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DESCRIPTION The post-translational attachment of the ubiquitin domain generates a targeting signal that localizes modified proteins to the proteasome, the nucleus, the cytoskeleton and the autophagic system. At least five distinct proteins are known to possess the ubiquitin domain and target modified proteins to different cellular locations. It has been shown that the accumulation of polymeric ubiquitin caused by mutation of deubiquitinating enzymes interferes with the targeting of polyubiquitinated proteins to the proteasome and thus, protein degradation. Polymeric ubiquitin can also arise as a result of the translation of the proubiquitin mRNA. This gene codes for tandem repeats of the ubiquitin domain, and the polyprotein that results must be processed to the monomer by proteolytic cleavage at the mature C-terminus of ubiquitin.Finally, ubiquitin-fusion proteins are also produced in eukaryotic cells and these must be processed at the C-terminus of ubiquitin to release ubiquitin and the C-terminal fusion proteins that are required for ribosome biogenesis and function. It is likely that the cell would have to avoid producing polymeric ubiquitin from these genes to prevent interference with the complex metabolism of post-translational ubiquitination. Based on this expectation, and the observation that the pro-ubiquitin protein is never observed we have hypothesized that processing of the pro-ubiquitin gene product is co-translational. Further, we suspect that the translation of this message is periodically stalled to allow recruitment of the processing enzyme and that this stalling is due to the presence of low abundance codons in each copy of ubiquitin coding sequence. Experiments proposed here will examine these hypotheses and further define cotranslational processing events.
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{
"pile_set_name": "NIH ExPorter"
}
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Project 3 Summary Sarcoma is a malignant cancer derived from transformed cells of mesenchymal origin. Progression and metastasis of sarcomas is regulated by microenvironmental cues. Low intratumoral O2 (hypoxia) most dramatically increases pulmonary metastasis, and poor clinical outcomes, though we do not yet completely understand the critical effects of hypoxia on sarcoma cells and the microenvironment. Thus, defining how primary tumor cells respond to O2 in their microenvironment is essential for understanding metastasis and identification of novel therapeutic targets. Our recent work has shown that hypoxia promotes sarcoma metastasis, through induction of HIF1??PLOD2 and the subsequent deposition of aberrant collagen that leads to distant metastasis. However, we do not yet know 1) how cell migration/invasion is altered in the presence of the O2 gradients that occur in tumors, 2) which collagen modifying-enzymes define the ECM, and 3) precisely how cells? motility is modified in response to altered collagen structure in the microenvironment. The proposed studies address each of these unknowns. To model the O2-gradients that develop in tumors as they outgrow their vascular supply we developed novel O2- controlling hydrogels that can serve as 3D hypoxic microenvironments. We hypothesize that sarcoma cell invasion and migration is guided by increased O2 tension and facilitated by hypoxia-induced ECM remodeling. We will determine if O2 gradients regulate the direction, speed and distance of migrating sarcoma cells, how these factors depend on hypoxic ECM remodeling, focusing on collagen microstructure. Our approach will integrate mathematical modeling and experimental in vitro and in vivo models, linking O2-ECM-cellular invasion and migration in sarcomas. The specific aims are: (1) To determine sarcoma cell and tumor graft responses to spatial oxygen gradients; (2) To characterize collagen remodeling during sarcoma invasion under hypoxic gradients; (3) To determine how collagen fiber organization regulates hypoxic invasion and migration. The results of the experiments proposed here will identify the molecular and physical mechanisms underlying the initial steps of metastasis, invasion and migration, and develop predictive models for these mechanism, all leading to novel therapeutic targets.
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{
"pile_set_name": "NIH ExPorter"
}
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During the past decade, it has become clear that the rate of aging, like many other processes in biology, is subject to regulation. In many animals, including mammals, longevity is regulated by a conserved insulin- and IGF-l-like signaling pathway. This longevity system was discovered in the genetically-tractable organism C. elegans, and a great deal about it has been learned by studying this animal. In C. elegans, DAF-2, an insulin/IGF-l-like receptor, activates a conserved signal transduction pathway that inhibits longevity, at least in part, by inhibiting the activity of the transcription factor DAF-16. In mutants defective in this signaling pathway, DAF-16 accumulates in the nucleus and coordinates expression of a battery of diverse downstream genes that together produce dramatic extensions of lifespan. This study addresses key, unsolved questions about this signaling pathway. Sensory neurons regulate the DAF-2 pathway, possibly in response to environmental cues, and this study tests the hypothesis that sensory neurons influence lifespan by controlling the production or release of specific insulin-like peptides. The C. elegans heat-shock transcription factor, HSF-1, which regulates the highly-conserved heat-shock response, acts with DAF-16 to regulate the expression of genes required for youthfulness and longevity. This study will determine how HSF-1 activity is regulated and how it functions in this system. Finally, a large number of genes with unknown functions act in this system to influence longevity. This study will better define the biochemical roles that these genes play in the aging process. Together these studies may ultimately define new therapeutic strategies for combating age-related disease and increasing the quality of old age.
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{
"pile_set_name": "NIH ExPorter"
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The aim of this research is to improve the understanding of a new cell surface regulatory process (Talk-Back regulation) in the "Mutants" group of oral streptococci. The physiological aspects of this group of streptococci that enable them to survive the environmental stresses of "feast or famine" and continuous assaults by host defense mechanisms inherent to the oral cavity, are basic to understanding the persistence and pathogenicity of these bacteria. Elucidation of the mechanism(s) by which the cell surface (in this instance, the Gram positive cell wall) communicates its growth status to the cytoplasm seems likely to be important to this regard. In addition to further describing a basic regulatory circuit, this study will also describe molecular events leading to the secondary inhibition of RNA and protein synthesis observed in the tolerant response to beta-lactam and other cell wall active antibiotics. This experimental approach will utilize established bacteriological and biochemical methods to analyze molecular events subsequent to exposure of growing bacterial cultures to benzylpenicillin and other inhibitors of cell wall synthesis. Analysis of putative regulatory nucleotides and stress proteins associated with the "Talk- Back" response will be coupled with parallel studies on cultures undergoing other environmental stresses. Compositional and structural changes in the cell wall of bacteria subsequent to treatment with antibiotics will be performed via HPLC analysis of enzymatic digestion products. Appropriate regulatory mutants (to be isolated) will greatly facilitate the proposed research and provide the basis for future genetic analysis of the regulatory circuits of these streptococci. The long range objectives of this research are to define the molecular aspects of "Talk-Back" regulation and the involvement of this pathway in the tolerant and nonlytic death responses to cell wall antibiotics and to improve the understanding of the basic physiology of the "Mutants" streptococci which enable them to survive in an contribute to diseases of the oral cavity.
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{
"pile_set_name": "NIH ExPorter"
}
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The carcinogenic and non-carcinogenic mechanisms of As are incompletely understood, but an emerging body of evidence suggests that As exposure leads to epigenetic dysregulation. In 4 independent studies in our Bangladesh cohort we have demonstrated that chronic As exposure is associated with increased global DNA methylation, contingent on adequate folate status. We hypothesize that the mechanism underlying this relates to As-induced alterations in histone modifications. Two potential mechanisms include a) in vitro, As induces G9a mRNA and protein expression; G9a is a central player in epigenetic regulation, and b) As is a very potent inhibitor of pyruvate dehydrogenase, an enzyme that catalyzes the final step in Acetyl CoA biosynthesis; inhibition of Acetyl CoA biosynthesis may decrease histone acetylation leading to chromatin condensation and increased DNA methylation. Folate is a key regulator of one-carbon metabolism mediated methylation reactions, including methylation of DNA and histones. A large randomized trial is currently underway in Bangladesh to evaluate the effects of folic acid (FA) supplementation on As methylation. We propose a cross-disciplinary collaboration that will take advantage of a unique repository of samples collected from this trial to carry out a set of aims related to nutrition/environment interactions. In these aims, we will characterize the influence of As exposure on histone modifications, relate changes in histone modifications to changes in DNA methylation, and characterize the impact of FA supplementation on these marks. Finally, using the Intinium Human Methylation450 array, we will identify a set of genes that are differentially methylated by As exposure and determine gene-specific histone modifications at these loci. Collectively, these aims will begin to elucidate the molecular events that underlie the effects of As and folate on DNA methylation. The implications of identifying an influence of FA supplementation on histone modifications are considerable, as this represents a simple, low-cost, low-risk intervention as a potential therapeutic approach to reverse As-induced epigenetic dysregulation.
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{
"pile_set_name": "NIH ExPorter"
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The conformations in aqueous solution at physiological pH and temperature of enkephalins and their analogs will be determined using absorption spectroscopy and circular dichroism. The main features that will be determined are: 1) the existence, if any, of ordered back-bone structures (S, or other, bends); 2) the relative position of aromatic side-chains; 3) the extent of intra- and/or intermolecular hydrogen bonding of the hydroxyl proton on the tyrosyl residue. With this data any conformation-activity correlations will be deduced. These experiments will be supplemented by calculations of conformational energies using branch-chain rotational isomeric state theory. These calculations of conformational energies using branch-chain rotational isomeric state theory. These calculations will allow a determination of any confirmations of significantly lower energy and, lacking that, give the percentage of molecules in those conformations that are attainable at body temperature. In vitro testing of the binding capacity of these molecules to brain homogonates and in vivo testing of analgesic activity will be compared to distinguish between these two functions. Results will be compared to known conformations of morphine and its analogs in solution.
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{
"pile_set_name": "NIH ExPorter"
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The zebrafish (Danio rerio) has proved to be an outstanding animal model to explore vertebrate development. Ongoing activities in the zebrafish research community have resulted in a multitude of resources available to researchers using this animal model. For example, (1) there is an International Zebrafish Stock Center and (2) a variety of antibodies and cDNA libraries are available. Furthermore, the extensive genetic characterization that zebrafish have undergone, including the creation of YAC genomic libraries and microsatellite genetic linkage maps, provide a cutting edge ability to rapidly take advantage of this animal model. This wealth of resources has been under exploited for evaluation of biology or pathophysiology of mature zebrafish. Most scientists evaluate zebrafish at the embryonic/larval stages after mutagenesis or toxin exposure. Thus, it is plausible that many mutations or toxin mediated effects that occur in late life will not be identified. It follows that a wealth of information with relevance to human biology and disease is being overlooked. The goal of the current project is to understand some aspects of the basic biology of aging zebrafish. To accomplish this, the following specific aims will be performed: 1) Characterize zebrafish demopraphy. Experienced zebrafish investigators estimate that the zebrafish lifespan is approximately two to four years. However, a critical evaluation of median and maximum lifespan has not been performed. These data are necessary for understanding the normal biology of zebrafish for comparative medicine purposes. The PI will evaluate lifespan parameters in wildtype and AB strains of zebrafish. Additionally, the effect of population density on lifespan will be evaluated. 2) Create an atlas of anatomy and age-associated histopathology of zebrafish. Evaluation of gross, histologic, and clinical pathologic changes that occur during aging in the zebrafish have not been systematically explored. To provide information about healthy animals and age-specific pathology, aging zebrafish will be euthanized at 6-month intervals for anatomic and histological, and pathological evaluation. These images will be collated into atlases that will be published and presented on the ZFIN Web site. 3) Demonstrate the utility of the mature zebrafish as an animal model for aging research. It is well established that the aging process diminishes mammal's ability to adapt to environmental stress. This is manifested, in part, by decreased heat shock protein (HSP) response to stress. Thus, the PI will test the hypothesis that HSP response is decreased in aging zebrafish. The PI will subject young, mature, and old zebrafish to both heat and ethanol stress and evaluate their mortality rates and their ability to activate HSP production. The PI also will assess the activation of the HSP70 promoter in fish transgenic for the HSP promoter driving the green fluorescent protein cDNA. When this project is completed, the PI will have established important baseline parameters that should allow mature zebrafish to be a powerful research tool for investigators whom explore research areas germane to a wide variety of NIH Institutes.
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{
"pile_set_name": "NIH ExPorter"
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Failure of cells to respond to DNA damage is a key mechanism of toxicity by environmental agents and a primary step in the onset of cancer. In this research program, we are using systematic approaches to map and model the genetic networks underlying the DNA damage response (DDR). Since DDR pathways are widely conserved, our studies bridge between Homo sapiens and the budding yeast Saccharomyces cerevisiae? impacting our knowledge of how genotoxic agents lead to pathogenesis in humans, coupled to a classic model organism for which new genetic technologies are readily developed and deployed. In the past five years of funding (NIH grant R01-ES14811), we made significant progress in identifying changes in yeast genetic, transcriptional, and signaling networks in response to DNA damage stress. We also developed innovative new technologies, including the experimental technique of ?differential? genetic interaction mapping and a computational approach to translate interaction networks into hierarchical, data-driven gene ontologies. In the next period of support, we will: (1) Significantly expand the yeast genetic interaction maps to include dynamic growth curves and specific DDR pathway readouts at high-throughput; (2) Develop and apply CRISPR technology to create parallel genetic network maps in human cell lines; and (3) Integrate all new and prior data to build comprehensive ontologies of DDR subsystems in yeast and human, which we will compare to systematically identify areas of conservation and divergence and validate specific DDR phenotypic predictions in mechanistic assays. This work moves us closer to a comprehensive structure/function model of the DDR. A growing set of DNA- damage-induced genetic networks and ontologies in model species and humans are important resources for understanding genetic polymorphisms that predispose an individual to environmental DNA damage and DDR- related diseases.
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{
"pile_set_name": "NIH ExPorter"
}
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Youth substance use and abuse in the U.S. is a critical public health issue and a risk factor for morbidity and mortality in adolescents. Examination of risk for substance use has identified that maltreatment in childhood is a relevant factor; research has indicated that youth exposed to maltreatment are more likely than their non- maltreated peers to have substance use problems. In some cases, children are removed from their homes due to maltreatment or neglect, and recent research revealed higher rates of substance use disorders in youth residing in out-of-home placements when compared to peers in the general population. Although the relation between maltreatment in childhood and subsequent substance use is recognized, pathways related to risk and protection for substance use behavior and other negative outcomes following maltreatment have yet to be fully delineated. One important factor, coping behavior, has been implicated in the relation between adverse life events and subsequent behavioral and emotional outcomes. The association between substance use and coping remains unclear; many have hypothesized that substance use may actually be an indicator of maladaptive coping. Further, significant methodological weaknesses exist in the literature investigating the link between maltreatment and substance use in youth including overuse of retrospective reports by adults about childhood maltreatment and substance use, failure of researchers to investigate the psychometric properties of their measures of substance use or maltreatment, exclusion of relevant covariates, failure to include diverse types of abuse such as neglect and psychological maltreatment, and use of small samples. Previous reviews of the literature have highlighted the need for attention to these methodological issues and more research conducted with youth enrolled in out-of-home care. To address this gap, this study will provide a comprehensive investigation into the role of maltreatment as a predictor of substance use and coping behaviors in a sample of youth in foster care. To achieve this, the proposed study will utilize a structural equation modeling framework to assess competing models of moderation: 1) a model with maltreatment predicting substance use with coping behaviors as a moderator and 2) a model with maltreatment predicting coping strategies with substance use as a moderator. Preliminary analyses will examine the measurement model of the substance use measure, as well as potential covariates of substance use in the foster sample such as child and placement characteristics. Model testing will assess overall fit of each moderation model, then Participants will be drawn from a larger, longitudinal study funded by NIH entitled Studying Pathways to Adjustment and Resilience in Kids (SPARK) and will include 150 children and adolescents aged 8 years and older who are enrolled in Jackson County foster care. Improved scientific knowledge of the complex interactions of these factors from this study will provide a more comprehensive model for investigating associations between child maltreatment, coping behaviors, and substance use in foster care youth.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this project is to apply current methods in molecular biology to study disseminated neurofibromatosis (NF). The project involves collaboration with several NF clinical centers to insure the proper collection of tumor tissue, family pedigrees, and the preparation of EBV transformed cell lines for DNA analyses. The specific aims of the project are: (1) To select highly informative families for linkage studies in NF; (2) To test these families with specific and random DNA probes with the aim of locating a close marker to the NF gene; (3) To use possibly specific NF probes to get closer to the NF gene after a linked marker is located. Various technical possibilities are considered for speeding up the search for linked polymorphisms.
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{
"pile_set_name": "NIH ExPorter"
}
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Nephrotic syndrome (NS) is a common kidney disease, but the molecular mechanisms underlying the disease remain unclear for the vast majority of cases. We previously reported that the small heat shock protein, hsp27, a known regulator of actin polymerization, is highly expressed in glomerular podocytes (the cells most affected in NS), and that glomerular hsp27 expression and phosphorylation are significantly induced in experimental NS. We also reported that hsp27 is able to dramatically regulate (i.e. hsp27 overexpression protects, while reduced expression sensitizes) the podocyte response to PAN-induced cellular injury and actin cytoskeletal disruption. More importantly, we recently confirmed that glomerular hsp27 expression is induced in both multiple animal models of NS, as well as in human NS, suggesting that induction of glomerular hsp27 represents a generalized podocyte stress response. Since hsp27 has not been confirmed to bind actin directly in vivo, we attempted to clarify the mechanism of hsp27-mediated regulation of podocyte structure by screening a glomerular yeast two-hybrid library, and identified two novel hsp27 binding proteins: 1) Hic-5, a known focal adhesion protein and paxillin homologue, and 2) Arpda, a known member of the Arp2/3 actin polymerization initiation complex. We confirmed hic-5 as a true hsp27 binding protein by co-IP, mapped its interaction domains with hsp27, and showed that hic-5 can inhibit hsp27-induced thermo-protection in an interaction-dependent manner. We also confirmed both ArpCIa and ArpClb as true hsp27 binding proteins by co-IP and quantitative FRET analyses. Based on the above, we hypothesize that hsp27 plays a critical role in the regulation of podocyte structure and response to injury, and that these actions are mediated via its novel binding proteins, hic-5 (a paxillin homologue with a role in focal adhesion dynamics), and ArpC1 (a member of the Arp2/3 actin initiation complex). To test this hypothesis we will complete development of podocyte-specific hspbl (hsp25) gene-targeted mice and perform phenotypic analyses to definitively determine the role of hsp25 in podocyte biology. We will also determine if induced alterations in hic-5 expression or interaction with hsp27 can regulate podocyte: 1) Focal adhesion formation rate, composition, and function (i.e. adhesion), and 2) Response to injury. Lastly, we will determine if induced alterations in ArpC1 expression or interaction with hsp27 can regulate podocyte: 1) Arp2/3 complex function (i.e. initiation of actin polymerization) and composition, and 2) Response to injury. Confirmation of hsp27's importance in the regulation of podocyte structure, and identification of its molecular mechanism(s) of action in podocytes, could not only improve our understanding of the molecular mechanism(s) underlying the development of NS, but also permit the development of more highly targeted and/or less toxic therapies for this very common kidney disease. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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The goal of our proposed study is to investigate whether a transient period of diabetes remission after roux-en- y gastric bypass surgery (RYGB) induces a sustained benefit in long-term microvascular and macrovascular disease outcomes. Our project would be an expansion of our recent research in this area in which we utilize electronic medical record data from four large, integrated health plans and care delivery systems in the HMO Research Network (HMORN) to examine short and long-term health outcomes of over 9,000 severely obese individuals with type 2 diabetes who have undergone RYGB over an 18-year period (1996-2014). The longitudinal cohorts and rich, clinical and administrative data available at these HMORN sites are crucial for conducting valid, robust longitudinal studies of bariatric surgery because extensive adjustments, stratification, and sub-setting that are required. For our new application presented here, we propose to extend and expand our prior cohort in terms of size, duration of follow-up, and the number of clinically-important outcomes addressed. We will increase the follow- up period for our initial cohort by six years to further improve our ability to characterize the longer-term complications of diabetes. We will increase the size of our cohort by tapping a previously unutilized data source - H1,000 adults with diabetes who received RYGB at Group Health Cooperative. Finally, our major innovation will be to explore the legacy effect of RYGB through a series of analyses that characterize long- term, micro- and macrovascular outcomes across three patient groups: those who do not remit their diabetes after RYGB; those who experience durable diabetes remission after RYGB; and those who relapse their diabetes after an initial remission. Microvascular outcomes will include the following: (i) renal disease: decreased glomerular filtration rate and micro/macroalbuminura; and (ii) eye disease: diabetic retinopathy and clinically significant macular edema. Macrovascular outcomes will include the following: (i) cardiac ischemia, infarction, angioplasty/stent, or coronary bypass surgery; (ii) stroke or transient ischemic attack and (iii) peripheral vascular disease-related ulceration, revascularization or amputation This study will test the following novel hypotheses: (1) severely obese patients who experience a durable remission of diabetes after RYGB will have fewer incident microvascular and macrovascular complications compared to those who do not remit their diabetes, (2) the beneficial effects of a transient period of diabetes remission after RYGB will persist after diabetes relapse (legacy effect); i.e., relapsing patients will experience fewer complications than those who do not remit their diabetes after RYGB, and (3) the duration of diabetes remission and/or control after RYGB will be significantly associated with the risk of incident microvascular and macrovascular complications.
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{
"pile_set_name": "NIH ExPorter"
}
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Using the resources of an epidemiologically defined, longitudinal twin study, we examine developmental course and biopsychosocial risks for childhood psychopathology. Building on existing screening at ages 2 and 7 years, we characterize risk factors at ages 2, 7, and 12 relevant to the development of internalizing (anxiety and depression), externalizing (oppositional and conduct disorder), and ADHD. Most of our at-risk cases qualify for a DSM-IV diagnosis, yet we include individuals with sub-threshold symptoms and a large control group. Specific aims include characterization of risk factors, analysis of genetic and environmental effects, and improved measurement and classification of childhood disorders. "Child-based" risk factors include earlier symptoms, temperament and stress reactivity, basal and reactive cortisol, testosterone and DHEA, cognitive abilities and attributional styles, and cognitive-affective processing skills. Family and other psychosocial risk factors include parental diagnosis and family history of psychopathology, twin-twin and twin-parent social interaction styles, multiple facets of family stress, and negative parenting. The research methods that we employ include structured diagnostic interviews with caregivers and children, medical records, observer ratings, child self-report and parent-report questionnaires, videotaped home-based behavioral batteries, and computer-based testing (mostly reaction time tasks). The study's significance lies in understanding how known risk factors interact and/or mediate each other's effects on child psychopathology in a genetically informative, longitudinal design. Twin methodology allows us to parse phenotypic variance and covariance among measures into genetic and environmental components, and the components are studied developmentally. The results should also enhance our understanding of comorbidity, heterogeneity within disorders, and the association of disorders with traits. The project's public health relevance lies in its identification of risk factors for common childhood disorders and its investigation of how they interact in the context of genetic and environmental factors. The knowledge gained should inform efforts toward early detection, improved interventions, and better classification of childhood disorders. [unreadable] [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The inverse association between HDL concentrations and coronary artery disease (CAD) risk has prompted a search for hygienic factors which elevate HDL levels. Physical activity is an attractive area for intervention since endurance athletes demonstrate HDL levels 10-20 mg/dl higher than their sedentary counterparts. Exercise training in previously sedentary subjects, however, has demonstrated a much more modest exercise effect, and the mechanism by which physical activity increases HDL levels is not defined. We hypothesize that exercise augments the transfer of lipids from lower density lipoproteins to HDL and that this lipid enrichment retards HDL degradation. The proposed studies will examine this hypothesis. We will compare HDL metabolism and fat tolerance in active distance runners and sedentary men under controlled dietary conditions. We will also examine the effects of two years of exercise training with or without weight loss on HDL metabolism in previously inactive men. Parameters of HDL metabolism including apoprotein synthetic and catabolic rates will be measured using radiolabelled HDL. HDL cholesterol and apoprotein concentrations, the ultracentrifugal distribution of HDL lipids, and the activity of lipoprotein lipase and hepatic triglyceride lipase in postheparin plasma will be quantified. The clearance of intravenous fat will be measured as an index of fat tolerance. Questions which we will address include: Do initial HDL levels determine the change in HDL with exercise training? What is the magnitude of the exercise effect? Are exercise- induced changes in HDL magnified by concurrent weight loss? How does exercise training alter HDL metabolism? This research should provide insight into factors regulating HDL levels and the metabolic alterations produced by physical activity. Because of the inverse association between HDL and coronary artery risk, the results may also suggest strategies for CAD prevention.
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{
"pile_set_name": "NIH ExPorter"
}
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HIV-1 infection and AIDS disproportionately affect people in resource-limited settings. Although the neurologic complications of HIV infection are well-documented in western countries, the neurologic impact of HIV infection in China is unclear. HIV-1 transmission via drug abuse and sexual intercourse is primarily responsible for the current HIV/AIDS epidemic in China. This current proposal is written in response to the RFA-DA-08-005 entitled "International research collaborations to study HIV/AIDS and drug abuse (R21) to explore the feasibility of addressing the relationship between drug abuse and NeuroAIDS in China, a resource-limited developing country with a rapidly increasing number of HIV-infected individuals in the world. Our long-term goal is to better characterize the neurologic complications associated with HIV infection in China and their response to treatment, and to better understand their viral and host correlates. This study will build upon an established collaboration between HIV NeuroAIDS investigators at Indiana University School of Medicine in the United States and a diverse group of physician scientists at the First Affiliated Hospital of Xi'an Jiaotong University Medical College in Xi'an, China. We have worked together and identified and recruited a cohort of HIV+ and HAART naive individuals. For this one-year R21 application, we propose: (1) To establish a repository of cerebrospinal fluid (CSF), plasma, and peripheral blood mononuclear cells (PBMCs) from the cohort of HIV+ and HAART naive individuals; (2) To evaluate neurologic manifestations of the cohort of HIV+ and HAART naive individuals; (3) To evaluate neurocognitive impairment (NCI) of the cohort of HIV+ and HAART naive individuals. Comparison of the neurology and neurobehaviors of these patients with or without a history of drug abuse will allow us to address the roles of drug abuse in NeuroAIDS in China. These studies will also help to train NeuroAIDS researchers and build the drug abuse/NeuroAIDS research capacity in China. Furthermore, these studies shall add to our understanding of the neurobehavioral complications of HIV diseases in China and the viral and host correlates to NeuroAIDS, and ultimately lead to better informed clinical management of the affected population. [unreadable] [unreadable] HIV-1 infection often causes a number of brain diseases and affects the ability of people to care for themselves and thus the quality of their daily life. The social and economic impact can not be overemphasized. The current study seeks to have a better understanding of the prevalence and nature of these diseases in the selected population. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this project is to develop an improved, minimally-invasive treatment for intracranial aneurysms. Intracranial aneurysms are spherical outpouchings of blood vessels in the head that result from weakness in the vessel wall. Unruptured aneurysms are present in approximately 3% of the general population, and rupture can be devastating with a high morbidity and mortality. In recent years a new type of minimally- invasive endovascular device to treat aneurysms has been introduced to the market. These devices, known as flow diverting stents, are composed of a tubular metal mesh and are placed in the parent artery to cover the neck of the aneurysm being treated. The small pores in the mesh prevent blood from entering the aneurysm, which causes blood trapped inside the aneurysm to become stagnant and clot. The aneurysm then undergoes a process of healing with the stent acting as a scaffold for reconstruction of the parent artery. Despite their success, first-generation flow diverters suffer from a number of limitations such as delayed aneurysm occlusion, often on the order of months, and the need for multiple devices per patient to achieve complete occlusion. In as many as 25% of patients complete occlusion is never achieved. NeuroSigma's next generation flow diverter is based on Thin Film Nitinol (TFN) technology. TFN is an advanced biomaterial fabricated on silicon wafers that overcomes many of the limitations associated with first-generation flow diverters. Results from animal studies suggest that the TFN flow diverter gives superior rates of aneurysm occlusion and faster parent artery repair than first-generation devices. The purpose of this project is to build upon these intriguing findings an develop a rapid occlusion flow diverter, where placement of a single device results in speedy, definitive aneurysm repair. At the completion of this project the TFN flow diverter will be ready t begin first- in-human studies as an improved treatment for the millions who suffer from intracranial aneurysms worldwide.
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{
"pile_set_name": "NIH ExPorter"
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Vascular development results in a stable pattern of blood vessels that can be modified only by specific physiological or pathological signals. Blood vessel formation is based on endothelial cell responses to these signals and other cell-cell interactions of endothelial cells that are mediated through receptor molecules. Tyrosine kinase receptors that bind angiogenic factors, and endothelial cell surface adhesion receptors have functions consistent with a role in blood vessel formation in vivo. The specific involvement of these molecules in developmental and pathological blood vessel formation, however, has not been demonstrated. The experiments described in this proposal will test the hypothesis that endothelial cell receptors are important in blood vessel formation. The role of known receptors in developmental and pathological blood vessel formation will be investigated, and novel receptors that may have a role in blood vessel formation will be identified using murine models that we have developed: cultured mouse endothelial cells and a unique in vitro developmental system using mouse embryonic stem cells. The regulation of receptor expression will be investigated in a mouse endothelial cell line that responds to cytokine stimulation and undergoes morphological alterations, and this regulation will be compared to that of embryonic endothelial cells. Mouse embryonic stem cells that differentiate in vitro will be used to study the role of the known endothelial cell adhesion receptors and the effects of genetic perturbations of tyrosine kinase receptors on vascular development. The endothelial cell cultures will be used to generate cDNA libraries and to isolate novel receptor genes that may be unique to vascular development. Finally, the relationship between vascular development and hematopoiesis will be examined both in terms of lineage and function using the embryonic stem cell differentiation system. The results of these experiments will help to determine the role of endothelial cell receptors in the regulation of blood vessel formation, and with these mouse models both addition and deletion of genetic material by targeted manipulation of endogenous genes will be available as tools for further studies of vasculogenesis and angiogenesis. A molecular understanding of blood vessel formation will also help to develop therapies that target tumor angiogenesis and vascular diseases such as atherosclerosis.
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{
"pile_set_name": "NIH ExPorter"
}
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Phencyclidine (PCP) is a widely abused drug. A major site of action of PCP in brain is the PCP receptor that is part of the N-methyl-D-aspartate (NMDA) receptor/ion channel complex. PCP, by binding to PCP receptors, blocks a cation channel that is opened by NMDA receptor agonists. Glycine, by binding to a strychnine-insensitive receptor on the channel complex, facilitates channel opening by NMDA agonists. There is an urgent need to identify and characterize the PCP/NMDA/glycine receptor/channel proteins. These are currently poorly characterized. In order to characterize the proteins, highly specific receptor ligands must be developed. IN this application we are proposing to develop novel PCP and NMDA receptor ligands and photoaffinity radiolabels. The novel PCP receptor active drugs and affinity ligands are derived from the potent and selective drug MK801 and from two novel drugs developed in our laboratories, IDDC and N,N'- dinaphthyl-guanidine (DNG). Computer modelling will be utilized to develop yet other novel PCP receptor (affinity) ligands. Proposed affinity labels for the NMDA binding site are based on the antagonist CPP. The novel, radiolabelled affinity probes will be used to characterize the PCP and NMDA binding protein(s) associated with the PCP/NMDA/glycine receptor ion channel complex. In order to characterize the [3H]-glycine binding protein subunit composition model of the PCP/NMDA/glycine receptor complex and to elucidate the pharmacophore of the PCP-site. The affinity receptor protein(s) thus laying the groundwork for eventual cloning of the genes coding for the receptor/ion channel complex. A detailed analysis of the molecular structure of PCP/NMDA/glycine receptors is necessary to better understand the molecular basis of PCP abuse and to devise treatment strategies.
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{
"pile_set_name": "NIH ExPorter"
}
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The proposed research program involves an extensive study of cyclic AMP-dependent chloride secretion by rabbit colon, in vitro, which will include (a) evaluation of a model for active chloride secretion elicited by cyclic AMP which is suggested by our preliminary observation; (b) studies of the requirement for serosal Na in the Cl secretory process; (c) determination of the unidirectional influx of Cl from serosal solution into the epithelial cells; (d) studies of the effects of pharmacologic agents on chloride secretion; (e) studies of the mechanism of Na transport which appears to be involved in Cl secretion; and (f) studies of the effect of cyclic AMP on cell Ca distribution and the kinetics of Ca exchange. The overall aim is to (1) characterize the mechanisms responsible for cyclic AMP-induced Cl secretion rabbit colon; (2) elucidate the role of Na in this process, especially with regard to the role of Na transport in the energetics of Cl secretion; and (3) provide a critial appraisal of the role of intracellular Ca as a mediator of the secretory response. The results of these studies should provide a basis for the understanding of secretory processes of a variety of epithelial tissues and should provide insight into the mechanisms responsible for salt and water losing enteropathies.
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{
"pile_set_name": "NIH ExPorter"
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The rates of oxidation of the alpha and beta subunits of hemoglobin by ferricyanide were found to be about equal. Reaction of hemoglobin with hydroxylamine between pH 7 and 9 produces two unidentified low-spin triplets in the electron spin resonance spectrum. Although these triplets do not correspond in position to those produced by reaction of several simple amines with ferric hemoglobin, one does correspond in position to a triplet produced by reaction of heme with a thiol and imidazole, suggesting that the ferric ion position ordinarily occupied by a water molecule or hydroxyl in methemoglobin is replaced by a cysteine sulf-hydryl when hydroxylamine reacts with hemoglobin. After venous injection of about 15 mg pentavalent antimony per liter, no trivalent antimony above the limits of polarographic detection (approximately 1 mg per liter) could be found, suggesting that conversion to the more toxic trivalent ion does not occur during the killing of the leishmaniasis trypanosome.
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{
"pile_set_name": "NIH ExPorter"
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ABSTRACT Medulloblastoma is the most common malignant brain tumor that afflicts children. Despite therapy with surgery, radiation and chemotherapy, outcomes of these highly toxic treatments are sub-optimal with significant long-term morbidity. Medulloblastoma consists of 4 distinct molecular subgroups (Wnt, Sonic Hedgehog, Group 3 and Group 4). Among these, Group 4 medulloblastoma is the most common subgroup but its underlying biology is the least characterized. We have recently demonstrated a critical role for Enhancer of Zeste Homolog 2 (EZH2) in Group 4 medulloblastoma. EZH2 is the catalytic core protein of the PRC2 chromatin-remodeling complex, which catalyzes the trimethylation of histone3 lysine27 (H3K27me3) and mediates epigenetic silencing of genes involved in cell fate decisions, differentiation and cancer. We demonstrated that EZH2 is overexpressed in Group 4 medulloblastoma patients, regulates the proliferation and self-renewal capacity of medulloblastoma cells and that the H3K27me3 mark is enriched in Group 4 medulloblastoma patients associated with adverse outcomes. However, the mechanisms underlying EZH2 mediated control of medulloblastoma tumorigenesis are poorly understood. Our preliminary data demonstrates that EZH2 suppresses expression of key regulators of neuronal differentiation and promotes transformation of neural stem cells, raising the intriguing possibility that aberrant EZH2 expression enforces a neuronal differentiation block and maintains pluripotent state in neural and tumor stem cells. The exact gene expression programs regulated by EZH2 in medulloblastoma are unknown and the impact of abnormal EZH2 expression in the cerebellum is undetermined. We hypothesize that EZH2 mediates medulloblastoma tumorigenesis by inhibiting differentiation of cerebellar stem cells and maintaining a pluripotent state. Our objective is to examine the biological impact of EZH2 on medulloblastoma tumorigenesis and test potential novel therapeutic molecules targeting EZH2. To pursue our hypothesis we will first investigate the proposition that EZH2 suppresses gene expression of neuronal differentiation programs in medulloblastoma by altering chromatin occupancy of the H3K27me3 histone core at key promoters. Next we will pursue concept that aberrantly increased EZH2 expression in Group 4 specific cerebellar stem cells will inhibit differentiation and induce tumor formation in the murine cerebellum using a novel mouse model of medulloblastoma. Finally we will test three clinically relevant inhibitors in vivo using patient derived xenograft models of medulloblastoma. Successful completion of the proposed work will determine the role of EZH2 in medulloblastoma and establish the potential of therapeutically targeting this enzyme in clinically relevant in vivo models.
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{
"pile_set_name": "NIH ExPorter"
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This Phase I proposal is a collaborative, multi-disciplinary effort involving researchers from TPL, Inc. (Albuquerque, New Mexico) and the Department of Bioengineering at the University of Washington. Our objective is to explore a new generation of miniature optical imaging probes for high-resolution intraluminal imaging of neoplasia at early stages. Development of a technology that enables in vivo imaging of biological tissues at or near cellular level in real time could permit diagnosis of neoplasia at early stages and precision guidance of biopsy. Optical coherence tomography (OCT) and confocal microscopy are capable of real-time imaging of biological tissues at or near cellular level, functioning in a form of "optical biopsy" but without the need for tissue removal. In vivo and endoscopic applications of OCT and confocal microscopy require a scanning miniature probe for real-time imaging and a mechanism for real-time focus tracking in order to maintain a high transverse resolution at varying depths. The proposed imaging probe integrates beam delivery, collection, focusing, and fast transverse scanning into a highly compact single unit. Unlike conventional endoscopic imaging probes, the proposed miniature probe performs rapid transverse beam scanning, which enables a new image acquisition scheme to achieve real-time-focus tracking and real-time ultrahigh-resolution optical coherence tomographic imaging in the forward direction. The specific aim of this Phase I exploratory study is to develop an extremely compact, rapid scanning, forward-imaging probe based on a novel fiber-optic resonant scanner and cutting-edge micro imaging optics. The imaging probe can integrate endoscopic OCT with confocal microscopy to achieve ultrahigh-resolution imaging in real time. A successful Phase I study will warrant further relevant technology development refinement as well as pre-clinical feasibility investigations planned for Phase II. If both Phase I and II are successful, this technology will lead to a new generation of miniature imaging devices for high-resolution, non-invasive optical biopsy of neoplasia at or near cellular level in vivo and in real time.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary/Abstract Stroke is the leading cause of long-term disability worldwide. However, despite extensive rehabilitation efforts, marked physical inactivity remains emblematic of individuals in the chronic phase of stroke recovery and contributes to poor health and quality of life. For poststroke rehabilitation to be better positioned to enable more physical activity, factors such as insufficient training dosage and poor ecological validity, which limit the effectiveness of clinic-based gait training, must be overcome. Community-based rehabilitation approaches can overcome these limitations; however, the amount of walking practice that they are able to leverage is inherently limited by the reduced walking capacity typical after stroke. Indeed, recent work has shown that despite producing some improvement in poststroke walking activity, individuals still remain largely sedentary after community-based walking programs. A critical need thus exists for rehabilitation technologies that bridge the gap between the clinic and the real world and facilitate the massed walking practice in salient contexts that is necessary to induce experience-dependent neuroplasticity. In response to this need, we are developing lightweight and unobtrusive soft wearable robots (exosuits) suitable for community-based rehabilitation after stroke. Exosuits use innovative force-transmitting, conformal textiles that anchor to the body and proximally-mounted, lightweight, cable-based actuation systems to generate assistive joint forces. Adaptive control algorithms that use signals from the minimum number of sensors direct the timing of assistance. These innovative wearable robots have the potential to increase an individual's capacity for community ambulation. Indeed, we have demonstrated through preliminary treadmill-based studies that an ankle-assisting exosuit produces paretic limb functional restoration in a manner that induces gait symmetry, reduces the energy cost of walking, and increases walking stability. In contrast to the rigid exoskeletons that have dominated the last half century, exosuits provide partial assistance through a lightweight platform (<3kg overall, including motors and batteries) that requires low power (<100W), can be used unilaterally, and does not impose an unnatural gait on the wearer. The exosuit technology is ideally suited to assisting the impaired gait of individuals poststroke who have residual walking capacity. This technology development-focused project seeks to build gait-restorative exosuits made of modular components that can be easily assembled to serve the heterogeneous needs of individuals poststroke. This project will optimize existing ankle, knee, and hip modules that have undergone >1000 hours of testing on >130 healthy and impaired subjects for use by persons poststroke in clinical- and community- settings. This work will be completed by a multidisciplinary team of engineers, clinicians, and researchers. Qualitative and quantitative data gathered from patients and other stakeholders will inform all development.
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{
"pile_set_name": "NIH ExPorter"
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Diabetic and non-diabetic nephropathies and the Metabolic Syndrome are associated with chronic kidney disease (CKD) - a program comprising proteinuria, accumulation of profibrotic collagen in the renal interstitium and relentless decline in renal function. CKD remains an unsolved challenge for the nephrologist, as it almost inevitably leads to end-stage renal disease, a life-threatening condition that necessitates renal replacement therapy. Currently approved therapies aimed at halting CKD are incompletely effective, evidenced by the dramatic and rising rates of requirement for renal replacement therapy. Scatter factor/hepatocyte growth factor (SF/HGF) is a renotrophic factor that is mitogenic for renal epithelial cells, exerts antifibrotic effects within the renal interstitium and stimulates renal repair. Clinical feasibility of SF/HGF administration is however limited by a number of logistical and financial issues relating to gene or protein therapy, especially in the setting of chronic diseases wherein orally bioavailable therapeutics are preferred. Using a drug-discovery engine comprising phage display, molecular modeling, rational drug design, targeted proteomics and preclinical biology, Angion Biomedica Corp. has identified BB3, an orally bioavailable small molecule SF/HGF mimetic. BB3 activates the SF/HGF receptor c-Met and recapitulates the bioactivities of SF/HGF in every assay tested to date. Similar to SF/HGF, BB3 downregulates the fibrotic program in activated astrocytes, hepatic stellate cells and renal fibroblasts. In preclinical models of dermal, pulmonary and hepatic fibrosis, BB3 is therapeutic, opposing profibrotic collagen deposition and preserving tissue microarchitecture. Our SBIR Phase I data indicate that orally administered BB3 is therapeutic in experimental CKD, attenuating mortality, limiting proteinuria and opposing renal interstitial collagen deposition. Additionally, data from a pilot study suggest that in a preclinical model of Metabolic Syndrome, oral administration of BB3 reduces proteinuria and attenuates renal interstitial collagen accumulation. From a drug development and regulatory perspective, a chronic (6- month), oral, escalating-dose, good laboratory practice safety study of BB3 in rat and dog is ongoing under the aegis of the NIH Rapid Access to Intervention Development program. Of significant clinical relevance, BB3 has successfully completed a Phase I acute safety clinical trial in healthy volunteers;a Phase IIA pharmacokinetics and tolerance study of BB3 in patients presenting with renal insufficiency is underway. The present SBIR Phase II application is designed to make a comprehensive and in-depth analysis of BB3 efficacy in preclinical models of CKD. Dose-dependence, dose schedule and therapeutic time window of BB3 will first be established in experimental CKD. Renal antifibrotic effects of BB3 will be evaluated together with standard-of-care and in models comprising co-morbid disease. Successful completion of the proposed work, together with the current clinical status of BB3, will enable us to enter this drug candidate into a safety and efficacy trial in CKD. PUBLIC HEALTH RELEVANCE An orally bioavailable small molecule antifibrotic has significant clinical potential for the treatment of chronic kidney disease.
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{
"pile_set_name": "NIH ExPorter"
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In cardiovascular research there is an urgent need for methods which would quantitatively identify: regions of high shear stresses in the flow (hemolytic factor), regions of high and low stresses at the will (atheromatous lesions), separated or reverse flow regions (thrombogenic factors), and functional (physiologic) severity of a stenosis. A computer model capable of simulating physiologically important features of blood flow in vessels would be an indispensable tool in research, education, and clinical practice. The objective of Phase II is to develop such computer software. The computer model will be able to calculate three-dimensional steady or time-dependent flow for an arbitrary vessel geometry including lesions and bifurcations, and a wide range of Reynolds numbers. In Phase I we developed a two-dimensional and axisysmmetric version of such a program. In Phase II the program will be extended to three dimensions and we plan to develop and/or upgrade: an ergonomically designed user-friendly menu system; a machine interface to read digitized arteriograms; color graphics; and a computer-aided geometry input system.
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{
"pile_set_name": "NIH ExPorter"
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The central thrust of this program is the study of the etiologic and pathogenetic factors involved in immunopathologic responses directed toward endogenous antigens. Among the endogenous antigens are included traditional autoantigens, viruses and tumor antigens since operationally the etiologic factors and pathogenic consequences of immune responses to each appear quite similar. Experiments are designed to investigate: 1) the control mechanisms which normally establish and maintain self tolerance, 2) the ways in which such controls may fail or be circumvented thereby permitting autoimmune responses, 3) the interaction of the immunologic effectors, antibodies and sensitized cells with endogenous or auto-antigens and 4) the pathogenic processes set in motion by the immunologic reactions. Specifically we will 1) analyze the behavior of lymphocyte surface glycoproteins during activation and cell-cell interactions as a means of studying regulatory processes, 2) investigate the processes of tolerance induction and termination and autoimmunity which may be associated with the latter, 3) make a comprehensive study of murine SLE concentrating on genetic and other etiologic factors and the critical pathogenetic processes leading to disease, 4) investigate the immunologic and nonimmunologic host factors which may influence the pathologic impact of immune complexes and 5) study mechanisms which permit viruses to remain latent and the effects of viral infection on the function of the immune system.
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{
"pile_set_name": "NIH ExPorter"
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Project 1 Cowley, Jr., Allen Wilson INDIVIDUAL PROJECTS: PROJECT 1 MOLECULAR &FUNCTIONAL REGULATORY NETWORK IN HYPERTENSION Project Leader: Allen W. Cowley, Jr., Ph.D., Professor of Physiology Co-lnvestigators: Mingyu Liang, M.B., Ph.D., Associate Professor of Physiology Howard Jacob, Ph.D., Professor of Physiology Andrew Greene, Ph.D., Professor of Physiology ABSTRACT Essential hypertension affects more than 50 million Americans and increased blood pressure salt-sensitivity is a prominent feature in certain populations of hypertensive patients, especially African Americans. Although it is evident that the common forms of hypertension are multiifactorial (polygenic and environmental), the genetic basis of the frequent forms of this disease remain elusive. The Dahl salt-sensitive (SS) rat is a widely used animal model that recapitulates many aspects of human salt-sensitive hypertension and associated renal injury. The goal of Proiect 1 is to study the interplay of gene and protein expression using integrative systems approaches to determine the functionality of a single cell type of the kidney (renal medullary thick ascending limb of Henle = mTAL). The mTAL is known to play an important role in renal medullary function, sodium excretion, and hypertension in the SS rat and in human hypertension. We hypothesize that gene(s) sequence variants within a congenic region of chromosome 13 (SS.IS^*^^(R);BN alleles substituted for SS) alter molecular regulatory networks affecting function of the mTAL thereby contributing to salt-sensitive hypertension and renal injury. In Aim 1, the congenic region (SS.IS^'^^(R);13.2 Mb) will be sequenced to obtain a finished high-quality assembly of this region and annotated to search for gene sequence variants. Subcongenic strains will be developed to identify a narrow region that attenuates salt-sensitivity to ~ 5 Mb and a sequence capture chip will then be utilized for follow-up sequencing of this region in 14 additional strains that have a different evolutionary history from the SS and BN. The sequence differences (SNPs and other genomic variants) will be filtered using the criteria that a casual variant in the SS should only be in common with other salt-sensitive strains. Aim 2 will construct a molecular and physiological regulatory network (BayeN) of the mTAL epithelial cell and use that model to identify pathways and genes that may contribute to salt-sensitive hypertension and renal injury in SS rats. Aim 3 will select a candidate gene based on results from Aims 1 and 2 and either knock the gene out using zinc finger nucleases (ZFN) and/or over-express the gene using transgenic approaches to study the impact of this gene upon the transcriptome/proteome/metabolome and associated functional network. Several technological leaps and conceptual approaches are unique to this Project. These include 1) Next generation sequencing to provide very high resolution sequencing of a 13.2 Mb region of SS and congenic SS.13^'^" strains coupled with a NimbleGen sequence capture chip for sequencing of additional rat strains;2) Transcriptome analysis (Affymetrix) and a mass spectrometry sub-proteome and metabolome analysis using isolated and purified mTAL cells comparing the SS and salt-resistant congenic rats;3) The use of a large-scale Bayesian analysis which is "knowledge-constrained" to seek yet unknown pathways hidden using the transcriptome and functional data;4) A novel technology (zinc finger nucleases) to efficiently knock out a gene found to contribute importantly to the mTAL regulatory network, providing a definitive way to validate and characterize functional relevance. Project 1 addresses a critical challenge facing the field of hypertension: to identify the complex components (genes, proteins and pathways) that are responsible for alterations in kidney function leading to salt-sensitive hypertension. It is highly synergistic with Project 2 which tests the innovative concept that non-protein-coding genes (microRNA) may play an important role in hypertension and renal injury and Project 3 which aims to Identify the mechanisms by which mutations distant from the renin gene on chr 13 regulate the activity of renin and angiogenesis in the SS rat. All projects benefit greatly from genomic, genetic, proteomic and computational technological advances provided by Cores 8 and C. 75 Program Director/Principal Investigator (Last, First, Middle);Cowley, Jr., Allen WllSOn Project 1 DETAILED BUDGET FOR INITIAL BUDGET PERIOD FROM THROUGH DIRECT COSTS ONLY 07/01/11 06/30/12 List
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{
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Project Summary The vestibular sensory epithelia encode dynamic and static head movements in trains of action potentials that project to the central nervous system along primary afferent neurons. These signals provide the principal drive for behaviors such as the vestibulo-ocular reflex, which functions to stabilize visual gaze during head movements associated with dynamic behaviors, particularly during locomotion. While the response dynamics of vestibular afferents have been widely studied in a variety of animal models, the input/output relations have been limited to preparations that are restrained and/or anesthetized. Recent investigations using lower vertebrates have shown that peripheral vestibular stimulus processing is modulated by centrifugal efferent feedback driven by rostrally-projecting efference copy originating in spinal locomotor pattern generators. Therefore, critical insight into behaviorally-relevant peripheral vestibular stimulus processing would be gleaned under conditions of awake, behaving preparations. At present, data collected under these conditions do not exist. Therefore, such information requires the development of preparations in which the dynamic response characteristics of vestibular afferent neurons are investigated under conditions of natural locomotion. The present proposal aims to achieve this goal through the development of methods to record from individual vestibular afferent neurons in behaving chinchillas, agile rodents commonly used in studies of peripheral vestibular neurophysiology. There are two factors that support the use of this animal model for these studies: 1) the superior vestibular nerve can be accessed from the middle ear, precluding the need for an intracranial approach to the afferents; and 2) these animals tolerate chronic preparations very well. Critical to this project is the availability of a miniature electrophysiology platform that incorporates a 9 degree-of-freedom movement sensor along with a multichannel electrode headstage and on-board data storage. This hardware will enable the direct correlation of afferent discharge during head movements that occur during unrestrained natural locomotion. The successful development of these methods in an agile animal model will provide the foundation for future investigations of vestibular afferent dynamics under a variety of behavioral and treatment conditions, and will transcend the limitations of imposed upon our understanding of head movement coding by the constraints of passive stimulus presentation in restrained and/or anesthetized conditions. In so doing, the research aims of this application address the Functional Connectivity topic of research Priority Area 1 of the NIDCD Strategic Plan. In addition, the methods developed are seminal to understanding the role of vestibular sensory contributions to spatial navigation and orientation behaviors, and as such address other critical research priorities as identified in the Strategic Plan.
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{
"pile_set_name": "NIH ExPorter"
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Physiologic concentrations of melatonin have been previously shown by us to increase estrogen receptor concentrations in vitro in a human breast cancer cell line. This in vitro model system is being investigated further to determine if this will be a reliable biological response and to assess what hormonal parameters affect the melatonin response of these cells to melatonin. The present data indicate that melatonin promotes estrogen-stimulated growth of MCF-7 cells implanted in nude mice and promotes tumor growth in vitro of insulin-stimulated MCF-7 cells.
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{
"pile_set_name": "NIH ExPorter"
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The Illinois Center for Occupational and Environmental Safety and Health exists to improve, promote, and maintain the health of workers and communities by applying innovative and interdisciplinary approaches to: 1) prepare professionals to be leaders in occupational and environmental safety and health who will direct and manage occupational and environmental safety and health programs, teach other occupational and environmental health professionals, and research issues pertinent to occupational and environmental safety and health;2) provide continuing education to occupational and environmental health and safety professionals and outreach to workers and communities to improve their knowledge, skills, and awareness of key issues in occupational and environmental safety and health, devoting special attention to the problems and needs of at risk and underserved workers and communities;3) contribute to the knowledge base in occupational and environmental safety and health by preparing doctoral students, performing faculty and student research on problems of regional, national, and global significance, and disseminating the results of their research;and 4) serve as a regional information resource. The Occupational and Environmental Health and Safety Education and Research Center (Illinois ERG) is comprised of 14 programs. There are 7 academic programs: Industrial Hygiene (IH), Hazardous Substances (HSAT), Occupational Medicine (OM), Occupational Health Nursing (OHN), Agricultural Safety and Health (ASH-A), Occupational Safety (OS), and Occupational Epidemiology (OE) There are three continuing education and outreach programs: Continuing Education in industrial hygiene, occupational medicine, occupational health nursing, and occupational safety (CE);Hazardous Substances (HST);and Agricultural Safety and Health (ASH-CE). The ERC also has Center Wide Activities including a Center Administrative Core, Outreach, Diversity Recruitment, and Interdisciplinary Coordination (CWA), as well as a Pilot Projects Research Training program (PPRT) and Targeted Research Training program (TRT). This application is the competitive renewal application for Occupational Health Nursing. The Illinois ERC has made a significant impact in meeting the regional and national needs for occupational safety and health training. As seen by the number of trainees, continuing education offerings, publications, presentations, and our relationships with other academic programs and community partners, we have advanced the field of occupational safety and health in our region and in the US. In the previous project period we have established an outstanding record in training and are a recognized entity in the field of occupational safety and health in our region and nationally.
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{
"pile_set_name": "NIH ExPorter"
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Autosomal dominant polycystic kidney disease (ADPKD) is a major cause of morbidity and is the fourth leading cause of ESRD In the world, affecting more than 500,000 U.S. citizens. Researchers at the University of Alabama, Emory University, University of Kansas, Mayo Clinic and Washington University joined together in 2000 to create the Consortium for Radiologic Studies of Polycystic Kidney Disease (CRISP I) and In 2006 included the University of Pittsburgh in place of Washington University for CRISP II. The primary objectives of CRISP I and II were to: establish accurate, reliable and reproducible magnetic resonance based measurements of total kidney volume (TKV), liver cyst volume (LCV), renal blood flow (RBF), and patterns of cyst growth and expansion. Based on 7.3 years of longitudinal follow-up in 200 CRISP I/I I participants, we can now 1) establish an unequivocal relationship between TKV and qualitative (patient reported outcomes) and quantitative (renal insufficiency) end-points;as well as 2) identify potential modifiable risk factors associating with TKV and LCV to intervene upon. TKV ultimately may be used as a surrogate marker of disease progression in clinical trials. The goals of CRISP III extend the observations of CRISP l/ll. The overarching Aim for CRISP III is to develop and enhance prediction models that best predict renal insufficiency in ADPKD. Specifically, Aim 1;Extend the serial quantification of TKV and LCV to develop and test new models for predicting the risk of developing renal insufficiency. Aim 2: Determine the extent to which age and sex-adjusted measurements of RBF predict the rate of change in TKV and determine if RBF and TKV independently predict the risk of developing renal insufficiency. Aim 3: Develop methods to quantify the influence of renal cyst number, volume, and topography at baseline on the subsequent course of TKV and GFR and the risk of developing renal insufficiency. Aim 4: Expand and analyze CRISP biological samples to improve genotype/phenotype and biomarker studies. Aim 5. Determine whether intensive dietary counseling and intervention in a small group of CRISP participants is successful in modifying the relatively fixed pattern of sodium intake observed in CRISP I and reducing the rate of growth of the polycystic kidneys. PUBLIC HEALTH RELEVANCE: The results of these studies will impact the lives of patients with ADPKD. Developing predictive markers of disease severity early, prior to loss of renal insufficiency will result in increased efficiency of theray/drug development and result in increased life expectancy and improved quality of life in patients with ADPKD.
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{
"pile_set_name": "NIH ExPorter"
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We have initiated studies to identify the oxidative cytotoxins of stimulated leukocytes and their cytocidal mechanisms. We have shown that the bactericidal reactions of leukocyte peroxidase- generated hypochlorous acid (HOCl) and chemically similar chloramines cause destruction of metabolic energy reserves within the cell and loss of energy-linked functions such as active transport. The locus of oxidative attack is the bacterial plasma membrane; the metabolic dysfunctions leading to massive net ATP hydrolysis can be traced primarily to selective oxidative inactivation of nutrient transport systems and the proton translocating ATP synthase that links respiration to oxidative phosphorylation. We are applying several analytical methods to identify the molecular sites of attack on the ATP synthase F1 complex that give rise to its inactivation. The common response that we observe by several bacteria with disparate metabolic capabilities and the need for all cells to maintain ATP- synthesizing capabilities suggests that these reactions are capable of accounting for the "universal" character of HOCl toxicity towards prokaryotes. In the proposed research, we will investigate inactivation of microbial eukaryotes, whose ATP- synthesizing enzymatic systems are mitochondrial, hence nominally protected from extracellularly-generated HOCl. We will initiate parallel studies to identify the lethal reactions of hydroxyl radical and similar active oxygen species (high-valent metal oxo ions) that might be formed in metal-catalyzed reactions involving H2O2. We will compare loss of viability with specific metabolic functions or individual biochemical components to identify the loci of oxidative attack associated with cellular death. For this purpose, we will use differing methods of forming hydroxyl radical to allow its generation in the extracellular medium, at the surface of the bacterial cell, or intracellulary. Following protocols that we have developed for the studies with HOCl, we will examine the consequences of OH and Fenton-type chemical systems upon bacterial transport and respiratory systems, biosynthetic capabilities and selected cytosolic enzymes, nucleotide phosphorylation, and lesions in nucleic acids. Organisms exhibiting cleanly differing mechanisms of oxidative inactivation by HOCl and OH will then be used to probe for the primacy of one or the other within neutrophil phagosomes. These studies will clarify leukocytic mechanisms comprising host cellular response to infection and should yield insights into the nature of chemotactic and inflammatory responses.
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{
"pile_set_name": "NIH ExPorter"
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Studies on metabolism, pharmacokinetics and clinical pharmacology of new medications.
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{
"pile_set_name": "NIH ExPorter"
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Obesity is increasingly recognized as a major public health problem that is showing an alarming rise in prevalence in young people. While environmental factors, such as high-fat diets and lack of exercise, account for much of this rise, genetic differences may make some people more prone to obesity than others, even when diet and activity levels are similar among them. Obesity often has its genesis during childhood and, especially, adolescence, when differences among individuals in body fat amount and distribution tend to develop during puberty. It is thus important to understand the factors that affect growth, development, and body composition during this period. We will investigate the genetic factors that influence growth and body fat deposition during adolescence by using data and biological samples previously collected during Project HeartBeat!, a longitudinal study of cardiovascular disease risk factor development during childhood and adolescence. In two Texas communities, schoolchildren from 8-18 years of age were measured up to 3 times annually over a 4-year period, encompassing the ages when puberty normally occurs and adult body habitus usually develops. Subjects were enrolled at 8, 11, or 14 years of age to establish 3 separate cohorts that would overlap in age as the study progressed and permit construction of a synthetic cohort encompassing the full 10-year age range of the participants. The sample included 136 African Americans (70 females) and 506 non-Hispanic whites (250 females), with a mean of 8.6 examinations per subject. In each individual, we will genotype multiple single-nucleotide polymorphisms (SNPs) in 80 genes from hormonal systems important in growth and development, emphasizing genes known or suspected to affect body size and body fat distribution. In each gene, all promoter region and exonic SNPs, together with haplotype-tagging SNPs identified in public databases by 3 different algorithms, will be typed. A 3-stage algorithm will be used to identify a subset of SNPs in which multilevel mixed models will be used to analyze the associations of both individual variants and multilocus haplotypes with variation in longitudinal growth curves for traits related to obesity and body fat distribution, such as body mass index, % body fat, and various skinfolds. By targeting genes known to be involved in the development of body size and composition, we will significantly increase our understanding of the genetic factors that contribute to interindividual variation in growth and body fat distribution. Our analyses may also help identify genetic variants that may predispose some individuals toward obesity.
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{
"pile_set_name": "NIH ExPorter"
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ABSTRACT: The goal of proposed project is to determine the molecular identity of myosin light chain phosphatase phosphatase (MYPT1 phosphatase) and clarify the regulatory role of this poorly investigated critical component to create a molecular and cellular basis for understanding of the physiology and pathophysiology of smooth muscle contraction. Smooth muscle contraction is regulated by the Ca2+ independent pathway in addition to the well known Ca2+ dependent pathway. The key component of the Ca2+ independent pathway is myosin light chain phosphatase (MLCP), whose activity is regulated by the phosphorylation of the regulatory subunit of MLCP, called myosin targeting subunit 1(MYPT1). The research in the past has centered on the RhoA/ROCK pathway, a protein kinase phosphorylating MYPT1. However, recent studies have suggested that the Ca2+ independent regulation of MLC phosphorylation cannot solely be explained by RhoA/ROCK. We propose that MYPT1 phosphatase is the missing regulatory component that explains the unsolved research problem for understanding smooth muscle contractile regulation. Nothing is known about this important regulatory component. Our recent results have suggested that MYPT1 phosphatase is regulated during the contraction-relaxation cycle in smooth muscle (Nakamura et al., 2007). Furthermore, MYPT1 phosphatase is not inhibited by CPI17,which potently inhibits MLCP activity, suggesting that MYPT1 phosphatase is a different molecule from MLCP. Based upon these findings, we propose the following hypothesis. External stimuli alters the MYPT1 phosphatase activity, which causes the change in the MYPT1 phosphorylation level, thus regulates MLCP activity concertedly with the regulation of the RhoA/ROCK pathway. The proposed project will address this hypothesis. First we will isolate MYPT1 phosphatase from smooth muscle and determine the partial amino acid sequence of the subunits of MYPT1 using a Mass Spectrometry technique. Based upon the sequence information, we will identify the genes encoding the MYPT1 phosphatase holoenzyme and functionally express this enzyme (Aim 1). We will then study the characteristics and the regulation of MYPT1 phosphatase at the molecular level. A key question is how MYPT1 phosphatase activity is regulated. We hypothesize that the non-catalytic subunits of MYPT1 phosphatase play a key role in the regulation, and we will study the regulatory function of the non-catalytic subunits including the effect of phosphorylation using Mass Spectrometry analysis (Aim 2). In Specific Aim 3, we will test the effect of elimination of the identified MYPT1 phosphatase on MLCP activity and MLC phosphorylation in smooth muscle to confirm the importance of the identified MYPT1 phosphatase. Finally we will examine the regulation of MYPT1 phosphatase in smooth muscle by external stimuli. It is anticipated that the obtained information of MYPT1 phosphatase will provide a clue to understand the physiology and pathophysiology of organs containing smooth muscle.
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{
"pile_set_name": "NIH ExPorter"
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In spite of the progress made in the understanding of the biology, genetics and immunology of melanoma, the outcome for patients with advanced-stage disease has remained poor. A step forward toward better therapies was recently provided by the improvement in overall survival observed in melanoma patients treated with an anti-CTLA4 antibody. Attempts to further augment the efficacy of this treatment would still face however a variety of immunosuppressive factors operative in melanoma-bearing hosts. Among those, one that has gained much attention is the ability of melanoma tumors to induce T-cell tolerance. Our studies to date of the epigenetic regulation of T-cell unresponsiveness point to histone deacetylase inhibitors (HDI) as promising immunomodulatory compounds given their dual ability to influence the immunogenicity of melanoma tumors and enhance T-cell function. These observations together with our additional findings that HDAC6 and HDAC11 regulates melanoma immunogenicity and T-cell responsiveness respectively, provided the rationale to mechanistically address the role of HDACs in melanoma immunobiology. The hypothesis to be tested is therefore whether epigenetic manipulation of specific HDACs might augment the immunogenicity of melanoma cells and/or augment T-cell responses leading to breaking of immune tolerance and enhancement of the efficacy of CTLA4 blockade. The animal models, molecular and pharmacological tools we have in hands together with the access (through the Pathology Core of this SPORE) to human melanoma samples would allow us to gain insights into the role of HDAC6 in melanoma proliferation, survival and immunogenicity (Aim 1), and the role of HDAC11 in T-cell anti-melanoma immunity (Aim 2). In addition, the expertise provided by the Clinical Core will allow the successful completion of a Phase I clinical trial aimed to assess the safety and immunologic effects of HDAC inhibition in combination with an anti-CTLA4 antibody in patients with stage IV melanoma (Aim 3). The new knowledge to be generated by this team effort would lead to novel epigenetic-based immunotherapy that by overcoming the remarkable barrier of melanoma-induced T-cell tolerance would improve the efficacy of CTLA4 blockade.
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{
"pile_set_name": "NIH ExPorter"
}
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Most asthma exacerbations in children and young adults result from rhinovirus (RV) infections. As the most important cause of asthma-related morbidity it is essential that clinical investigations be performed in humans to define the underlying mechanisms. We hypothesize that the immune responses generated in the nose of asthmatics underlie subsequent systemic modulation of the immune system, and that - in susceptible individuals (those with pre-existing asthma) - this modified nasal milieu is responsible for the asthma exacerbation. Specifically, we propose that this modification produces a distinct pattern of immune responsiveness to RV in the upper airway of asthmatics, which triggers the development of a Th2 cytokine signature state that drives the adverse outcome of RV infection in the lower airway of asthmatics. In addition, we propose that the intensity of this Th2-inducing nasal airway milieu is further exaggerated in these allergic asthmatics, by a concomitant defect in the development and expression of effective anti-RV immune responses, leading to greater susceptibility to RV. Determining the etiology of rhinovirus-induced asthma exacerbations will identify specific targets to prevent and treat these episodes. Specific Aim 1 will address the hypothesis that epigenetic changes develop in nasal epithelial cells (EC) during the evolution of allergic airway disease as a result of which nasal EC are programmed to produce cytokines central to orchestrating an allergic inflammatory immune response. We will primarily determine whether nasal epithelium from allergic asthmatics, when infected with RV, is programmed to secrete cytokines that promote a Th2 cytokine signature (IL-25, IL- 33, and TSLP). Specific Aim 2 will interrogate the complementary hypothesis that increased susceptibility to RV and extent of nasal infections in asthmatics amplifies the consequences of this Th2-inducing bias. Specifically we propose that over time nasal epithelium in asthmatics is epigenetically re-programmed, resulting in the greater susceptibility of EC to RV infection. Initially we will analyze nasal EC ex vivo to test the hypothesis that EC from asthmatics will be more susceptible to RV infection as manifested by a greater magnitude of RV replication, a more rapid tempo of infection and overall greater death of RV-infected cells. We will then corroborate this in vitro analysis with in vivo studies by infecting asthmatics and controls with RV, monitoring viral load over time as a measure of the pace of infection. Most importantly, we will perform nasal biopsies at the peak of infection to determine the extent of viral infection and whether this represents cytopathic (necrotic) or apoptotic cell death. And, finally, Specific Aim 3 will address the molecular and cellular basis for the defect in anti-viral immunity in asthma. We will establish primary cultures of EC from control and asthmatic individuals prior to RV infection and examine them for baseline- and RV infection- induced expression of cytokines central to anti-viral immunity (IFNs-?, -, and ? and IL-15). We expect that anti-viral mediator expression will correlate inversely with the susceptibility, tempo, and severity of RV infection.
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{
"pile_set_name": "NIH ExPorter"
}
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Core A, the Administrative Core, is a central convergence point for all faculty, staff, postdoctoral fellows and graduate students affiliated with the program. This core provides secretarial and accounting support for participants in all projects and cores, in accordance with institutional and federal regulations. It processes manuscripts, letters, reviews, overnight packages and other professional activities. Core A coordinates the program review by Internal and external advisors. This core is directed by Dr. Troy Stevens, and operated by Ms. Charlene Jordan (Program Coordinator) and Judi Naylor (Accountant). The administrative activities of this core maintain program efficiency.
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{
"pile_set_name": "NIH ExPorter"
}
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Candidiasisis is the most common opportunistic infection in patients with AIDS. Fluconazole treatment is effective, but resistance can occur. L-743,872 is an investigational antimicrobial agent which has demonstrated potent antifungal activity. This is a multicenter study to 1) determine the efficacy and safety of IV L-743,872, and 2) select the optimal dose of the drug for the treatment of candida esophagitis. Eighteen subjects will be studied nationwide. As of January, 1998, the drug is now named MK-0991. Retreatment for relapse after 28 days of followup is now allowed with a new amendment to the protocol.
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{
"pile_set_name": "NIH ExPorter"
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1. Administrative Core Project Summary/Abstract We propose no change to the NN/LM PSR RML staffing level for the Cooperative Agreement. We are organized into the positions of Director, who will exercise overall governance; Associate Director, responsible for day-to-day operations; four program coordinators; and one office coordinator. With the advent of the NN/LM DOCLINE Coordination Office (NDCO) in the Cooperative Agreement, staffing efforts in that area will be redirected to emergency preparedness activities and other aspects of Network member services. All staff members will continue to participate in the ongoing process of recruiting new Network members in the course of various RML work activities, particularly those involving outreach and educational services. Recruitment initiatives will build on efforts established during our participation in the NN/LM Task Forces convened during the 2011-2016 RML contract period, particularly the Community College Outreach Task Force and the K-12 Outreach Task Force. Communication is vitally important to our Network members, and we have developed a well-rounded set of communication tools to meet the needs of a variety of audiences. Much reliance is still placed on the traditional mechanisms of email and telephone, but as increasing numbers of new Network members join the field, social media are becoming increasingly popular. We established Facebook and Twitter presences during the last several years, and in coming years we will explore other social media platforms that may be appropriate to adopt. We are cognizant of the fact that in some cases institutional firewalls or other blockages prevent Network members from utilizing these tools. We will build on the groundwork laid by the NN/LM Disaster Ready Initiative, formerly the Emergency Preparedness & Response Initiative, headed by Dan Wilson at the University of Virginia. We will conduct training sessions for Network members, and promote disaster information resources from NLM, the Medical Library Association, and other organizations as appropriate. We have always maintained a history of monitoring disasters in the Region, and determining Network members who may be in need of assistance as a result of the disaster. Wildfires and tropical Pacific storms are generally the most disruptive events in the Region. We will also maintain our ?buddy? relationship with the NN/LM Greater Midwest Region, and we recently established back-up personnel to manage their web site in case of disaster. In addition, in the event of national disaster affecting NLM, we have developed contingency plans for providing assistance as needed.
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{
"pile_set_name": "NIH ExPorter"
}
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The long term objective is to deepen our understanding of the pathogenesis of diabetic retinopathy and of the basis for treating this disease by photocoagulation. A widely believed, but unsubstantiated, working hypothesis is the retinal ischemia or hypoxia is responsible for retinal neovascularization. Direct noninvasive measurement of tissue oxygenation in the human retina could provide the data necessary to test this hypothesis. The technique of redox fluorimetry has been used to assess oxygenation of non-ocular tissues. We have adapted this technique for use in the eye and have built an instrument for noninvasively measuring the fluorescence of the flavin adenine dinucleotide (FAD) moiety of flavoprotein. FAD is an electron carrier in the mitochondrial electron transport chain whose fluorescence intensity decreases during hypoxia. The purpose of the proposed work is to do the critical experiments that will tell us how to best use measurements of retinal flavoprotein fluorescence to determine the oxygenation of the retina. We would like to produce a map of the retina that shows its oxygenation, and which gives the location and severity of focal patches of retinal hypoxia. Experiments with monkeys and with patients with known defects in the retinal circulation are designed to validate the use of redox fluorimetry in the retina. Comparisons between normals and patients with proliferative retinopathy will be used to test the hypoxia hypothesis above. Comparison of maps of retinal flavoprotein fluorescence with fluorescein angiograms in patients at different stages of diabetic retinopathy may establish the relationship between anatomic and functional abnormalities in the course of the disease. The clinical significance of this work is that it may facilitate more selective application of photocoagulation in the treatment of diabetic retinopathy.
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{
"pile_set_name": "NIH ExPorter"
}
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Respiratory syncytial virus is an important cause of severe lower respiratory tract illness in young children. Those at particular risk for serious RSV illness are children with bronchopulmonary dysplasia, the chronic lung disease of prematurity with respiratory distress and an oxygen requirement lasting at least 30 days, and those with complex congenital heart disease. We plan to test the hypothesis that PFP-2 vaccine is safe, immunogenic and will prevent or significantly ameliorate serious RSV illness in high-risk children.
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{
"pile_set_name": "NIH ExPorter"
}
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The goal of this project is to understand lymphoid organ development and function in ontogeny and inflammation. Directed over expression of lymphotoxin (LT) (either LTa or LTab) results in ectopic lymphoid aggregates with the characteristics of lymphoid organs. These "tertiary lymphoid organs" (TLOs) are also seen in many situations of chronic inflammation in autoimmunity, including the early stages of insulin dependent diabetes mellitus (IDDM). High endothelial venule (HEV) genes are regulated in development, in TLOs, and in acute inflammation. The mature HEV phenotype, characterized by expression of PNAd, LTbR, and a HEV suIfotransferase (HEC-6ST) depends on signaling through the LTbR in normal and HEC6ST- lacZ mice. After immunization, HEVs revert transiently to an immature phenotype and then recover. The recovery is dependent on B cells and LTbR. Lymphangiogenesis also occurs in TLOs and in acute inflammation. At early times after immunization, the occurrence of vessels that are positive for both LV and HEV markers is dependent on LTbR. Results with a novel bimodal dendrimer that visualizes lymphatic vessels by magnetic resonance imaging (MRI) and fluorescence suggest that mice deficient in LT have defective LVs. It is not known how LT contributes to lymphangiogenesis. Two hypotheses will be tested. Hypothesis I. Lymphatic vessels are regulated by LT in inflammation and development; Hypothesis II. Continual signaling through LT is necessary for maintenance and function of lymphoid organs particularly in respect to HEVs and LVs. The specific aims are to: 1. Determine how HEVs and LVs are regulated in acute inflammation by determining if B cells produce the LT ligand that regulates HEVs and LVs, by evaluating the role of macrophages in lymphangiogenesis, and by determining if the double positive vessels represent LVs budding from HEVs; 2. Determine how HEVs and LVs are regulated in chronic inflammation by evaluating vessel phenotype, density, and function, the role of macrophages, and LTbR and TNFR signaling; 3. Determine if and how LT influences constitutive LVs by evaluating LV function and phenotype 4. Determine if continual LT signaling is necessary to maintain HEVs and LVs in TLOs by treatment with LTbR and TNFR inhibitors and by turning LT on and off with a tetracycline inducible system. Elucidation of HEV and LV regulation will provide strategies for their inhibition in autoimmunity and allow control of insulitis. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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The Duluth Clinic has worked for the past thirteen years to develop a multi-disciplinary organization involved in the care of oncology patients and to function as a clinical cancer research unit for the geographic area. We have been joined by the Thunder Bay Regional Cancer Centre in Thunder Bay, Ontario which has a large geographic area also. The Duluth Clinic was a founding member of the North Central Cancer Treatment Group in 1978 and has functioned as a CCOP organization for the past five years. During this time, the Duluth CCOP has actively entered patients onto clinical group protocols (762 cancer control patients, 1,765 cancer treatment patients), has assisted in the development of new programs, functioned as study chairs on numerous protocols, and has presented frequently at national meetings. All of the disciplines in oncology are brought together under the direction of this CCOP. An excellent working relationship is established with the NCCTG, and data of unexcelled quality is generated, as shown by our past performance. Geographically, the Duluth CCOP is the only source providing the benefits of clinical cancer research to this large geographic area. The plan is to continue to bring the advantages of clinical research to patients, to bring new programs (such as through the National Surgical Adjuvant Breast Program) to the area, and to continue to increase efforts in cancer prevention, cancer control, and oncology rehabilitation. The North Central Cancer Treatment Group functions as the primary research base with the secondary research bases being the Eastern Cooperative Oncology Group, the Children's Cancer Study Group, and the National Surgical Adjuvant Breast Program. The goal of the Duluth CCOP is to accrue over 200 patients per year onto cancer treatment protocols and 400 patients onto cancer control programs. By virtue of the already established programs and the commitment to quality, it is believed that a high level of productivity and quality can be maintained.
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{
"pile_set_name": "NIH ExPorter"
}
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Alcoholic liver disease is caused by chronic heavy drinking, and is one of the major causes of liver cirrhosis. Thus, it is important to assess drinking behavior in all patients who present with liver disease. However, obtaining an accurate drinking history can be difficult, as the specificity of alcohol biomarkers is low in this population, and obtaining an accurate history by self-report or through collaterals is prone to error. Blood phosphatidylethanol (PEth) is an ethanol metabolite and novel biomarker that has undergone several small validation studies in substance abuse settings. While this early validation work appears promising, the characteristics of PEth for detecting clinically important levels of drinking in a more representative clinical population are largely unknown. Our pilot work began to address this issue in patients with liver disease, and suggests that PEth is highly sensitive for at least moderate to heavy drinking in this group. The studies described in this application further characterize the relationship of PEth to the amount of alcohol consumed in patients with liver disease, determine the accuracy of PEth for differentiating clinically important levels of recent drinking, and determine if PEth is superior to a well-studied alcohol marker (%CDT) in this population. Pertinent to the ultimate clinical utility of PEth, a secondary goal is to assess the association of baseline PEth with subsequent clinical outcomes. If highly valid, the clinical use of PEth will improve many facets of care for liver disease. This includes diagnosis of alcohol-related liver disease, assessment of response to treatment for heavy drinking and alcohol use disorders, early identification of relapse, and selection of the most appropriate candidates for liver transplantation. Findings may also have implications for other clinical populations (e.g., obstetrics, primary care, trauma), for evaluating the epidemiology of alcohol-related problems, and as a surrogate end-point in treatment outcomes research. PUBLIC HEALTH RELEVANCE: RELEVANCE Chronic heavy drinking is a common cause of serious liver disease, and abstinence or substantial reduction in drinking is the only effective treatment;however, physicians do not have clinically useful, objective measures of heavy drinking in patients with liver disease. This research will evaluate blood phosphatidylethanol, a novel marker of heavy drinking, in liver disease patients. Results may provide health care providers with an important tool for detecting heavy drinking, for monitoring response to alcohol treatment, and for early detection of relapse.
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{
"pile_set_name": "NIH ExPorter"
}
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Prevention of the angiopathic consequences of long-term human diabetes remains the primary goal of diabetes therapy. One possible approach to this problem is the development of a totally implantable, artificial "beta cell", to provide "physiologic" glucose homeostasis in the diabetic patient. The main goals of the proposed research were formulated in anticipation of the development of such a device, and in view of the progress being made in several centers toward the development of an implantable glucose sensor. The primary aim of the present study is the modification and testing of our implantable infusion pump to include a flow regulating capability; the entire package would then be suitable for inclusion into a artificial beta cell. The first phase of the project was to assess and solve the problems of insulin's heat lability, binding to inert pump components, and possible effects on local blood coagulation during chronic in vivo insulin infusion. The next phase will involve the design and in vitro testing of two types of flow regulators, to provide both continuously variable flow and two rates of constant infusion, one of the rates being controlled by an on-off valve. The final project phase will be the long-term in vivo testing of pumps plus flow regulators in experimentally diabetic animals.
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{
"pile_set_name": "NIH ExPorter"
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The proposed research continues an interdisciplinary effort to assess the role of bacterial toxins and the host response in determining the course and severity of endophthalmitis. Based on these determinations and characterization of the molecular properties of the toxins involved, novel therapeutic strategies specifically targeting these toxins have been and will be derived and tested. During the initial period of support, a plasmid encoded cytolysin elaborated by many clinical isolates of Enterococcus faecalis was observed to influence significantly the severity of endophthalmitis. Based on a molecular understanding of the structure, activity and activation mechanism of this toxin, two experimental therapies involving the use of specifically inhibitory peptides have been designed for proposed analysis. One of these therapies targets the activation step required for the cytolysin to be converted from a biologically inactive precursor to the active cytolytic species. The second peptide based strategy involves the use of"RGD" based peptides to block bacterial attachment to sensitive tissues of the retina, a phenomenon believed to result in concentration of the active lysin in proximity to the retina. Because of the emerging importance of "RGD" domain/integrin interactions in a broad variety of intercellular processes, observations made during this aspect of the study will be of wide ranging interest for a variety of intraocular therapies. Additionally, a novel application of submicroscopic, optically transparent liposomes will be tested for the ability to redirect membrane active toxins away from sensitive ocular structures during endophthalmitis. These experimental therapies are possible because of the application of state- of-the-art molecular biology techniques to characterize the respective toxin and its activities. Molecular characterization of the toxins elaborated by the highly virulent eye pathogen, Bacillus cereus, will be initiated during this research to determine which among a constellation of toxins contribute measurably to endophthalmitis caused by this organism. Based upon comparative studies of the molecular pathogenicity of B. cereus and E. faecalis for the eye, new general principles on the host/parasite interaction in intraocular infection will emerge forming the basis for the design and testing of additional experimental therapies for salvaging vision in these sight threatening infections.
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{
"pile_set_name": "NIH ExPorter"
}
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This application requests funding for the establishment of a Specialized Cooperative Center in Reproductive Research at Yale and Vanderbilt Universities to be named "The Center for Endometrial Biology and Endometriosis." Disorders of the endometrium and endometriosis result in significant morbidity including infertility, abnormal uterine bleeding, pelvic pain and dysmenorrhea. Endometriosis affects 10-15% of reproductive aged women and up to 50% of women with infertility. Endometrial disorders are a common problem with wide-spread public health implications. Despite the prevalence of these disorders, little progress had been made in understanding these conditions or their treatment. Therapy relies on hormonal manipulation and is fraught with side effects and treatment failures. The central theme of this application is discovery of novel mechanisms and therapeutic targets for endometrial disease and endometriosis. Each project, supported by preliminary data, hypothesizes a novel regulatory mechanism involved in the etiology or growth of this tissue, as well as suggests or tests novel therapies. The Center is collaborative and supported by three cores. Five interactive research projects address the central elements of this hypothesis. Project I will characterize the engraftment of endometrium and endometriosis with transdifferentiated bone marrow-derived stem cells. This Project will determine the extent to which bone marrow gives rise to the endometrium, the underlying mechanisms and functional consequences. Project II will explore the role of tissue factor in regulation of endometrial growth. Macrophage derived tissue factor contributes to endometrial growth and can be blocked by a novel immunoconjugated molecule (ICON) that targets tissue factor. Project III will look at failure of progesterone action in endometriosis due to disruption of a novel pathway involving epithelial- dominant cell-cell communication and TGF beta as well as retinoid signaling. Project IV will further determine the role of the mevalonate pathway on endometrial stromal cell growth and tests the novel use of statins as inhibitors of endometrial growth. Project V is a pilot project that explores the chemokines that recruit macrophages in endometriosis and tests the effects of traditional and novel treatments discovered here on signal transduction. The Center also contains three Cores. An Administrative Core, a Cell and Tissue Core and Mouse Modeling Core will be used by each of the investigators and complete a comprehensive Center.
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{
"pile_set_name": "NIH ExPorter"
}
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A set of important papers were completed that demonstrate the power, elegance and usefulness of information theory for a variety of molecular systems: We discovered a number of p53, p63 and p73 genes; We analyzed splice site mutations in Xeroderma Pigmentosum variant patients; We characterized sigma 70 promoters in E. coli; We discovered clusters of Fur binding sites in E. coli; We determined the relationship between DNA protein binding on- and off- rates with information of the binding site. This revealed the evidence that DNA binding is a coded function instead of a simple chemical reaction. Further details can be found in the papers, all of which are availabe from http://www.ccrnp.ncifcrf.gov/toms/ Z01 BC08396-14
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{
"pile_set_name": "NIH ExPorter"
}
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The WHO estimates gonorrhea infections occur in 78 million people globally every year. The extensive spread of antimicrobial resistant Ng has prompted the CDC to designate it as an Urgent Threat pathogen. Alarmingly, resistance is now emerging to the remaining current standard of care (SOC) dual therapy of ceftriaxone (CRO) and azithromycin (AZM). Despite this global health crisis, few new therapeutic agents are currently under clinical development to treat AMR Ng. Thus, new agents with novel mechanisms of action (MOA) not cross resistant to existing drug classes and not themselves susceptible to rapid resistance selection are needed to address the clinical spread of AMR Ng. Our proposal aims to develop a new AMR Ng therapeutic with a novel MOA not previously exploited in a clinical setting to treat GC, thereby replacing the SOC agent, CRO, and in doing so address the most serious threat of CRO-resistant Ng. We recently identified a novel and patentable analog of the natural product Moenomycin A (MoeA), we name Medinamycin (MedM). Whereas ?-lactams like CRO inhibit penicillin binding protein (PBP)-mediated transpeptidation of peptidoglycan (PG) polymers, MedM acts as a PBP transglycosylation (TG) inhibitor that abolishes PG synthesis. MedM displays exceptional Ng activity (MIC range, 0.0005-0.004 ug/ml) comparable to CRO, potent bactericidal activity, and a low frequency of resistance (FOR <1.14x10-9) similar to MoeA. MedM also exhibits an advantageous pharmacokinetic profile, highlighted by good subcutaneous exposure and long half-life anticipating single dose efficacy against AMR- Ng. Building upon a solid foundation of preliminary data, our Aims are: Aim 1 (Phase 1; Ph1). Establish MedM MOA in Ng and development potential as a novel GC agent. Milestone 1. Obtain 50 mg of MedM and demonstrate directly in Ng that MedM + AZM FICI < 4, MedM FOR < 1 x10-9, MedMR mutants map to Ng PBP-TG active site, and MIC90 < 0.125 ug/ml across 25 clinical isolates. Acceptable in vitro toxicity and minimal off-target activity. Establish dose-ranging for in vivo models. Efficacy POC is achieved with favorable 50% protective dose for survival (< 10 mg/kg) for 4 days. Aim 2 (Phase 2; Ph2). Establish MedM suitability for critical in vivo modeling. Milestone 2. Develop fermentation to provide 0.5 g MedM, demonstrate MIC90 equal or superior to ETX0914 (< 0.25 ug/ml) in 100 diverse AMR-Ng clinical isolates, and identify a formulation using a safety approved vehicle that achieves 10x MIC90 exposure in relevant species for in vivo efficacy models described in Aim 3. Aim 3 (Ph2). Establish MedM as a drug development candidate to treat GC. Evaluate compound efficacy in a murine female gonococcal lower genital tract infection (FGLGTI) and S. aureus deep thigh infection model. Milestone 3. Demonstrate that MedM achieves 100% clearance in FGLGTI models < 5 days IM treatment and >3 log reduction in murine deep thigh infection model within 24 h IM treatment.
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{
"pile_set_name": "NIH ExPorter"
}
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The focus of this program project has been the analysis of mechanisms leading to therapy-related acute myeloid leukemia (t-AML) in patients who previously received a variety of chemotherapeutic agents and/or radiation. Initially we focussed on mapping genes on the long are of chromosome 5 (5q) because deletions of 5q are one of the most common abnormalities in t-AML following treatment with alkylating agents and/or radiation and we defined a 2 to 4 mega base segment on 5q31 as the critical region. Loss of chromosome 7 is the most common change in t- AML; we have mapped two loci on 7q (7q22 and 7q32-33) as containing the critical genes. We will define the critical regions on chromosomes 5 and 7 more precisely and search for the involved genes. Drugs that target topoisomerase II are sometime associated in t-AML with balanced translocations especially involving the MLL gene at 11q23. We are analyzing the structure of MLL and the sequence of the MLL genomic breakpoints to search for potential mechanisms for breakage and translocation of MLL following exposure to the epipodophyllotoxins. We will analyze the cellular localization, phosphorylation state and the binding potential of the most common MLL partner gene, namely AF4 (4q21).
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{
"pile_set_name": "NIH ExPorter"
}
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The central objective of the proposed study is to identify and characterize the reaction products of the polycyclic aromatic carcinogens with DNA, both in vivo and in vitro. During the project period it is proposed to use sensitive spectroscopic and chromatographic techniques to characterize the DNA-carcinogen adducts produced in vivo in tissue cultures and in mouse skin. The in vivo reactions are thought to proceed by way of reactive metabolic intermediates of the carcinogens, but very little is known in detail about the structure of these intermediates. It is proposed to identify the major intermediates by comparison of the products of in vivo reactions with the in vitro reaction products of DNA with suspected likely intermediates, synthesized separately. It will then be possible to synthesize, in vitro, large quantitites of DNA adducts which are similar to the adducts produced in vivo. The in vitro reactions and their products will be subjected to detailed kinetic studies and physicochemical analyses. A variety of spectroscopic techniques will utilize the excited singlet and triplet states of the polycyclic aromatic moeities in the DNA-carcinogen adducts as probes of the structure of these complexes. These techniques will include fluorescence and triplet-triplet absorption spectroscopy, optically detected magnetic resonance, and electric-field-induced linear dichroism measurements. To obtain guidelines for the application of these techniques to the covalent adducts, analogous experiments will also be performed with physical intercalation-type complexes of DNA with polycyclic carcinogens and with acridine-type dyes, the structures of which are comparatively better understood.
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{
"pile_set_name": "NIH ExPorter"
}
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1 Project Summary 2 Cerebral Palsy (CP) is a group of disorders attributed to non-progressive alterations occurring in the brain dur- 3 ing fetal and newborn development. These alterations affect movement, development and posture, causing 4 significant lifetime functional limitations. CP is the most common motor disorder in children, affecting 500,000 5 individuals in the US alone. Children with CP are commonly affected by gait impairments. One of the most 6 common impairments is equinus gait, or ?toe-walking?, which is characterized by predominantly plantarflexed 7 feet during the whole gait cycle. Different interventions are available to treat equinus gait in children with CP 8 and include physical therapy, orthoses, botulinum toxin injections and surgery. To determine which intervention 9 is most appropriate to restore mobility for a given patient, periodic gait evaluations are performed in the clinic. 10 This Phase I SBIR proposal will develop a wearable system (Home Evaluation, Assessment, Rating and Train- 11 ing of Gait, HEART-Gait system) that continuously monitors and analyses simple gait parameters in children 12 with CP outside the clinical laboratory and during normal daily activities, which will provide clinically useful out- 13 comes data not currently available and critical diagnostic data for determining intervention timing. With access 14 to continuous data, the medical team is better informed and can effectively plan treatment, thus improving the 15 quality of care and the lives of children with CP and their families. Specific Aims are 16 Aim 1. Develop working prototype of HEART-Gait: Milestones are: 17 MS1: Veristride will optimize sensor selection and placement; construct prototype data acquisition hardware, 18 firmware and software. Months 1-2. 19 Aim 2. Iteratively design HEART-Gait system: Milestones are: 20 MS2: Veristride and the Spaulding team will work closely to modify the prototype system to make it suitable for 21 monitoring toe-walking in children with CP. Months 2-5. 22 MS3: The Spaulding team will interview 10 clinicians (unaffiliated with the project) and 10 families (guardians 23 and children) to gather their feedback about the HEART-Gait system. Months 2-5. 24 Aim 3. Evaluate the feasibility of HEART-Gait: Milestones are: 25 MS4: The Spaulding team will recruit 20 children (5-12 years) with CP and toe-walking pattern. Data will be 26 collected simultaneously from HEART-Gait and camera-based motion capture. Months 5-10. 27 MS5. The Spaulding team will derive a clinical index of the severity of toe-walking based on ankle biomechan- 28 ics data collected using the camera-based motion capture system. Months 6-11. 29 MS6. Veristride will collaborate with Spaulding team to develop machine-learning algorithms to estimate toe- 30 walking severity and characterize accuracy. Months 7-12. 31
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{
"pile_set_name": "NIH ExPorter"
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Project Summary Pseudomonas aeruginosa causes a variety of acute infections, such as ventilator-associated pneumonias, burn superinfections, neutropenic bacteremia, and medical device-related infections. In addition, P. aeruginosa chronically infects cystic fibrosis patients and causes significant morbidity and mortality in this population. With the increasing prevalence of antibiotic resistant organisms, developing agents that will modulate host immune responses will be key adjunctive treatments to conventional antibiotics in combating pathogens such as P. aeruginosa. In order to do this a detailed understanding of the innate immune pathways involved in recognition of pathogens such as P. aeruginosa are required. In this proposal we will examine the role of the NOD-like receptor (NLR) family member NLRC4 in recognition and response to infection with P. aeruginosa. In macrophages NLRC4 is activated in a multiprotein complex called the inflammasome upon infection with a wide variety of Gram-negative bacteria. The activation of the NLRC4 inflammasome ultimately results in the activation of the cysteine protease caspase-1 and its processing and secretion of proinflammatory cytokines. P. aeruginosa activates caspase-1 in an NLRC4-dependent manner leading to macrophage cell death and the release of interleukin (IL)-12 and IL-18. NLRC4-deficient mice also demonstrate increased susceptibility to infection with P. aeruginosa in vivo. In addition to Pseudomonas, other Gram-negative bacteria such as Salmonella, Legionella and Shigella have also been found to activate the NLRC4 inflammasome. One factor that is common to all these pathogens, and required for NLRC4 inflammasome activation, is a functional bacterial type III (T3SS) or type IV (T4SS) secretion system. However, it still remains unclear if NLRC4 serves as a cytosolic pattern recognition receptor for bacterial compounds that gain entry into the cytosol of the host cell through T3SS or T4SS, or if it indirectly senses the consequences of plasma membrane damage caused by the bacterial secretion systems. This proposal outlines three novel aims that will examine the molecular mechanism involved in activation of NLRC4: i) Defining the molecular components and cellular location for assembly of the NLRC4 inflammasome. ii) Elucidating the role of Toll-like receptors and reactive oxygen species in the upstream signaling pathways leading to NLRC4 inflammasome activation. iii) Determining if NLRC4 recognizes host plasma membrane components following their disruption by bacterial T3SS. PUBLIC HEALTH RELEVANCE: Project Narrative Pseudomonas aeruginosa causes a wide variety of opportunistic infections, particularly in hospitalized patient populations where it is responsibly for significant morbidity and mortality. P. aeruginosa is among the most common causes of ventilator associated pneumonia and amongst hospital acquired infections carries one of the highest mortality rates. With the increasing prevalence of multi-drug resistant strains on P. aeruginosa our treatment options are diminishing rapidly. Understanding how this pathogen interacts with and evades the immune system will help us identify new therapeutic targets to treat pseudomonal infections and is hence directly relevant to the VA patient care mission. Our studies will focus on how the innate immune pattern recognition receptor NLRC4 is activated by P. aeruginosa. In addition the information gained from the proposed studies will also shed light on the pathogenesis of related Gram-negative organisms.
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{
"pile_set_name": "NIH ExPorter"
}
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Understanding the role of chromatin modifying factors in response to ionizing radiation (IR) will provide invaluable insights into questions of both how cancers start and how to cure cancers. The ATM (ataxia-telangiectasia mutated) protein is a major participant in .all cellular responses to IR. We have been studying the role of ATM in DNA damage repair, telomere chromatin structure and oncogenic transformation. Cells deficient in ATM have defects in DNA repair, display altered telomere chromatin structure and have a higher frequency of spontaneous as well as IR-induced oncogenic transformation. Recently, we identified a chromatin-modifying factor "hMOF", the human ortholog of the Drosophila MOF gene (Males absent On the First) which interacts with ATM. hMOF has histone acetyltransferase (HAT) activity. Cellular exposure to IR enhances hMOF-dependent acetylation of its target substrate, lysine 16 of histone H4 (H4- K16), independent of ATM function. Inactivation of hMOF results in abrogation of ATM function. Based on the facts that hMOF is involved in ATM function, expression of MOF fragment enhances oncogenic transformation in vitro and that tumors show loss of H4-K16 acetylation (H4-K16Ac), we hypothesize that hMOF is involved in tumorigenesis. In the proposed work, we will determine the link between MOF and tumorigenesis by the functional characterization of mouse Mof (mMof) in a murine cell system and pathobiology of mMof haploinsufficient and mMof conditional knockout mice. The in vitro studies will determine Atm dependent and independent mMof functions in response to IR for cell killing, DNA damage repair and oncogenic transformation. The impact of the loss of H4-K16Ac on the predisposition of mouse tumor development will be determined in mMof haploinsufficiency or mMof conditional knockout mice. Experiments described in this proposal will investigate the functional links among mMof, H4-K16Ac and spontaneous as well as IR-induced tumor formation. These studies will improve our understanding of the role of mMof in the IR response and tumorigenesis. Ultimately, understanding the basis for biochemical differences in chromatin structure between normal and tumor tissue could provide strategies for modifying the response to IR that could be useful in clinical radiation therapy.
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{
"pile_set_name": "NIH ExPorter"
}
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PROJECT SUMMARY/ABSTRACT HIV+ adults who drink are already physiologically frail due to HIV infection, comorbidity (including hepatitis C infection), polypharmacy and associated substance use. In this setting, biomedical consequences of alcohol use can occur with moderate use and are often unappreciated or misattributed. The ?Consortium to improve OutcoMes in HIV/Aids, Alcohol, Aging & multi-Substance? (COMpAAAS) is supported by NIH/NIAAA award U24AA020794 to study this issue in a single sample, the Veterans Aging Cohort Study (VACS) (~50,000 HIV+ US veterans demographically matched to ~100,000 uninfected comparators). VACS will employ a direct alcohol biomarker (Phosphatidyl-ethanol [PEth] and a validated measure of physiologic frailty (VACS Index). In this set of three applications, the Antiretroviral Therapy Cohort Collaboration (ART-CC) and Kaiser Permanente (KP) teams join the Veterans Healthcare System (VA) team as COMpAAAS Tripartite: ART-CC, KP, and VA. Our long term goal is to inform alcohol intervention design and implementation. Together we propose to study biomedical consequences of alcohol and associated substance use in HIV, extending the scope and generalizability of VACS to multiple healthcare systems in North America and Europe and substantially increasing sample size and diversity of HIV+ subjects. Importantly, COMpAAAS Tripartite also expands the sample of uninfected comparators, a critically important group if we are to understand how alcohol differentially affects biomedical outcomes in HIV. KP will be able to identify demographically-matched uninfected comparators from their Northern California region. A new VA sample of veterans born in 1945-1965 (Birth Cohort) expands access to Hepatitis C infected (HCV+) and women comparators. The tripartite group will also participate in an HIV+ substudy (n=2250), The Medications, Alcohol, Substance Use in HIV Study (MASH), in which new data on potentially inappropriate medications (PIMS) and biomarkers for alcohol and substances (tobacco, marijuana, opioids, cocaine, and methamphetamine) will be collected. As the lead site for Aim 2, KP will examine the impact of alcohol and smoking on HIV outcomes, preventive care, and medical comorbidities. We anticipate that among HIV+ individuals, hazardous alcohol use, alcohol use disorders and smoking will negatively impact each of these outcomes; and that these effects will be amplified in HIV+ individuals compared with uninfected individuals. Initially, analyses will use electronic health record data including self-reported alcohol and substance use. Analyses will be repeated in the final year correcting for biases in self-reported alcohol and substance use based upon MASH results. Consistent with the RFA, all grants contribute data for all aims, have identical aims and protocols. This application addresses key interactions between alcohol and tobacco use, antiretrovirals, and medications that may increase mortality, hospitalization, frailty, and adversely impact preventive health care. We anticipate findings may lead to innovative intervention development, such as combined alcohol, smoking and preventive care interventions.
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{
"pile_set_name": "NIH ExPorter"
}
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Our laboratory has identified a critical role for the acetyltransferase p300 in the regulation of postnatal cardiac myocyte growth. We find that acetyltransferase p300 is rapidly upregulated in most or all forms of cardiac hypertrophy. We have shown that the cellular concentration of p300 is directly and stoichometrically related to cardiac myocyte growth capacity, such that even small changes in p300 levels have large effects on myocyte growth. Loss of a single p300 allele is sufficient to impair cardiac growth with age and in response to pressure overload. We also find that p300 is also rapidly induced during ischemic and oxidative stress, where it conveys a cytoprotective signal. The rapid induction of p300 thus appears to be a critical cardiac stress response. Despite the likely importance of dynamic control of p300 levels, nothing is known about the mechanisms that control upregulation of p300 mRNA and protein levels during stress. Based on these findings, we hypothesize that (1)"immediate-early", myocyte-autonomous induction of p300 during cardiac stress is the nodal event in the induction of cardiac hypertrophy by other identified effectors, including calcineurin, and that (2) induction of p300 is mediated by a sequence of signal-responsive pre- and post- transcriptional events, including phosphorylation, acetylation, ubiquitination, and removal of repression by non-coding RNAs. We propose to test the impact of modulators of acetylation and phosphorylation, and the role of specific p300-regulated miRs, on the accumulation and acetyltransferase activity of p300 during hypertrophic signaling. We will carry out cardiac-specific deletion of p300 and CBP to determine whether these proteins have distinct or myocyte-autonomous roles in hypertrophy. Finally, we will perform genetic complementation experiments between p300tg and HDAC5- or 9-deficient mice, and between p300-deficient and calcineurin tg mice. These experiments will definitively establish the position of p300 relative to the class II histone deacetylases and calcineurin in the transduction of hypertrophic signals in vivo. PUBLIC HEALTH RELEVANCE More than three million Americans are living with heart failure. Despite significant clinical advances, mortality remains extremely high;fewer than 40% will survive 5 years after their first episode of heart failure. The heart begins to undergo hypertrophy, or enlargement, months or years before it fails. Hypertrophy by itself is an independent risk factor for death. No therapy exists to prevent hypertrophy or delay its progression to heart failure. By revealing the specific molecular signals for hypertrophy, our studies may help lead to new treatments for these common and lethal forms of heart disease.
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{
"pile_set_name": "NIH ExPorter"
}
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The ability to raise broadly neutralizing antibodies is likely to be an important component of any effective vaccine against HIV-1. One class of these antibodies has special properties that merit more intensive study: (l) These antibodies have epitopes that are further exposed by binding of gp120 to CD4. (2) They compete with gp120 binding to CCR5 and to CCR5-derived peptides. (3) As we show here, many of these antibodies are, like CCR5, modified by tyrosine sulfate in their gp120-binding domains, and, in the single case characterized, sulfate groups are critical for gp120 binding. (4) As we also demonstrate here, members of this latter group bind with high affinity to envelope glycoproteins of isolates that utilize CCR5, but not those that utilize CXCR4. (5) As we observe here, this class of antibodies has a significant bias toward the use of a single variable chain gene (VH1-69) from approximately forty in the human germline. Collectively, these data suggest that these antibodies can function as mimics of CCR5, and thus may have the potential to neutralize a broad range of primary isolates. Their common variable chain suggests that they associate in a manner similar to HIV-1 gp120, despite differences in their complementarity determining regions (CDRs). Here, we propose to document the presence and functional role of sulfate groups on these antibodies, determine the degree of overlap between the epitopes of these antibodies and the CCR5-binding region of gp120, and explore the basis of observed preference for the VH1-69 variable gene. These studies will provide further insight into the association of gp120 with CCR5 and with this important class of antibodies, and will lay the groundwork for future studies of tyrosine-sulfated antibodies in HIV-1 infected individuals and infected or vaccinated animals.
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{
"pile_set_name": "NIH ExPorter"
}
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Plasmodium falciparum, a protozoan parasite, is the major causative agent of human malaria. Alarming increase in malaria-related deaths world-wide specially tropical countries calls for novel therapeutic stratgies to overcome this disease. Insights into the signal transduction pathways of malaria parasite in its development may prove useful for devising newer methods to stop its propogation. Our long term goal is to dissect signal transduction pathways at molecular and cellular level to understand their function in the development of this parasite. Recently, we have identified a novel signalling pathway in P. falciparum which involves PfPKB, a protein kinase B (PKB) like enzyme, which is very different from the manner in which host PKB is regulated. PfPKB is activated by calmodulin in a calcium dependent manner and phosphopliase C (PLC) activity is responsible for the required calcium release. These findings suggest that PfPLC, the homologue of PLC, works as an upstream activator of PfPKB in this pathway. We intend to address the following issues to help understand the function of PfPKB, PfPLC and the regulation of this pathway: 1. What is the role of PfPKB in parasite life cycle? Pharmacological and peptide inhibitors of PfPKB will be used to answer these questions. Some of these inhibitors have already been developed by us and preliminary studies using these have suggested that PfPKB may be an important player in parasite life cycle. Genetic studies of PfPKB to disrupt its function will be performed to establish its function. 2. Identification of PfPKB substrates. To understand the role of PfPKB, identity of its target proteins is essential, which will provide crucial information about its function. A proteomics based or a chemical genetics approach will be used for this purpose. 3. Regulation of parasite signalling by PfPKB-substrate interaction. Once the identity of PfPKB targets is known, biochemical and cell biology studies directed at understanding PfPKB-target interplay in the context of parasite signalling will be studied. 4. Role of PfPLC as a regulator of PfPKB and other signaling events. PLC inhibitors attenuate PfPKB activation in the parasite. Understanding of PfPLC regulation will be crucial to follow the upstream events involved in the signaling of PfPKB as well as other pathways. We intend to study i) how PfPLG is regulated in the parasite and what factors infulence its activity;ii) the mechanism of it's targeting to its site-of-action in the parasite.
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{
"pile_set_name": "NIH ExPorter"
}
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Herpes viruses are significant human pathogens causing a wide range of clinical diseases ranging from relatively benign reactivated infections such as cold sores to more deadly cancers. The very young, the elderly and immunocompromised populations, such as transplant patients and those with HIV-1 infections, are susceptible to more serious disease outcomes. Epstein Barr Virus (EBV) and Kaposi's sarcoma associated herpesvirus (KSHV) are associated with AIDS related cancers and human cytomegalovirus (HCMV) with severe retinitis and pneumonitis in HIV-1 positive individuals. In transplant patients, herpes simplex virus (HSV) and varicella zoster virus can reactivate as disseminated life-threatening infections, HCMV causes an increased incidence of graft rejection and EBV is associated with post-transplant lymphoproliferative disease. There are a limited number of drugs available to treat herpesvirus infections. A problem arising from this limited repertoire is the development of drug resistant virus strains. New therapeutic approaches for treating herpesvirus infections are therefore needed. The human herpesviruses each encode a serine/threonine protein kinase. The dependence on these protein kinases for efficient virus replication and spread, their conservation across the herpesvirus family and their enzymatic nature make these proteins attractive targets for anti-viral drug development. We also propose that identifying key cellular proteins or pathways activated by the virus protein kinases would permit the development of combinatorial anti-cell protein plus anti-protein kinase therapeutic strategies that would limit the development of drug resistant virus variants. We present two approaches designed to obtain basic information that would facilitate the development of anti-herpesvirus treatments targeted at the viral protein kinases. Viral protein kinase substrate recognition overlaps with that of cellular cdc2/CDK1 kinase but is extended beyond cdc2 sites. The degree to which the extended site recognition is unique to, or common to, the different herpesvirus protein kinases is not known. A bioinformatic approach to protein kinase site identification will be undertaken using an algorithm developed by the Co-Investigator and a database of 644 EBV, HCMV, KSHV and HSV-1 substrates identified in my laboratory using human proteomic arrays. The predicted motifs will be validated in in vitro phosphorylation assays and in transfected and infected cell extracts. We recently identified TIP60 as a cell substrate of the protein kinases that is critical for herpesvirus replication. Using the EBV system as the model, we will identify cell genes whose expression is regulated by BGLF4/TIP60 mediated chromatin remodeling by interrogating genome wide CpG methylation of Akata B cell DNA after doxycycline induction of BGLF4 and control kinase dead BGLF4. Data from the screen will be mined using bioinformatic analyses. Selected targets will be validated and the effect of their knockdown on virus replication determined. These experiments will uncover additional cell proteins or networks that could be targets for anti-viral therapeutic strategies.
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{
"pile_set_name": "NIH ExPorter"
}
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Epidemiological studies provide strong evidence that diet can modify cancer incidence. Populations concuming legume-derived products exhibit significantly decreased rates of breast, colon and prostate cancer. Legume products contain an array of anticarcinogens of diversified structures. Biochemical evidence correlating their presence due to legume consumption and cancer reduction in human populations is lacking. Our long term objective is io understand the quantitative relationship between soybean consumption and cancer reduction. In this proposal, we will investigate the pharmacokinetics of soybean-derived anticarcinogens in humans. The hypothesis to be tested is that continuous consumption of soybean products is necessary to maintain constant blood and tissue levels of isoflavones. This will be tested as follows: 1. The influence of food processing on isoflavone levels will be determined in different commercial soybean products. 2. The influences of age, sex, dietary habit, and disease state on absorption, disposition, and clearance of isoflavones will be determined prior to and/or after a shot term and chronic soymilk diets. The levels of isoflavones while on an unrestricted diet, at 1 week off a soybean-derived diet, at 1-4 weeks on a soybean diet and at 1-2 weeks after withdrawal from a 4 week-soybean diet will be analyzed. These will be measured in blood, urine as well as tumorous and non-tumorous breast and colon tissues. The levels of isoflavones will be compared before and after soymilk consumption, between different age or sex groups, between Asians and non-Asians, among cancer subjects, normal vegetarians, and normal omnivores, or between tumor and non-tumorous tissues. The Greater Houston-Galveston area has a large Asian population (Chinese, Vietnamese, and Indians) accustomed to a high soybean content diet. Many individuals in this population are also vegetarians. Furthermore, large Hispanic and Black populations are also available in this area. Thus, the population composition is uniqudly suited for current pilot and future large scale dietary intervention trials. This study will provide a rational basis for future dietary studies of cancer prevention by soybean meals.
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{
"pile_set_name": "NIH ExPorter"
}
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The objective of the project is to examine the hypothesis that cellular differentiation and malignant transformation are accompanied by expression of specific cell surface proteins. The surface proteins will be analyzed by a newly developed two-dimensional gel electrophoresis system. The final goal of the proposal during the period of grant support is to analyze and compare surface proteins of the normal and T/t locus mutant mouse embryos. The recessive t mutants of homozygous lethal type are uniquely suited for this project since: 1) they interfere with development of embyros at six specific stages, 2) they interfere with the further differentiation of ectoderm, 3) they are expressed as the surface antigens, and 4) a number of mutant strains are available. I propose to analyze proteins of sperm, unfertilized eggs, fertilized eggs and embryos. Analysis of sperm proteins is feasible by techniques available in this laboratory. However, sensitivity of protein detection needs to be improved for the analysis of proteins in eggs and embryos. An approximately 10 to the 4th power-fold increase of sensitivity seems possible by utilizing horseradish peroxidase-catalyzed reaction as follows: Proteins are tagged with biotin by reacting with succinimidated biotin. Biotin molecules are then complexed with avidin-horseradish peroxidase conjugate. Finally, horseradish peroxidase will be detected by catalytic polymerization of a radioactive substrate such as tyrosine, aniline derivatives or diaminobenzidine or its derivatives. A cell surface protein of MW 135,000 is inducible in Chinese hamster ovary cells by cyclic AMP derivatives or prostaglandin E1. This is an ideal model system for the study of rate of synthesis and turnover of surface proteins, pathway of protein expression on cell surfaces, and mode of protein-protein interaction on plasma membrane.
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{
"pile_set_name": "NIH ExPorter"
}
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With the aging of the American workforce, the largest percentage increase in employment is occurring in the 55-64 year old age group. This research is designed to assess effects of workplace biomechanical, psychosocial, organizational and non-workplace factors on normal age related changes in musculoskeletal function and performance. There are particular emphases on the role of gender and on identifying sub- populations that may be at increased risk of exceeding age expected changes in musculoskeletal function and performance. We use the term musculoskeletal health (MSH) to define the complex of physiological performance and effective function. An estimated 1200-1400 workers representing both genders, and meeting a broad age demographic, will be studied in Connecticut manufacturing facilities. Their evaluation will include questionnaires that survey individual, organizational and psychosocial factors; direct measures of physical performance, strength and muscle power; and instrumented and observed exposure measurement. There are cross-sectional and longitudinal components employed in a mixed design that accommodates the relatively short study period, and the longer time span of many health outcomes relevant to the musculoskeletal system. A key aspect of this study is the influence of the so-called "healthy worker survivor effect", the extent to which musculoskeletal symptoms and function influence withdrawal from the workforce. This is addressed through several imputative models. We will also estimate the interactive effect of risk factors (extrinsic and intrinsic) on the distribution of subjective and objective findings and undertake the compilation of broader indices of MSH. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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Neurotransmitter release is one of the most regulated membrane fusion events. Unlike constitutive vesicle trafficking, synaptic vesicles are recruited to the presynaptic membrane, but do not readily fuse. Instead, an average of ten vesicles are stably docked at a region of the synapse termed the active zone awaiting an action potential. Synaptic vesicle fusion is closely associated with the Ca2+ influx that follows arrival of an action potential. Exocytosis is triggered within approximately 0.2 msec of the Ca2+ arrival. Although many key factors have been found to be essential for Ca2+-dependent neurotransmitter release, such as SNAREs, synaptotagmin, complexin, Munc18, and Munc13, the molecular mechanism of synaptic vesicle membrane fusion and Ca2+- triggering remains unclear. In this renewal application we wish to continue our efforts to decipher the molecular mechanism by using a combination of single molecule and single particle fluorescence imaging microscopy experiments. Driven by our hypothesis that SNAREs and auxiliary proteins do not fully assemble and, thus, do not trigger fusion until a Ca2+ signal arrives, we have designed labeling strategies to monitor distinct conformational states of the protein and protein-complexes involved in the process. We have also developed fluorescent reporters that provide information about content and lipid exchange as the system proceeds from vesicle docking, to membrane contact, and, finally, to fusion. These labeling strategies and the single molecule and single vesicle experimental setups are the cornerstones for the proposed research. Our assays are a breakthrough, since for the first time, we observe fast fusion kinetics upon Ca2+ triggering. Our new assay now enables us to determine the role of the factors involved, their stoichiometry (e.g., how many SNARE complexes and synaptotagmin molecules are required for fast Ca2+ triggered fusion?), to propose and test mutants of these factors that affect their function in the assay, and then to use these mutants as tools for experiments with neuronal cultures to dissect their role in vivo. Furthermore, we have recently demonstrated the ability of super-resolution imaging of synaptic proteins by using labeled antibodies to an unprecedented degree of localization accuracy. This in vivo imaging approach will enable us to image the distribution and correlation of synaptic proteins along axons, and so it will complement our reconstitution and single molecule biophysical studies.
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{
"pile_set_name": "NIH ExPorter"
}
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Specific aims: The aim of this project is to identify the molecular defect responsible for X-linked reticulate pigmentary disorder with systemic manifestations in males (XLPDR, MIM301220), a rare Mendelian genetic disorder.
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{
"pile_set_name": "NIH ExPorter"
}
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The proposed studies examine how children acquire counterintuitive concepts - how they learn and come to believe in ideas that defy their intuitions about how the world works. Being able to sort through and selectively accept certain counterintuitive information has profound implications for children's acquisition of a host of culturally-transmitted ideas, ranging from the scientific to the spiritual, including ideas relevant to children's health. For example, children who are first learning about biological evolution must override their idea that species do not change across generations, and for children to understand heliocentrism, they must override their first-hand perceptions (every day, the sun appears to revolve around the earth). Moreover, for children to understand the importance of bacteria and viruses, they must believe that powerful entities exist that cannot be perceived with the naked eye. Thus, counterintuitive concepts play pivotal roles in many domains of thought. Though recent research has examined how children represent counterintuitive concepts, and how much children trust information that others provide, these two lines of research have yet to be merged. The proposed experimental studies are designed to address three specific aims with regard to how children come to believe counterintuitive concepts: Aim 1: Assess how various aspects of the context--including the immediate learning context and the broader cultural context--influence children's willingness to accept counterintuitive ideas. Aim 2: Determine what types of people children trust when learning counterintuitive information; in particular, examine whether and when children consider informants' traits and their expertise Aim 3: Identify qualities of the information itself (e.g., explanatory depth) that affect children's willingness to accept the counterintuitive information as true. To address these aims, children (4- to 8-year-olds) attending schools and museums in the Boston area will engage in tasks in which they learn counterintuitive information from different types of people and in different contexts. Results promise to contribute both to basic science and to children's formal and informal education. Results will inform and inspire research on science education in both formal settings (i.e., the classroom) and informal settings (e.g., science museums and educational media). Results will also inform research on the transmission, pervasiveness, and development of counterintuitive concepts embedded in children's spiritual and religious beliefs - beliefs that can contribute to a sense of meaning and mental well- being. Moreover, these results have implications for children's physical health - understanding how best to convey counterintuitive ideas to children, educators and parents can better teach children about important health-related entities, such as bacteria and viruses. Intervention studies can then capitalize upon the factors that are identified as best promoting children's acquisition and acceptance of counterintuitive concepts.
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{
"pile_set_name": "NIH ExPorter"
}
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This Asthma and Allergic Diseases Center Grant application is an effort to continue a long-standing fruitful collaboration among 4 UCSF faculty members with a long-standing interest in the mechanisms underlying allergic airway inflammation and asthma. The central goals of this application are to determine critical mechanisms underlying the initiation and persistence of allergic airway inflammation and airway hyperresponsiveness. The application includes two projects that will utilize murine models and a third that will examine the relevance of molecular targets and pathways identified in these models to asthma prevalence, severity and drug responsiveness in humans. The Center Principal Investigator and leader of Project 1, Dean Sheppard, has identified critical roles for activation of transforming growth factor ? (TGF??) by two different integrins (?v?6 and ?v?8) in airway hyperresponsiveness in a chronic model of allergic asthma and in modulation of cognate immune responses. This proposal will take advantage of a series of lines of genetically modified mice to directly examine the role of TGF?? in these effects, to determine the critical cells types responsible, and to identify the molecular mechanisms underlying these responses. Project 2 is based on recent data from Richard Locksley, the project leader, that chitin, a prominent structural component of fungi, parasites and crustaceans, activates macrophages and primes innate immune cells for initiation of type 2 immune responses. Work proposed in this project will utilize a series of novel reporter lines to examine the critical chitin-responsive cells and the mechanisms and relevance of this pathway for initiation of allergic inflammation. Project 3, co-led by Esteban Burchard and John Fahy will follow-up on preliminary observations about genetic associations between sequence variants in chitin-degrading enzymes and TGF?? and allergic sensitization and asthma to more deeply interrogate associations and gene-gene interactions for sequence variants in chitinases and multiple components of the TGF? activation and signaling pathways, including the integrin subunits examined in Project 1. This project will also evaluate the functional significance of associated genes using bronchoalveolar lavage samples and tissue from asthmatic patients and healthy control subjects. These projects will be supported by a Physiology and Tissue Analysis core that will provide extensive support for all 3 projects, and by a centralized Administrative core. Lay summary - This Center will evaluate the mechanisms underlying initiation and persistence of asthma. By identifying novel pathways and molecular targets and testing their relevance to asthma, asthma severity and drug response in humans, the work in the Center should provide clues for the development of new treatments for this common and often devastating disease. PROJECT 1: ?v Integrins in Cognate Immunity and Airway Hyperresponsiveness (SHEPPARD, D) PROJECT 1 DESCRIPTION (provided by applicant): Mice lacking the epithelial integrin, ?v?6, that we have shown activates latent TGF-?, are protected from the persistent airway hyperresponsiveness (AHR) that follows chronic allergen challenge. Surprisingly, this protection is not associated with any decrease in sub-epithelial airway fibrosis, a central TGF-? -dependent feature of this model. Mice with leukocyte specific knockout of the related integrin, ?v?8, which also activates TGF-? show evidence of enhanced adaptive immunity. In this proposal, we will determine whether these altered responses in ?v?6 subunit knockout mice are a direct consequence of loss of the ?v?6 integrin and/or of TGF-? activation from conducting airway epithelial cells using "rescue" mice expressing either the wild type integrin or active TGF-? in airway epithelial cells. We will evaluate the effects of ?v?6 antibodies and a TGF-? RII-lg chimera on these same endpoints to further confirm the importance of this pathway and evaluate the feasibility of targeting this pathway for therapeutic intervention. To determine the mechanisms by which loss of ?v?6 protects from induction of AHR, we will evaluate the relationship between airway responsiveness and expression of a small number of candidate genes identified as linked to this phenotype in preliminary experiments utilizing expression microarrays. We will also determine the cellular distribution of expression by immunostaining and/or in situ hybridization, and will evaluate functional significance using commercially available lines of mice expressing null mutations of specific candidates. Because the cytokine IL-13 is known to play a central role in induction of AHR in multiple models, and because two of the most promising candidates identified by microarrays, leukotrienes C4 synthase and interleukin-18 have been suggested to be upstream of IL-13 induction in the airways, we will also examine the cellular sources of IL-13 in chronically challenged wild type and ?v?6 knockout mice. Finally, we will determine how loss of leukocyte ?v?8 leads to enhancement of adaptive immunity and examine the relevance of this pathway to allergic airway inflammation and its consequences. Lay summary - This project will examine how a single growth factor, transforming growth factor ?, can either contribute to development of chronic asthma or inhibit allergic sensitization and its consequences, depending on where and how this growth factor is activated.
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{
"pile_set_name": "NIH ExPorter"
}
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This Phase I trial will identify the maximum tolerated dose of Thymataq in patients with solid tumors, asses the toxicity of Thymitaq administered sequentially with cisplatin, and determine whether the administration of cisplatin alters the clearance of Thymitaq.
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{
"pile_set_name": "NIH ExPorter"
}
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Description (adapted from the application): This pilot project deals with elucidation of mechanisms involved in the DNA/lipid complex-induced toxicity in CF patient during cationic liposome-mediated gene therapy. The symptoms observed include fever, myalgia and arthralgia, occurring at six hours with full recovery between 24-48 hours after the administration of DNA/lipid 67 complexes. The applicant believes that the toxic effects of DNA/lipid complexes are caused by unmethylated CpG motif in plasmid DNA. It is hypothesized that CpG motif activates T lymphocytes and macrophages to release inflammatory cytokines (TNF-a and IL-1), Th1-inducing cytokines (IL-12 and IL-18), Th-1 cytokine (IFN-r), leading to fever, myalgia, arthralgia and activation of NK and B cells. The applicant has proposed two Specific Aims to test his hypothesis. Aim 1 is to explore the effect of methylated and unmethylated CpG motifs of plasmid DNA in the presence or absence of lipid 67 on the production of specific cytokines in human peripheral blood cells (PBMCs) in vitro, and to test the effect of anti-inflammatory cytokines IL-10 and IL-4 on the production of the specific cytokines. Aim 2 deals with similar experiments in vivo using IL-12 or IFN-r knock out mice.
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{
"pile_set_name": "NIH ExPorter"
}
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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We are refining our affinity isolation techniques to enable increasingly rapid purifications of protein complexes. One tactic that we have shown to work well involves the use of small diameter magnetic beads. We are planning some further experiments to help elicidate a theoretical model for the dependence of the isolation time on the bead diameter. Other tactics involve use of small volume cleanup apparatus, including low volume size exclusion chromatography.
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{
"pile_set_name": "NIH ExPorter"
}
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Experimental autoimmune encephalomyelitis (EAE) is an animal model that reproduces many of the clinical and pathological features of multiple sclerosis (MS). The development and the progression of EAE, like other autoimmune diseases, results from the pathogenicity of effector cells and the negative regulation imposed by regulatory T cells (Tregs). Since Tregs express the Fox-P3 transcription factor, we have generated a Foxp3- EGFP "knock-in" mouse strain in order to track CD4+ CD25+ Fox-P3+ Tregs by GFP-expression. The Fox-P3- EGFP reporter mice readily develop EAE following immunization with myelin oligodendrocyte glycoprotein (MOG). With the help of an I-Ab/MOG35-55 tetramer, we are able to track the development and effector functions of MOG specific effector and Fox-P3+ regulatory T cells. Of all the encephalitogenic effector T cells, in recent years IL-17 producing Th17 cells have gained a lot of attention and are implicated in mediating tissue inflammation including EAE. We have found that IL-6, an acute phase protein, not only suppresses the generation of Fox-P3+ Treg cells induced by TGF-[unreadable], but in turn induces Th17 effector T cells. Based on this observation, we hypothesize that there is a reciprocal relationship between pathogenic Th17 cells and protective Fox-P3+ Treg cells, and cytokines like IL-6 produced by the innate immune system dictate the balance between pathogenic/regulatory T cells and the development of autoimmunity. In addition to IL-6, we have now observed that IL-4 can also suppress induction of Fox-P3 and together with TGF-[unreadable] can induce a unique population of T cells that produces IL-9 and IL-10. These IL-9/IL-10 producing Th9 cells are not regulatory T cells but effector T cells that induce colitis and peripheral neuritis upon adoptive transfer into immunodeficient mice, however, what is their role in the induction of EAE has not been evaluated. We propose to: 1) Test the role of IL-6 in promoting tissue inflammation and in regulating the balance between pathogenic Th17 and Treg cells;2) Study the role of Th9 cells in the induction and regulation of EAE;and 3) Study the role of IL-23, a cytokine that expands and stabilizes Th17 cells, in generating pathogenic effector T cells and induction of tissue inflammation in the CNS. For studying the role of IL-23 in EAE, we have generated a novel IL-23R.GFP "knock-in" mouse strain, which in a heterozygous state becomes a reporter for IL-23 receptor expression and in the homozygous state becomes a "knock-out" line. With the availability of reporter mouse strains for Fox-P3+ and IL-23R+ cells, we are in a unique position to study how inflammatory cytokines particularly IL-6 regulate the balance between pathogenic and regulatory T cells and promote tissue inflammation and EAE. PUBLIC HEALTH RELEVANCE: This project deals with studying the effects of proinflammatory cytokines (IL-6 and IL-23) on regulating the balance between protective regulatory T cells (Fox-P3+ Treg) and pathogenic effector T cells, including Th17 cells and a novel T cell subset named Th9 cells, during an autoimmune disease of the central nervous system called Experimental Autoimmune Encephalomyelitis (EAE). Using "reporter" mice for effector and regulatory T cells and autoantigen specific tetramers, we will be able to study the balance and interplay between antigen- specific effector and regulatory T cells during the autoimmune reaction.
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{
"pile_set_name": "NIH ExPorter"
}
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The purpose of this proposal is to create, at the University of Washington, a university-wide Center for Health Services Research with emphasis on health policy research and analysis and implementation of the findings of these activities. The training of a select number of doctoral students is an integral part of this process. The focus of the Center will be to meet the needs of local and regional groups involved in or affected by the health services policy-making process. Because of the nature of the issues addressed, findings are likely to be of national import as well. In order to meet the above objectives, Center staff will rely on a combination of outside and inside advisory groups and will build on the existing research and analysis activities of the Department of Health Services. These activities currently focus on issues of access, cost, and quality of health care services delivered. Needs will be identified by legislators members of the executive branch of state government, providers, third parties and consumers. These needs will be translated into a program of research and analysis by Center staff working cooperatively with an internal advisory group of current faculty and principal investigators. Existing data bases will be adapted to address specific issues which are raised and, where necessary, consideration will be given to the development of new research and analysis efforts. Results of the research and analysis activities will be implemented through a variety of methods including testimony to congressional committees, written reports, working papers, newsletters, consultations, technical assistance and continuing education. An explicit program of evaluation of Center performance will be developed to help ensure both the scientific and technical quality of the research and analysis and its responsiveness to users' needs.
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{
"pile_set_name": "NIH ExPorter"
}
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This application is to meet the needs of 12 NIH-funded research projects on the BU Medical Campus (BUMC) and surrounding area for mass spectrometry instrumentation. These NIH funded-projects require state-of-the- art proteomics, glycomics or glycoproteomics analysis capabilities. The needs of the community of NIH funded investigators include (i) bottom-up proteomics of cross-linked proteins and post-translationally modified proteins, (ii) combined glycomics and proteomics from histological tissue slides; (iii) bottom-up glycoproteomics. There are no instruments available on the Boston University Medical Campus with the combination of sensitivity, speed and resolution necessary for such analyses. Accordingly, the present request is to replace existing obsolete instrumentation with a new Thermo-Fisher Scientific Q-Exactive HF LC/MS system. This new instrument produces high resolution, high mass accuracy tandem mass spectra that is essential for interpreting cross-linked proteins and post-translationally modified proteins. It extends the capability of traditional triple quadrupole mass spectrometers by producing high resolution and mass accuracy targeted mass spectral data that are crucial for quantifying changes in abundances of modified proteins in biological systems. It has unique tandem MS multiplexing capability that leverages ultra-performance chromatography systems now available. It has powerful data independent analysis features, including precursor ion multiplexing, that enable non-targeted quantification of modified peptides. It is very robust and well-suited to high-throughput analyses. Major user project areas include: (1-3) bottom-up proteomics analysis of cross-lined proteins or post-translationally modified proteins; (4-7) combined glycomics and proteomics of histological tissue; (8, 9) bottom-up glycoproteomics. In summary, the requested instrument system is necessary for the success of the described group of major and minor users on the BU Medical Campus and surrounding area.
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{
"pile_set_name": "NIH ExPorter"
}
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In the Genomics for Cardiovascular Disease section 7 new staff were hired, administrative assistant, research fellows, two post-doctoral fellow, post-bachelorette, nurse, physician assistant and an additional two will start in November 2014. Dr. Gibbons lab has successfully presented our research at many national and international conferences and has published over 12 manuscripts in peer review journals. Over 15 manuscripts are currently being developed. Within FY13-14 Dr. Gibbons has successfully established a new protocol to recruit over 1800 African-Americans for clinical evaluation and research testing called GENE-FORECASTSM (GENomics, Environmental FactORs and the Social DEterminants of Cardiovascular Disease in African-Americans STudy). Recruitment efforts include the District of Columbia, Montgomery and Prince Georges County. Our goal is to recruit 1800 volunteers for evaluation in the NIH Clinical Center over the next 3 to 5 years. We have exceeded our recruitment goals this year with 373 volunteers recruited and many more events still being added to our recruitment calendar. Our recruitment began in January 2014 and our Outreach and Recruitment Coordinators strategy involves attending and presenting at community health fairs and events (264) that have spanned across the Washington Metropolitan area, in conjunction with a cohort of volunteers recruited during our telephone surveys, (109) conducted by a third party. We are currently exploring marketing and advertisement vendors to develop a campaign to increase recruitment and visibility of the GENE-FORECAST research study. Additionally, Dr. Gibbons has successfully established a send protocol the Minority Health Genomics and Translational Research BIorepository Database (MH-GRID) project infrastructure facilitated the collection of biospecimens and related multidimensional data elements within a consortium of minority-serving clinics. MH-GRID Project was designed as a case-control study to assess both the genetic and social-behavioral determinants of severe hypertension in AA. Accordingly, the MH-GRID data set includes social-behavioral variables, clinical phenotype characterization and whole exome sequence data on cases with severe hypertension as well as normotensive controls. Dr. Gibbons has successfully establish a robust computer infrastructure and sequencing pipelines to support the analyses through actively computing, imputing, and simulating genomic and phenotypic models developed by dimensionality reduction techniques to gain useful insights from large genomic data sets. Within FY13-14, Dr. Gibbons has also established a cryo-repository facility with 100% tracing of biological samples utilizing Lab-Matrix (laboratory information management system), and setup two functional wet-bench labs. A total of six publications were generated for 2014 and 2015 by the Social Epidemiology Research Unit.
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{
"pile_set_name": "NIH ExPorter"
}
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E. coli aspartate transcarbamoylase (ATCase) is a classical allosteric protein whose function is thought to be controlled primarily by a quaternary structure transition between two states. The E50A mutant is one of a few ATCase mutants for which a third state different from T or R state in the two state model has been suggested by biochemical studies. We conducted time-resolved solution scattering studies on kinetics of the quaternary structure transition of the mutant immediately after mixing with a substrate solution. The structure change of this mutant was found to take place on the order of seconds, a rate much slower than that of wild-type ATCase thus eliminating the use of the cryoenzymology technique. The substrate-bound form of E50A appeared to be in a quaternary structure intermediary between T and R states, and effects of an allosteric effector ATP and bisubstrate analog PALA on the kinetics were studied.
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{
"pile_set_name": "NIH ExPorter"
}
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Coronary heart disease is a major cause of mortality in the US and in our Veterans. Biological approaches to treat the diseased heart using cell delivery have shown only modest therapeutic benefit, in part due to poor cell survival and ineffective electromechanical coupling to the host myocardium. Our previous research demonstrates that delivery of therapeutic cells cultured in an extracellular matrix (ECM) can improve the survival and functionality of the transplanted cells, owing to the structural support of the ECM as a scaffold and the signaling cues they impart to the cells. In particular, we have previously demonstrated the potency of anisotropic nanofibrillar scaffolds in guiding cellular alignment along the direction of the nanofibrils, enhancing cell survival in ischemic tissues, and imparting signaling cues that confer cell function consistent with a non- diseased state. Anisotropic scaffolds may be well-suited for engineering cardiac tissue which also has highly organized cellular structure. In addition, induced pluripotent stem cells (iPSCs) may be a candidate cell source for the generation of autologous therapeutic cardiovascular cells. The long-term objectives of this research are to engineer a three-dimensional vascularized cardiac patch with pre-formed physiological cellular organization to repair ischemic heart disease; and to investigate the basic biological mechanisms underlying ECM-mediated cell-cell interactions that enhance cardioprotection under conditions of ischemia. We hypothesize that a three- dimensionally aligned iPSC-derived cardiac patch with endothelial interactions will provide more functional and viable engineered tissues for repair of myocardial infarction, due to more effective electrical coupling and organized tissue morphology, as well as the activation of cardioprotective nitric oxide signaling imparted by ECM nanopatterning. Accordingly, our specific aims are: 1) To engineer a vascularized aligned iPSC-derived cardiomyocyte (CM) patch and elucidating the molecular mechanisms of ECM-mediated nitric oxide signaling in enhancing iPSC-CM survival and phenotype. The iPSC-derived CMs (iPSC- CMs) and iPSC-derived endothelial cells (iPSC-ECs) will be co-cultured on three-dimensional oriented nanofibrillar collagen scaffolds with culture conditions optimized to promote iPSC-CM cell survival and function within the patch, when compared to patches that lack oriented nanopatterning or vascular interactions. The role of ECM-mediated nitric oxide signaling in enhancing iPSC-CM survival and phenotype will be investigated in the context of gain- and loss-of-function assays. 2) To determine the therapeutic effect of a vascularized aligned iPSC-CM patch for treatment of myocardial infarction. The vascularized oriented cardiac patch will be transplanted onto the epicardium of rats after myocardial infarction. The animals will be monitored over time for functional improvement in cardiac function, wall thickness, and electromechanical coupling. The role of nitric oxide in mediating the functional effects of the aligned vascularized cardiac patch will be quantified by measuring production of nitric oxide and related signaling molecules. The knowledge gained from these studies will provide a stronger foundation of basic knowledge and improved methods for clinical development of engineered cardiac patches, ultimately with the goal of restoring myocardial function to the diseased hearts of our Veterans.
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{
"pile_set_name": "NIH ExPorter"
}
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We have requested funds to establish a confocal image analysis center to be shared by eight research groups in the Biology Department at Yale University. The user group has a broad range of microscopic requirements which include imaging of nuclear proteins in yeast, cytoskeletal proteins and structures in tissue culture cells, analysis of antigens involved in the patterning of vertebrate neuronal projections and a spatial analysis of cells in developing plant leaves, Drosophila, and mouse embryos. The research groups propose to use a wide range of fluorescence techniques to detect both gene transcripts and protein products. A confocal image analysis system will greatly enhance the level of detail which these studies can attain. In some cases, confocal microscopy appears essential. In comparison to conventional microscopy, confocal microscopy provides dramatic enhancements in contrast and resolution, and promises to become a standard biological technique. This is particularly true for immunofluorescence applications. Equipment of this type is not currently available at Yale.
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{
"pile_set_name": "NIH ExPorter"
}
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Robust technologies are lacking for genome-wide profiling of single cell transcriptomic and epigenomic changes. This situation constitutes a bottleneck and hinders the progress of many important fields where single cell analysis is required. Our recently developed procedure, whole DNA pool amplification (WPA), offers highly specific amplification of a complex DNA mixture with high efficiency and minimized bias. WPA can amplify as little as sub-femtogram quantities of DNA, generating micrograms of specific product. Recently we have adapted WPA toward developing a method for whole transcriptome amplification of the entire mRNA transcript profile at full lengths and with strandedness, and preliminarily tested it on 1 nanogram of cDNA, 10 cells and single cells. In this proposal, using single cells isolated from different preimplantation stage embryos as a proof of principle model, we will focus on the development of single cell research tools for genome-wide quantitative assessment of mRNA transcriptome and CpG methylation by deep sequencing analysis. Specifically, we plan to: 1). optimize a procedure for cell lysis, gDNA removal, and cDNA generation from single cells;2). develop an approach for generating highly specific materials suitable for high throughput sequencing by circularizing first strand cDNA from the intact cells treated as above, amplifying this cDNA in full lengths and followed by olig-dT/rU selection when necessary;3). develop a procedure that combines enzymatic discrimination of DNA methylation, DNA circularization, and isolation of CpG-rich DNA fragments, which will enable high-throughput sequencing to identify differentially methylated HpaII or other restriction sites for a single cell;and finally 4). analyze the mRNA transcriptome (covering expression, splicing form, allelic specific expression, etc) and CpG methylation patterns of 3 types of single cells isolated from mouse preimplantation embryos. This study will enable us to study the genome-wide changes of gene expression (mRNA), DNA methylation, and in the future other epigenomic elements, at single cell level and at 4-dimmersions for preimplantation embryos, and allow for unprecedented characterization of this critical period of development and differentiation. The technological and theoretical achievements from this project will contribute significantly to the study of the mechanism for human development and diseases in terms of its epigenomic regulation, and will eventually benefit the understanding, diagnosis and treatment of human diseases such as those related to children development, cancer, neuron/mental, and immune system. PUBLIC HEALTH RELEVANCE: The technological and theoretical achievements from this project will contribute significantly to the study of the mechanism for human development and diseases in terms of its epigenomic regulation, and will eventually benefit the understanding, diagnosis and treatment of human diseases such as children development disorders, cancer, neuron/mental, and immune system related clinical problems.
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{
"pile_set_name": "NIH ExPorter"
}
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The major epithelial permeability barriers are controlled and regulated by a cascade of events that is triggered by cell-cell and cell-extracellular matrix (ECM) contacts. Anomalies of integrin regulation and function have been implicated in the etiology of various pathologic conditions, including inflammatory disorders such as inflammatory bowel disease. In addition to integrins, several classes of cell surface glycoproteins, including CD98, have been shown to participate in integrin-mediated events. Through research funded by my CCFA research award, we have demonstrated the following: i) CD98 appears to associate with beta 1 integrin, disrupting outside-in integrin signaling, which normally confers cytoskeletal organization, a critical element of barrier function (J Biol. Chem. 2001, 276(42):39282-9); and ii) a novel mechanism appears to integrate cellular events, such as adhesion and amino acid transport, and the direct binding of cell surface molecules, including CD98 and ICAM-1 (J Biol. Chem. 2003, 278(26):23672-7). In the present RO1 application we propose to explore the molecular mechanisms of these phenomena. Thus, the general aim of this proposal is to better understand the expression of glycoprotein CD98 in the intestinal epithelium and its function in the epithelial-epithelial and epithelial-matrix interactions that regulate intestinal epithelial permeability. Specific aims 1 and 2 will examine the hypothesis that CD98 is a key molecule in the regulation of intestinal epithelial permeability barriers. Specific aims 3 and 4 will examine the hypothesis CD98 and beta1 integrin cluster signaling molecules that regulate the paracellular pathway in Caco2-BBE monolayers. The project will involve a variety of biochemical and biophysical methods with an emphasis on molecular approaches. The completion of the proposal should molecularly define the functional role of CD98 in the regulation of the intestinal barrier permeability. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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Identifying which interventions work best for who is a critical, yet unresolved, issue in prevention science. There has been a paucity of research on underlying mechanisms by which change occurs in prevention models with early school-aged children in spite of growing evidence that a high level of brain plasticity characterizes the early stages of development, providing an optimal window of opportunity. Moreover, no school-based prevention trials have utilized a model of intervention responsively that incorporates the new generation of scientific disciplines, thus limiting the ability to address these issues in an integrated and translational fashion. Existing studies also lack measures reflective of processes (e.g., neurocognition and physiology) shown to modulate skills (e.g., inhibitory control and emotion regulation) that have repeatedly been implicated in high-risk behaviors. To fill these gaps in knowledge, the proposed 5-year R01 study would be the first in-depth school-based intervention trial that infuses relevant aspects of cognitive neuroscience with existing theories of developmental psychology and prevention science. The ultimate goal is to identify underlying conditions (inhibitory control and emotional regulation) that may serve to both mediate and moderate intervention effects in the early years of schooling. Accordingly, we propose to conduct a theory-driven examination of Promoting Alternative Thinking Strategies (PATHS), an evidence- based model with replicated outcome effects, to understand how the intervention works and for whom. The PATHS program will be implemented in kindergarten classrooms through first grade in 2-3 schools (Ne150) with an additional 2-3 schools as an active control condition (Ne150) using a randomized trial design. These Baltimore City schools serve a demographically mixed community with relatively low academic achievement and a high level of behavioral problems. Two cohorts of kindergartners will receive PATHS for two consecutive years, while the control school will receive professional development workshops as well as incentives, testing, and observations similar to the experimental schools. Pretesting will be conducted in the fall of K to assess background and behavioral factors, cognitive functioning, and emotional regulation (measured by physiological stress reactivity). Measures will be repeated once at the completion of the school year, again at the end of first grade, and then a follow-up at 6 months following program completion during second grade for each cohort. Directly following from the recent conceptual and empirical work of the study team and others, this study represents a substantial innovation with implications for reducing behavioral disorders in youths. It is critical to identify underlying bases for individual differences in intervention responsively to tailor preventive approaches to specific needs of subgroups, thereby enhancing efficiency, cost-effectiveness, and maximization of impact. PUBLIC HEALTH RELEVANCE: Collaborative transdisciplinary work integrating developmental psychopathology, neuroscience, and prevention science promises to significantly advance not only an etiological understanding of risk behaviors but also of variability in intervention efficacy via the use of a multiple-levels approach. There is a clear scientific need to examine both mediators and moderators of preventive intervention effects to better understand their mechanisms of action. The proposed study will identify underlying conditions that mediate ultimate outcomes of a model preventive intervention to substantially improve our ability to monitor program effects and target program components. Use of a multiple-levels approach such as the one proposed herein offers optimism that preventive intervention research will continue to advance in terms of its sophistication and comprehensiveness, and translational efforts to develop interventions that promote resilient functioning will be fostered. Ultimately, knowledge generated from this study will highlight areas in need of improvement for a class of preventive interventions that are used in many U.S. elementary schools, as well as the design of new components that are better suited to various subgroups.
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{
"pile_set_name": "NIH ExPorter"
}
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This study is determining the effects of different liquid formula diets when given as the only source of food on diarrhea and nutritional status in people with AIDS.
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{
"pile_set_name": "NIH ExPorter"
}
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This application is for a K08 Mentored Clinical Scientist Research Career Development Award entitled ?Degradation of Toll-Like Receptor 8 by RNF216 in response to plasma MicroRNA?s ? A Novel Mechanism Regulating Inflammation in Acute Lung Injury?. I am a physician in pulmonary and critical care medicine at the University of Pittsburgh. I am applying for this award to acquire advanced training in cell biology, translational research methods, and bioinformatics to develop my career as a physician scientist focused on the study of acute respiratory distress syndrome (ARDS). The main objective of my proposal is to determine how a novel Toll-Like Receptor 8 (TLR8) degradation pathway in monocytes regulates severe lung injury. TLR8 is a pattern recognition receptor that senses immunogenic RNA, including some host-derived plasma microRNA?s. TLR8 activation initiates signaling leading to the secretion of cytokines, contributing to excessive inflammation that is characteristic of severe ARDS. My preliminary data indicate that TLR8 is degraded in monocytes in a mechanism dependent on the post-translational modification of ubiquitination. Further, I have identified a candidate ubiquitin- transferring E3 ligase termed RNF216 responsible for targeting TLR8 for degradation, and I observe that RNF216 mRNA expression is decreased in patients with ARDS. Lastly, I have identified a subset of circulating plasma microRNA?s in ARDS subjects that may function as novel TLR8 ligands. The aims of this study are: i) to define the mechanism regulating TLR8 protein levels by the ubiquitin/proteasome system in monocytes ii) to determine if RNF216 modulates inflammatory signaling by directing TLR8 degradation and define RNF216 expression in ARDS, and iii) to examine plasma miRNA?s in ARDS subjects as TLR8 ligands. These studies will provide insight into a novel pathobiologic model whereby TLR8 degradation, regulated by RNF216 mediated protein ubiquitination, controls inflammation in response to host-derived plasma microRNA?s in ARDS. Plasma miRNA-induced TLR8 activation, augmented by reduced RNF216 mediated ubiquitination and degradation of TLR8, may drive excessive inflammation. Thus, modulating TLR8 degradation may be a novel strategy to reduce excessive inflammatory responses in ARDS. This project will provide me advanced skills in cell biology and bioinformatic approaches to analyze molecular datasets. I will be trained in translational research methodologies to strengthen my development into an independent investigator. I have committed mentoring from our Division Chief, Dr. Rama Mallampalli and a PhD comentor in Dr. Bill Chen. Additionally, my mentoring committee includes Dr. Robert Lafyatis ? an international expert in innate immunity, Dr. Stephen Chan ? an authority in translational miRNA biology, and Dr. Bryan McVerry ? a superb translational scientist and member of the Acute Lung Injury Center of Excellence overseeing the clinical Acute Lung Injury program. My work will be completed within the Division of Pulmonary, Allergy, and Critical Care Medicine at the University of Pittsburgh, which is committed to the development of physician scientists.
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{
"pile_set_name": "NIH ExPorter"
}
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Mammalian cells have evolved an intricate defense network to maintain genomic integrity by preventing the fixation of permanent DNA damage from endogenous and exogenous mutagens. A major genomic surveillance mechanism involves cell cycle checkpoints that exist at the G1-S and G2/M transitions and are regulated in response to DNA damage. Defects in these stages may result in a mutator phenotype that is associated with tumorigenesis. p53 safeguards the genome during cellular stress by activating both G1/S and G2/M cell cycle checkpoints. Two p53 downstream targets, p21waf1 and Gadd45, appear to be involved in these pathways. p53-mediated G1/S checkpoint is at least, in part, due to the activation of p21waf1. Recently, we discovered that Gadd45 is essential for one of the G2/M checkpoints activated in response to ultraviolet radiation or the alkylating agent methyl methanesulfonate in a p53-dependent manner. DNA damage activates Gadd45, which, in turn, binds to a G2-specific kinase Cdc2 and prevents the association with its regulatory subunit cyclin B1 and the inactivation of its kinase activity. Blocking Gadd45 expression can sensitize tumor cells to killing by cisplatin, a DNA-damaging cancer chemotherapy drug. This finding may offer a novel strategy to identify inhibitors that will provide new means of cancer treatment. In human cells, two additional Gadd45 family members, Gadd45b and Gadd45g, have been identified based on their extensive sequence homology. Although both Gadd45b and Gadd45g also bind to Cdc2 in vivo, they do not inhibit Cdc2 kinase and induce a G2/M arrest. To further define the functional domain, we have constructed a series of Gadd45 deletion or missense mutants. We have identified that the region between 50-76 is essential for its ability to bind to Cdc2, PCNA and p21waf1 in vivo, and to induce a G2/M arrest. The unique effect of Gadd45 on the G2/M arrest may be due to the presence of a region containing DEDDDR residues, which differs from the DEEEED residues in Gadd45b and the GEEDEG residues in Gadd45g. Therefore, the binding of Gadd45 to Cdc2 is insufficient to induce a G2/M arrest and additional activity contributed by the DEDDDR residues may be necessary to regulate the G2/M checkpoint. Interestingly, Ran, a small nuclear GTPase implicated in both cell cycle progression and nuclear export, also contains this motif. Forced expression of Ran also induces a G2/M arrest, whereas the deletion of this motif abolishes such activity. These data suggest that Gadd45 and Ran may utilize a similar pathway to regulate cell cycle progression from the G2 phase to mitosis, and this motif may serve as a common structural entity to activate the G2/M checkpoint. Although Gadd45-mediated G2/M arrest is dependent on p53, it does not require p21waf1 and 14-3-3s (two proteins that have been proposed to be involved in the ionizing radiation-induced G2/M checkpoint). While Cdc25C and cyclin B1 overexpression can override Gadd45-induced G2/M arrest, induction of p53 causes a downregulation of Cdc25C and cyclin B1 (two rate-limiting factors essential for transition from G2 to mitosis). Therefore, we propose that Gadd45 may activate the G2/M checkpoint through two mechanisms: a direct binding and inhibition of the Cdc2/cyclin B1 kinase, and a direct activation of p53 during the G2 phase to eliminate the abundance of Cdc25C and cyclin B1, thereby insuring a persistent G2/M arrest. Currently, we are constructing a normal human fibroblast cell line with a somatic "knock-out" of the Gadd45 gene. In addition, we have made an adenovirus expressing Gadd45. These reagents will be useful to test futher the role of Gadd45 in the G2/M cell cycle checkpoint.
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{
"pile_set_name": "NIH ExPorter"
}
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K562 human erythroleukemia cells constitutively express epsilon and gamma, but not beta-globin genes. The different expression of globin genes observed in intact K562 cells could be simulated in vitro as K562 nuclear extract (NE) actively transcribes the epsilon and gamma-globin gene DNA templates, but not the beta-globin gene. We have used the K562 in vitro transcription system to examine a transcriptional control element between - 392 and -177bp 5' of the canonical cap site for the epsilon-globin gene which has been reported to function as a silencer. We find that K562 NE cannot actively synthesize RNA in vitro from the epsilon-globin gene DNA deletion templates which contain the entire or a part of the silencer sequence but no positive regulatory region. Separating the K562 NE by ion exchange chromatography, we isolated a transcriptionally active fraction (F175) for globin genes and a fraction (F50) which contains the trans- acting factors associated with the silencer activity. The F175 was transcriptionally active for all tested globin genes including the epsilon- globin gene containing the silencer sequence. In contrast, F50 showed a strong dose dependent inhibitory effect on epsilon-globin gene transcription directed by either unfractionated K562 NE or F175. This suppression by F50 was not observed on transcription of adenovirus 2 major late promoter. In electrophoretic mobility shift assays using the epsilon- globin gene silencer region as probe, F50 and F175 exhibited different DNA binding protein patterns: a specific protein band in F50 appears to be associated with the silencer activity. These studies suggest that this protein may be specifically responsible for the activity of the silencer element of the epsilon-globin gene and that the expression and silencing of the epsilon-globin gene during development may be modulated by the interactions of this protein with the cis-acting DNA silencer.
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{
"pile_set_name": "NIH ExPorter"
}
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The long term objectives of this program are to develop an improved membrane artificial lung system for long term extra corporeal gas exchange support of infants and adults. The major design goals are: 1) to optimize gas exchange efficiency using secondary flows at the blood-membrane interface, thus minimizing the surface area requirements for gas exchange; 2) to reduce prime volume and use new polymeric surfaces which reduce blood trauma and thrombosis, thus allowing reduced heparinization; and 3) to provide a limited suction level and self-pumping capability to the membrane lung, eliminating the need for the gravity-fed bladder/roller pump assembly and to further reduce priming volume. The specific aims of the Phase I project are to test the feasibility of a high-efficiency self-pumping membrane lung by developing a bench prototype to perform a series of tests aimed at assessing the degree of gas transfer enhancement and the self- pumping characteristics of the system. The minimal contacting region between blood and membrane surfaces, the reduced priming volume, and the avoidance of stagnation points in this new membrane lung, in combination with the use of improved heparin coated microporous membranes, are expected to result in minimizing platelet injury, and the requirements for blood heparinization, the major causes of hemorrhagic complications during ECMO.
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{
"pile_set_name": "NIH ExPorter"
}
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The objectives of the study are to continue the ongoing Phase III randomized polycythemia vera study and the associated "efficacy" studies. By means of the major PVSG protocol the therapy amongst phlebotomy, radioactive phosphorus and chorambucil will be selected as being optimum resulting in the least morbidity and the longest survival. The natural history of polycythemia vera will be characterized and the modifying effects of myelo-suppressive therapy determined. Cytogenetic abnormalities are being detected by studies of the bone marrow and histopathologic marrow examinations are being performed.
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{
"pile_set_name": "NIH ExPorter"
}
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The NCI mandates that all CCSG-funded cancer centers establish a protocol review and monitoring system for oversight of the clinical research conducted at that institution. At the USC Norris Comprehensive Cancer Center (NCCC), the Clinical Investigations Committee (CIC) has this responsibility. The CIC performs the scientific review of proposed cancer clinical studies, and oversees the conduct of all clinical studies open at the NCCC. To facilitate the oversight of open trials, the internal Data and Safety Monitoring Committee (DSMC) will meet monthly to review institutional clinical trials (i.e., cancer-related trials involving an intervention where a USC investigator is the overall principal investigator (PI) of the study and the trial is not part of a national cooperative group or the CCCP Consortium, or sponsored by a pharmaceutical company that will responsible for the data management and oversight of the progress of the trial). Each in-house trial will be reviewed annually or more frequently as determined by the CIC. The reviews of the DSMC complement those of the QAMC, a long-standing Cancer Center committee which is charged with (among other responsibilities) reviewing accrual, and SAE's and major violations in a timely fashion, for all open protocols, as well as conducting audits of patients on institutional clinical trials. The DSMC will work in close coordination with the Clinical Investigations Support Office, which plays the main role of coordinating the conduct of clinical research in the NCCC. The objectives of the DSMC are to: 1. review, at established intervals, safety and aspects of the protocol and study conduct that may impact on safety of current or future patients 2. review all serious adverse events or other adverse events that require expedited reporting 3. review the progress of the study 4. review efficacy results that trigger early closure or early reporting 5. review the completeness and the quality of the research data 6. report the results of each review and make recommendations to the study PI, IRB, the Quality Assurance Monitoring Committee and the CIC.
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{
"pile_set_name": "NIH ExPorter"
}
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The overall purpose of the experiments in this proposal is to evaluate the role of androgens and estrogens in the development of ischemia heart disease (IHD) in males. To accomplish this objective, we will use a multifactorial approach. Studies have demonstrated diverse effects of gonadal steroids on specific parameters related to cardiovascular integrity, but the basic question of whether androgens or estrogens have deleterious or protective effects in IHD remains controversial. Thus, we will simultaneously employ rat models of arteriosclerosis, vascular reactivity and thromboembolism. In addition, the effects of the hormones on plasma lipid (TG, total cholesterol, HDL cholesterol) and apoprotein patterns in selected cases will be evaluated. The effect of hormonal manipulation will be qualitatively and quantitatively evaluated, so that both the nature of the effect and dose-dependency are determined. For each group of studies, the results will be examined for the likely net effect of the exogenous or endogenous steroid in ischemica heart disease. We do not necessarily expect that results in the three models will be in concordance: a specific manipulation might have opposite effects on thrombosis and vasoreactivity. It is to facilitate comparisons between models that the methods used are identical wherever possible. Some questions we expect to address in this final analysis are: i. Can either androgens or estrogens be considered general risk factors or protective agents in male IHD? ii. Is the observed sex difference in IHD attributed to the endocrine status of the male? iii. Is manipulation of the endocrine system, for example by anti-adnrogens, a potential therapeutic measure in male IHD. iv. Is endocrine manipulation especially deleterious or protective with respect to a specific aspect of IHD, for example thrombosis, but not to other aspects? The study will clarify the endocrine mechanisms of the higher incidence of IHD in males and suggest possible preventive or therapeutic measures. Furthermore, the studies should help greatly in clarifying the confusion in the literature regarding estrogens, androgens and gender in IHD in general as well as in specific aspects of IHD such as thromboembolism, lipoprotein metabolism, alteriosclerosis, and vasoreactivity.
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{
"pile_set_name": "NIH ExPorter"
}
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DNA ligases are critical enzymes for virtually all DNA transactions, including DNA replication, repair and recombination. Vertebrates have three DNA ligase genes: Lig1, Lig3 and Lig4; none can be directly deleted from the mouse germline. The Lig3 gene encodes two isoforms: mitochondrial Lig3 (which is cell essential) and the more abundant nuclear Lig3. Recently, we established a mutant mouse strain harboring a knock-in mutation that specifically ablates nuclear Lig3 using a one-step CRISPR/Cas9- mediated genome editing strategy in mouse embryos. Nuclear Lig3 is widely considered to be the primary ligase for DNA single strand break (SSB) repair due to its strong interaction with an essential SSBR factor called X-ray cross complementation factor 1 (XRCC1). Our unique mouse model will allow for the first time, a thorough dissection of the function of Lig3 in nuclear DNA repair in vivo. Experiments proposed in this application will provide the initial characterization of these mice. A comprehensive pathological phenotype analysis will be performed. Because DNA repair defects are often associated with genomic instability and neurodegenerative diseases, additional experiments will focus on the impact of Lig3 deficiency on spontaneous tumor development and neuropathology. Successful completion of the proposed studies will provide unprecedented insight into Lig3's normal physiologic role. Conversely, Lig3 overexpression is found in a number of tumor cell lines and tumor samples from human patients; this impacts DNA double strand break (DSB) repair such that the Lig4-dependent canonical non-homologous end-joining (NHEJ) is, to varying extents, replaced by the Lig3-dependent alternative end-joining (A-EJ), which has been implicated in chromosomal translocations. We will address the impact of nuclear Lig3 deficiency on IgH/c-myc translocation that occurs during immunoglobulin (Ig) heavy (H) chain class switch recombination (CSR). These particular translocations are critical to the development of many human and mouse B cell tumors. Furthermore, characterizing the dependence of these translocations on Lig3 may be relevant to many other tumor types. Successful completion of these studies may elucidate a pathological role of Lig3 in tumorigenesis.
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{
"pile_set_name": "NIH ExPorter"
}
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The long term goals are to identify the changes in the program of cellular gene expression induced by the 12S gene that results in primary-epithelial cell immortalization and the mechanism(s) by which they occur. Knowing how a cell becomes immortalized is important for our ultimate understanding of the multistep conversion of a normal cell to the tumorigenic transformed phenotype. Although most human neoplasms are of epithelial origin, very little is known about their transformation. Since the 12S gene specifically affects this cell type, it provides a means to study then progression at the cellular and molecular level. Much is known about the first exon of 12S, therefore the investigator will concentrate on the second exon, whose expression is also required for immortalization. The investigator will perform a detailed genetic analysis of the second exon to determine which region(s) and function(s) are important for immortalization. Cellular genes whose expression is modulated by the presence of this region, will be identified.
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{
"pile_set_name": "NIH ExPorter"
}
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Scale up the purification of three human placental enzymes to homogeneity suitable for monoclonal antibody production. Particular attention will be paid to the specificity of the antisera.
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{
"pile_set_name": "NIH ExPorter"
}
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The Graduate Program in Biochemistry and Molecular Biology involves seven departments in Robert Wood Johnson Medical School (RWJMS) and Rutgers University (RU). Graduate Training is currently supported by five training awards from the NIH. Continued support for these 5 traineeships is requested. RWJMS and RU represent the primary facilities for research and graduate (and undergraduate) training in the State of New Jersey. The Busch Campus in Piscataway is home to RWJMS and RU, as well as the Center for Advanced Biotechnology and Medicine (CABM), The Waksman Institute, Rutgers School of Pharmacy, The School of Public Health, and the Environmental and Occupational Health Sciences Institute (EOHSI). This concentration of research capital has fostered strong links between the various institutes, and among the faculty. The Joint Program in Molecular Biosciences (MolBioSci) took advantage of this resource to form a collective program to recruit graduate students, with the specific objective of enhancing their training potential. The Program in Biochemistry and Molecular Biology is administered by the Department of Biochemistry (RWJMS). The quality of incoming graduate students is excellent, as is the retention and graduation rate. A significant pool of candidates is available, since approximately 50% of the incoming class represents American citizens, and 25% are underrepresented minority students. Trainees are supported for two years during their third and fourth years of training. The number of participating faculty is currently 33, with 23 members from RWJMS and 10 from RU. The campus retains notable strengths in structural biology, enzymology of DNA replication, biochemistry of transcription and DMA repair, and regulation of development. Various model systems are explored, including rodent, fly, nematode, frog, plant, yeast and bacterial systems. Graduate training in the Program is accompanied by significant oversight, as formal mechanisms are in place to closely monitor the progress of the student, to remediate deficiencies, and insure completion of the dissertation. Similarly, the Program is itself subject to evaluation, to provide the strongest environment for graduate education and training. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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Core D: Neuropathology Core Project Summary/Abstract The UCI ADRC Neuropathology (NP) Core links clinical evaluations with definitive neuropathological diagnoses. Neuropathology remains the definitive method for diagnosing Alzheimer's disease (AD). The UCI ADRC cohorts consist of a longitudinal cohort, and two unique patient populations: adults with Down syndrome and individuals over 90 years of age ( 90+ ), representing cohorts at high risk of developing dementia. Down syndrome represents the single most prevalent cause of early-onset AD, whereas the 90+ cohort represents subjects with a high rate of conversion to dementia. Both cohorts provide unique opportunities to study the transition of `cognitively normal' to dementia. The overarching goal of the NP Core is to provide the infrastructure to support research on all three ADRC cohorts that aim to elucidate the underlying mechanisms that define normal aging, the transition to MCI, and the subsequent transition from MCI to AD/dementia. As part of its mission, the UCI NP Core disseminates well-characterized tissues, biospecimens and reagents to basic scientists at UCI and abroad to stimulate and facilitate research in AD and other age-associated neurodegenerative diseases. Using standardized methods for processing, the NP Core supports multi-center collaborative studies. Additionally, the NP Core seeks to innovate by establishing infrastructure to support novel technologies, techniques, and data collection to increase the value of the stored tissue and biospecimens. Lastly, the NP Core reports important AD research findings in collaboration with the Education Core. In order to achieve the goals of the NP Core, we propose seven specific aims: (1) Perform an accurate and timely autopsy of the ADRC cohorts (Longitudinal, Down syndrome, and 90+) and provide a standardized neuropathological report to the Data Management and Statistics Core for deposition with NACC, and to the Clinical Core for dissemination to the families of the deceased participants and their physicians; (2) Store, catalog, and disseminate brain tissue and biospecimens from ADRC subjects; (3) Store, catalog, and disseminate novel reagents made by UCI investigators; (4) Support and consult on ADRC projects, ADRC pilot studies, and studies on AD mechanisms by investigators funded by other means, including ADC collaborative projects; (5) Develop and establish new core functions to facilitate innovative research to study the transition of MCI to AD; (6) Participate in the public education of Chinese-Americans and others; (7) Train pathologists, neurologists, neurosurgeons, neuroscientists, neuropsychologists, and students on the neuropathological features of AD and other dementing disorders.
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{
"pile_set_name": "NIH ExPorter"
}
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The University of Maryland Baltimore submits this conference grant application for support of a Maryland statewide Summit on Childhood Obesity. The overall goal for the Summit is to ultimately prevent childhood obesity. The aims are developed in partnership with a broad range of stakeholders, health care professionals, researchers and community partners with expertise and commitment to addressing childhood obesity and related health disparities. The specific aims for the Summit are: To develop/implement a statewide Summit on Childhood Obesity to foster discussion, networking, identification of needs, exchange of strategies, and dissemination of evidence based information on childhood obesity;To define and discuss Maryland disparities in obesity morbidity and mortality, disparity populations and geographic regions, To identify existing and support the development of new multilevel strategies to prevent childhood obesity including systems changes, community engagement, policy and cultural influences on interventions, and;To disseminate an inventory of resources and programs in Maryland on childhood obesity which will support an action framework and implementation timeline. Childhood obesity represents a significant health issue for America's children and families. In Maryland, childhood obesity trends show an increase in prevalence including in urban, rural, African American and Latino children. These rates lead to increasing health care costs, increasing rates of significant chronic illness including type 2 diabetes, low school productivity, increased missed days from school and parental days missed from work. The solutions for this epidemic must be approached from multiple levels with cultural competence and sensitivity and in partnership with stakeholders at all levels. The planners and attendees include: multidisciplinary teams of health care professionals, advocates, food industry and systems, policy makers, school systems, parental groups and children. Working in collaboration with these stakeholders to assure not only changes in behavior, but also access to quality and affordable food, and engagement of parents, children, public health, policy makers and health care professionals in addressing this significant health issue. Innovative aspects of this Summit include a participatory planning process, a pre conference inventory, a conference and post conference state and local action plan and timeline to support sustainability. PUBLIC HEALTH RELEVANCE: The University of Maryland statewide Summit on Childhood Obesity addresses a major public health problem affecting Maryland's children, especially African American, rural and Latino children. This is a forum for networking, program planning and action steps to address disparities in this significant public health problem. Planners and participants include health care professionals, school systems and educators, parents and children, farmers and food industry, advocates and state/local policy makers, faith based leaders and researchers. At the Summit, an inventory of programs and resources and a plan for action with a timeline will be developed, disseminated, and monitored.
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{
"pile_set_name": "NIH ExPorter"
}
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A study of the shucture and function of a microtubule-nucleating complex from Drosophild embryo extracts was initiated. The y-TuRC (y-Tubulin Ring Complex) appeared as disc-shaped particles in the STEM when the specimen was fi=ze-dried, simila to previous TEM image& The preparation was relatively clean and the complexes were approximately 2.5 MDa. Many additional experiments are planned. It turns out that their preparations routinely have 2kDa peptides (to elute the complex from a column) which has'caused bad background problems. Work is underway to avoid this.
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{
"pile_set_name": "NIH ExPorter"
}
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Abstract Mammalian microRNAs are critical regulators of several human diseases. As a result, therapeutic manipulation of miRNAs in diseased tissues has emerged as a promising approach to combating human disorders, and has had success in some settings. Yet, effective delivery of miRNAs or their inhibitors to many cell and tissue types continues to be a major challenge, and this underscores the need for improved methods of small RNA delivery to relevant cell types. Recent reports have demonstrated that miRNAs can be released from cells in small lipid vesicles called exosomes. In turn, the exosomes can deliver miRNAs to recipient cells, and this system is thought to constitute a novel form of intercellular communication. This process includes an endogenous miRNA delivery mechanism that is largely uncharacterized, yet could provide valuable insights into how therapeutic miRNA delivery can be improved and how inflammatory responses are regulated. Our preliminary data demonstrate that mouse bone marrow derived dendritic cells (BMDCs) produce exosomes that contain specific miRNAs, including the inflammatory regulators miR-155 and mIR-146a, that can be delivered to recipient immune cells and subsequently mediate target knockdown both in vitro and in vivo. This results in an altered response to endotoxin both in vitro and in vivo, which provides experimental evidence that exosome-transferred miRNAs provide a novel layer of regulating inflammation. Further, we have also been able to successfully load specific miRNA mimics into exosomes and demonstrate that they can be delivered to recipient cells in a functionally relevant manner. As a result of these findings, we hypothesize that exosomal miRNAs play novel regulatory roles during physiologically relevant inflammatory responses, and that through an improved understanding of this process, exosomes can ultimately be coopted to deliver specific miRNA cocktails in a therapeutically relevant manner. We will carry out the following specific aims to test these predictions. First, we will define the process of exosomal miRNA delivery to immune cells. Next, we will determine the functional relevance of exosomal miRNAs during inflammation using radiation chimeras and Rab27a/b DKO mice with impaired exosome production. Finally, we will engineer custom exosomes and test their impact on inflammatory disease. Together, this project will shed light on how exosome transfer of miRNAs is regulated, determine the role of this system during physiologically relevant inflammation, and begin to understand the translational potential of this novel process.
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{
"pile_set_name": "NIH ExPorter"
}
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My work investigates the impact of the diffusion of genomic information into clinical and public health contexts. I am particularly interested in studying genomic communication relevant to diseases or populations that are stigmatized and/or underserved. I examine these issues both in simulated clinical contexts, and online through social media platforms. I currently have several ongoing or recently completed studies that contribute to these aims. In the clinical context, my colleagues and I have mounted a study to assess the benefits and/or risks of a physician introducing obesity genomics information into clinical weight-management encounters (Weight Management Interaction Study; NHGRI protocol # 11-HG-0238). We are interested in how this information will influence patients' health-related attitudes, beliefs, behavior, and patient perceptions of stigmatization. The project also aims to determine whether a physician's patient-centered versus doctor-centered approach to a health care encounter could mitigate or amplify the potential negative and positive consequences of genomic information provision. This study is being conducted in an immersive virtual reality-based clinical simulation in the Immersive Virtual Environment Testing Unit. During this reporting year, we have drafted two manuscripts which are currently under review, and are in the process of drafting additional manuscripts. In continuing to examine and inform provision of obesity-related genomic information in clinical encounters, colleagues and I have developed a study that assesses the impact of patient emotion on these processes. Emotions such as fear and anger arise in these clinical encounters from sources like receipt of risk information and perceptions of interpersonal treatment. Theoretically, the experience of emotion is expected to influence patient interpretation and reaction to genomic information provided by a physician. We have designed a two-pronged approach using both an internet-based and an immersive virtual reality-based simulation experiment to address these processes. Data collection for the virtual reality-based simulation study has been completed. Three manuscripts are currently in preparation stemming from this project. In addition to my experimental research related to communication about genomic information in the clinic, I also investigate issues around dissemination of genomic information in online and public health-related contexts. Together with Colleen McBride, I have experimentally investigated the effect on mothers of providing family history-based obesity risk feedback with respect to their young children (NHGRI protocol # 10-HG-0076), which is described in a separate report. I have also prepared and am currently fielding a survey assessing individuals' interest in and information seeking behaviors with respect to weight management and genetics information.
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{
"pile_set_name": "NIH ExPorter"
}
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PROJECT SUMMARY/ABSTRACT ? COMPUTATIONAL CORE We propose to establish a Center of Biomedical Research Excellence (COBRE) in Pharmaceutical Research and Innovation at the University of Kentucky (UK). CPRI will serve as a comprehensive multidisciplinary center focused on translational chemical biology [the nexus of chemical biology (the application of chemical biology principles to develop validated probe/models to advance our understanding of biology) and pharmaceutical science (the application of pharmaceutical principles to advance leads/materials/devices that address unmet clinical needs)]. The COBRE will leverage and develop unique translational chemical biology research support infrastructure/expertise to facilitate junior faculty mentorship and career development, innovative biomedical research probe/tool/model/materials development and validation, and the early advancement of potential ?translatable? assets. Within this context, the CPRI Computational Core, directed by Dr. Chang-Guo Zhan, will leverage unique computational expertise, commercial and in-house software/methods, and UK?s formidable computational processing network to support a range of computational services to advance COBRE project and pilot objectives. The Core will provide services individually tailored to project/pilot needs that facilitate the development of new hypotheses/models and streamline the prioritization of experimental studies. Specifically, the Core will enable molecular modeling/simulation of macromolecular targets and target-ligand complexes, ADMET prediction studies, structure-based virtual screening for ligand discovery, de novo ligand design, and artificial intelligence-based methods. This suite of services has been strategically designed to integrate with, and complement, the support provided by the CPRI Translational Core and the CPRI-affiliated COBRE for Molecular Medicine Organic Synthesis Core. Together, CPRI?s corresponding fully integrated suite of research support capabilities and expertise will accelerate innovative preclinical translational research and facilitate junior investigator career development.
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{
"pile_set_name": "NIH ExPorter"
}
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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The projected need for Indian rhesus macaques for AIDS-related research exceeds availability from current domestic breeding programs and there is an urgent need to expand breeding programs for future AIDS vaccine and pathogenesis studies. Rhesus macaques with defined major histocompatability complex (MHC) genotypes and known pedigree are becoming increasingly important for research to understand biologic variation in host immune responses and its effects on vaccine strategies and the pathogenesis of AIDS. The objective of this project is to expand the Oregon National Primate Research Center's specific pathogen-free (SPF) Indian rhesus macaque resource and sufficiently characterize their MHC haplotype to permit selected pedigree breeding for MHC class I alleles useful in AIDS research. Specific aims for accomplishing this objective include intensively managing a subpopulation of the Center's SPF Indian rhesus macaque breeding colony to maximize production of genetically diverse females to expand the breeding capacity of the colony. The breeding colony is typed for ten MHC alleles and managed for the production of MHCdefined offspring of known parentage. Both selective breeding of MHC-typed animals and assisted reproduction technology are used to enhance production of future breeder males that are homozygous for the MAMU-A*01 and other alleles important for assessing virus-specific cell-mediated immune function in simian immunodeficiency virus vaccine models for preventing AIDS virus infection. The colony is maintained in state-of-the-art sheltered field cage housing and is comprised of approximately 660 adult and juvenile animals and produces approximately 130 offspring annually.
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{
"pile_set_name": "NIH ExPorter"
}
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This project continues to focus on the formulation, analysis, and biophysical interpretation of mathematical models which describe various aspects of neuroelectric signaling for individual neurons. Among the topics of current interest are: (i) integration of synaptic input delivered to the soma and dendritic branches of a neuron; (ii) steady propagation of action potentials along axons; (iii) stimulus-response and threshold properties for repetitive-firing of action potentials. Mathematical models of these phenomena involve systems of linear and nonlinear ordinary differential equations and parabolic partial differential equations. Solutions and their mathematical stability are determined by analytical and numerical methods. One aspect in the approach of this project is to expose the qualitative mathematical structure for classes of models by exploiting simple, yet reasonable, equations. Because qualititatively related mathematical problems arise in other biological and chemical context, e.g. chemical and biochemical oscillations, this project may consider models from such applications.
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{
"pile_set_name": "NIH ExPorter"
}
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