: Parameter Estimation in Epidemiology: from Simple to Complex Dynamics

text: We revisit the parameter estimation framework for population biological dynamical systems, and apply it to calibrate various models in epidemiology with empirical time series, namely influenza and dengue fever. When it comes to more complex models like multi‐strain dynamics to describe the virus‐host interaction in dengue fever, even most recently developed parameter estimation techniques, like maximum likelihood iterated filtering, come to their computational limits. However, the first results of parameter estimation with data on dengue fever from Thailand indicate a subtle interplay between stochasticity and deterministic skeleton. The deterministic system on its own already displays complex dynamics up to deterministic chaos and coexistence of multiple attractors.


title: Understanding the effect of vector dynamics in epidemic models using center manifold analysis

text: In vector borne diseases the human hosts' epidemiology often acts on a much slower time scales than the one of the mosquitos which transmit the disease as a vector from human to human, due to their vastly different life cycles. We investigate in a model with susceptible (S), infected (I) and recovered (R) humans and susceptible (U) and infected (V) mosquitoes in how far the fast time scale of the mosquito epidemiology can be slaved by the slower human epidemiology, so that for the understanding of human disease data mainly the dynamics of the human time scale is essential and only slightly perturbed by the mosquito dynamics. This analysis of the SIRUV model is qualitatively in agreement with a previously investigated simpler SISUV model, hence a feature of vector-borne diseases in general.


title: On nonstandard finite difference schemes in biosciences

text: We design, analyze and implement nonstandard finite difference (NSFD) schemes for some differential models in biosciences. The NSFD schemes are reliable in three directions. They are topologically dynamically consistent for onedimensional models. They can replicate the global asymptotic stability of the disease-free equilibrium of the MSEIR model in epidemiology whenever the basic reproduction number is less than 1. They preserve the positivity and boundedness property of solutions of advection-reaction and reaction-diffusion equations.


title: Effects of dynamic quarantine and nonlinear infection rate in a model for computer worms propagation

text: We propose a new model for computer worms propagation, using dynamic quarantine and a nonlinear infection rate. The dynamic quarantine is based in epidemic disease control methods and in the principle ‘assume guilty before proven inocent’. This means that the host is blocked whenever its behavior looks suspicious. After a short time, the quarantined computer is released. The nonlinear infection rate is used to capture the dynamics of overcrowded infectious networks and high viral loads. We simulate numerically the model for distinct values of the quarantine times. We observe that increasing the quarantine time decreases the number of infectious hosts in the network.


title: A model for HIV/AIDS pandemic with optimal control

text: Human immunodeficiency virus and acquired immune deficiency syndrome (HIV/AIDS) is pandemic. It has affected nearly 60 million people since the detection of the disease in 1981 to date. In this paper basic deterministic HIV/AIDS model with mass action incidence function are developed. Stability analysis is carried out. And the disease free equilibrium of the basic model was found to be locally asymptotically stable whenever the threshold parameter (RO) value is less than one, and unstable otherwise. The model is extended by introducing two optimal control strategies namely, CD4 counts and treatment for the infective using optimal control theory. Numerical simulation was carried out in order to illustrate the analytic results.


title: A Simple Scoring Method for Predicting the Low Risk of Persistent Acute Kidney Injury in Critically Ill Adult Patients

text: The renal angina index has been proposed to identify patients at high risk of persistent AKI, based on slight changes in serum creatinine and patient conditions. However, a concise scoring method has only been proposed for pediatric patients, and not for adult patients yet. Here, we developed and validated a concise scoring method using data on patients admitted to ICUs in 21 Japanese hospitals from 2012 to 2014. We randomly assigned to either discovery or validation cohorts, identified the factors significantly associated with persistent AKI using a multivariable logistic regression model in the discovery cohort to establish a scoring system, and assessed the validity of the scoring in the validation cohort using receiver operating characteristic analysis and the calibration slope. Among 8,320 patients admitted to the ICUs, persistent AKI was present in 1,064 (12.8%) patients. In the discovery cohort (n = 4,151), ‘hyperbilirubinemia’, ‘sepsis’ and ‘ventilator and/or vasoactive’ with small changes in serum creatinine were selected to establish the scoring. In the validation cohort (n = 4,169), the predicting model based on this scoring had a c-statistic of 0.79 (95%CI, 0.77–0.81) and was well calibrated. In conclusion, we established a concise scoring method to identify potential patients with persistent AKI, which performed well in the validation cohort.


title: TG01/GM-CSF and adjuvant gemcitabine in patients with resected RAS-mutant adenocarcinoma of the pancreas (CT TG01-01): a single-arm, phase 1/2 trial

text: BACKGROUND: TG01 is the first cancer immunotherapy targeting KRAS oncogenic mutations. This study assessed the safety and efficacy of TG01/GM-CSF in patients with resected pancreatic adenocarcinoma. METHODS: Patients with stage I or II pancreatic adenocarcinoma who had undergone surgical resection (R0 or R1) received adjuvant gemcitabine with TG01/GM-CSF using two schedules of vaccination. Immune response was defined as a positive delayed-type hypersensitivity (DTH) response and/or positive T-cell proliferation assay. RESULTS: Thirty-two patients were enrolled between February 2013 and May 2016. Nineteen were treated with the high antigen burden, with four serious adverse reactions considered possibly related to TG01 treatment, including three allergic reactions. On this basis, a further 13 patients received a modified vaccination schedule with reduced antigen burden, with no serious adverse events related to TG01. Ninety-five percent patients in the main cohort and 92% in the modified cohort had a positive immune response. Median overall survival (OS) was 33.1 months, and median disease-free survival (DFS) was 13.9 months for the main cohort. For the modified cohort, the median OS was 34.3 months and median DFS was 19.5 months. CONCLUSIONS: TG01/GM-CSF with gemcitabine was well tolerated, with high levels of immune activation. OS and DFS compare favourably with published data for adjuvant gemcitabine. CLINICAL TRIAL REGISTRATION: This clinical trial was registered at ClinicalTrials.gov (NCT02261714).


title: The function and clinical application of extracellular vesicles in innate immune regulation

text: The innate immune system plays a crucial role in the host defense against viral and microbial infection. Exosomes constitute a subset of extracellular vesicles (EVs) that can be released by almost all cell types. Owing to their capacity to shield the payload from degradation and to evade recognition and subsequent removal by the immune system, exosomes efficiently transport functional components to recipient cells. Accumulating evidence has recently shown that exosomes derived from tumor cells, host cells and even bacteria and parasites mediate the communication between the invader and innate immune cells and thus play an irreplaceable function in the dissemination of pathogens and donor cell-derived molecules, modulating the innate immune responses of the host. In this review, we describe the current understanding of EVs (mainly focusing on exosomes) and summarize and discuss their crucial roles in determining innate immune responses. Additionally, we discuss the potential of using exosomes as biomarkers and cancer vaccines in diagnostic and therapeutic applications.


title: Physiology of gangliosides and the role of antiganglioside antibodies in human diseases

text: Gangliosides are structurally and functionally polymorphic sialic acid containing glycosphingolipids that are widely distributed in the human body. They play important roles in protecting us against immune attacks, yet they can become targets for autoimmunity and act as receptors for microbes, like the influenza viruses, and toxins, such as the cholera toxin. The expression patterns of gangliosides vary in different tissues, during different life periods, as well as in different animals. Antibodies against gangliosides (AGA) can target immune attack e.g., against neuronal cells and neutralize their complement inhibitory activity. AGAs are important especially in acquired demyelinating immune-mediated neuropathies, like Guillain–Barré syndrome (GBS) and its variant, the Miller–Fisher syndrome (MFS). They can emerge in response to different microbial agents and immunological insults. Thereby, they can be involved in a variety of diseases. In addition, antibodies against GM3 were found in the sera of patients vaccinated with Pandemrix®, who developed secondary narcolepsy, strongly supporting the autoimmune etiology of the disease.


title: BOARD-INVITED REVIEW: Fate and transport of bioaerosols associated with livestock operations and manures

text: Airborne microorganisms and microbial by-products from intensive livestock and manure management systems are a potential health risk to workers and individuals in nearby communities. This report presents information on zoonotic pathogens in animal wastes and the generation, fate, and transport of bioaerosols associated with animal feeding operations and land applied manures. Though many bioaerosol studies have been conducted at animal production facilities, few have investigated the transport of bioaerosols during the land application of animal manures. As communities in rural areas converge with land application sites, concerns over bioaerosol exposure will certainly increase. Although most studies at animal operations and wastewater spray irrigation sites suggest a decreased risk of bioaerosol exposure with increasing distance from the source, many challenges remain in evaluating the health effects of aerosolized pathogens and allergens in outdoor environments. To improve our ability to understand the off-site transport and diffusion of human and livestock diseases, various dispersion models have been utilized. Most studies investigating the transport of bioaerosols during land application events have used a modified Gaussian plume model. Because of the disparity among collection and analytical techniques utilized in outdoor studies, it is often difficult to evaluate health effects associated with aerosolized pathogens and allergens. Invaluable improvements in assessing the health effects from intensive livestock practices could be made if standardized bioaerosol collection and analytical techniques, as well as the use of specific target microorganisms, were adopted.


title: Decreases of Metallothionein and Aminopeptidase N in Renal Cancer Tissues(1)

text: Good molecular markers for investigating the biochemical differences between renal cancer and surrounding tissues have not yet been developed. Sixteen kidney samples (clear cell RCC) were investigated to determine the differences in the protein components between renal cancer and surrounding tissues, using HPLC analysis. The metallothionein (MT) and zinc levels were consistently lower in renal cancer tissues compared with in surrounding tissues. The mean concentration of MT in normal tissues surrounding renal tumors was about 15 times higher than that in cancer tissues. An immunohistochemical study confirmed that the expression of MT in renal cancer tissues was lower than that in adjacent normal tissues. The activities of aminopeptidases (APs) were significantly decreased in renal cancer tissues compared with in adjacent normal tissues. An immunohistochemical study and Western blot analysis confirmed that the expression of AP-N in renal cancer tissues was also lower than in adjacent normal tissues. These results suggest that the immunohistochemical detection of MT and AP-N could provide useful information as a pathological diagnostic tool for classifying renal cancer and surrounding tissues.


title: VIRUSES AND THE VERSATILE MACROPHAGE

text: Mononuclear phagocytes, including circulating monocytes and tissue macrophages, play a central role in resistance to viruses. This resitance can be expressed both non-specifically, and specifically in indiction, regulation and amplification of humoral and cell mediated immune responses to viruses. These lead to the extrinsic effect of macrophages on other virus-infected cells or free virus, and the intrinsic effect on viruses within macrophages. While these interactions usually appear to be protective, immunopathologic consequences as well as macrophage dysfunctions have also been noted. The outcome of any given interaction (viral elimination, peristance, latency or transformation) varies markedly with the type of macrophage. The molecular mechanisms involved in these very diverse macrophage-virus interactions are currently under study.


title: Epitope specificity and longevity of a vaccine-induced human T cell response against HPV18

text: Persistent human papillomavirus (HPV) type 16 and 18 infection can lead to pre-malignant and malignant diseases of the lower genital tract. Several lines of evidence suggest that T cell responses can control HPV infection. However, relative to other human viruses, strong effector memory T cell responses against HPV have been difficult to detect. We used an in vitro stimulation step prior to enzyme-linked immunospot assays to identify IFN-γ-secreting T cells specific for HPV16 and 18 E6/E7 peptides. This allowed the detection of HPV-specific CD4(+) T cells that were not evident in direct ex vivo assays. T cell responses against HPV16 or 18 peptides were detected in healthy volunteers (7/9) and patients with lower genital tract neoplasia (10/20). Importantly, this assay allowed tracking of vaccine-induced T cell responses in nine patients, following inoculation with a live recombinant vaccinia virus (HPV16 and 18 E6/E7, TA-HPV). Novel vaccine-induced T cell responses were demonstrated in five patients, but no clinical responses (lesion regressions) were seen. For one vaccinated patient, the T cell response was mapped to a single dominant HPV18 E7 epitope and this response was sustained for >3 years. Our data suggest that systemic memory T cells against HPV16 and 18, induced naturally or by TA-HPV vaccination, are relatively rare. Nevertheless, the assay system developed allowed estimation of magnitude, epitope specificity, and longevity of vaccine-induced CD4(+) T cell responses. This will be useful for vaccine design and measurement of immunological endpoints in clinical trials.


title: Functional Tumor Necrosis Factor–Related Apoptosis-Inducing Ligand Production by Avian Influenza Virus–Infected Macrophages

text: Severe human disease associated with influenza A H5N1 virus was first detected in Hong Kong in 1997. Its recent reemergence in Asia and high associated mortality highlight the need to understand its pathogenesis. We investigated the roles of death receptor ligands (DRLs) in H5N1 infection. Significant up-regulation of tumor necrosis factor (TNF)–related apoptosis-inducing ligand (TRAIL) and TNF-α, but not Fas ligand (FasL) mRNA, was detected in human monocyte–derived macrophages (MDMs) infected with avian influenza viruses A/Hong Kong/483/97 (H5N1/97) or its precursor, A/Quail/Hong Kong/G1/97. H5N1/97-infected MDMs exhibited the strongest induction of apoptosis in Jurkat T cells, and it could be reduced by TRAIL–receptor 2 blocking antibody. Furthermore, influenza virus infection enhanced the sensitivity of Jurkat T cells to apoptosis induced by TNF-α, TRAIL, and FasL. Our data suggested that functional TRAIL produced by influenza virus–infected MDMs was related to their cytotoxicity and that the enhanced sensitization to DRL-induced apoptosis detected in avian influenza may contribute to disease pathogenesis


Viral Taxonomy


title: Global importance of RNA secondary structures in protein-coding sequences

text: MOTIVATION: The protein-coding sequences of messenger RNAs are the linear template for translation of the gene sequence into protein. Nevertheless, the RNA can also form secondary structures by intramolecular base-pairing. RESULTS: We show that the nucleotide distribution within codons is biased in all taxa of life on a global scale. Thereby, RNA secondary structures that require base-pairing between the position 1 of a codon with the position 1 of an opposing codon (here named RNA secondary structure class c(1)) are under-represented. We conclude that this bias may result from the co-evolution of codon sequence and mRNA secondary structure, suggesting that RNA secondary structures are generally important in protein-coding regions of mRNAs. The above result also implies that codon position 2 has a smaller influence on the amino acid choice than codon position 1. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.


title: CCR5 Plays a Critical Role in the Development of Myocarditis and Host Protection in Mice Infected with Trypanosoma cruzi

text: The pathogenesis of myocarditis during Trypanosoma cruzi infection is poorly understood. We investigated the role played by chemokine receptor 5 (CCR5) in the influx of T cells to the cardiac tissue of T. cruzi—infected mice. mRNA and protein for the CCR5 ligands CCL3, CCL4, and CCL5 were detected in the hearts of infected mice in association with CD4(+) and CD8(+) T cells. There was a high level of CCR5 expression on CD8(+) T cells in the hearts of infected mice. Moreover, CCR5 expression on CD8(+) T cells was positively modulated by T. cruzi infection. CCR5-deficient mice infected with T. cruzi experienced a dramatically inhibited migration of T cells to the heart and were also more susceptible to infection. These results suggest that CCR5 and its ligands play a central role in the control of T cell influx in T. cruzi-infected mice. Knowledge of the mechanisms that trigger and control the migration of cells to the heart in patients with Chagas disease may help in the design of drugs that prevent myocarditis and protect against the development of severe disease.


title: Gastroenteritis in Children: A Two-Year Review in Manitoba. I. Etiology

text: During two years, 1,217 children hospitalized with gastroenteritis at the Children's Centre in Winnipeg, Manitoba, Canada were studied. Bacterial pathogens were present in 25% of these children: enteropathogenic Escherichia coli in 120, Shigella in 139, Salmonella in 24, and multiple pathogens in 18. Rotavirus was detected in 54 (II %) of 472 patients examined. Rotavirus and enteropathogenic E. coli were the most common pathogens in infants, and Shigella was the most common in older children. Bacterial diarrhea occurred more commonly in summer, whereas rotavirus infection occurred more commonly in winter. Among 276 children screened, enterotoxigenic E. coli was found in three, and Aeromonas shigelloides that produced a similar toxin in two others. Enteroinvasive E. coli was not detected in 70 children. Organisms producing toxins "cytotoxic" to HeLa cells were isolated from three of 90 children. Screening for enterotoxigenic or enteroinvasive organisms was not productive of a significant number of pathogens, and, although screening for rotavirus did improve the number of etiologic diagnoses, the etiology of the majority of cases of diarrhea remained unknown.


title: RAPID DIAGNOSIS OF VIRUS DISEASES

text: Virus infections which may cause severe or life-threatening illnesses meriting chemotherapy are surveyed. Experience has shown that effective antiviral chemotherapy must be given early in the illness, and so this paper concentrates on the earliest symptoms of the various diseases and on methods leading to rapid diagnosis. This can be made by recognizing virus particles of characteristic morphology, if they occur in large numbers. e.g. rotavirus in diarrhoea. Sensitivity may be enhanced by using immune serum to trap viruses. A variety of very sensitive immunological methods is now available for detecting virus-specific proteins in secretions of the respiratory and alimentary tracts. Monoclonal antibodies help to make these more specific. Virus nucleic acids can also be detected in secretions and cells in situ by hybridization with purified radio-labelled nucleic acid. Virus diseases specifically surveyed include varicella zoster and measles, both very important in immuno-suppressed patients; epidemic influenza; and the human papilloma viruses, of which 33 different types are now known, and which cause not common warts on hands and feet, but also venereally transmitted inapparent papillomas upon the surface of the uterine cervix, often leading to cervical carcinoma. The problems of diagnosis of acquired immunodeficiency syndrome are reviewed. The aetiology of chronic non-A, non-B hepatitis is still largely obscure, but a retrovirus may be involved. The diagnosis of rabies is also considered.


title: THE LABORATORY SELECTION OF ANTIVIRAL AGENTS

text: The major disease targets for antiviral chemotherapy are influenza, rhinovirus colds and herpetic infections Some success has been achieved against influenza with 1-amino adamantane (amantadine), and several new alicyclic amines are at the development stage. There are many organic compounds, covering a wide range of chemical types, which will inhibit very effectively most or all strains of rhinovirus in vitro, but all have failed to inhibit significantly disease development in infected human volunteers. Most success has been achieved with anti-herpetic drugs, and a variety of nucleoside analogues ar now available for clinical use. Primary testing methods for potential antiviral agents rely on high-through-put methods for detecting inhibition of virus replication in cell cultures. Secondary screening of active compounds is often done in organ culture e.g. tracheal tissue explants. In-vivo evaluation is done in animal models of the human disease, where available. Prolonged safety testing is required before human trials can be started.


title: Influenza Vaccination of Health Care Workers in Long-Term-Care Hospitals Reduces the Mortality of Elderly Patients

text: Vaccination of health care workers (HCWs) is recommended as a strategy for preventing influenza in elderly patients in long-term care. However, there have been no controlled studies to show whether this approach is effective. During the winter of 1994–1995, 1059 patients in 12 geriatric medical long-term-care sites, randomized for vac.cination of HCWs, were studied. In hospitals where HCWs were offered vaccination, 653 (61%) of 1078 were vaccinated. Vaccination of HCWs was associated with reductions in total patient mortality from 17% to 10% (odds ratio [OR], 0.56; 95% confidence interval [CI], 0.40–0.80) and in influenza-like illness (OR, 0.57; 95% CI, 0.34–0.94). Vaccination of patients was not associated with significant effects on mortality (OR, 1.15; 95% CI, 0.81–1.64). Results of this study support recommendations for vaccination against influenza of HCWs in long-term geriatric care. Vaccination of frail elderly long-term-care patients may not give clinically worthwhile benefits.


title: Inhibition of Japanese encephalitis virus replication in cultured cells and mice by a peptide-conjugated morpholino oligomer

text: BACKGROUND: Japanese encephalitis virus (JEV) has a significant impact on public health throughout Asia, and there is a pressing need for development of new therapeutics against it. METHODS: Peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs) are antisense agents that enter cells readily and interfere with gene expression. Four PPMOs, targeting various locations in the JEV genome, were evaluated for antiviral activity against JEV in cultured cells and the mouse model of JEV infection. RESULTS: A PPMO (P10882) targeting the JEV 3′ cyclization sequence (3′CSI) had significant antiviral activity in Vero (epithelial), Neuro2A (neuronal) and J774E (macrophage) cells at concentrations that were not cytotoxic. P10882 added before infection suppressed JEV replication to an undetectable level in Vero cells and produced a 93% and 66% reduction in titre in J774E and Neuro2A cells, respectively, when measured at 24 h post-infection. In uninfected cells, fluorescein-labelled PPMOs entered J774E cells most efficiently, followed by Vero and Neuro2A cells. The antiviral effect of P10882 was also demonstrated in vivo, where 60%–80% of 1-week-old mice treated intracerebrally with a 20 mg/kg dose of P10882 every 12 h for 5 days were protected from a lethal dose of JEV and showed an undetectable level of virus in brain tissue at 2 days post-infection. CONCLUSIONS: P10882, which targets sequence that is highly conserved across members of the JEV serocomplex, was previously shown to be effective in a mouse model of West Nile disease, and represents a candidate antiviral agent against members of the JEV serocomplex.


title: Elevation of hepatic microsomal epoxide hydrolase activity by 2-acetylaminofluorene: strain and species differences

text: Hepatocarcinogens have been shown to cause marked elevation of hepatic microsomal epoxide hydrobase activity in the rat at short intervals after administration. The present studies were designed to characterize 2-acetylaminofluorene (AAF) mediated epoxide hydrolase elevation and to investigate the relationship between epoxide hydrolase increases, AAF metabolism, and hepatocarcinogenicity. Oral or i.p. administration of AAF to F-344 rats produced log-linear doseresponse curves for epoxide hydrolase elevation, measured with either benzo[a]pyrene-4, 5-oxide or styrene oxide substrate. Following a single dose of AAF (35 mg/kg), epoxide hydrolase activity was maximally increased (560% of control) within 48 h, and the activity declined slowly, with a halflife of 17.5 days. Co-treatment with actinomycin D effectively blocked the AAF dependent increase in epoxide hydrolase, suggesblng that de novo protein synthesis is associated with the increase in enzyme activity. Dose-response curves for epoxide hydrolase induction by AAF, N-hydroxy-2-acetylaminofluorene (N-OH-AAF), and aminofluorene were compared, and the potencies for increasing epoxide hydrolase activity reflected the relative hepatocarcinogenic potentials of these agents. In mice, which are resistant to the hepatocarcinogenic action of AAF and deficient in AAF-N-hydroxylase activity, AAF caused no significant increase in hepatic microsomal epoxide hydrolase activity. Similarly, in Cotton rats and guinea pigs, which are lacking in ability to form the sulfate conjugate of N-OH-AAF, neither i.p. nor dietary administration of AAF elicited increases in epoxide hydrolase activity at doses which were maximally effective in F-344 rats. These results support the hypothesis that the ability of compounds to increase epoxide hydrolase activity is related to their carcinogenic potency. Furthermore, the results suggest that increases in epoxide hydrolase activity are associated with metabolism of AAF to the putative proximate carcinogen N-OH-AAF, and the subsequent conversion of this compound to the N-O-sulfate conjugate.


title: Molecular-based isothermal tests for field diagnosis of malaria and their potential contribution to malaria elimination

text: In countries where malaria transmission has decreased substantially, thanks to the scale-up of control interventions, malaria elimination may be feasible. Nevertheless, this goal requires new strategies such as the active detection and treatment of infected individuals. As the detection threshold for the currently used diagnostic methods is 100 parasites/μL, most low-density, asymptomatic infections able to maintain transmission cannot be detected. Identifying them by molecular methods such as PCR is a possible option but the field deployment of these tests is problematic. Isothermal amplification of nucleic acids (at a constant temperature) offers the opportunity of addressing some of the challenges related to the field deployment of molecular diagnostic methods. One of the novel isothermal amplification methods for which a substantial amount of work has been done is the loop-mediated isothermal amplification (LAMP) assay. The present review describes LAMP and several other isothermal nucleic acid amplification methods, such as thermophilic helicase-dependent amplification, strand displacement amplification, recombinase polymerase amplification and nucleic acid sequence-based amplification, and explores their potential use as high-throughput, field-based molecular tests for malaria diagnosis.


title: IFITM1 Is Superior to CD10 as a Marker of Endometrial Stroma in the Evaluation of Myometrial Invasion by Endometrioid Adenocarcinoma

text: Background: Distinguishing myometrial invasion from adenomyosis involvement is important for staging of endometrial endometrioid adenocarcinoma. We aimed to compare CD10, which has limited value in this scenario, with interferon-induced transmembrane protein 1 (IFITM1), a recently described sensitive and specific marker of endometrial stroma. Methods: We reviewed 25 hysterectomies containing endometrial endometrioid adenocarcinoma and adenomyosis. Tumor areas were classified as unequivocally myoinvasive or unequivocally noninvasive. Foci equivocal for invasion were also recorded. Immunohistochemistry for IFITM1 and CD10 was performed and scored in terms of intensity and distribution and classified as negative or positive. Results: Unlike CD10, IFITM1 staining showed significant differences in mean intensity (P < .0001) and distribution (P < .0001) between invasive vs noninvasive areas. Sixteen (84.2%) invasive and 34 (97.1%) noninvasive areas were positive for CD10 (P = .22). In contrast, none of the invasive vs 25 (71.4%) noninvasive areas were positive for IFITM1 (P < .0001). IFITM1 had 71.4% sensitivity and 100% specificity in detecting stroma surrounding endometrioid adenocarcinoma, hence excluding myoinvasion. Eleven (45.8%) of 24 foci designated as equivocal stained with IFITM1. Conclusions: Compared with CD10, IFITM1 has superior performance distinguishing endometrial stroma of adenomyosis from mesenchyma surrounding invasive endometrial adenocarcinoma. IFITM1 expression is highly predictive of the absence of invasion and may be valuable in cases in which determining myoinvasion has staging implications.


Germs on a Plane: Aircraft, International Travel, and the Global Spread of Disease


Hunting Common Cold Viruses by Some New Methods


title: AN ANALYSIS OF HOSPITALIZATIONS FOR ACUTE RESPIRATORY DISEASE IN RECRUITS IMMUNIZED WITH ADENOVIRUS TYPE 4 AND TYPE 7 VACCINES(1)

text: Dudding, B. A., F. H. Top, Jr., R. M. Scott, P. K. Russell and E. L. Buescher (Walter Reed Army Institute of Research, Washington, D. C. 20012). An analysis of hospitalizations for acute respiratory disease in recruits immunized with adenovirus type 4 and type 7 vaccines. Am J Epidemiol 95: 140–147, 1972.—Among 911 recruits immunized with live, oral adenovirus types 4 and 7 (ADV-4 and 7) vaccines, there were 149 hospital admissions to the acute respiratory disease (ARD) wards during the study. Eighteen admissions were not associated with ARD (15 rubella infections, 3 immunization reactions). Of the remaining 131 ARD admissions, 17 were complicated by pneumonia and 114 were judged to have uncomplicated ARD. Within the latter group, evidence for infection by a single respiratory pathogen was obtained from 68 admissions; 11 admissions were associated with multiple infectious agents; and 35 admissions yielded no infectious agent(s). Despite immunization, adenoviruses, primarily ADV-4, were still the most common cause of ARD. Among non-adenovirus associated ARD admissions, rhinoviruses were the most common (10), followed by influenza A2 virus (7), group A streptococci (2) and herpesvirus hominis (2). Thus, in this initial study of ARD among recruits immunized with ADV-4 and ADV-7 vaccines, no other respiratory pathogens emerged to replace adenovirus types 4 and 7 as the major causes of ARD in military trainees.


title: IFN-γ establishes interferon-stimulated gene-mediated antiviral state against Newcastle disease virus in chicken fibroblasts

text: Newcastle disease virus (NDV) causes severe economic losses through severe morbidity and mortality and poses a significant threat to the global poultry industry. Significant efforts have been made to develop novel vaccines and therapeutics; however, the interaction of NDV with the host is not yet fully understood. Interferons (IFNs), an integral component of innate immune signaling, act as the first line of defense against invading viruses. Compared with the mammalian repertoire of IFNs, limited information is available on the antiviral potential of IFNs in chickens. Here, we expressed chicken IFN-γ (chIFN-γ) using a baculovirus expression vector system, characterized its antiviral potential against NDV, and determined its antiviral potential. Priming of chicken embryo fibroblasts with chIFN-γ elicited an antiviral environment in primary cells, which was mainly due to interferon-stimulated genes (ISGs). A genome-wide transcriptomics approach was used to elucidate the possible signaling pathways associated with IFN-γ-induced immune responses. RNA-sequencing (RNA-seq) data revealed significant induction of ISG-associated pathways, activated temporal expression of ISGs, antiviral mediators, and transcriptional regulators in a cascade of antiviral responses. Collectively, we found that IFN-γ significantly elicited an antiviral response against NDV infection. These data provide a foundation for chIFN-γ-mediated antiviral responses and underpin functional annotation of these important chIFN-γ-induced antiviral influencers.


title: Detection by Immune Electron Microscopy of 27-nm Viral Particles Associated with Community-Acquired Diarrhea in Children

text: The proportion of diarrheal illnesses of unknown origin that were associated with small round virus (SRV, 23-38 nm) particles among children <2 years old attending an outpatient clinic in Baltimore was determined. During a 9-month period, stool specimens from 188 patients with acute diarrhea and 108healthy age-matched control children were examined for enteric bacterial pathogens, protozoa, enteric adenovirus, and rotavirus. An enteropathogen was identified in 75 patients (40%) and in 21 controls (20%). A random sample of specimens without an identifiable pathogen wasthen examined for SRV particles by immune electron microscopy (IBM) using commercial human gamma globulin. Viruses of 26-30 nm diameter that were not enteroviruses were detected in specimens from 9 (12.5%) of the 7l patients and 1 (1.8%) of the 53 control subjects (P < .04). Of6 patients with available acute and convalescent sera, 4 demonstrated a significant immune response when tested by IBM. All patients experienced a mild, self-limited (1–3 days) illness. These findings suggest that SRV may be endemic in the Baltimore community and may result in clinically significant diarrheal illnesses.


title: Murine CD4(+) T cell clones vary in function in vitro and in influenza infection in vivo

text: Several CD4(+) T(h)1 clones specific for Influenza haemagglutinln or nucleoproteln were transferred into syngeneic mice after Intranasal Influenza infection to examine whether they accelerate viral clearance in vivo similarly to CD8(+) cytotoxlc T cells. We observed changes in functional properties of the CD4(+) clones in vitro and variable effects on the course of Infection in vivo. While some clones resulted In more rapid virus clearance, others had no protective effect, but rather exacerbated Illness symptoms. Our results reflect problems in the in vivo use of CD4(+) T cell clones maintained in long-term culture. Their IL-2 and IL-5 release and cytolytic activity varied, while IL-3 and γ-IFN production as well as DTH induction were more stable. CD4(+) T cells primed by infection became cytolytic only after prolonged culture. The data point to the fine balance between exacerbation of disease and protection by CD4(+) T cells.


title: Universal evolutionary selection for high dimensional silent patterns of information hidden in the redundancy of viral genetic code

text: MOTIVATION: Understanding how viruses co-evolve with their hosts and adapt various genomic level strategies in order to ensure their fitness may have essential implications in unveiling the secrets of viral evolution, and in developing new vaccines and therapeutic approaches. Here, based on a novel genomic analysis of 2625 different viruses and 439 corresponding host organisms, we provide evidence of universal evolutionary selection for high dimensional ‘silent’ patterns of information hidden in the redundancy of viral genetic code. RESULTS: Our model suggests that long substrings of nucleotides in the coding regions of viruses from all classes, often also repeat in the corresponding viral hosts from all domains of life. Selection for these substrings cannot be explained only by such phenomena as codon usage bias, horizontal gene transfer and the encoded proteins. Genes encoding structural proteins responsible for building the core of the viral particles were found to include more host-repeating substrings, and these substrings tend to appear in the middle parts of the viral coding regions. In addition, in human viruses these substrings tend to be enriched with motives related to transcription factors and RNA binding proteins. The host-repeating substrings are possibly related to the evolutionary pressure on the viruses to effectively interact with host's intracellular factors and to efficiently escape from the host's immune system. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.


title: Infection with Borna Disease Virus: Molecular and Immunobiological Characterization of the Agent

text: Borna disease virus (BDV), which seems to be distinct from all other known viruses, exhibits a unique mechanism of pathogenesis. This review highlights several aspects of the biology of infection with this virus and summarizes the preliminary characterization of the agent. Studies on BDV may help to illuminate several important areas of neurobiology, including the mechanisms regulating the replication of a new type of RNA virus in the nuclei of neural cells, the neuroinvasiveness and neurotropism of such viruses, their T cell-mediated immunopathology, tolerance in newborn animals to persistent viral infection of the central nervous system, and behavioral diseases and eating disorders induced by such agents.


title: Identification of Mycobacterium species by comparative analysis of the dnaA gene

text: For the establishment of a diagnostic tool for mycobacterial species, a part of the dnaA gene was amplified and sequenced from clinically relevant 27 mycobacterial species as well as 49 clinical isolates. Sequence variability in the amplified segment of the dnaA gene allowed the differentiation of all species except for Mycobacterium tuberculosis, Mycobacterium africanum and Mycobacterium microti, which had identical sequences. Partial sequences of dnaA from clinical isolates belonging to three frequently isolated species revealed a very high intraspecies similarity, with a range of 96.0–100%. Based on the dnaA sequences, a species-specific primer set for Mycobacterium kansasii and Mycobacterium gastri was successfully designed for a simple loop-mediated isothermal amplification method. These results demonstrate that the variable sequences in the dnaA gene were species specific and were sufficient for the development of an accurate and rapid diagnosis of Mycobacterium species.


title: Transmission Dynamics and Prospective Environmental Sampling of Adenovirus in a Military Recruit Setting

text: BackgroundHigh levels of morbidity caused by adenovirus among US military recruits have returned since the loss of adenovirus vaccines in 1999. The transmission dynamics of adenovirus have never been well understood, which complicates prevention efforts MethodsEnrollment and end-of-study samples were obtained and active surveillance for febrile respiratory illnesses (FRIs) was performed for 341 recruits and support personnel. Environmental samples were collected simultaneously. Classic and advanced diagnostic techniques were used ResultsSeventy-nine percent (213/271) of new recruits were seronegative for either adenovirus serotype 4 (Ad-4) or adenovirus serotype 7 (Ad-7). FRI caused by Ad-4 was observed in 25% (67/271) of enrolled recruits, with 100% of them occurring in individuals with enrollment titers <1:4. The percentage of recruits seropositive for Ad-4 increased from 34% at enrollment to 97% by the end of the study. Adenovirus was most commonly detected in the environment on pillows, lockers, and rifles ConclusionsPotential sources of adenovirus transmission among US military recruits included the presence of adenovirus on surfaces in living quarters and extended pharyngeal viral shedding over the course of several days. The introduction of new recruits, who were still shedding adenovirus, into new training groups was documented. Serological screening could identify susceptible recruits for the optimal use of available vaccines. New high-throughput technologies show promise in providing valuable data for clinical and research applications