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The investigators propose to determine the age-specific incidence of dementia and Alzheimer's disease and the clinical course of thee conditions in Chinese subjects over age 65 in Shanghai by carrying out a five year reevaluation of subjects first studied in 1987. The initial community survey of 5,055 elderly was carried out in the Jing-An district of Shanghai in 1987 by a collaborative effort of the Shanghai Institute of Mental Health and scientists then at the University of Illinois in Chicago and the University of California at San Diego. During this first survey the age- specific prevalence of dementia was determined and found to be similar to, although at the lower limits of, that found in other countries. Two-thirds of the cases met the accepted clinical criteria for Alzheimer's disease. Of particular interest was the finding of the higher prevalence of dementia in women than in men over the age of 75 and a higher prevalence among those who had received no formal education early in life. The 1992 follow-up of those who were age 65 or older in 1987 will provide data on accuracy of initial diagnosis, age-specific incidence and the effect of education, gender, and differential mortality on prevalence figures, and will provide information concerning other putative risk factors, the clinical characteristics and rate of decline observed in Alzheimer's disease in Shanghai and caregiver burden in this society.	{ "pile_set_name": "NIH ExPorter" }
The long term goal of this research project is to understand the role of nuclear translocated proteins in ehrlichial pathobiology. Ehrlichia chaffeensis, an obligate intracellular bacterium, is the agent of human monocytotropic ehrlichiosis (HME), a life-threatening emerging tick-borne zoonosis. During E. chaffeensis infection, transcription levels of numerous monocyte genes are altered including those involved in innate and cell-mediated immune responses, apoptosis, membrane trafficking, signal transduction and cell cycle. The E. chaffeensis gp200, the largest ehrlichial glycoprotein characterized in our laboratory, has been identified within the nuclei of Ehrlichia-infected monocytes. Although macrophage gene expression appears to be altered to favor ehrlichial survival, the role played in gene regulation by nuclear translocated proteins of ehrlichial origin is unknown. This gap in scientific knowledge regarding the role of gp200 in the macrophage nucleus is an impediment towards understanding ehrlichial pathobiology and manipulating these medically important bacteria. The objective of this proposal is to determine the mechanism involved in gp200 nuclear translocation and to identify gp200 nuclear targets. We hypothesize that E. chaffeensis gp200 is translocated to the nucleus via a glycan-dependant nuclear import pathway, where it acts as a transcriptional repressor, activator and/or regulator to promote intracellular ehrlichial survival. We propose the following specific aims: 1) determine the host cell nuclear import pathway involved in the translocation of gp200 to the monocyte nucleus, and 2) identify the monocyte nuclear proteins and DNA motifs that interact with gp200 during infection. The nuclear import pathway will be mapped by observing the nuclear translocation of fluorescent gp200 in the presence of numerous pathway inhibitors. A protein/DNA array and SELEX analysis will be performed to identify the host genomic sequences bound by gp200, and the nuclear protein binding partners of gp200 will be identified with a protein/protein array and co- immunoprecipitation. Together, these results will further our understanding of pathogenic bacterial glycoprotein function and identify a role for protein glycosylation in ehrlichial virulence. HME is an emerging public health concern, and this research will facilitate development of new therapeutic interventions against one of the most prevalent life-threatening tick-borne diseases in North America. [unreadable] [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The objective of the proposed research is to examine the impact of a cancer diagnosis on physical and cognitive functioning among older cancer survivors based on longitudinal data from the NIA-funded Health and Retirement Study. Public use HRS data from ten waves (1992-2010) will be linked to restricted HRS cancer data to identify respondents newly diagnosed with cancer and to form cohorts by major types of cancer. Data on sociodemographics, co-morbidities, and health behaviors, will be used to assess differences in physical and cognitive functioning exhibited at subsequent time points among cancer survivors and compared to persons with no history of cancer, to help separate the effects of cancer from the normal effects of aging. These data will be linked to the HRS Medicare Claims and Summary Data (2007) to identify extent of disease and treatment modality(s). Multilevel modeling and logistic regression models will be used to examine change over time and the association between predictors and patterns of change. This research will test the hypothesis that the diagnosis of cancer has a significant negative effect on physical and cognitive functioning separate from the normal effects of aging. We hypothesize that the risk of adverse changes in physical and cognitive functioning after a cancer diagnosis is influenced by various treatment-related factors as well as by socioeconomic and health factors present before and/or after diagnosis. Insight gained from the proposed project will improve our understanding of the adverse physical and cognitive health outcomes following a cancer diagnosis in older adults and will inform interventions targeted at addressing both age-related and cancer-related consequences to improve the quality of life for long-term cancer survivors.	{ "pile_set_name": "NIH ExPorter" }
The Capacity Development for Research into HIV-Associated Malignancies in Nigeria (CADRE-Nigeria) training program targets development of clinical trials and cancer research expertise at the Institute of Human Virology, Nigeria (IHV-Nigeria) and its academic research partners. This research training implements four aims: clinical trials capacity, cancer registration, cancer epidemiology and general research capacity building. CADRE-Nigeria will conduct 13 short-term training courses, intermediate training for 41, and in depth long-range training for 3 trainees over 3 years. CADRE-Nigeria is anchored by the mentor-the-mentor training strategy pioneered by the Fogarty-sponsored UM-IHV AITRP (D43 TW001041) that builds sustainable capacity by engaging an international faculty mentor, a Nigerian faculty mentor and a more junior faculty or non-faculty trainee in a training triad that builds a critical mass of investigative staff at the Nigerian research partner site and creates an academic bond between the international and Nigerian faculty to promote sustained research collaboration. The major outcome of this training is a Nigerian HIV-malignancy center of excellence the AIDS Malignancy Clinical Trials Consortium-Nigeria (AMC-Nigeria) site in Abuja, Nigeria that implements NCI funded AMC clinical trials, HIV-malignancy research and serves as a national training center for south to south HIV-associated malignancy capacity development at other sites in Nigeria. To expand HIV-malignancy detection tumor registration and cancer epidemiology training will exploit the 140,000 in HIV care in IHV-Nigeria's PEPFAR program providing a rich resource for case referral for the AMC-Nigeria trials for NCI Cancer Specimen Resource networks and the cancer research agenda of the NIAID/NCI-funded West Africa International Epidemiologic Database to Evaluate AIDS (leDEA) network both linked to IHV-Nigeria. The expected impact of CADRE-Nigeria is participation in an increasing number of epidemiology and clinical trials protocols, improvement in cancer registration, HIV-Cancer linkage studies and management of AIDS Associated Cancers in Nigeria.	{ "pile_set_name": "NIH ExPorter" }
The aim of this research is to better define the interaction of acetylcholinesterase and nerve membranes. The possible orientation of this enzyme with respect to the membrane surface will be studied. In addition, changes in orientation and/or conformation upon chemical or electrical perturbation (stimulation) of the membrane will be studied. Direct physical data relevant to the geometrical orientation of membrane associated proteins is sparse in general, and non- existant for acetylcholinesterase. The techniques of spin-labeling combined with EPR, NMR and flourescence spectroscopy will be used. Spin-label and flourescent probes will be used to explore both the enzyme and lipid contributions to the enzyme-membrane interaction. These studies will increase our understanding of the molecular mechanism by which nerves function.	{ "pile_set_name": "NIH ExPorter" }
Our data show that RB plays a critical role on controlling thymus size by suppressing TECs proliferation cell-autonomously, and also thymocytes proliferation cell non-autonomously. T cell differentiation and maturation were not affected by RB-TS inactivation in TECs, suggesting that RB-TS inactivation in TECs is not sufficient to disrupt T cell development. Thus, it is very attempting to manipulate TECs as a therapeutic strategy in order to enhance the immune function in a wide variety of patients. However, long term inactivating RB-TS in TECs will have detrimental effect on animals. We have finished this project and are currently in the process of preparing the manuscript for publication. Y. Song, T.Sullivan, K.Klarmann, D.Gilbert, T. N. O'Sullivan, L.Lu, D.C. Haines, J. Keller, and T. Van Dyke. Inactivation of RB in thymic cytokeratin 18 cells results in lymphoid and stromal proliferative disorder in genetically engineered mice (in preparation)	{ "pile_set_name": "NIH ExPorter" }
In response to signals of either endogenous or exogenous origins, mammalian cells implement changes in gene expression patterns that profoundly influence the global response of the cell. While the transcriptional events regulating changes in gene expression have been thoroughly studied, post-transcriptional processes, which are less well understood, are emerging as major gene regulatory mechanisms. Post-transcriptional gene regulation includes pre-mRNA processing and maturation, mRNA transport, stability and translation, as well as protein processing, modification and degradation. With respect to mRNA stability, we are investigating the mechanisms that regulate the expression of various proliferation-associated, cell cycle regulatory, and stress-response gene products. We previously showed that the function of the RNA-binding protein HuR, which stabilizes target mRNAs, is strongly regulated by the AMP-activated kinase (AMPK). In follow-up studies, we recently found that AMPK promotes the nuclear localization of HuR by directly phosphorylating and indirectly acetylating importin a1. In separate investigations, 'en mass' analysis of HuR target transcripts using cDNA arrays recently revealed a ~20-b RNA motif present in HuR targets, which allows the successful prediction of novel HuR target transcripts. Other work has recently shown that multiple RNA-binding proteins influencing post-transcriptional fate in opposite directions (mRNA-stabilizing HuR and mRNA-destabilizing AUF1) were capable of binding target mRNAs simultaneously as well as competitively. With respect to translation, a series of systematic, cDNA-based studies have shed light on important links between enhanced mRNA stability and translational inhibition during the endoplasmic stress response.	{ "pile_set_name": "NIH ExPorter" }
This project is a test of the efficacy of a new diagnostic method for imaging the airways known as virtual bronchoscopy. Virtual bronchoscopy is performed by acquiring thin-section computed tomography (CT) images of the chest. These images are used to generate a three-dimensional model of the tracheal and bronchial walls on a graphics workstation. The model can be manipulated to allow the viewer to fly-through the tracheobronchial tree providing views similar to those obtained using bronchoscopy. The technique produces a display of the human bronchial system in a readily understood format. Moreover, it allows investigation of post-stenotic portions of the bronchial tree that are beyond the reach of fiberoptic bronchoscopy. Further, virtual bronchoscopy may be used to guide interventional procedures. The patients who will be studied in this proto-col will be those having inflammatory, infectious, or neoplastic pulmonary processes who would have had chest CT for clinical reasons. These patients will be recruited from current NIH protocols. The study design consists of scanning of the thorax using thin-section heli-cal CT, followed by three-dimensional surface rendering of the airways and transfer of the digital data to videotape. In one of four parts of the protocol, the virtual bronchoscopy will be compared with results from fiberoptic bronchoscopy in a blinded study. In a second part of the protocol, the virtual bronchoscopy will be used to perform a descriptive analysis of cavity lung lesions. In the third part, the utility of virtual bronchoscopy in diagnosis of neo-plastic lesions of the chest will be studied. In the fourth part, certain technical problems in the virtual bronchoscopy procedure will be investigated. The patients will only have fiber-optic bronchoscopy for clinically indicated purposes. We anticipate that virtual bronchoscopy will be diagnostically efficacious for disorders that produce a morphologic alteration in bronchial anatomy. There have been no complications. Virtual bronchoscopy has been shown to be useful for detecting stenoses. We now have access to a CT scanner with higher Z-axis resolution and are investigating its efficacy for virtual bronchoscopy.	{ "pile_set_name": "NIH ExPorter" }
The process of vision begins with the absorption of light by the visual pigment, rhodopsin. In a carefully choreographed process, rhodopsin transmits this information in the form of a conformational change to the G-protein transducin. However, in order for the eye to achieve reproducibility and rapid response, the action of rhodopsin is antagonized by arrestin which selectively binds light-activated rhodopsin that has been phosphorylated, and thus sterically occludes further activation of transducin. This selective binding of rhodopsin by arrestin is at the core of the entire visual process. Despite arrestin's discovery nearly thirty years ago, we know relatively little about the mechanism by which arrestin regulates its selectivity for rhodopsin, binding rhodopsin only after it has been both light activated and phosphorylated. In this application, we seek to understand the regulation of the binding interaction between photoactivated, phosphorylated rhodopsin and arrestin, both in terms of the molecular dynamics of the arrestin structure and in terms of the arrestin/rhodopsin complex. Understanding the regulation of this interaction between arrestin and rhodopsin is central not only to vision, but also to other G protein-coupled receptor systems that are regulated by a similar arrestin-like mechanism. We will specifically address the following questions: 1) For arrestin to bind rhodopsin, what structural changes need to occur? 2) What are the effects of the phosphorylated C-terminus of rhodopsin on the molecular dynamics of arrestin? 3) What is the supramolecular nature of the arrestin/rhodopsin complex? The answers to these questions will provide an important advance in our understanding of the visual process and will provide important information on which to build therapies in diseases caused by deficiencies in the inactivation of the visual pigment. PUBLIC HEATH RELEVANCE: The arrestin protein is at the core of vision, serving to quench the visual pigment after it has absorbed light. Diseases such as congenital stationary night blindness and retinitis pigmentosa (a form of retinal degeneration) result from mutations in arrestin that interfere with the function of this protein. Understanding how these mutations affect the function of the protein will help direct the development of therapeutic interventions for diseases that result from arrestin defects. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
This report described patients along with previous echocardiographic evidence indicate an association between aneurysm of the fossa ovale membrane of the atrial septum and prolapse of the mitral valve. The presence of aneurysm of a sinus of Valsalva and congenital anomalies of both aortic and pulmonic valves in 1 of the 2 patients suggests a developmental association of these additional anomalies with atrial septal aneurysm and MVP.	{ "pile_set_name": "NIH ExPorter" }
Ear infections are among the most common reasons for sick child visits to medical providers. Risk factors for otitis media have been identified in prior clinical studies, but epidemiologic studies of sufficient size to allow generalization to the wider community or national population have been less common. Our goal has been to extend this research by analyzing several U.S. national data sets, collected under the auspices of the National Center for Health Statistics, with support from NIH, and other available population-based datasets. A paper was published this year on the prevalence and risk factors for placement of tympanostomy ear tubes in U.S. children. We have also examined recent trends in physician office, emergency room, and hospital visits for otitis media and treatments in the U.S., and analyzed data from the Third National Health and Nutrition Examination Survey (NHANES III). Findings of these and other studies were presented at the Seventh International Symposium on Recent Advances in Otitis Media (OM) in Ft. Lauderdale, FL, last summer and will be published in the Proceedings. Using four yearly surveys, the National Ambulatory Medical Care Survey, the National Hospital Ambulatory Medical Care Survey, the National Survey of Ambulatory Surgery, and the National Hospital Discharge Survey, we have been able to assess the burden of disease due to OM in recent years in the U.S. Using 1996 data, we found a national rate of OM of 40 visits per 100 children under the age of 15 years, or 33 visits per 100 children if restricted to physician offices only. Hispanic children under the age of 6 were more likely to be diagnosed with OM compared to non-Hispanic white or black children. Also, children in the West were more likely to be diagnosed with OM than children from other regions of the U.S. However, this higher prevalence of diagnosis did not translate into higher rates of surgical intervention with myringotomy and tube placement. Instead, the South had the highest rate of tube placement. In NHANES III and other studies, for example, the Children in Focus sample of the Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC), we have studied the prevalence of otitis media with effusion (OME) based on published criteria for "flat" tympanograms. We have correlated results of hearing exams and other developmental outcomes for children with OME and presented this information at the OM meetings as well. In addition, we have examined outcomes at age 5 for children with otitis media from a longitudinal study, the Scandinavian Successive Small-for-Gestational Age Birth Study. This study provides further support for poorer developmental outcomes of children with frequent ear infections early in childhood and for those with fluctuating hearing loss associated with OME. This study, based on middle-income families in Norway and Sweden with universal access to health care, suggests that even among these relatively advantaged children important differences in developmental outcomes are found, especially for children with fluctuating hearing loss in early childhood.	{ "pile_set_name": "NIH ExPorter" }
Continuously emerging new and hard-to-treat microbes, and the growing incidence of multi-drug resistant infections pose formidable challenges to human health. Innovative approaches are urgently needed to speed up the discovery of new anti-infectives. Our aim is to achieve a paradigm shift in antimicrobial drug discovery by finding next generation anti-infectives that prevent disease by blocking pathogen adaptation to host physiology. To this end we propose using whole live animals for high throughput screening of small molecules. We have developed infection models in the nematode Caenorhabditis elegans that can be used to identify drugs that cure otherwise lethal infections. High throughput screening of nematodes in 384-well plates is followed by secondary screening in a more highly evolved model host, the fruit fly Drosophila melanogaster, increasing the likelihood of isolating drugs that will work in humans. Our approach is applicable to many different classes of microorganisms, including bacteria, viruses, fungi and parasites. It has several advantages over traditional drug discovery: (i) In addition to identifying conventional antibiotics, it will uncover entirely new classes of anti-infectives that only exhibit in vivo activity. Examples are "virulence blockers" and "immune escape blockers". (ii) Our approach is unbiased and requires no prior knowledge of potential drug targets or pathways. (iii) It bypasses the current bottleneck of toxicity/efficacy testing by automatically eliminating toxic compounds (because they would kill the nematodes), yielding quality hits with in vivo activity. (iv) It will identify compounds that prevent or mitigate microbial resistance development, or can be combined with antibiotic therapy, thereby increasing antibiotic efficacy. We predict that our approach can identify compounds that inhibit diverse aspects of virulence: (i) adhesion and colonization, (ii) epithelial barrier disruption, (iii) deep tissue invasion, (iv) biofilm formation, (v) avoidance of immune recognition, and (vi) modulation of immune signaling. Some of the molecular mechanisms underlying these processes are conserved across bacterial species. To establish proof-of-principle, we seek funding for discovering new anti-infectives against Pseudomonas aeruginosa, one of several gram-negative bacteria that have recently emerged in a multi-drug resistant form for which efficient antibiotics are either limited or not available. We plan to screen a large number of chemical compounds (250,000) to maximize the discovery of new classes of anti-infectives. Promising compounds will undergo characterization, efficacy testing in other gram-negative bacteria (Klebsiella, Acinetobacter, Enterobacter) and testing in mouse models of infection. For highly promising candidates we will attempt molecular target identification. PUBLIC HEALTH RELEVANCE: Microbes that cause disease are becoming resistant to antibiotics faster than we can find new ones, making many common infections untreatable and life threatening. The goal of our project is to find a way to identify a new generation of antibiotics. Rather than simply preventing bacteria from growing, these new sophisticated drugs will prevent disease by interfering with a microbe's ability to interact with the human body.	{ "pile_set_name": "NIH ExPorter" }
The Indiana University School of Medicine Initiative for Maximizing Graduate Student Diversity (IUSOM IMGSD) program addresses the critical shortage of underrepresented ethnic minorities (UEM) in tenure-track faculty positions at medical schools in the United States. The long-term goals of the IUSOM IMGSD are to: (1) increase the number of UEMs who matriculate in and graduate from Ph.D. programs in the basic medical sciences (BMS) and (2) increase the number of UEMs, in general, who pursue careers as independent biomedical research scientists and faculty members. IUSOM IMGSD will accomplish these goals by (1) recruiting four new UEM graduate students into the basic science departments each year and supporting a total of 8 annually (each student will receive two years of IMGSD stipend support);(2) providing sustained multi-tiered mentoring through all years of doctoral studies, but particularly the initial two years of doctoral studies;(3) providing enhanced curriculum and development sessions during the first two years of doctoral study, including personal one-on-one tutoring when needed;(4) providing faculty mentoring for grant-writing with the goal of obtaining extramural funding for the remaining four years of doctoral education. These goals will result in four specific outcomes: (1) annually, four UEMs will be funded and ready to perform state-of-the-art research under the direction of a nationally known research mentor;(2) UEMs will be publishing in peer-reviewed journals;(3) at graduation with a Ph.D., UEMs will be placed in postdoctoral research positions which will (4) lead them to careers as independent biomedical research scientists or faculty members at a University in the U.S. where they will do biomedical research. The success of IUSOM IMGSD achieving its goals is based upon the theory that minority students with sufficient financial support, rigorous curricula, strong research experiences and multi-tiered mentors will complete their Ph.D. degrees and pursue careers as independent academic biomedical research scientists/faculty members.	{ "pile_set_name": "NIH ExPorter" }
The management of diabetic patients is often complicated by concomitant high blood pressure (hypertension) and high levels of LDL-cholesterol and triglycerides, often coupled with low HDL- cholesterol (dyslipidemia).The majority of diabetes related mortality is due to cardiovascular events, and epidemiological studies have shown that cardiovascular risk increases with increasing levels of blood sugar, blood pressure, and blood lipids. A variety of drugs are available to treat each of these conditions, and some have been shown to have an effect on cardiovascular risk. For example, controlling LDL-cholesterol with statin therapy reduces the rate of cardiovascular events in diabetic patients, but not to the level characteristic of non-diabetic individuals. The ACCORD trial investigated whether intensive pharmacological therapy in diabetic patients, with the goal of normalizing glycemia, blood pressure, and blood lipids, would further reduce cardiovascular events. However, no additional effect was seen with intensive blood pressure or lipid therapy, and intensive glycemia management actually increased mortality. These failures of seemingly rational treatment approaches could be the result of differential response of individuals to particular therapeutic regimens due to genetic polymorphism in genes relating to the metabolism or mechanism of action of the medicines used. Many candidate genes could be advanced as possible sources of this genetic variation, but our knowledge of all genes contributing to metabolic and cardiovascular phenotypes is incomplete, and therefore a candidate gene approach cannot be assured of identifying the relevant genes. We therefore propose a genetic study of the ACCORD trial that looks at functionally significant genetic variation in all genes in the human genome to investigate the following specific aims: 1) Identify genetic variants in patients from the ACCORD Lipid Trial that predict responses to treatment with fenofibrate. 2) Identify genetic variants in patients from the ACCORD Lipid Trial that predict responses to treatment with statins. 3) Identify genetic variants in patients from the ACCORD Glycemia Trial that predict acute responses to treatment with specific anti-hyperglycemic agents, and long-term responses to intensive vs. standard treatment strategies. Identification of genetic variants affecting outcomes of glycemia and lipid modifying therapies would enable the targeting of particular interventions to patients most likely to benefit and least likely to be harmed, improving cardiovascular outcomes and reducing the burden of morbidity and mortality attributable to diabetes. The genes containing these variants may prove to be novel targets for drug development, leading to new medicines for improving outcomes for diabetic patients in the future.	{ "pile_set_name": "NIH ExPorter" }
This is a phase II/III study to compare the efficacy and toxicity of 2'3'- Dideoxyinosine (DDI) (BMY-40900) and zidovudine (ADV) in patients with AIDS or advanced ARC who have tolerated ZDV therapy for twelve months or longer.	{ "pile_set_name": "NIH ExPorter" }
Developing an in-depth understanding of the factors that regulate the induction, quality, and longevity of anti- viral T cell responses is essential for devising rational strategies to combat viral infections. A hallmark of robust anti-viral immunity is the elaboration of potent CD4 and CD8 T cell responses which act cooperatively to bring about control of the infection. The consequences of ineffective responses can be catastrophic, resulting in viral persistence or the loss of long-lived immunological memory. Nevertheless, the signals which drive the development of the most robust polyfunctional effector responses and dictate the emergence of memory T cells are not fully understood. The purpose of this proposal is to capitalize on compelling preliminary findings which indicate that IL-21 plays a vital role in anti-viral immunity. Our initial studies clearly show that without sufficient levels of IL-21 the generation of polyfunctional effector CD8 T cells is compromised, and quite strikingly T cell exhaustion is exacerbated during chronic infections. Significantly, many of the phenotypic and functional impairments in anti-viral CDS T cells which manifest in the absence of IL-21 resemble the defects observed in CD4 T cell deficient hosts. A principal producer of IL-21 are CD4 follicular helper cells (Tfh) which function to help antibody responses, therefore Tfh derived IL-21 may be the critical factor that helps CD8 T cell responses. It is, however, unclear whether IL-21 is acting directly or indirectly on anti-viral CD8 T cells to promote their functionality, how T cell heterogeneity is shaped by IL-21, whether Tfh are the essential cellular sources of IL-21, and whether IL-21 based treatments have therapeutic benefits on viral control both during acute and chronic infection. We therefore propose the following specific aims: 1). Determine the direct effects of IL-21 on polyfunctional anti-viral CDS T cells. 2). Elucidate the role of CD4 T cell-derived IL-21 in promoting anti-viral immunity. 3). Define the significance of IL-21-producing CD4 T cells during viral infections of humans 4). Determine the therapeutic effects of IL-21 on anti-viral immunity and control.	{ "pile_set_name": "NIH ExPorter" }
Acute lung injury (ALl) is frequently associated with sepsis. Activation of procoagulant and fibrinolytic cascades occurs in ALl, and includes increased expression of urokinase plasminogen activator (uPA) and its inhibitors, such as plasminogen activator inhibitor (PAl-1), in the lungs. ALl is also characterized by the accumulation of large numbers of activated neutrophils in the lungs. Our preliminary data demonstrate a novel pro-inflammatory role for uPA. These results show that uPA contributes to acute lung injury produced by endotoxemia and also potentiates LPS and PGN induced neutrophil activation. Determination of the molecular mechanisms and receptors through which uPA enhances neutrophil activation and contributes to the development of ALl is the major goal of the proposed experiments. Our hypothesis is that processes involving the balance between uPA and its inhibitors contribute to neutrophil activation and pulmonary inflammation in sepsis induced ALl. The specific aims of this project are: 1) To elucidate the effects of uPA and the kringle domain of uPA on neutrophil activation, the receptors involved in uPA induced potentiation of neutrophil activation, and the role of uPA in contributing to the development of LPS and P. aeruginosa induced ALl; 2) To determine the importance of uPA generated plasmin in contributing to neutrophil activation and to the development of ALl; and 3) To examine how interactions between uPA and PAl modulate neutrophil activation and development of ALl. Our goal is to characterize mechanisms involved in neutrophil activation that are modulated by exposure to uPA, plasmin, and PAl-1, and that contribute to the development and severity of ALl. Delineation of such uPA dependent pro-inflammatory mechanisms, that may not involve proteolytic properties of uPA, will provide novel insights into interactions between fibrinolytic cascades and neutrophil activation, particularly in the setting of neutrophil dependent inflammatory responses, such as ALl. The proposed experiments are likely to suggest new therapeutic approaches to improve outcome from important clinical conditions, such as sepsis induced acute lung injury in which both fibrinolytic and neutrophil dependent pathways play major roles.	{ "pile_set_name": "NIH ExPorter" }
Nitrogen fixation, the reduction of atmospheric dinitrogen to ammonia catalyzed by the enzyme nitrogenase, is the sole biological process for replenishing the nitrogen that is used in the biosynthesis of cellular materials. This enzyme system consists of two metalloproteins, the Fe-protein and MoFe-protein, that mediate the coupling of ATP hydrolysis to substrate reduction. Nitrogenase is a prototypic example of an enzyme with multiple and varied iron-sulfur clusters that participate in electron transfer and substrate reduction, as well as providing an excellent model for energy transduction of ATP hydrolysis. We will utilize crystallographic, biochemical and spectroscopic approaches to investigate the enzymatic and metallocluster assembly mechanisms of nitrogenase. Special emphasis will be placed on establishing the atomic identity and mechanistic significance of the light-atom ligand we recently observed in the center of the FeMo-cofactor, and in assessing the structural framework for the nucleotide-mediated gating of electron transfer processes in nitrogenase. Towards these objectives, we will address: 1. Very high resolution (<1.2A) structures of the nitrogenase proteins, and particularly the associated metalloclusters, in defined oxidation states. This will include not only establishing the metric parameters of the clusters, but also the use of diffraction-based methods to assign the oxidation states to individual metal sites within each cluster, and to determine the orientation of the relevant EPR g-tensors with respect to the metallocluster structures. Intermediates in the incorporation of the FeMo-cofactor into the MoFe-protein will also be studied to address aspects of the biosynthetic mechanism of the nitrogenase metalloclusters. 2. The interactions of Fe-protein with MgATP and the site(s) of binding to the physiological electron donors for the reduction of substrates, ferredoxin and flavodoxin. 3. The mode of substrate and inhibitor binding to nitrogenase under turnover conditions, initially through spectroscopic methods and ultimately by crystallography. From these studies, we seek to establish molecular details of mechanistically relevant states of the nitrogenase proteins that are essential to defining a chemically explicit mechanism for substrate reduction. Broader implications will address nucleotide dependent transduction processes based on the similarities between Fe-protein and other nucleotide switch proteins involved in signal and energy transduction processes.	{ "pile_set_name": "NIH ExPorter" }
The National Survey of Oral Health in Seniors was conducted by NIDR in cooperation with Westat, Inc. during 1985-86 and provides the baseline for the oral health status of U.S. non-institutionalized persons aged 65 and older. The sampling frame was provided by the Area Agencies on Aging and the sample consisted of all older persons who attended selected multi- purpose senior centers within a designated 12 month interval. The sample of 5,649 seniors represented over 4 million older persons aged 65-103 who attend senior centers in the U.S. Clinical assessments were made for coronal caries, root caries and periodontal destruction. Demographic and health histories were collected for each older person participating in the clinical examination. The specific aims of this effort were to document the survey design for the senior sample and to produce a public use data file including clinical protocols and the statistical methodologies for calculating national or regional estimates, weights, and sampling errors. The documented file generated by this effort and the accompanying hard copy documentation entitled "Public use data file documentation and survey methodology--Part II Seniors" is pending release by the National Archives.	{ "pile_set_name": "NIH ExPorter" }
This protocol examines fat digestion and absorption and considers the possibility that there may be another transport form of these fatty acids other than chylomicrons in lymph. An alternative transport route of absorbed omega-3 fatty acids could be via the portal vein as free fatty acids of short-chain length. To investigate these possibilities, we are studying the digestibility and absorbability of omega-3 fatty acids of fish oil in human subjects.	{ "pile_set_name": "NIH ExPorter" }
Mast cells adhere to fibronectin following aggregation of high affinity receptors for IgE (Fc epsilon IR) on the membrane surface. This process is mediated by an alpha-5 containing integrin. Fc epsilon RI-induced adhesion is transient (compared to stem cell factor-induced adhesion), and occurs through 'inside-out signaling' which results in a conformational change in the surface integrins but does not alter their number. Mast cells spontaneously adhere to vitronectin. This event is followed by the phosphorylation of multiple intracellular proteins, a process which can be detected as early as 5 minutes after adhesion. Among the phosphorylated proteins is focal adhesion kinase (FAK). FAK is also phosphorylated after aggregation of Fc epsilon RI, or after the addition of c-kit ligand, also termed stem cell factor (SCF). IL-3 dependent mast cells undergo apoptosis (programmed cell death) following removal of IL-3. This is prevented by the addition of SCF. Addition of TGF-beta prevents this SCF-mediated rescue, thus demonstrating one way in which the microenvironment can control mast cell numbers. Mouse bone marrow-derived mast cells respond to both IL-3 and c-kit ligand and are inhibited by m-CSF, GM-CSF, and gamma-IFN. Mouse peripheral blood mononuclear cell-derived mast cells respond only to c- kit ligand. Human peripheral blood-derived human mast cells are derived from a CD34+ Fc epsilon RI- cell population. CD34 positivity is rapidly lost in culture, while the cell population becomes Fc epsilon RI+. The culture of human mast cells is enhanced if human serum is used instead of fetal calf serum. Fibrinoblasts in vitro selectively support the survival of mast cells, while internalizing and degrading basophils.	{ "pile_set_name": "NIH ExPorter" }
Arthropod-borne viral diseases have re-emerged as major health problems in most tropical countries. For example, the incidence of dengue fever has increased greatly over the past 20 years, recently including its hemorrhagic form with associated human fatalities. The mosquito Aedes aegypti is the primary vector. In addition, A. aegypti is an excellent laboratory model for examining vector interaction with a malaria parasite (Plasmodium gallinaceum) and a lymphatic filarioid nematode (Brugia malayi). Whole genome sequence data is available for humans, Drosophila melanogaster, the malaria parasite vector Anopheles gambiae, the malaria parasite Plasmodium falciparum, and several other parasite genome projects are underway. This proposal seeks to initiate an A. aegypti genome project, with an expectation that complete knowledge of the mosquito vector, parasite and human genomes will promote and foster new disease control strategies. The immediate objectives of this project are to: (1) sequence both ends of 40,000 cDNAs derived from normalized libraries produced from 4 specific mRNA sources (from pooled mRNAs isolated from various developmental stages and adult tissues and from females exposed to infected blood meals containing dengue virus, a malaria parasite, and a filarial worm parasite), (2) to sequence both ends of 50,000 genomic DNA Bacterial Artificial Chromosome (BAC) clones, and (3) to use fluorescent in situ hybridization (FISH) technology to physically map 1000 A. aegypti BAC clones to metaphase chromosomes. These aims will develop a body of genome information including EST sequences, genomic sequences, and the physical map positions of large genomic clones that will enhance gene discovery, and also provide critical tools for an eventual complete genome sequencing project. The proposal complements a nearly completed genome project to examine the primary malaria vector, Anopheles gambiae, as the 2 species reflect the range of genome size and organization among mosquito species. The long range goal is to promote and foster collaboration between funding agencies to support an effort to sequence the entire A. aegypti genome.	{ "pile_set_name": "NIH ExPorter" }
The purpose of the research is to examine and evaluate electrophysiological measures which are related to the developing sensory, perceptual and cognitive processes of infants and young children. The general method is that of recording of computer averaged evoked EEG responses to visual and auditory stimuli in normal children and in children with abnormalities of sensory and mental development. The relationship between sensory evoked potentials, electroencephalographic, behavioral and biochemical parameters of development are being examined. The applications of evoked response recording to clinical diagnosis, i.e. in children with sensory, mental, and perceptual abnormalities, are being investigated and evaluated. The method is being applied as a tool in audiologic diagnosis. Research on the EEG and evoked potentials in patients with Down's syndrome, degenerative CNS diseases of early childhood, cortical blindness, and malnutrition is being done. The research will provide basic data on the maturation of the EEG and sensory evoked potentials of human infants during the course of development and will attempt to relate data obtained from psychological and neurological testing with the neurophysiological data. It will provide normative data for comparison with that obtained from patients with sensory and neurological handicaps.	{ "pile_set_name": "NIH ExPorter" }
This proposal is to request support for a Keystone Symposia meeting entitled "Cancer Genomics and Epigenomics", organized by Andrew P. Feinberg, Greg J. Hannon and Rolf Ohlsson, which will be held in Taos, New Mexico from February 19 - 24, 2008. In the "post-genome" era, in which the human DNA sequence is available, as are those of many animal models, cancer genetics is being transformed both by the access to this sequence and by the technology that the genome project facilitated. It is now clear that many genetic changes occur in tumors, including mutations and copy number changes, as well as an even larger number of epigenetic alterations, such as altered DNA methylation, chromatin modification, and loss of genomic imprinting. High-throughput genomic and epigenomic approaches are now available, but are also being developed to assess the scope of these genomic/epigenomic changes. There is also a deeper evolution of our understanding that the genome and epigenome may be intrinsically disrupted in tumors, for example by palindrome formation and whole-band methylation. These advances raise several major questions, including: Which genetic and epigenetic changes play a causal role in tumor initiation and progression? What is the genetic and epigenetic basis of cancer predisposition? Is there large scale disruption of the cancer genome and epigenome and how does this arise? And how can we exploit advances in cancer genomics and epigenomics? What are the cutting edge technologies that will advance the sensitivity of detection of subpopulations such as cancer stem cells? This symposium will explore the technological developments, large scale alterations, and gene-specific changes emerging in the genome-wide approach to cancer, as well as the implications of these discoveries for cancer prevention and therapy. [unreadable] [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
This application requests support for a scientific workshop focusing on achieving healthy weights in African American communities under PA-10-071. The proposed two-day scientific workshop would engage 80-100 academic researchers and community research partners to achieve the following objectives: 1) assess current knowledge of effective weight control interventions relevant to black communities, examining current evidence across the life course and from multiple disciplinary perspectives~ and 2) develop synthesis recommendations for research, practice, and policy. Participants will include academic scholars and scholars-in- training from public health, nutrition and food studies, exercise science, public policy, community development, marketing, economics, and other fields, as well as community research partners. The rationale for the conference relates to the lack of evidence on effective interventions in African American communities in the face of much higher than average obesity- related risks. However, studies published during the last 10 years, combined with the recent surge in community-based programming in this area, can support a fruitful dialogue and help to refine the focus of future initiatives. This focus directy addresses NIH strategic goals for elimination of racial/ethnic disparities in obesity and cardiovascular diseases. Given the urgency of controlling the obesity epidemic, a timely workshop that will engage researchers, community research partners, and others to extract and synthesize insights from current research and practice is warranted. The 2012 workshop will be the 5th in a successful series of biennial conferences convened under the auspices of the African American Collaborative Obesity Research Network (AACORN), a national research network that is based at the University of Pennsylvania. AACORN links scholars, researchers and community-based research partners nationally in efforts to increase the quality, quantity, and effective translation of research to address weight issues in African American communities. Workshop outcomes will be disseminated through the internet, academic journals, and other channels.	{ "pile_set_name": "NIH ExPorter" }
We propose to study cellular alterations in the lung which might occur and accompany the well-known pulmonary congestion and edema with endotoxemia. Endotoxin has been shown to affect cell membranes and to interfere with oxidative cell metabolism in organs other than the lung. We would study membrane functions such as permeability to Na ion, K ion and Cl minus, active Na minus K ions transport and membrane-linked enzymes, Na ion, K ion-ATPase, adenylate cyclase. Cellular energy yielding processes will be assessed by measuring contents of energy-related adenine nucleotides, studying mitochondrial respiratory functions, and by assaying enzymes related to aerobic lactate production. We have previously studied these cellular functions in liver and in lung by control animals and/or animals subjected to hemorrhagic shock. We therefore have, in most instances methodologies worked out for a detailed study of lung cells with endotoxemia. In order to distinguish effects of endotoxemia on the endothelial cells from those on other cells of the lung, we propose to isolate and culture the pulmonary endothelial cells prior to cellular measurements. We would also study endotoxin-related changes in the type II alveolar epithelial cellls isolated from lungs. We plan further to study possible effects of endotoxemia on the cellular ion and water transport capability of the tracheobronchaial epithelial cells. This would be studied by measuring the short-circulating current and ion fluxes across the tracheal wall. Such measurements would allow us to evaluate changes in the ability of the tracheobronchial epithelial membranes to actively secrete water into the tracheobronchial lumen, a function which could determine the fluidity of the mucosal secretions in the airways. Cellular studies of effects of endotoxing would be carried out in intact animals as well as in isolated perfused lungs to possibly separate the direct effects of endotoxin from those that are mediated via the formed elements of the blood. The studies will contribute to our understanding of cellular problems that might be occurring in the lung with sepsis.	{ "pile_set_name": "NIH ExPorter" }
The long term goal of the research is to elucidate the cellular and molecular basis of excess scar formation during wound repair. Keloids are tumor-like skin scars that affect 10 to 20% of people of African decent, Asians, and Hispanics without appropriate treatments. During the past funding period, we used 14 freshly isolated and low passages (<3) strains of normal and keloid fibroblasts from human patients and provided new evidence that (PAI-1) overexpression and elevated collagen accumulation are intrinsic features of keloid fibroblasts. We also provided new and different evidence of a causal relationship between PAI-1 expression and collagen accumulation: adenoviral overexpression and siRNA and shRNAmir suppression demonstrate that PAI-1 produces elevated collagen accumulation in normal and keloid fibroblasts, respectively. Finally, by testing protease-inhibitory and vitronectin-binding mutants of PAI-1 for their capacity to induce collagen accumulation, we found that the latter was equipotent with wild-type PAI-1 and the former was only ~50% effective. Thus, PAI-1 utilizes protease inhibition as well as another of its functions to control collagen accumulation (Tuan et al., 2008, Am J Pathol). The goals of the renewal application are 1) to advance a therapeutic strategy that targets plasminogen activator inhibitor-1 (PAI-1) to control keloid collagen accumulation and prevent or treat keloid formation;2) to further define and expand the mechanisms utilized by PAI-1 to regulate collagen accumulation in keloid fibroblasts.	{ "pile_set_name": "NIH ExPorter" }
The proposed MDRC brings together experienced and talented investigators, unique resources, and innovative techniques to develop medications for the treatment of cocaine and heroin abuse. Existing medications, primarily to treat heroin addiction, all have serious flaws. A major theme of this Center is that the addict population is heterogeneous and effective medications will most likely emerge for subgroups rather than as universal antidotes. The second major theme is the search for better methods to identify these subgroups and to screen possible medications for effectiveness prior to instituting large scale clinical trials. We will use tools as diverse as improved psychiatric screening, cue reactivity paradigms, laboratory self-administration protocols, brain imaging techniques, and staggered clinical trials. The proposed Center consists of 3 projects. Project I will develop a smoked heroin paradigm and then use it in the Residential Laboratory to evaluate both new medications for opiate addiction, e.g., depot naltrexone and methoclocinnamox, as well as improving outcome and acceptability of existing medications such as methadone and naltrexone. Project 2 will use an improved SPECT imaging technique to demonstrate the correlation between regional brain activation and different phases of cocaine withdrawal, developing markers for evaluating new medications. A promising medication, whether to treat withdrawal, craving, or act as a blocker, could be studied for its effect on the relevant brain markers. The quantitative nature of the findings could cut through the thicket of variables that confound integration of controlled clinical trials. In keeping with the theme of targeting subgroups, Projects 3 will evaluate treatment seeking cocaine (3) abusers and then use existing psychiatric medications to see whether treating specific disorders such as depression or anxiety will improve outcome and retention. Project 3, in addition, will use both a cue reaction paradigm and a novel clinical trial design to pilot test new medications for cocaine treatment. Overall, the Center has the potential to become an important resource in the search for better medications for heroin and cocaine abuse that will improve our treatment capability.	{ "pile_set_name": "NIH ExPorter" }
All animals have evolved behaviors that result in innate responses to the external world. These responses can often be observed without prior learning or experience, suggesting that the neural circuits that generate them are developmentally programmed. In mice sexually dimorphic behaviors represent a set of innate behaviors that are controlled by odorant cues and by internal regulators such as gonadal hormones. Sexually dimorphic behaviors are qualitative or quantitative differences in behavior between the sexes, and much work remains to be done to characterize the neural circuits that mediates these behavioral responses. Such innate behavioral differences between the sexes result from sexually differentiated neural circuits. Testosterone and its receptor, the androgen receptor (AR), are required for male-specific behaviors. We and others observe sexual dimorphism in AR expression in a pool of neurons within the bed nucleus of the stria terminalis. This research proposal takes a genetic approach to characterize the role of this AR+ dimorphic subpopulation of neurons in sexually dimorphic behaviors in mice. This project will examine whether the AR+ BNST neurons are activated during mating and aggression, whether odorant cues are sufficient to activate them, and whether vomeronasal odorant detection is utilized to relay these odorant cues. Using gene targeting this project examines the behavioral consequences of ablating the dimorphic BNST neurons in the adult animal. Finally, using a genetic strategy this project will examine the function of AR in the dimorphic BNST by deleting the AR gene in the adult animal. An inherited loss of function of AR in humans manifests as physical and behavioral feminization (androgen insensitivity syndrome). In adult humans anti-androgen therapy or low levels of testosterone may be associated with a loss of libido and emotional well-being. Our examination of animals with a deletion of AR in BNST neurons should shed some light, in principle, on how neurons respond to a loss of testosterone signaling. More generally, our studies should further our understanding of how discrete brain regions integrate external sensory cues with internal physiological states to generate meaningful behavior.	{ "pile_set_name": "NIH ExPorter" }
The ability of various receptor agonists and antagonists were used to characterize the discriminative stimulus effects of ethanol. Antagonists at the 5HT3 receptor/ion complex were found to block the discriminative stimulus properties of ethanol in pigeons given orally administered ethanol. These data confirm data from electrophysiological studies which demonstrated that ethanol was potentiating the effects of serotonin at the 5-HT3 receptor. The discriminative stimulus properties of ethanol were blocked in a dose dependent manner by two antagonists acting at the 5-HT3 receptor. Increasing the dose of ethanol could overcome this blockade. The specificity of the blockade by 5-HT3 antagonists was tested using antagonists of the 5-HT2 receptor and a dopaminergic antagonist. Neither of these compounds blocked ethanol's discrimination. The ability of the 5-HT3 antagonists to block other behavioral effects of ethanol was tested using an hypnotic dose of ethanol (measuring loss of righting reflex), and a moderate dose of ethanol (measuring motor coordination). The 5-HT3 antagonists did not block either of these effects of ethanol, suggesting that the action of ethanol at the 5-HT3 receptor is not related to the motor effects of ethanol. The role of the 5-HT3 receptor in mediating the reinforcing effects of ethanol was also tested in a preparation using a rhesus monkey self-administering ethanol intravenously.	{ "pile_set_name": "NIH ExPorter" }
Nucleoside reverse transcriptase inhibitors (NRTIs) are an important component of long-term AIDS therapy and are associated with a host of mild to lethal side effects that are caused by mitochondrial toxicity of the drugs, often localized to adult non-replicating tissues. These drugs must be converted by cellular enzymes to the triphosphate to be active as inhibitors of the viral reverse transcriptase. Two mechanisms of toxicity have been proposed. The analog triphosphates inhibit the mitochondrial DNA polymerase leading to mtDNA depletion; or for at least one NRTI, AZT, toxicity may be caused by the inhibition of thymidine phosphorylation with alterations of the dNTP precursor pools supporting mtDNA replication. While the rates and levels of phosphorylated NRTI analogs have been measured in dividing cells in culture, the degree to which these drugs are modified or phosphorylated in adult non-replicating tissues in which the expression of salvage enzymes are expected to be different has not been studied. The goal of this proposal is to obtain a better understanding of the toxicities of the recently released and actively used NRTIs by studying their metabolism in non-replicating tissues and isolated mitochondria. This goal will be accomplished by: 1) Determining the conversion of radioactive NRTIs to their tri-phosphates in mitochondria isolated from a variety of rat tissues, in the perfused heart, and in vivo in a variety of rat tissues; 2) Using NRTIs and tissues identified in (1) that have high levels of NRTI-triphosphate, perform long term treatment studies to determine if NRTI triphosphates are associated with mtDNA depletion, and if so; 3) study the effect of exogenous addition of competing deoxynucleosides and/or uridine on NRTI phosphorylation and mtDNA depletion; 4) Determine if any of NRTIs inhibit phosphorylation of the typical deoxynucleosides in mitochondria, the perfused heart, or in vivo in other non-replicating rat tissues; 5) Using NRTIs identified in (4), determine if dNTP pools are disrupted and if this disruption leads to decreases in mtDNA level, and if so; 6) Study the effect of addition of the naturally occurring competing deoxynucleosides and/or uridine on correcting the dNTP defect. To broaden this analysis, mRNA levels of enzymes of the deoxynucleoside salvage and synthesis pathways will be measured by RT-PCR to capture changes in gene expression that may be caused by long-term drug therapy. Further, microarray and proteomic analysis will be included to correlate with the RT-PCR results and to capture genome wide gene expression changes that may provide important information relative to toxicity. Finally, as NRTI toxicities are reported to be different between males and females in both humans and rodents, and because these differences may be related to sex specific differences in metabolism, all of the above studies will be done in male versus female cohorts. The successful completion of the aims of this proposal will not only provide significant essential data in understanding NRTI toxicity, but will also address sex specific differences and potential treatments including uridine supplementation, which can only improve rational therapy to limit toxicity.	{ "pile_set_name": "NIH ExPorter" }
Identifying and intervening molecular pathways that directly contribute to vascular complications will have a significant clinical impact in preventing insulin dependent diabetes mellitus (IDDM)-associated vascular complications. We hypothesize that impaired physiological repair mechanisms due to altered endothelium- derived relaxation factor (nitric oxide) signaling pathway mediated by sustained high glucose is responsible for vascular complications. Under normal physiological condition, interaction of heat shock protein (Hsp-86) with endothelial nitric oxide synthase (NOS III) is responsible for nitric oxide-dependent endothelial functions. However, under high glucose condition, there is an enhanced inhibitor kB kinase activity (IKK-2), which competes out NOS lll from binding to Hsp-86. Further IKK-2 binding phosphorylates Hsp-86. This sequence of events lead to inadequate availability of Hsp-86 to NOS III, which decelerates NOS lll activity with a final outcome of integrin-focal adhesion disassembly, delayed endothelial migration and poor vascular repair. Blocking IKK-2 with genetic or pharmacological inhibitors in combination with agents, which are already in clinical practice, may open new avenues to improve nitric oxide production and reduce vascular damage in IDDM patients. The proposed study constitutes innovative approaches with the use of in vitro cell culture based assays using mammalian aortic endothelial cells. Use of genetically-induced diabetic animal model and optimized arterial injury procedures further complement the in vitro mechanistic approach. The specific aims are: Aim 1 will investigate the competitive cross talk between NOS III and IKK-2 with Hsp-86 under the influence of high glucose. The effect of sequentially altered nitric oxide signaling pathway on endothelial migration will be evaluated. Aim 2 will determine in vivo, the contribution of Hsp-86 - IKK-2 cross talk in NOS lll dysregulation in Type-1 mouse model. Aim 3 will evaluate the potentials of blocking IKK-2 independently or in combination with L-arginine in improving endothelial repair mechanism in response to arterial injury. The knowledge gained will help to identify new avenues that will improve the current treatment modalities aimed to prevent or minimize the severity of vascular complications of IDDM patients.	{ "pile_set_name": "NIH ExPorter" }
The acquisition of a pulsed liquid phase protein sequencer, high sensitivity amino acid analyser and a microbore HPLC is proposed to equip a centralized protein chemistry laboratory in the Wistar Institute. This equipment will be vitally important for at least 17 projects which are described in this proposal. In several cases specific projects represent the protein chemistry needs of relatively large research groups and these 17 projects will therefore directly or indirectly benefit the research efforts of about half of the 80 professional scientific research staff members of the Wistar Institute. Acquisition of this equipment is part of a major commitment of this institute to develop a modern fully equipped facility which can provide essential protein chemistry services and collaborative resources to diverse biomedical problems. The facility will be largely self-sustaining via charge backs to investigators after the first year. It will be available to all potential users at the Wistar Institute with priority given to the user groups described in this proposal. The P.I. of this proposal will assume direct administrative and scientific responsibility of the facility. An internal Advisory Committee comprised of users and administrative officials will aid in establishing priorities and policies to ensure maximum utilization of this resource. The administration of the Wistar Institute has allocated space and has made the necessary staff commitments for efficient establishment of this facility. The Institute is also committed to maintaining these instruments for their useful lifetime.	{ "pile_set_name": "NIH ExPorter" }
This Center assists investigators in the U.S.A. and abroad in the identification of arboviruses. The Center maintains a complete collection of virus seeds, antigens, and immune fluids which it uses for its own work as well as distributing them on request to specialized laboratories. During 1975, the Center received virus and/or antiserum to 84 strains from 18 countries. Viruses were identified from Venezuela, U.S.S.R., Senegal, Scotland, Norway, Malaysia, Indonesia, Ecuador, Brazil, Australia, Ethiopia, Thailand, Portugal, Kenya, and several areas of the U.S.A. At least 20 of these are either new viruses or new to the geographic region or host in which they were found. Serologic surveys of Turkey, Liberia, and Ghana were done. Serological tests to determine the etiology of Korean hemorrhagic fever, juvenile arthritis in Connecticut, and chromosome breakage in the Yanomama Indians were undertaken. New techniques for study of arboviruses were developed including type-specific identification of California group viruses by CF with one injection hamster sera, and trypsinized human O cells for the HI test. The goal for next year is the identification of 80 viruses from the Americas and other parts of the world. New techniques for study of the antigenic, biologic, and physico chemical properties of viruses of major human disease potential such as Crimean hemorrhagic fever-Congo, yellow fever, dengue, and Venezuelan encephalitis are being developed and will be stressed. BIBLIOGRAPHIC REFERENCES: Bowen, G.S., Tomori, O., Wulff, H., Casals, J., Noonan, A., and Downs, W.G. Lassa fever in Onitsha, East Central State, Nigeria, 1974. In press, Bull. Wld. Hlth. Org., 1976. Casals, J. Arenaviruses, Yale J. Biol. & Med. 48: 115-140, 1975.	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY/ABSTRACT The Cancer Cell Biology (CCB) Program of Winship Cancer Institute of Emory University is a laboratory-based translational program focused on understanding the changes in function of human cells as a result of cell transformation. Deregulation of the intra- and extra-cellular functional interactions of the cell's proteome leads to abnormal cell phenotypes, and these processes are at the root of the cancer process. The research within the CCB Program is organized into three scientific themes: (1) Cell Survival and Death Mechanisms, which focuses on intrinsic and extrinsic regulation of cell death through understanding and targeting both the Bcl-2 family and of death receptors; (2) Tumor?Stroma Interactions, which seeks to understand how tumor cells communicate and adhere, the biology of cancer stem cells, angiogenesis and immune interactions, and the process of EMT that is important for cell motility, invasion, and metastasis; and (3) Cancer Cell Metabolism, which focuses on how genetic alterations affecting signaling pathways impact the posttranslational modifications of key metabolic enzymes and redirect metabolic intermediates toward the synthesis of biomolecules of cell growth and proliferation. Under the leadership of Erwin Van Meir, PhD (leader) and Lawrence Boise, PhD (co-leader) the CCB Program has 36 core members from 16 different departments in the School of Medicine or Emory College. Between 2012 and the present, this highly collaborative group of researchers published 541 cancer- relevant scientific articles. Of these, 94 (17%) were intra- and 163 (30%) were inter-programmatic collaborations, and 214 (40%) involved a collaboration with another cancer center or academic organization. As of March 31, 2016, CCB held $17.7 million in annual total cancer-relevant research funding, of which $12.2 million (69%) was awarded directly from the NCI. The scientific advances driven by the CCB Program are highly significant in that they unravel novel mechanisms underlying cancer formation and growth and, in this process, unveil potential therapeutic targets.	{ "pile_set_name": "NIH ExPorter" }
The major goal of the proposed research is to employ modern functional neuroimaging in normal subjects to characterize brain areas related to specific aspects of explicit memory and skill learning. This proposal focuses on three specific areas of research including: 1) experiments designed to identify brain regions used during various encoding and retrieval tasks. Models of explicit (or declarative) memory often include processes related to selection of information to be stored, encoding of that information, information storage, and information retrieval. An extension of previous work on the functional anatomy of retrieval processes and a beginning exploration of brain regions related to encoding processes are proposed. 2. experiments designed to determine what task demands modulate blood flow in medial temporal lobe regions particularly in the hippocampus. Because damage to the medial temporal lobe, particularly the hippocampus and adjacent structures, produces specific deficits in explicit (or declarative) memory, much attention has been focused on the hippocampal contribution to memory. Experiments are proposed to explore the contribution the hippocampal regions might make to memory, focusing on ideas related to specific aspects of encoding. 3. experiments designed to explore the functional anatomy of skill learning. Skill learning appears to be intact in amnesic individuals. Skill learning also shows several properties that may be implemented by different neural systems. For example, evidence gathered from this and other laboratories supports the hypothesis that different brain regions are used in the performance of very similar tasks in skilled and unskilled versions. Experiments are proposed to assess the generality of these effects. Further experiments are proposed to explore other properties of skill learning, including passage through multiple stages of skill acquisition, chunking of simple skills Into more complex units, and hierarchical encoding of high-level motor programs. While this proposal focuses on memory and learning in normal subjects, the knowledge gained from such studies should be relevant across a wide range of health issues. Memory impairment is one of the most tragic consequences of degenerative illnesses, such as Alzheimer's disease and other dementias, as well as being a frequent complaint even in normal aging. An understanding of the normal human biology of memory can not help but to deepen our understanding of these health problems and lead to better approaches to their solution.	{ "pile_set_name": "NIH ExPorter" }
Unlike eukaryotes, most bacteria do not have a proteasome, but instead, use other protease complexes such as ClpAP and HslUV for their ATP-dependent proteolysis needs. In Mycobacterium tuberculosis (Mtb), however, proteasome is not only present, but its activity is essential for the pathogen to persist in the macrophages o mammalian lung epithelium. Recent research has firmly established that the mycobacterial proteasome is a druggable target and that inhibiting its activity could kill the bacteria. Surprisingly, Mtb uses a ubiquitin (Ub)-like tag, the prokaryotic ubiquitin-like protein (Pup), for delivering the doomed protein for proteasomal degradation. Despite functional similarities between Pup and Ub systems, recent work has revealed fundamental differences between these two conjugation pathways, and between Ub and Pup themselves. Therefore, the prokaryotes and eukaryotes have developed parallel but distinct mechanisms to regulate the protein stability by proteasomes. During the previous funding period, we have successfully addressed several important questions centered on the Mtb 20S proteasome assembly, gate closure and opening mechanism, and how the proteolytic activity is inhibited by the general or Mtb-specific proteasomal inhibitors. We have also revealed that the proteasomal ATPase Mpa recognizes and recruits the pupylated protein substrates via a binding-induced folding mechanism. In the next funding cycle, we will continue to study Mtb proteasome inhibition by novel compounds, and to understand structure and function of the Mtb Pup-proteasome pathway. PUBLIC HEALTH RELEVANCE: Mycobacterium tuberculosis (Mtb) is the causative agent for TB. The biological pathway Pup- proteasome is essential for Mtb to resist killing by the mammalian host macrophage. Our research on Mtb Pup-proteasome pathway will improve our knowledge on the unique system and facilitate anti-TB drug development.	{ "pile_set_name": "NIH ExPorter" }
One of the objectives of the proposed work is the evaluation of the biochemical effects of known and new anticancer agents on colon tumors and on normal tissues in mice. Study of inhibitors of pyrimidine biosynthesis on metabolism in tumors and normal tissues will be continued and effects of such inhibitors on the metabolism and action of pyrimidine analogs will be examined. Combinations of inhibitors of pyrimidine biosynthesis, such as N-(phosphonacetyl)-L-aspartate (PALA) and pyrazofurin, with fluorinated pyrimidines and with cytidine analogs are of particular interest. Metabolism of purine analogs, such as 6-thiopurines and 9-beta-D-arabino-furanosyl-2-fluoroadenine (2-F-AraA), their metabolic effects, and effects of nitrosoureas in combination with these analogs in murine colon tumors is another area of proposed research. We propose to continue studies on levels of enzymes of purine and pyrimidine metabolism in murine colon tumors and to extend these studies to human colon tumors and normal tissues. Results obtained will be of value in understanding effects of inhibitors; they provide a link between experimental studies in experimental neoplasms and clinical cancer. Alkylating agents (MeCCNU, cyclophosphamide) that are known to produce therapeutic responses in murine and human colon carcinoma will be studied for their effects in vivo on DNA of murine colon carcinoma and normal tissues. Proposed studies will include extent of alkylation, chain scission, cross-linking and repair.	{ "pile_set_name": "NIH ExPorter" }
. Ixodes ticks serve as the vector for 3 human infections in the U.S.-- Lyme disease, Babesiosis and human granulocytic ehrlichiosis. The incidence of all three diseases has been increasing over the last decade. Despite the availability of antibiotic therapy after infection, each of these diseases is associated with significant morbidity or, in some cases, even mortality. Preventative measures targeted towards human disease have been largely limited to tick avoidance methods and there are currently no human vaccines available for any of the diseases. An alternative strategy for prevention of human disease would be to develop treatments to reduce carriage and transmission of these pathogens in their wild reservoirs, Ixodes ticks and small rodents such as white-footed mice. Our laboratory has been involved in developing a vaccinia virus (VV) based anti-Borrelia burgdorferi vaccine for white-footed mice based on the outer surface A (OspA) protein of B. burgdorferi. While this strategy has shown promise, protection from an OspA vaccine for mice is limited to Lyme disease. Recent progress by other investigators has shown the potential for the development of anti-tick vaccines. Vaccination of animals with tick antigens has been shown to decrease feeding and infestation by ticks and/or to decrease transmission and acquisition of infectious agents by ticks. In this proposal, we will test tick antigens expressed in a VV vector for their ability to protect against tick infestation and disease transmission when administered orally to mice. Identification of anti-tick targets will allow us to develop a multivalent vaccine containing both tick and pathogen antigens for improved reduction of tick borne diseases in reservoir animals. The overall goal of this project is translational, in that, we will attempt to make use of recent advances in scientific understanding and apply them towards the development of a vaccine that could have a direct impact on the incidence of human, tick transmitted diseases through reduction of disease in wildlife reservoirs. The tick-borne infections, Lyme disease, Ehrlichiosis and Babesiosis, are significant public health problems in the U.S. One potential approach to the control of these diseases is to reduce carriage of the organisms in their wild-life reservoirs. In this proposal, we outline a strategy to develop an ecologically targeted vaccine that blocks transmission of these pathogens between tick and mammalian reservoirs through inhibition of tick feeding, thus reducing human infections by reducing the numbers of infected ticks.	{ "pile_set_name": "NIH ExPorter" }
Shigellosis is a global human health problem and a biodefense area of concern. The disease, caused by Shigella species, is a significant cause of morbidity and mortality accounting for 164 million cases worldwide and 1.1 million deaths annually, most notably amongst children. The disease is extremely infectious, requiring only 10-100 bacterial cells to initiate infection. Moreover, there is a global rise in the occurrence of muti-drug resistant isolates, such as the epidemic and pandemic Shigella dysenteriae type 1 strain. Despite this trend, the standard methods for the confirmed identification of the disease and the determination of antibiotic susceptibility are traditional culturing methods, followed by serologicl and biochemical tests; these assays require 24-48 h to complete in a lab environment. The long-term goal of this research is to develop a rapid, and portable diagnostic assay technology that can detect the causative agents of shigellosis. Importantly, the diagnostic technology will be able to simultaneously provide an antibiotic susceptibility profile, which will enable appropriate treatment options and thus lead to improved patient prognosis. The R21 phase will generate the proof-of-principle results for the development of light-tagged Shigella reporter phages that can detect Shigella spp. by specifically conferring a bioluminescent signal response. Aim 1 will identify and prioritize Shigella phages which display species specificity and broad strain infectivity. Aim 2 will integrate the bacterial luxAB reporter genes into non-essential sites of th prioritized Shigella phage genomes to create luxAB-tagged reporter phages. In the presence of target bacteria, the reporter phage bind to specific cell receptors, inject their phage DNA and use the host's transcriptional and translational machinery to produce the luciferase enzyme. Upon substrate addition, the ensuing bioluminescent response can be readily detected. Following the demonstration that the Shigella reporter phages have the necessary detection attributes, the R33 grant will develop the reporter phage technology into a clinical diagnostic for shigellosis. We hypothesize that the bioluminescent phage detection system will be able to: (i) rapidly (within minutes) detect the presence of Shigella and differentiate between the species; (i) require significantly fewer cells to achieve a positive signal (more sensitive); (iii) function dirctly with clinical specimens (does not require the isolation of pure bacterial cultures); (iv) provide concurrent antibiotic susceptibility data (and help patient prognosis), and (v) function with a simple handheld detection device (field appropriate/battery operated system). The cost of producing the reporter phage and the consumable costs are minimal. The assay does not require technical expertise or processing. Consequently, we believe the technology is particularly well suited to resource-limited settings and will be able to function in a non-laboratory environment as per the requirements set forth in this RFA.	{ "pile_set_name": "NIH ExPorter" }
The purpose of this study is aimed at evaluating EEG based online monitoring as a practical attainable tool to predict and prevent critical decrements in performance and alertness usually occurring after sleep loss or sleep curtailment typically encountered on shuttle missions.	{ "pile_set_name": "NIH ExPorter" }
Segmentation of the embryonic vertebrate hindbrain into rhombomeres ensures proper positioning of hindbrain derivatives and is essential for normal neural activity. Rhombomere formation is regulated by hox genes whose expression is carefully coordinated to ensure correct timing and spatial distribution of transcripts. hox expression is regulated by a combination of retinoic acid (RA) mediated signaling and by hox genes regulating each other's expression. Our preliminary data reveal a nucleosome-depleted region (NDR) forming at hox promoters in response to RA. We also find that Prep and Pbx cofactors occupy hox promoters well before onset of transcription and that the presence of these cofactors correlates with 'active' histone modifications and recruitment of RNA Polymerase II. We find this constellation of factors to be insufficient to drive transcription - instead, Hox proteins appear required for initiation of transcription. Based on these data we hypothesize that activation of hox expression is achieved by: 1) RA inducing an NDR at hox promoters, 2) Prep and Pbx cofactors binding at the NDR, 3) recruitment of histone-modifying enzymes and RNA Polymerase by Pbx and Prep, 4) binding of Hox proteins to activate transcription. We have developed two aims to test this hypothesis: 1) Define sequential action of cofactors and Hox proteins in initiation of hox gene transcription. In this aim we will establish te order of action and specific roles for each factor required for hox gene expression, 2) Define the role of nucleosome-depleted regions in the activation of hindbrain hox gene expression. In this aim we will determine how NDRs are formed and what role they play in control of hox gene expression. Our work will impact human health by exploring how the embryonic nervous system forms. The importance is underscored by the fact that human hox mutations produce developmental disruptions of the brainstem and cause cognitive defects such as autism. Since our work will improve our understanding of how neural gene expression is initiated in undifferentiated cells in general, our results will also inform studies aimed at driving the differentiation of precursor cells (such as ES and iPS cells) towards a neural fate for basic and clinical applications.	{ "pile_set_name": "NIH ExPorter" }
The long-term objective of the proposed research is to learn more about: 1) the effects of anesthetic agents on cerebral blood flow (CBF) and metabolism (CMR); 2) the relationships of these effects to basic pathomechanisms of focal ischemia; and 3) ways of modifying outcome from such ischemia insults occurring during anesthesia and surgery. Our specific aims are to test the hypotheses that: 1) the extent of permanent injury resulting from focal ischemia is related to the anesthetic present during the insult; 2) acute anesthetic- induced changes in CBF and CMR can be related to outcome: 3) the volume of brain tissue at risk for infarction is related to the manner in which different anesthetics preserve or alter the matching of CBF and CMR; 4) anesthetic effects on CBF and CMR relationships vary with time; and 5) the duration of anesthetic exposure prior to the ischemic insult will influence the volume of tissue "at risk" and this may vary with anesthetic agent. The following methodology will be used to test our hypotheses. Physiologically stable rats will be anesthetized with specific doses of isoflurane, halothane, or pentobarbital with duration of anesthetic exposure carefully controlled. Anesthetic effects on regional CBF and CMR will be simultaneously quantitated by 14C- iodoantipyrine and 14C-deoxglucose autoradiography respectively. Volume of ischemic tissue at risk for infarction as a result of middle cerebral artery occlusion will be determined by computer- assisted image analysis. This volume will then be related to anesthetic dose/response effects on CBF/CMR ratios. Outcome will be evaluated by: 1) computed quantitative 3-dimensional reconstruction of infarct volumes; 2) behavioral patterns; and 3) densitometric determination of brain water content. The health relatedness of this proposal derives from the fact that numerous conditions requiring surgery predispose patients to potentially disastrous cerebral ischemic insults while receiving anesthesia (e.g. carotid endarterectomy, cardiopulmonary bypass). Significant advances have been made in understanding basic pathomechanisms of cerebral ischemia and anesthetic effects on cerebral physiology, yet indications for application or avoidance of various anesthetic techniques during potentially ischemic clinical situations remain poorly defined. The proposed studies will contribute to our understanding of these problems.	{ "pile_set_name": "NIH ExPorter" }
This HIVRAD program builds upon our recent successes in generating stable, proteolytically mature gp140 trimers (SOSIP gp140s) that mimic virion-associated envelope (Env) in topology and antigenicity. The overall goals of this HIVRAD program are reflected in three major milestones: 1) determine the structure of cleaved Env trimers at <4A resolution, 2) demonstrate methods to overcome HIV-1 Env's immunosuppressive properties, and 3) identify a SOSIP trimer vaccine that elicits heterologous neutralization of diverse HIV-1 isolates. These goals are reliant on the preparative production of novel SOSIP trimers in highly purified, homogeneous form. SOSIP trimers are exceptionally complex biomolecules whose production requires a high level of technical expertise, and Core B lends this expertise to the HIVRAD in order to support the goals of Project 1 (HIV-1 Env Vaccine Design, Dr. Moore) and Project 2 (HIV-1 Env Trimer Crystallography, Dr. Wilson). The overall role of Core B is expressed in terms of four Specific Aims: 1) Produce and characterize Env proteins for crystallography studies;2) Produce and characterize Env proteins for immunogenicity studies;3) Evaluate the immunogenicity of Env proteins in small animals, 4) Evaluate Env proteins for effects on human dendritic cells and antigen presentation in vitro. In addition to producing SOSIP trimers, Core B provides a central resource for additional high-quality materials (e.g., gp120 monomers, control antigen, CD4-based proteins, antibody Fabs) that are required by Project 1 and Project 2. Core B also possesses significant experience in vaccine testing and thus will be responsible for performing small-animal immunogenicity studies in collaboration with Project 1. Lastly, Core B will collaborate with external groups to explore and understand the interactions of HIV-1 Env with human dendritic cells in vitro. In addition, Core B will serve as a central point of contact and will manage consortial/contractual agreements with several leading academic and corporate collaborators who lend specialized expertise in the areas of vaccine delivery (Aldevron, Fargo, ND), NAb analyses (Monogram Biosciences, South San Francisco, CA), NAb specificity analyses (Dr. James Binley, Torrey Pines Institute for Molecular Studies, San Diego, CA), in vitro immunogenicity studies (VaxDesign, Orlando, FL), and exploratory immunogenetics (Dr. Sunil Ahuja, University of Texas). In fulfilling these roles, Core B represents an integral element of this innovative, multidisciplinary HIVRAD program. RELEVANCE (See instructions): Nearly 1% of the world's population is infected with HIV, and a preventive vaccine is urgently needed. Most efficacious vaccines elicit antibodies that can neutralize the pathogen, but current-generation HIV vaccines are not effective in this regard. Obstacles include our limited understanding of the structure and immunology of HIV-1 envelope trimers. This HIVRAD represents an innovative approach to addressing these challenges in order to provide a fundamental advance in our ability to elicit HIV-neutralizing antibodies with a vaccine.	{ "pile_set_name": "NIH ExPorter" }
This proposal requests funds to support the 4th NIH Roadmap Membrane Protein Structure and Technologies Conference. The NIH Roadmap Initiative was created to focus short term on critically important, transformative, high impact, an strategically important areas of biomedical importance. Since membrane proteins play such a critical role in so many cellular and physiological processes and are so difficult to express, solubilize and purify, the production of membrane proteins was identified as a serious roadblock to understanding their structure and function and chosen as one of the top Roadmap Initiatives. This initiative is currently funded by NIH-wide support via the NIH Common Fund. The first phase of the Structural Biology Roadmap Initiative focused on creating Centers for Innovation in Membrane Protein Production. Two Centers, each with ~8 PIs, one at University of California at San Francisco (UCSF) and the other at The Scripps Research Institute (TSRI), were funded and established in 2004. Because of the potentially transformative information that could be generated and shared, NIH staff initially decided that there should be an annual inter-Center meeting between the Centers. The first Inter- Center meeting was hosted by Robert Stroud at UCSF in May, 2006. Since then the NIH Common Fund membrane protein focus has been expanded. To date, 41 grants have been funded through this mechanism. The 2006 meeting was followed by 3 meetings with all Roadmap awardees contributing, hosted alternately by Ray Stevens at TSRI (11/2007) by Robert Stroud at UCSF (3/2009) and by Ray Stevens at LaJolla Shores (11/2010). These NIH Roadmap Membrane Protein Structure and Technologies Conferences became the primary forum for exchanging technologies for membrane proteins in a critical phase of a rapidly developing field. New families of membrane protein structures emerged through the program. The conference became extremely popular and numbers of attendees had to be limited in 2010. The 4th conference, hosted by Robert Stroud, is planned for November 28-30, 2012 at the Westin Market Street Hotel in San Francisco with workshops at UCSF on November 27th. This venue allows expansion of the number of participants to 250 attendees, and provides conference facilities where talks and poster presentations, as well as breakfast, lunch and coffee breaks are in adjacent spaces to encourage discussion and conversation during breaks. The length of the meeting is also extended from 1 1/2 days to 2 1/2 days to allow more participants to attend and present. The meeting will provide a unique opportunity for membrane protein biologists, members of all the projects funded through the RFA Membrane Protein Production and Structure Determination and members of the 9 Membrane Protein Centers funded by the Protein Structure initiative to discuss the current status of their projects. The meeting will also be open to other NIH funded researchers for attendance and abstract consideration for the program. PUBLIC HEALTH RELEVANCE: This proposal requests funds to support the 4th NIH Roadmap/Common Fund Membrane Protein Structure and Technologies Conference planned for November 28-30, 2012 in San Francisco. The length of the conference has been extended and the number of attendees has been expanded to accommodate all membrane protein researchers funded by the NIH Common Fund and Protein Structure Initiative and to open the meeting to other NIH funded PIs. Presentation and discussion will deal with progress as well as barriers to progress that could transform membrane protein structure determination.	{ "pile_set_name": "NIH ExPorter" }
These studies are primarily aimed at transplantation of catecholamine-containing tissues, including adrenal medulla, tumor cells, and embryonic brain tissue into the brain. The purpose of these experiments is to elucidate the properties of these tissues after transplantation and the response of the host brain to the transplanted tissues. Specifically, these experiments employ non-primate animal models to (1) develop the techniques of brain tissue transplantation for clinical use in Parkinson's disease; (2) develop brain tissue transplantation techniques so that they eventually may be applicable to other disorders, such as schizophrenia or Alzheimer's disease if and when these disorders become well enough understood to permit such applications; and (3) elucidate factors that control the development of the brain and responses of the brain to injury or impairment, with particular emphasis on the nigrostriatal dopamine system. During the past reporting year, significant progress has been made in these areas.	{ "pile_set_name": "NIH ExPorter" }
This study will survey the long-term patterns of hospitalization of 650 schizophrenic outpatients admitted to a research clinic between 1958 and 1963. The sample includes 250 patients with no previous history of state hospitalization. The study will be concerned with determining: (1) the clinical course of the entire cohort of 650 patients with specific emphasis on the frequency and duration of hospitalization, as well as duration of time between hospitalizations, separately for age, sex, race, and marital status; (2) the clinical course of the 250 patients in our sample who had no previous history of state hospitalization; (3) validation of the Hospitalization Proneness Scale (HPS) as a predictor of long-term clinical course; (4) the impact of changing patterns in mental health care (the advent of pharmacotherapy and community mental health care) on the patterns of hospitalization of schizophrenic patients; (5) the interaction between hospitalization proneness (HPS) and changing treatment patterns on clinical course; (6) a final objective will be to confirm the reliablility of the "clinic" diagnosis of schizoprenia with diagnoses made during hospitalizations.	{ "pile_set_name": "NIH ExPorter" }
This proposal is for a renewal of our Alcohol Research Center. It provides only for core activities. We are asking for support for eight core activities: an animal breeding colony; continuation of our twin registry; the secondary analysis of the many extensive data bases currently in the department; a molecular neurobiology facility; the training of interviewers for the many clinical, follow-up, family, and epidemiologic studies; biostatistical consultation; a tissue culture facility; and the continuation of our office to assist in the follow-up of patients and other subjects. Each of these core activities will provide support and consultation to individual investigators whose research could benefit from one of the core activities. The investigators will be expected to obtain their own support through the R01 mechanism. In addition to the eight core activities, the proposal requests support for two to three pilot projects at any one time, designed to enable investigators to carry out preliminary studies and to acquire a sufficient data base to permit them to submit applications for independent R01's. Each investigator seeking to carry out a pilot project will confirm to the university's rules governing human studies, animal experimentation, etc. The core budget provides partial support for a number of faculty members related to their responsibility for directing one or another core activity or for their participation in the overall scientific direction for alcoholism-related research.	{ "pile_set_name": "NIH ExPorter" }
We propose to develop a miniature real-time volumetric ultrasound imaging system employing two-dimensional (2D) capacitive micromachined ultrasonic transducer (CMUT) arrays. We will design and implement a miniature probe fitting into the tip of an endoscope that will be connected to an external beamforming unit using a small number of cables. The probe will contain a 2D CMUT array and integrated circuits to handle the data acquisition and front-end pre-processing. A small-sized active phased subarray will be multiplexed over a large transducer array in consecutive firing steps to allow for real-time data acquisition. The acquired data will be transferred to the external unit and processed to form real-time pyramidal volumetric images. CMUT technology provides excellent element sensitivity, large temporal bandwidth, and potential for integrating the transducer with front-end electronic circuitry. The image reconstruction algorithm is based on phased subarray processing which allows for a low-cost front-end architecture while providing high-quality real-time images. The combination of CMUT technology and phased subarray (PSA) imaging will enable us to realize a small ultrasound scanner where the size and cost of the front-end circuitry is critical. We have achieved the following preliminary results: (1) Identified several image-guided surgical applications for cancer treatment, (2) specified the probe parameters based on physician input, (3) fabricated 128-element 1D CMUT array and (4) used it to form a high-quality 2D image, (4) fabricated a prototype 16x16-element 2D CMUT array, (5) developed PSA theory for 2D and 3D imaging, (6) tested 2D PSA theory with simulations and experimental data, and (7) shown that the system specs allow for real-time 3D ultrasound imaging. We propose to take the next major developmental step and demonstrate real-time 3D imaging from a miniature probe by accomplishing the following specific aims: (1) form preliminary 3D images using the prototype array and existing acquisition system, (2) design and fabricate new 16x16 2D CMUT arrays, (3) implement real-time beamforming and visualization, (4) design and build integrated circuits for the front-end probe electronics, (5) perform final integration into a fully function real-time 3D ultrasound system, and (6) perform preliminary clinical evaluation.	{ "pile_set_name": "NIH ExPorter" }
Not provided	{ "pile_set_name": "NIH ExPorter" }
Selective serotonin reuptake inhibitors (SSRIs) are important drugs in the treatment of child and adolescent psychiatric disorders. SSRIs achieve their therapeutic effect by increasing the extracellular level of 5-HT. Animal experiments indicate that supra-physiological 5-HT levels during brain development can lead to irreversible cytoarchitectural and behavioral abnormalities. Our data also indicate that postnatal administration of fluoxetine in mice results in permanent brain abnormalities including neuronal hyperexcitability in a sensory pathway and audiogenic seizures. There have been no longitudinal studies to assess the long-term effects of SSRI-administration in adults who received treatment during childhood. Such studies require many years and even decades. Developmental processes are much faster in rodents allowing the evaluation of postnatal SSRI-treatment on adult behavior in a relatively short time scale. The goals of this proposal are to better characterize the long-term adverse effects associated with the postnatal administration of SSRI in clinically relevant doses and to specify some of the mechanisms that underlie these effects. [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The temporal resolution in fMRI studies is limited not only by imaging speed but also by a relatively slow intrinsic hemodynamic response and a finite signal-to-noise ratio. The temporal characteristics have been investigated in our laboratory. To determine the upper limit of temporal resolution in a single area during repeated tasks, motor cortex activity was investigated during visually instructed finger movements. In a separate study taking advantage of the temporal resolvability of fMRI, activity in the human primary motor cortex, the premotor cortex, and the supplementary motor area during a delayed cued finger movement task was measured by functional magnetic resonance imaging. Activity during movement preparation was resolved from activity during movement execution in a single trial.	{ "pile_set_name": "NIH ExPorter" }
[unreadable] [unreadable] Five to ten percent of premature infants will sustain significant neurological injuries. Periventricular white matter injury (PWMI) is now the most common cause of brain injury in preterm infants and is characterized by a loss of white matter and secondary ventricular enlargement. Unfortunately, strategies aimed at directly preventing PWMI are not available. Recently we discovered that adenosine, acting through A1 adenosine receptors (A1ARs), plays a major role in the pathogenesis of PWMI. We find that hypoxic rearing is associated with abnormal maturation of oligodendrocytes and delayed myelination, as in PWMI. Importantly, blockade of A1 ARs prevents hypoxia-induced ventriculomegaly and reduced myelination. Based on these observations we believe that it is possible to prevent hypoxia-induced brain injury and PWMI by blocking adenosine action using drug therapy. To develop a new clinical strategy for PWMI prevention, we propose the following Specific Aims: [unreadable] SA1. Test four different approaches for blocking adenosine action in hypoxia, including PEG-adenosine deaminase, which degrades adenosine and is an FDA approved drug; caffeine, which is a non-specific adenosine antagonist and is an FDA approved drug; 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), which is a potent and highly selective A1AR antagonist; SCH58261, which is an A2aAR antagonist. [unreadable] SA2. Test the effectiveness of the above approaches in preventing hypoxia-induced brain injury. [unreadable] SA3. Test the effectiveness of anti-adenosine therapy in preventing hypoxia-induced behavior disturbances. It is anticipated that these studies will lead to novel and practical strategies for directly preventing brain injury. [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The long-term objectives of this research plan are to determine how the vascular smooth muscle cell and mechanical characteristics of microvessels adapt to various pathological and physiological stresses, and to identify the regulatory mechanisms responsible for these adaptations. Physical work and exercise are constant physiological stresses, and exercise training is often prescribed in the treatment and management of various pathological conditions such as hypertension, diabetes, and obesity. The focus of the current proposal is to investigate the mechanisms responsible for the changes in vascular smooth muscle function and regulation of peripheral vascular resistance which result from aerobic exercise training in normotensive and hypertensive animals. Specifically, the goals of this proposal are to determine 1) if the vascular reactivity of skeletal muscle arterioles to various vasodilator and vasoconstrictor stimuli is altered as a result of exercise training, 2) if the enhanced functional hyperemia which is known to occur in trained skeletal muscle results in an excess delivery of oxygen to the contracting skeletal muscle tissue, 3) if arteriolar reactivity to constrictor stimuli is altered in a non-muscle tissue, the small intestine, as a result of aerobic training, 4) if the changes in vascular function resulting from training are similar between normotensive and hypertensive animals, and 5) if anatomical or mechanical changes contribute to a decline in peripheral vascular resistance in trained skeletal muscle of hypertensive animals. Arteriolar properties will be evaluated in the spinotrapezius muscle and small intestine of sedentary and aerobically trained normotensive Wistar-Kyoto and Spontaneously Hypertensive rats. Vascular reactivity will be assessed by measurement of arteriolar diameter and pressure during superfusion of adenosine and norepinephrine, application of a myogenic stimulus using the "box" technique, arterial hypoxia induced by respiration of a nitrogen-room air mixture, and stimulation of endothelial derived relaxant factor release by local application of acetylcholine. Arteriolar and venular hemoglobin oxygen saturation and the relative change in tissue NADH concentration will be evaluated in the resting and contracting muscle using image analysis techniques to evaluate the relative relationship of oxygen delivery to metabolic demand. Microvascular blood flow will be determined by measuring labeled red cell flux. Functional arteriolar and capillary density will be measured in the resting and contracting muscle and anatomical vessel density will be measured in fixed tissues. The passive mechanical characteristics of the arteriole wall will be evaluated in vivo and will be used to determine the active and passive contributions to total wall tension during the various perturbations. Total wall and vascular smooth muscle cross-sectional areas will be determined in fixed tissues and used to calculate changes in total and active wall stress during in vivo perturbations. The results of this study will elucidate the mechanisms responsible for the decline in peripheral vascular resistance known to result from aerobic exercise training in both normotensive and hypertensive animals, and may provide insight to the inherent physiological efficacy of exercise training in the treatment and prevention of various diseases which involve microvascular pathologies.	{ "pile_set_name": "NIH ExPorter" }
This proposed project aims to validate and refine a novel Magnetic Resonance Imaging (MRI) technique that uses targeting MR contrast probes to detect altered endogenous gene expression resulting from amphetamine exposure in living animals. We will use an animal model of addiction in C57Black6 mice. The targets to be investigated in this project are mRNA of a transcription factor fosB and the activator protein 1 (AP-1) whose altered expression is strongly implicated in drug addiction. Most current molecular imaging methods detect cell surface proteins as markers of gene expression. Our proposed technique detects intracellular products of gene expression, such as mRNA and its products that may have DNA binding ability in the brains of living mice. This method uses DNA with antisense sequence to the target mRNA or DNA with consensus sequence for AP-1 binding. In preliminary studies, we labeled these DNA with MR contrast agent and showed in a proof-of-concept study that the probe is capable of reporting elevation of c-fos mRNA live animals using MRI after acute amphetamine injection. Our goals are to extend and improve the sensitivity of this technique and establish the correlate between MR signal and histological assessment of intracellular targets. By achieving these goals, we will be able to apply this novel MR technique as a quantitative tool for the detection of endogenous gene expression of other genes such as cyclic AMP responsive elements (CREB) and a host of other products of gene expression that are implicated in drug addiction. PUBLIC HEALTH RELEVANCE: In this project, we will extend and refine a novel Magnetic Resonance Imaging (MRI) technique that we have developed to assess the efficacy of acupuncture treatment for substance abuse phenotype by detect altered gene expression in the brains of live animal affected by addictive drugs. The goal of this project is assess and correlate the gene and gene expression product modifications as the result of acupuncture treatment for drug addiction. This application will further our understanding of how acupuncture treatment impacts the progression of drug addiction. Our studies will open new avenues for pharmacotherapeutics by determining whether agents directed against fosB itself, or against any of its many target genes, would be useful in the treatment of diverse types of neuropsychiatric disorders.	{ "pile_set_name": "NIH ExPorter" }
The long term objective of this competing renewal grant application is to develop novel prodrug strategies to improve corneal permeability of topically applied antimicrobial agents employed in the treatment of both bacterial and viral corneal stromal and epithelial keratitis. In this grant proposal we aim to characterize the ascorbate carrier systems expressed on the corneal membrane. We also propose to synthesize amino acid, peptide and ascorbate ester prodrugs of erythromycin, levofloxacin and ofloxacin and ascorbate ester prodrugs of ACV, targeted towards the respective transporters expressed on the corneal membrane, and to evaluate their ability to circumvent P-gp and MRP mediated efflux. The specific aims of this renewal application are a) To synthesize derivatives of erythromycin, levofloxacin and ofloxacin targeting amino acid (phenylalanine and glutamate prodrugs), peptide (valine, valine-valine and glycine-valine prodrugs) and ascorbate (ascorbic acid prodrugs) transporters expressed on the cornea and to evaluate these conjugates with respect to their b) physiochemical properties and cytotoxicity; c) bio- reversion in ocular fluids and cell homogenates, d) uptake, transport and simultaneous bioreversion across isolated rabbit cornea, and human and rabbit primary corneal epithelial cell cultures, e) affinity for P-gp and MRP efflux pumps, f) in vitro antibacterial efficacy and g) in vivo antibacterial efficacy against bacterial keratitis in the rabbit model. In vitro and in vivo studies will be conducted by Dr. William Suling at the Southern Research Institute, Birmingham, AL and by Dr. James Hill at the LSU Eye Center, New Orleans, LA, respectively. Uptake and transport of erythromycin, levofloxacin and ofloxacin prodrugs will be studied across isolated rabbit cornea, human and rabbit primary corneal epithelial cell cultures (rPCEC), alone, in combination and in the presence of topically applied anti-inflammatory steroids (prednisolone, prednisone and 6-alpha-methylprednisolone). We propose to synthesize ascorbate prodrug of ACV study transport and simultaneous bio-reversion across isolated rabbit corneas, and human and rabbit primary corneal epithelial cell cultures; antiviral efficacy against in vitro viral screens of HSV-1, HSV-2, CMV, VZV and EBV and in vivo antiviral efficacy against HSV-1 (McKrae strain) induced acute epithelial and stromal keratitis in the rabbit model. In vitro and in vivo studies will be conducted by Dr. Sam Ananthan and Dr. Earl Kern under NIAID funded research contract and by Dr. James Hill. \n vivo ocular bioavailability of erythromycin, levofloxacin, ofloxacin and ACV and their prodrugs will be examined following topical application (alone, in combination or in the presence of prednisolone, prednisone and 6-alpha-methylprednisolone as inhibitors of corneal efflux pumps) in both anesthetized and conscious rabbit models by employing ocular microdialysis technique.	{ "pile_set_name": "NIH ExPorter" }
The Program Project that this Bioanalytical Core Proposal is part of intends to develop new opioid analgesic drugs that will replace existing narcotics. It is anticipated that these new drugs will be as effective as existing opiate drugs but will be safer and have less abuse potential. The Bioanalytical Core Proposal provides essential analytical services to other parts of the associated Program Project responsible for the development of new opioid peptide and peptide-like compounds. These services include measurements of binding afinity at the different opioid receptor types, measurements of potency using isolated tissue assays, determination stability against degradation by tissue, studies of the ability of these compounds to pass membrane barriers and analgestic activity after direct injection into the brain and after systemic injection. The basic information on opioid receptor binding affinity and potency will be obtained for all compounds under development. Compounds selected from these studies will be further tested using the other three approaches. Stability, membrane permeation and analgesic activity studies will identify new compounds most likely to have clinical utility as new opioid drugs.	{ "pile_set_name": "NIH ExPorter" }
Joubert Syndrome and related disorders (JSRD) are a group of autosomal recessive conditions characterized by a distinctive hindbrain malformation (the molar tooth sign - MTS) combined with intellectual disability (mental retardation), hypotonia, ataxia, and variably, cystic renal disease, retinal dystrophy and hepatic fibrosis. The overall context of the project is to explore the role of the primary cilium/basal body (PC/BB) in the development/function of the brain and retina. Specifically, we propose to study the molecular function of CC2D2A, a newly discovered gene responsible for JS, and to identify additional components of the genetic/protein network underlying JSRD using several highly informative families. In Aim 1, we will investigate CC2D2A function by determining temporal, spatial and subcellular expression as well as identifying additional proteins that interact with CC2D2A. In Aim 2, we will use the cc2d2a photoreceptor phenotype in zebrafish to dissect the molecular function of Cc2d2a and model oligogenic inheritance by examining the effects of JSRD gene mutants/morphant combinations. Given that <50% of JSRD patients have mutations in the known genes, the goal of Aim 3 is to identify additional components of the genetic/protein network responsible for JSRD. This work has broad implications for human disease. Most specifically, it will enhance our understanding of the development/function of the brain, retina and kidney, provide improved diagnostic and prognostic information for patients with JSRD and potentially identify molecular targets for therapies to prevent or delay the progressive retinal, kidney and liver disease seen in JSRD and other ciliopathies. Given the protean role of the PC/BB in cellular function and Mendelian diseases, combined with the emerging associations between PC/BB genes and more common neurological diseases such as schizophrenia and autism, this work is also likely to reveal mechanisms underlying common diseases of the brain, retina, kidney and other tissues.	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY Bronchopulmonary dysplasia (BPD) affects up to half of extremely preterm infants, and is associated with significant adverse respiratory, developmental, educational, and health economic outcomes. The NICHD Neonatal Research Network (NRN) Hydrocortisone for BPD Trial has enrolled 652 of a planned total of 800 preterm infants at high risk for BPD in a placebo-controlled trial of hydrocortisone (HC). The proposed ?Hydrocortisone for BPD Respiratory and Developmental (HYBRiD) Outcomes Study? builds on this ongoing clinical trial to characterize the functional respiratory and developmental outcomes of this established trial population at 5 years corrected age, or early school age. Functional assessments, which cannot be performed before early school age, provide parents and schools with a realistic picture of a child's strengths and challenges in every day scenarios, so that appropriate services can be provided to prevent school failure. This proposed trial follow-up takes advantage of (1) a large, well-described, trial population of children who had severe neonatal lung disease and are being followed until 2 years corrected age; (2) the support of the NRN infrastructure, (3) the NRN's superb track records in school age trial follow-up and collaboration with NHLBI, and (4) the long-standing successful relationship between the NRN Data Coordinating Center (DCC) at RTI and the Clinical Coordinating Center (CCC) and NRN investigators. The CCC and DCC both have extensive expertise and experience in the conduct and follow-up of large neonatal clinical trials. The HYBRiD Outcomes Study will use novel functional assessments of early school age developmental and respiratory outcomes to assess the impact of both HC exposure and severity of lung disease. The primary outcome of functional impairment is defined as any one of four outcomes that can be measured directly in the child: cognitive delay, motor impairment, academic delay, or poor functional exercise capacity as measured by the 6-minute walk test (6MWT). The HYBRiD Outcome Study specific aims are, within the NRN HC for BPD Trial at 5 years corrected age, to evaluate: (1) the impact of HC exposure on rates of functional impairment; and (2) the relationship between severity of BPD (based on the categorical NIH consensus definition of BPD) and functional impairment. The HYBRiD Outcome Study's functional respiratory outcomes, which are novel in this population, will be validated with impulse oscillometry in the participants at a subset of centers. Supported unanimously by the NRN, this study will address several critical knowledge gaps about the early school age outcomes of children with neonatal respiratory failure, based on both the severity of BPD and exposure to HC, as well as the relationships between respiratory and developmental outcomes. These results will have immediate impact on counseling of parents in the neonatal intensive care unit and management of neonates in the unit and after discharge. In addition, these data will be essential for the development of future intervention studies aimed to improve the long-term outcomes of preterm infants with respiratory failure.	{ "pile_set_name": "NIH ExPorter" }
Autism spectrum disorders (ASD), depression and schizophrenia are complex psychiatric conditions involving serotonin (5-HT) dysfunction with a range of symptoms which prominently include impaired social behavior. Drug interventions for these disorders, such as risperidone (Risperdal) and fluoxetine (Prozac), commonly enhance 5-HT neurotransmission or mimic its downstream effects, but these interventions often do not improve social behavior, particularly in patients with gene polymorphisms impairing 5-HT transporter (SERT) function. SERT tightly controls the strength and duration of 5-HT neurotransmission by high affinity uptake of 5-HT from extracellular fluid in brain, but a role for organic cation transporter 3 (OCT3) in 5-HT uptake is emerging as an important mechanism contributing to regulation of extracellular levels of 5-HT in brain. These new findings suggest OCT3 might be a novel target for therapeutic intervention to improve social behavior. Consistent with this idea, we found that acute OCT3 blockade increased social behavior in socially-impaired BTBR mice. The studies we propose here will build on these new findings using SERT knock-out (-/-) mice, which like BTBR mice, are less social than their wild-type (SERT +/+) counterpart. Importantly, SERT-/- mice have increased OCT3 expression and function relative to +/+ mice, making them a unique and useful tool for exploring the effects of OCT3 blockade on 5-HT neurotransmission and social behavior. We hypothesize that the OCT3 blocker, 1,12-diethyl-2,22-cyanine iodide (decynium-22 (D-22)), will improve social behavior in SERT-/- mice more effectively than risperidone, while fluoxetine, which blocks SERT, will have no effect in SERT -/- mice. We expect, however, that D-22 administered in combination with risperidone will produce the greatest increase in social behavior in SERT -/- mice. D-22's effects on SERT +/+ mice, which express both SERT and OCT3 and are highly sociable relative to other strains, will be of interest for use of OCT3 blockade to other ends in broader patient populations. Behavioral data will be correlated with D-22 inhibition of [3H] 5-HT uptake into synaptosomes prepared from hippocampus. Serotonin is a substrate for several biogenic amine transporters, so measuring its uptake in SERT -/- mice, along with selective blockade and control experiments to account for contributions from other transporters expressed in hippocampus, will provide important new information about the relative roles of SERT, OCT3 and other monoamine transporters in 5-HT uptake. Finally, to confirm that D- 22 is exerting its effects centrally, we will measure brain concentration of behaviorally active doses of D-22. Extremely little is known about the role of OCT3 in brain and even less about the effects of systemically administered D-22. Therefore, these fundamental experiments are an essential first step in establishing the potential of OCT3 as a novel target for therapeutic intervention in the treatment of social dysfunction prominent in many psychiatric disorders. PUBLIC HEALTH RELEVANCE: Dysfunction of the serotonin neurotransmitter system has been strongly implicated in the etiology of autism, schizophrenia and depression, and may also underlie impaired social interaction common to these disorders. The proposed studies examine a novel target for pharmaceutical intervention to regulate 5-HT neurotransmission and improve sociability, the organic cation transporter 3 (OCT3), which like the serotonin transporter (SERT, the target of Prozac), removes serotonin from extracellular fluid in the brain to stop neurotransmission. Blockade of OCT3 may prove to be a more uniformly useful mechanism to enhance 5-HT transmission, particularly in patients with SERT gene polymorphisms that make drugs like Prozac less effective.	{ "pile_set_name": "NIH ExPorter" }
Ionotropic glutamate receptors (iGluRs) are ligand-gated ion channels that form transmembrane, cation- permeable channels. The (S)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazole) propionic acid (AMPA) subtype of iGluRs (AMPAR) is essential for the fast excitatory neurotransmission in the central nervous system (CNS). Malfunction of AMPARs has been implicated in several neurodegenerative diseases such as Alzheimer's disease (AD) and amyotrophic lateral sclerosis (ALS), as well as other neurological diseases such as cognitive deficits, epilepsy, schizophrenia, and mood disorders. For AMPARs as well as other iGluR subfamilies, active channels are tetramers exclusively formed by assembly of subunits within the same subfamily, a molecular process principally controlled by the extracellular amino-terminal domain (ATD). This phenomenon serves to control the permeation and kinetic properties of iGluR ion channels and is thus critical for maintaining normal physiological function of iGluRs. The goals of this proposal are to understand the molecular mechanisms by which the ATD guides subfamily-specific iGluR assembly. The specific aims are: (1) we will determine the crystal structures of the ATD of AMPARs; (2) we will characterize the role of the ATD in functional assembly of homomeric and heteromeric AMPAR channels; and (3) we will characterize the underlying mechanism by which ATDs guide subfamily-specific dimer-dimer association of iGluRs. The proposed study should provide a better understanding of the molecular principles governing iGluR assembly and function which could ultimately lay groundwork for future therapeutic interventions. Furthermore, molecular mechanisms governing iGluR assembly could be applicable for studying other multimeric ion channels/receptors, such as potassium channels, cyclic nucleotide-gated channels, nicotinic acetylcholine receptors, GABA receptors and others. Aberrant structure or function of these receptors/channels has been linked to many human neurological and psychiatric diseases. PUBLIC HEALTH RELEVANCE: Malfunction of the AMPA subtype glutamate receptors (AMPARs) has been implicated in neurodegenerative diseases such as Alzheimer's disease (AD) and amyotrophic lateral sclerosis (ALS), as well as other neurological diseases. The normal physiological functions of AMPARs are actively controlled by subfamily-specific channel assembly between different subunits which serves to control the permeation and kinetic properties of ion channels. Our proposed study should provide a better understanding of the molecular principles governing iGluR assembly, which could ultimately lay groundwork for future therapeutic interventions.	{ "pile_set_name": "NIH ExPorter" }
In order to understand the molecular basis of diseases, particularly those involving single amino acid substitutions, we need to elucidate the relationship between protein structure and function at the molecular level. Therefore, the long-term goals of this project are to acquire a deeper understanding of the relationship between protein structure and function by using alkaline phosphatase as a model system. The function of this enzyme is to catalyze the nonspecific hydrolysis of phosphate esters, and the lack of the activity of this enzyme results in the fatal hereditary disease hypophosphatasia, which is due to insufficient phosphate for bone calcification. Alkaline phosphatase from mammals is closely related to the corresponding bacterial enzyme, and the enzyme from Escherichia coli has become the model for the study of all alkaline phosphatases. The specific aims of this proposal are to answer fundamental questions concerning the relationship between the structure and function of alkaline phosphatase. We will concentrate on the molecular details of the catalytic mechanism, the need to maintain a dimeric structure for the correct function of the enzyme, the mode by which information is passed between the subunits, a molecular explanation of intergenic complementation, the factors critical for correct secondary structure formation, the function of the metals in catalysis, and the contribution of the electrostatic field around the active site for catalysis. In order to accomplish this goal, we will take advantage of a set of thousands of point mutations created within the alkaline phosphatase gene more than 30 years ago in an early effort to prove that the gene and the protein produced from it were colinear. This set of mutants today provides a unique resource for the investigation of the interrelationship between the structure and function of alkaline phosphatase. Analysis of this set of mutants as well as mutants created during the last grant period will be accomplished by a variety of techniques including x-ray crystallography, 31P and 113Cd NMR, 18(O) isotope effects, and kinetic studies with phosphonates, phosphorothioates and the chiral (Rp) [O16,O17, O18] p-nitrophenyl phosphate. Correlations will be made between the functional changes induced by the amino acid substitution and the three-dimensional structure of the mutant enzymes. This work will not only be important for the understanding of this particular system, but more importantly for formulating general concepts about enzyme catalysis, and the function of metals in proteins, and providing a molecular explanation for intergenic complementation.	{ "pile_set_name": "NIH ExPorter" }
Despite their widespread use in the treatment of glaucoma, much information is still needed regarding the clinical and biologic effects of carbonic anhydrase inhibitors. This project will use clinical studies of dose-response and side effects to obtain information regarding the effective range of drug dosage, frequency of administration, and the nature and extent of drug side effects as a function of individual drugs and their dosages. The relationship of side effects and dosage to the therapeutic response of reduced intraocular pressure will also be studied. Effects of carbonic anhydrase inhibitors on clinical biochemical findings will be related to dose-response and side effects, and will help to develop a clinically valuable test for patient compliance in taking these drugs. Laboratory biochemical studies will measure levels of drug in blood and urine of patients in the study in order to correlate such levels with drug dosage. These studies will be combined with studies of carbonic anhydrase inhibition in blood cells to help delineate the disposition, transport, and cellular effects of carbonic anhydrase inhibitors.	{ "pile_set_name": "NIH ExPorter" }
High density lipoproteins (HDL) are heterogeneous particles, and have been classified based on their apolipoprotein composition. ApoA-l and apoA-Il are the two major apolipoproteins associated with HDL. The two major classes of HDL particles include LpA-I, which contains apoA-l but not apoA-ll, and LpA-I,A-Il, which contains both apoA-l and apoA-ll. LPA-I, but not LpA-I,A-II, has been found to be associated with coronary heart disease risk. The major goal of this project is to determine the factors which regulate the plasma levels of LpA-I and LpA-I,A-11 in both normal subjects as well as patients with low levels of HDL, who may be at increased risk for premature coronary disease. The metabolism of these two major HDL particles in normal subjects was determined to be different, with LpA-I having a significantly faster catabolic rate than LpA-I,A-ll. Several subjects with significantly low levels of HDL (less than 25) but no evidence of coronary disease were studied in order to determine the etiology of the low HDL utilizing radiolabeled apoA-l and apoA-Il. All subjects were shown to have rapid catabolism of both apolipoproteins. Therefore, hypercatabolism of HDL apolipoproteins may be the major cause of significantly decreased HDL levels. This metabolic defect does not appear to markedly predispose to premature coronary artery disease. These studies included subjects of age 21 to 57 years. 56 percent of the subjects were women. The studies included two Asian and one East Indian subject.	{ "pile_set_name": "NIH ExPorter" }
Nucleic acids will be sequenced and nucleic acid sequences will be analyzed for constraints on nucleotide order that may be a consequence of Genotypic Selection. Genotypic Selection is a novel form of selection postulated to exist at the intracellular level and acting directly on the nucleic acids before their expression. The constraints imposed on actual nucleotide sequences will be examined for patterns of internal terminator sequences, true palindromic sequences, self-complementary sequences, and frequencies of 2-8 long nucleotide groups and their contexts. These patterns will also be analyzed for their relationship to each other. For example, the gene V ribosome binding site of f1 bacteriophage sequence, AUG.AUU.AAA.GUU.GAA.AUU.AAPu, is composed of internal terminators palindromically arranged. In addition the nature of tRNA-mRNA interaction will be examined at the nucleotide level to determine the actual number of base pairs required per interaction. The allowable conformations for tRNA anti-codon loops can then be determined as well as the question of messenger RNA species specificity. A model of tRNA interaction has recently been proposed by Crick, Klug, Brenner, and myself. I have found by computer analysis of certain known sets of tRNA and mRNA sequences, particular ones which are uniquely suited for testing this hypothesis. I have demonstrated tRNA-tRNA complexes that are dimers, trimers, tetramers and pentamers of a given tRNA. These multimers will be isolated and characterized by nucleotide blocking and nuclease digestion. Complexes of tRNA-mRNA will be generated and analyzed for the nature of the tRNA-mRNA interaction. Also, the direct sequencing of certain DNA restriction fragments of phi80 and f1 bacteriophages and the indirect sequencing of adjacent regions will be done to determine transcription starts, attenuators and stop sequences as well as additional coding sequences. The significance of such studies is that they allow a test of the predictions of Genotypic Selection; a perspective where the genotype is a phenotype for syntactical and structural selection by the replication, transcriptional and translational machinery.	{ "pile_set_name": "NIH ExPorter" }
The translational machinery of the ribosome performs the essential task of protein synthesis. In order to do this it must properly coordinate the interaction of the anticodon of each tRNA with the appropriate mRNA codon and catalyze peptidyl bond formation at a site more than 70 A away. The relatively constant rate of protein synthesis implies that each tRNA must interact with the ribosome similarly, despite significant differences in tRNA sequence, amino acid identity, and anticodon identity. It has been suggested that small differences tRNA structure and the presence of modified residues play a role in modulating the interactions of tRNA with the ribosome. Novel methods of tRNA construction, in concert with new thermodynamic and kinetic assays, will be applied to (1) determine how different tRNAs are built to maintain uniform ground-state thermodynamic interactions with the ribosome, and (2) to establish how tRNA structure modulates the movement of tRNAs on the ribosome during translation. The experiments are designed to understand the dependence of protein synthesis on tRNA-ribosome interactions.	{ "pile_set_name": "NIH ExPorter" }
The long term goal of this research is to understand the mechanisms of antiarrhythmic drug modification of the cardiac sodium channel. The experimental approach in this grant proposal combines the measurement of Na channel gating current, which are a direct measure of the channel's molecular conformational states, with modification of cardiac Na channels by toxins or agents to help understand antiarrhythmic drug interactions with the cardiac Na channel. In our present grant period we have found the applying this approach has made important advances in the understanding of antiarrhythmic drug modification of cardiac Na channels by QX-222 and lidocaine, and int he understanding of cardiac Na channel inactivation by the use of the site-3 toxin, Anthopleurin-A. For the next grant period we propose to continue our studies of cardiac Na inactivation, which has been shown to be important in the action of antiarrhythmic drugs, and of lidocaine's modification of cardiac Na channel gating currents. This grant proposal will use both native cardiac Na channels in single heart cell preparations and mutated cardiac Na channels expressed in large, fused tsA201 cells to study Na channels modified by lidocaine, by Na channel toxins and agents, and by site-directed mutagenesis. The specific aims are; 1. Determine if batrachotoxin (BTX) modification of cardiac Na channels alters movement of the channels's voltage sensors similar to changes caused by the antiarrhythmic drug, lidocaine, and determine if either lidocaine or BTX inhibit movement of the voltage-sensor associated with cardiac Na channel inactivation from the open-state (OyieldsI inactivation). 2. Determine if the voltage-sensor associated with O yields I inactivation in cardiac Na channels is separate from the putative inactivation gate formed by the intracellular linker region between domains III and IV. 3. Determine if immobilization of off-gating charge in cardiac Na channels is associated with the putative inactivation gate. 4. Determine if the voltage-sensor for O yieldsI activation in cardiac Na channels is associated with the S4 segment of domain IV. 5. Determine if the cardiac Na channel gating charge-voltage relationship measured during step depolarizations to test potentials near INa threshold underestimates the total gating charge. The results of the proposed studies in the this grant should advance our understanding of the molecular mechanisms of cardiac Na channel inactivation, and our understanding of which components of the cardiac Na channel's voltage sensors are modified by lidocaine. In combination, they should contribute to the formation of a better molecular model of antiarrhythmic drug interactions with the cardiac Na channel, and form the foundation for future studies of antiarrhythmic drug-Na channel interactions.	{ "pile_set_name": "NIH ExPorter" }
Asthma is the most common chronic disease in childhood, and prevalence, hospitalization and mortality are increasing. This increase is most commonly seen in non-white, poor, inner city children. Although the causes of this epidemic are not clearly understood, lack of access to medical care, indoor and outdoor pollutants, and indoor and outdoor antigens seem important. Recent small-scale community intervention to reduce exposure to dust mites and other antigens has been shown to result in clinical improvement. Further, research is needed to evaluate community based interventions which will control a variety of antigens and pollutants. Inner city, primarily minority, children with asthma are being identified through a school based mobile asthmatic clinic, the Breathmobile, which delivers high quality, continuous care to these children. Working with school nurses and community organizations and the 3 Breathmobile units, we propose a comprehensive community-based intervention aimed at reducing asthma triggers in the home. The major goal of this study is to determine whether a comprehensive environmental health education program, enhanced by least toxic integrated pest management for cockroach control, will result in reduction in concentrations of antigens in household dust and/or improvement in clinical status among these children. The study population will consist of 300 children with chronic persistent asthma randomly selected from the 3,000 asthmatics identified by the school-based Breathmobile program. Children will be randomized into three groups: (1) 100 children will continue to receive usual care from the Breathmobile; (2) 100 children will receive usual care plus a standardized antigen-reduction strategy (STARS), a community based, family oriented environmental health training program; and (3) 100 children children will receive usual Breathmobile care plus STARS, enhanced by professional pest control and home cleaning. Exposure to environmental asthma triggers in the home will be assessed by measuring concentrations of dust mite antigen, and cockroach antigen, in house dust. Outcomes of interest include change in knowledge, change in concentration of antigen in house dust, school absence, clinical assessment of asthma severity, and medication use. Co-variates to be considered include exposure to environmental tobacco smoke, indoor and outdoor air pollutants, housing characteristics (such as molds, mildew, air conditioning) and demographics (such as family size, gender, ethnicity). In years 4 and 5 of the intervention, the community based infrastructure developed in years 1-3 to support the evaluation of the strategies for reducing exposure to environmental asthma triggers will be transferred to cooperating community groups and greatly expanded to provide household asthma audits, education, and intervention service at low cost upon referral from school nurses and the Breathmobile program. To evaluate efficacy, 200 asthmatic children will be randomized into 2 groups. One will receive usual care, the other, community intervention services.	{ "pile_set_name": "NIH ExPorter" }
HIV-1 infection of women is a relatively rare outcome of sexual exposure to HIV-1. The risk may vary for each woman, and it may also vary for the same woman during different exposures. Susceptibility is determined by a complex set of factors, including the amount and properties of the virus to which a woman is exposed, individual host genetic factors, as well as multiple modifiable biological cofactors present at the time of exposure. In this program project grant, we proposed to examine the mechanisms by which biological cofactors increase susceptibility of women to HIV-1. The proposed studies will include women in multiple high-risk groups: sex workers, pregnant/postpartum women and women in HIV-1 discordant relationships. Specifically, we will examine the role of: 1) pregnancy and the postpartum effects (Project 1);2) vaginal flora and specific bacterial infections (Project 2);3) innate immune factors, including as they relate to hormones and genital tract infections (Project 3);and 4) mucosal HIV-1 antibodies induced in response to repeated HIV-1 exposure (Project 4) on susceptibility to HIV-1 infection in women. For these studies, we will take advantage of cohort studies, collaborative partnerships and infrastructure developed by our research team, the Seattle/Kenya collaborative research group, over the past ~ 15 years. Our research group, which includes virologists, immunologists, bacteriologists, epidemiologist, statisticians and clinical scientists, has a long and productive history of collaboration, and is known for its focus on translational research in women. Here we will build on that foundation to conduct multiple collaborative projects within this program grant. We expect that each of these projects will draw on the expertise of others in the team, and in some cases, will include parallel studies that address synergistic biological questions in the same populations. RELEVANCE: Collectively, these studies are designed to provide a comprehensive picture of the biological factors that contribute to increased risk of HIV-1 infection in women. Such studies will be critical for understanding unique aspects of HIV-1 transmission in women, and for finding approaches to decrease transmission that consider issues that are relevant to women. PROJECT 1: Incidence, Timing and Cofactors that Contribute to the High Risk of HIV-1 Infection in Peri and Post Partum Women (Project Leader: John-Stewart, G) PROJECT 1 DESCRIPTION (provided by applicant): Pregnancy and the postpartum period have been associated with increased risk of HIV-1. In Africa, where both HIV-1 seroprevalence and fertility rates are high, the pregnancy/postpartum period may be one in which HIV-1 acquisition contributes substantially to HIV-1 in women. Pregnancy, delivery, and the postpartum period are associated with hormonal, genital mucosal, and genital flora changes that could predispose to acquisition of HIV-1. Project 1 will enroll 2,000 HIV-1 uninfected women identified during pregnancy and followed to 9 months postpartum to determine risk and cofactors of HIV-1 incidence. Women who acquire HIV-1 will be compared to women who do not in order to determine the role of genital coinfections, ulcers, delivery practice, lactation, partner characteristics, vaginal flora changes, HSV-2, genital innate immune factors, and systemic immune activation on HIV-1 transmission. In addition, changes in cofactors over the course of pregnancy and postpartum may influence susceptibility to HIV-1. Thus, in a subset of 100 women we will compare genital innate immune factors, and systemic cellular immune activation longitudinally during pregnancy, early postpartum, and later postpartum. These comparisons will provide opportunity to determine patterns of change in the genital tract during this dynamic period of change in women. In quantifying co-infections, mucosal innate immune responses, and systemic cellular immune activation, we will be able to determine interactions between these important determinants in 3 different but inter-related areas (co-infection, mucosal innate milieu, systemic cellular) that affect susceptibility to HIV-1. Project 1 will have scientific links to Project 2 (effects of changes in vaginal flora on HIV-1 transmission), Project 3 (which will involve cytokine profile analyses within women from the Project 1 cohort) and Project 4 (adaptive humoral mucosal responses in HIV-1 uninfected women with HIV-1 infected partners). Because of the complexity of the female genital ecosystem and the variety of factors that could alter susceptibility to HIV-1, each Project will focus on complementary factors that may modify transmission of HIV-1. Together, these will lead to a multi-faceted evaluation of transmission in women in different important groups of women at risk for HIV-1, (pregnant/postpartum, female sex workers, and discordant couples), that conceivably share some cofactors for HIV-1 susceptibility but are distinct in others. RELEVANCE: Women may be at increased risk for HIV-1 during and after pregnancy. This study will determine the risk of and cofactors for acquiring HIV-1 during and after pregnancy in a cohort of women in Kenya. Factors including genital infections and immunity will be assessed. These data will be directly relevant to developing appropriate strategies to protect women from HIV-1 during and after pregnancy.	{ "pile_set_name": "NIH ExPorter" }
ABSTRACT A health workforce trained to conduct Sexual and Gender Minority (SGM)-inclusive cancer research is needed to meet the needs of SGM persons seeking comprehensive cancer care, including prevention, screening, treatment, and survivorship care. Research on SGM cancer care has described many SGM cancer disparities, and a growing number of studies of SGM individuals have documented increased exposure to cancer-risk factors and certain cancers, as well as poorer cancer outcomes. Yet SGM patients are extremely diverse, presenting with unique and varied care needs. Although the body of research focused on SGM groups continues to grow, gaps persist in the knowledge of specific SGM cancer risk factors and cancer treatment experiences that is critical for developing evidence-based clinical practice guidelines in oncology care across the cancer care continuum ? from prevention to survivorship. As no nationally available resource routinely trains and prepares the oncology and biomedical workforce for SGM cancer research, our short-term goal is to design an up-to-date, competency-based 1.5 day workshop for early career researchers: titled SGM Cancer CARE (Sexual and Gender Minority Cancer: Curricular Advances for Research and Education). Informed by our needs assessment, the curriculum will be designed by a multi-disciplinary and multi- institutional leadership team and advisory committee that has national perspective and experience with SGM populations, and in-depth knowledge and expertise in multiple key areas, including cancer research, clinical care, curriculum development, and health care advocacy. This project has the following specific aims: 1) Refine curriculum topics and learning objectives; 2) Design the curricular modules and materials; and 3) Pilot test and evaluate the curriculum. It will be created for health and research professionals with MD, DO, PhD, or DrPH degrees, targeting individuals in the process of shaping their research agendas and interested in SGM cancer issues. It will be presented in a 1.5 day workshop through five modules: 1) clinical cancer research; 2) behavioral science and interventions; 3) epidemiology and population-level research; 4) community research, vulnerable populations, and responsible conduct of research, and 5) opportunities and challenges across modules (i.e. research strategy, funding opportunities, mentorship, and collaborations). The workshop will be piloted as a pre-conference opportunity for the annual LGBT Health Workforce Conference sponsored by the BNGAP organization. The workshop will provide participants with knowledge and skills to conduct quality and culturally appropriate research to improve SGM care across the cancer care continuum, and will facilitate participant access to a national research community and health care provider network dedicated to shared goals in SGM cancer research. Long-term, the resulting curriculum can be expanded and adapted for integration into existing curricula, stimulating training and education of additional professionals to increase SGM cancer research in order to create health equity for vulnerable SGM populations.	{ "pile_set_name": "NIH ExPorter" }
The purpose of this study is to identify the undesirable side effects of HMG-COA reductase inhibitors on skeletal muscle metabolism and fatigue in hypercholesterolemic subjects. Also, this study will provide credence to hypothesis that highly lipid soluble drugs such as lovastatin and simvastatin may exacerbate the myalgic effects, caused by strenuous eccentric exercise; whereas highly hydrophilic drugs such as pravastatin may be relatively safer.	{ "pile_set_name": "NIH ExPorter" }
DAT18-01. Abuse of stimulants, such as cocaine and methamphetamine, results in a variety of serious health conditions, and drug abusers have poorer health outcomes than non-drug using, demographically similar populations. In the central nervous system, the use of stimulants induces neuroinflammation through a greater release of inflammatory factors and recruitment of additional leukocytes. This predisposes drug abusers to a higher incidence of neuropsychiatric, cerebrovascular and motor disorders, and can also exacerbate the neuropathogenic impact of infection with HIV, HCV and a number of bacterial pathogens. The precise pathways by which stimulants mediate these effects are not clear, but many of these effects could be induced by drug-associated activation of specific inflammatory triggers such as NF- kB. However, direct links between stimulants and these neuroinflammatory mechanisms have not been described. The premise of this proposal is that dopamine acts as a common mechanism by which stimulants activate myeloid cell NF-kB and thereby initiate or exacerbate neuroinflammation. Use of all stimulants acutely increases CNS dopamine levels, exposing cells in dopamine-rich brain regions to aberrantly high dopamine concentrations. Among these cell populations are myeloid cells, such as perivascular macrophages and microglia, which are the primary immune cells in the CNS. Our published research shows that acute exposure to elevated dopamine increases myeloid production of inflammatory cytokines, such as IL-1b, IL-6, CXCL8 and CXCL10. Our preliminary data suggest that dopamine acts by activating the NF- kB pathway and priming the NLRP3 inflammasome, a complex that regulates the release of IL-1b, a master regulator of inflammation. The specific pathways mediating this effect are not clear, and therefore these studies will generate detailed information about specific dopamine receptors, gene and protein targets mediating dopamine activation of NF-kB and NLRP3 in human macrophages. Determining the specific signaling mechanisms and genes involved in dopamine induced increases in NF-kB activity will indicate pathways that could be targeted to ameliorate the neuroinflammatory effects of stimulant use, significantly improving the long-term health outcomes of stimulant users. The data developed in this proposal will serve as a basis for future projects examining the modulation of the myeloid dopaminergic system as a therapeutic strategy for limiting the increased incidence of neurologic disease and inflammation associated with drug abuse. These projects will examine both novel effectors and the repurposing of existing dopaminergic therapeutics to ameliorate inflammation in the stimulant abusing population.	{ "pile_set_name": "NIH ExPorter" }
Motivation, the desire to act, is thought to arise within the limbic system. We are studying the biological basis of motivated behavior and reward expectancy in monkeys using a task that manipulates their motivation. The monkeys work faster and with fewer errors when a cue indicates that a juice reward will be delivered immediately after the next correct response than when the cue indicates that additional trials will be needed. Previously we learned that single neuronal responses in the ventral striatum and perirhinal are directly related to the associatively learned meaning of the cue in this complex behavioral task. Neurons in both regions keep track of whether the animal is at the beginning of or somewhere in the course of a behavioral sequence that ultimately leads to reward. Some neurons signal that a new series of trials is starting, others signal that a series of trials is in progress, and still others signal the rewarded trial is starting when it follows one or more unrewarded trials. Thus populations in both brain regions provide neural signals that could reinforce complex reward-seeking behavior. However, neither brain region showed a signal related to directly to reward expectancy. Now we have shown that neurons of the anterior cingulate cortex respond more strongly as more trials have elapsed, thus showing a signal related to expectancy. This is a provocative finding because the cingulate cortex shows abnormal activation in imaging with humans having disorders of reward expectancy such as obssessive-compulsived disorder and drug abuse. Because of its close connections with the visual system, the rhinal cortex has been shown to be important for normal pattern recognition behavior. Rhinal cortex and ventral striatum are strongly connected, and both show highly structured organization of dopaminergic input leading us to speculate that perirhinal cortex might show reward-related activity similar to the activity we had seen in the ventral striatum. Given the emphasis on the relation between perirhinal cortex and pattern-recognition behavior in the past, the prominence of the perirhinal activity related to the reward schdules was unexpected. To investigate whether perirhinal cortex plays a critical role in this associately learned reward schedule related behavior, normal monkeys and monkeys with bilateral rhinal cortex lesions were studied using the reward schedule task described above. Normal monkeys associate new visual cues with the schedule starting within a single training session. In contrast, animals with bilateral rhinal cortex ablations are severely impaired in making this association, being unable to do so even after six weeks of daily training. Thus, perirhinal cortex is a critical structure for developing the associative relation between a visual cue and its meaning for reward schedules. We hypothesize that dopaminergic input provides signals sensitive to long-term progress through a planned or expected series of tasks which culminated in reward. To test this, we injected material that temporarily should block production of D2 and NMDA receptor proteins into the remaining rhinal cortex of monkeys with a unilateral rhinal cortex lesion. For a period of about 10 weeks these animals are unable to associate new visual cues with reward schedules, thus mimicing the results after a bilateral ablation. Animals with control injections of nonspecific DNA are unimpaired. These data suggest that rhinal cortex is critical for forming the associations between stimuli and their motivational/emotional meaning for predicting reward in reward schedules, and that either D2 and/or NMDA receptors are must be intact for this function.	{ "pile_set_name": "NIH ExPorter" }
Neurodegenerative diseases currently have few treatments and no cures. This stems from the lack of understanding of the causes of neuron death in neurodegeneration. Recent research focusing on the proteins discovered to accumulate in the brains of patients, including TDP-43 in amyotrophic lateral sclerosis (ALS), has taken research towards understanding the mechanisms that underlie disease. Modeling disease using pathogenic disease proteins has proved useful for identifying potential pathways, interactions and mechanisms involved in neurodegeneration. In this proposal I will study TDP-43, a pathological protein involved in ALS and frontotemporal dementia, both devastating neurodegenerative disorders. Through modeling TDP-43 related disease in yeast and flies, we have identified a novel interaction between TDP-43 and Ataxin-2, a protein mutated in another neurodegenerative disease, spinocerebellar ataxia type 2 (SCA2). We identified mutations in Ataxin-2 as a novel genetic risk factor for ALS, potentially the most common discovered to date. The TDP-43/Ataxin-2 interaction sits poised as a potential pathogenic mechanism that could provide not only knowledge of disease processes, but also as a potential therapeutic target. Therefore, I will first define the domains of Ataxin-2 and TDP-43 required for the proteins to interact with each other. Second, I will determine if mutations in Ataxin-2 affect the pathological signature of TDP-43, which includes downstream biochemical alterations of the protein, to learn if these mutations influence known disease processes. This will involve training in neuropathology and biochemical methods used to analyze human tissue. Lastly, I will examine if mutations in Ataxin-2 contribute to other neurodegenerative diseases related to ALS that also show TDP-43 involvement. This aspect of the proposal will require training in human genetics methods and analyses, as well as histopathological techniques. Our discovery of a role for Ataxin- 2 in ALS via an interaction with TDP-43 opens up many new avenues for investigation. These experiments will expand our knowledge of the TDP-43/Ataxin-2 interaction and, more broadly, give insight into the similarities and differences of mechanisms underlying different related neurodegenerative diseases.	{ "pile_set_name": "NIH ExPorter" }
The transcriptome of male germ cells at different stages of development, i.e. mitosis, meiosis and post-meiosis as represented by type A spermatogonia, pachytene spermatocytes and round spermatids, respectively, was profiled by Serial Analysis of Gene Expression (SAGE) and cDNA microarray hybridization previously. The SAGE data were deposited in our publicly accessible website (URL:http://www.nichddirsage.nichd.nih.gov/publicsage/). A number of genes with similar profile of change among the germ cells using both techniques were selected for further studies. We hypothesize the distinct expression pattern of a gene reflects the specific role of the respective gene in different stages of spermatogenesis. Currently we are investigating three such genes, namely, Testis expressed gene 13 (Tex13), the mouse Lin-28 homolog, and a novel isoform of the mouse Ard-1 gene which encodes a putative co-subunit of the murine N-terminal acetyltransferase 1 complex. Anti-sense Transcription in Differentiating Germ Cells Anti-sense transcripts have been shown to be involved in transcriptional and post-transcriptional gene regulation including genomic imprinting, X-inactivation, RNAi, RNA editing, and mRNA processing, splicing, stability, transport, and translation. Computational analysis had identified substantial number of antisense transcripts in the mouse and the human genome recently. However, with the exception of the anti-sense transcript of the SPEER2 gene, no other example of sense-antisense transcript pairs in mammalian germ cell has been reported. Examination of 64 differentially expressed genes identified in mouse type A spermatogonia, pachytene spermatocytes, and round spermatids showed the presence of anti-sense transcripts of 41 genes (66%). Among the 41 genes, 29 genes with appreciable number of SAGE tags in the germ cells were picked for confirmation using orientation specific RT-PCR. The presence of anti-sense transcripts was confirmed experimentally for 17 genes. Examination of tissue distribution of 9 selected genes including Uba52, Calm2, Ppp1cc, Ppic, Tsg1, Tcte3, Pdcl2, Prm 1 and Prm2 showed a wide spectrum of tissue-specificity of the expression of the sense and the anti-sense transcripts. The antisense transcripts of four genes, namely Uba52, Tcte3, Prm1 and Prm2 were cloned and characterized. Alignment of the nucleotide sequence of the anti-sense transcripts with the genomic sequence of the genes encoding the sense transcripts allowed localization of the anti-sense transcripts to exons and introns of the sense gene, psuedogene, intronic as well as intergenic sequences. This study shows that anti-sense transcription occurs more frequently in differentiating germ cells than in somatic cells. We are in the process of identifying in vitro and in vivo systems suitable for testing functional activities of the anti-sense transcripts cloned. Novel Activity of Cytochrome C Oxidase in Germ Cell Development Extensive apoptosis occurs during spermatogenesis, particularly in spermatogonia and spermatocytes. The mechanism of apoptosis in germ cells has not been totally worked out. Recent studies suggested that cytochrome c plays a critical role in apoptosis occurring in germ cells. A search of the germ cell SAGE libraries revealed that all 13 subunits of cytochrome c oxidase, complex IV of the respiratory chain are expressed at appreciable levels and follow a comparable differential expression pattern in these cells. On the other hand, some of the subunits of complexes II, III, and V of the respiratory chain are absent and only 12 of the 43 subunits of complex I have the same differential expression pattern. These results imply that cytochrome c oxidase may not function as a component of the respiratory chain in the germ cells. Thus, we hypothesize that cytochrome c oxidase oxidizes cytochrome c causing its release from the inner mitochrondrial membbrane . Transfer of the solubilized cytochrome c into the cytosol results in the amplification of calcium-dependent apoptosis. We are in the process of proving this hypothesis using in vitro cell models. Functional Genomic Studies of Gonad Development and Sexual Dimorphism of the Brain In order to understand the mechanism that regulates the transition of primordial germ cells to gonocytes and the initiation of sexual dimorphism, we have undertaken the task of profiling the expressed genes in embryonic gonads of the mouse. We apply SAGE to male and female embryonic gonads at E10.5 (embryonic day 10.5), E11.5, E12.5, E13.5, E15.5, and E17.5 and mesonephros at E13.5, E15.5 and E17.5. We have so far completed the analysis of ~152,000 SAGE tags for each of the male E10.5, E11.5, and E12.5 gonads. These tags identify 24,460, 214,762, and 26,378 genes in the E10.5, E11.5, and E12.5 gonads, respectively. The 10 most abundant tags represent Cybb, Cyp2e1, Cox5b, Tctp1, Hbb-y, Tuba2, alpha 2, 4 ribosomal proteins (X-linked S4, L26, 29, and L10A) and one uncharacterized cDNA. Among the 10 most abundant tags that are present in the embryonic gonads and absent in germ cells, 4 represent genes encoding hemoglobin chains, namely Hba-X, Hbb-b1, and Hbb-Y, and Hba-a1. The roles of the hemoglobin genes in early embryonic gonad development are not clear at this point. In spite of the fact that less tags were sequenced for the germ cell SAGE libraries (~ 111,000 tags for each germ cell type versus ~152,000 tags for each embryonic gonad stage), more specific genes are found in germ cells than in embryonic gonads (4,946 germ cell specific-gene tags versus 4,755 embryonic gonad specific-gene tags). These preliminary observations have important implication on the regulation of gonad development and germ cell differentiation. We will continue to analyze expressed genes in male gonads at later embryonic ages as well as female gonads and mesonephros. We are also interested in the role of sex chromosome-linked genes in sexual dimorphism of the brain. To that end, we have generated human sex-linked gene cDNA microarrays on which the cDNA of 724 X-linked and 28 Y-linked human genes were spotted on a glass slide. These microarrays will be used to profile expressed genes in brains and gonads of male and female mice at E10.5, E13.5, E15.5, E17.5, newborn mice, and adult mice. The expression profile of male and female brain and gonad at different time point will be compared. The relationship between gonad development and onset of sexual dimorphism of the brain will be examined. The Role of Dby in Mouse Gonad and Germ Cell Development Dby (also known as Ddx3y) has been considered as a strong candidate mediating the function of the Y chromosome in spermatogonial proliferation. In the mouse the Dby gene gave rise to two alternative transcripts that differ only in the length of 3' untranslated region as a consequence of alternative polyadenylation signals. Both transcripts were ubiquitously expressed and were present in male germ cells and Sertoli cells. They were more abundant in type A spermatogonia compared to pachytene spermatocytes and round spermatids. Expression of Dby in the embryonic gonad increased from E10.5 and reached a peak at E17.5. The expression level of Dby decreased after birth and remained low in adult male gonads. Testicular expression of Dby was comparable to its X chromosome homolog, Ddx3. In contrast, D1Pas1, the autosomal homolog of Dby, was predominantly expressed in pachytene spermatocytes and round spermatids. The differential expression patterns of Dby and its X- and autosome-homologs suggests that though the three genes are highly homologous they participate in different regulatory pathways. Dby in mouse, unlike that in humans, may only affect earlier stages of spermatogenesis.	{ "pile_set_name": "NIH ExPorter" }
We propose pilot experiments to explore segregation and mixing in thin layers of granular material. This area is germane to a current hot topic in physics, math and engineering, namely granular materials. It is also an area of considerable technical interest, with relevance to the production of drugs. Segregation in granular materials has been made famous by the "Brazil nut effect", but is important in many different settings, including shaking of thin horizontal layers of material. The pilot project would involve the construction of a simple experimental shaker container and coil. We would use organic materials of different sizes (poppy seeds and similar) that have been successfully imaged using MRI (i.e. Fukushima). The container would be vibrated in Bob Behringer's lab using existing equipment. As soon as the vibration is stopped, the grains effectively "freeze", i.e., maintain their relative positions. We would then place the shaker in the magnet and carry out a 3D scan to determine the degree of internal segregation. We envision carrying out a modest number, - half a dozen, such scans in order to establish the validity of the technique. We would then actively pursue funding from the sources noted above, using the pilot scans as a demonstration of feasibility. At the same time, Igor Aronson and his collaborators would proceed with the development of new theoretical models.	{ "pile_set_name": "NIH ExPorter" }
For years, interactions between double stranded (duplex) DNA were presumed to be independent of the DNA structure and base pair sequence because the nucleotides are buried inside the double helix and shielded by the highly charged sugar-phosphate backbone. In discussion of such interactions, duplex DNA was explicitly or implicitly modeled as a uniformly charged cylinder. However, this concept was based on intuitive perception rather than experiments or rigorous theory. In reality, the experimental evidence, e.g., transformation of duplex DNA from a non-ideal helix with 10.5 base pairs/turn in solution into a nearly ideal helix with 10.0 bp/turn in aggregates, suggested that this concept may be wrong. Starting from the classical paper of Rhodes and Klug published in 1980, it became clear that interactions between duplex DNA not only depend on but also affect the double helix structure.[unreadable] [unreadable] To account for possible effects of the structure of the sugar phosphate backbone on DNA-DNA interactions, over the last decade we have been developing a theory of electrostatic interactions between macromolecules with helical patterns of surface charges. Even the simplest models, which do not account for dynamic variations in the structure, e.g., due to the thermal motion, already suggest possible explanations for many observations. The latter include the torsional deformation of the double helix upon aggregation mentioned above, counterion-specificity of DNA condensation, multiple liquid crystalline phases in DNA aggregates, and measured intermolecular forces. We, therefore, continue development of this theory and its applications to various phenomena. [unreadable] [unreadable] Most importantly, this theory predicts that the dependence of the backbone structure on the nucleotide sequence may be sufficiently strong to affect DNA-DNA interactions. The resulting effects, e.g., direct recognition of sequence homology between 100 base pair (bp) or longer fragments of duplex DNA, may obviously have significant biological implications. In particular, 100-300 bp sequence homology recognition is essential for avoiding recombination mistakes that lead to cancer, genetic disorders, etc. Experiments recently reported in the literature suggested that local, transient pairing of homologous sequences in intact DNAs may precede double strand breaks, further recognition by protein-covered single strands, and strand crossover. Direct interactions between duplex DNAs in nucleosome-free regions were proposed to be involved, but the mechanism and possibility of sequence homology recognition in such interactions remained unknown.[unreadable] [unreadable] During the last year, we completed the first series of purely physical experiments probing sequence-dependent DNA-DNA interactions in vitro. We imaged a mixture of two fluorescently tagged, double helical DNA molecules with identical nucleotide composition and length (294 bp), but different sequences. In electrolytic solution at minor osmotic stress these DNAs formed discrete liquid-crystalline aggregates (spherulites). We observed spontaneous segregation of the two kinds of DNA within each spherulite, revealing nucleotide sequence recognition between double helices separated by water in the absence of proteins, consistent with our theoretical predictions. While these experiments unequivocally demonstrate the possibility of sequence homology recognition without unzipping the double helix, much work remains to be done to test whether the mechanism of this recognition is indeed as predicted by the theory and whether such recognition plays any role in DNA pairing in vivo. Further experiments are currently in progress.	{ "pile_set_name": "NIH ExPorter" }
This research is aimed at uncovering the cellular mechanisms of behavioral heirarchies (choice) and associative learning in a "simple" model organism, the marine gastropod Pleurobranchaea californica. Corollary aims of the research are to determine the neural organization of the motor system that controls feeding behavior in this organism, since the feeding behavior is involved in both choice and learning. Specific projects we are undertaking are 1) a study of the synaptic organization of the feeding system in the brain of this organism; 2) an ultrastructural analysis of putative chemoreceptors (scanning and transmission electron microscopy); 3) a physiological study of chemoreception; 4) a cellular analysis of behavorial "switching" between feeding and rejection in this organism.	{ "pile_set_name": "NIH ExPorter" }
The proposed experiments are designed to identify the energy transducing enzyme(s) for mitosis and other microtubule-dependent movements through microinjection into living cells of active site specific monoclonal antibodies to various myosins and dyneins, to characterize the molecular interactions responsible for actin filament binding to micro-tubules and to determine the morphological relationships among actin, myosin and micro-tubules in the mitotic spindle and actin and myosin in the contractile ring. We will complete the characterization of monoclonal antibodies to Acanthamoeba myosins-I and II, identify the mechanism by which some interfere with the interaction of purified actin and myosin, use the antibodies to establish that antagonists have been identified for all classes of myosin in the cell and then microinject individual and combinations of these active site specific antibodies into dividing cells to test which myosins (if any) are required for cytokinesis and mitosis. Identical studies will be carried out using human cytoplasmic myosins isolated from platelets to elicit and test monoclonal antibodies that will then be microinjected into dividing HeLa cells. We will complete the characterization of our collection of monoclonal antibodies to sea urchin cytoplasmic dynein in collaboration with Drs. Begg and Pratt (Harvard Medical School) and test them for effects on mitosis by microinjection. We will raise and characterize monoclonal antibodies to Tetrahymena ciliary dynein. They will be used for studies in collaboration with Dr. Ken Johnson (Penn State University) on the mechanism of dynein and to test by microinjection whether protiens with active sites similar to ciliary dynein are required for mitosis. The biochemical studies on actin-microtubule interaction will focus on how the microtubule associated proteins cross-link the two fibers and how this interaction is regulated by phosphorylation and other factors. We will study by electron microscopy the distribution of actin filaments and myosin in dividing HeLa cells using ferritin-labeled monoclonal antibodies to myosin and 2 improved fixation techniques for actin filaments.	{ "pile_set_name": "NIH ExPorter" }
Genetic factors play a major role in the etiology of autoimmune thyroid diseases (AITD), Hashimoto's thyroiditis (HT) & Graves' disease (GD). Our hypothesis is that the etiology of AITD depends on interactions between immune regulatory genes and thyroid specific genes. Our goals are to identify the AITD susceptibility genes and to dissect the mechanisms by which they cause disease. Our findings during the last grant period, which are the starting point for the specific aims of this proposal, included: (1) Identifying novel AITD loci which are specific for subsets of AITD (Italian patients, & childhood AITD); (2) Narrowing down two major replicated AITD loci (12q & 14q), setting the stage for gene-identification; (3) Discovering a new Kozak sequence SNP in the CD40 gene that predisposes to GD by increasing the translational efficiency of CD40. For the next grant period we propose to build on these finding, and our specific aims are: (1) To identify the AITD susceptibility genes in the two subset specific loci by fine mapping in Italian GD patients, linked to a locus on 3q, and childhood AITD, linked to loci on Xp & 10q; the subset-specific AITD genes may represent novel therapeutic targets specific to subgroups of AITD patients; (2) To identify the AITD susceptibility genes in the 2 major replicated loci on 12q & 14q by linkage disequilibrium mapping, haplotype analysis, and gene sequencing. (3) To test the hypothesis that the CD40 Kozak SNP predisposes to GD by inducing over-expression of CD40 on antigen presenting cells (APC's), causing augmented immune responsiveness, as well as increasing CD40 expression on thyrocytes, thereby focusing the immune response to the thyroid. In vitro studies: we will examine the effects of the CD40 SNP genotypes on CD40 expression, signaling, and function in thyrocytes and APC's, in order to dissect the effects of the SNP on CD40 expression & function in these cells. In vivo studies: We are generating transgenic mice over-expressing CD40 in the thyroid. These mice will enable us to test, in vivo, the effects of CD40 over-expression in the thyroid on susceptibility to the induction of an experimental GD model. We will also use these mice to test CD40 blockade as a novel therapy for experimental GD. In summary, the current proposal builds directly on the knowledge gained in the previous grant period. Our approach has already been successful in identifying novel disease-associated genes. Establishing the immunogenetic pathways causing thyroid autoimmunity will lead to a better understanding of the basic etiology of AITD. This could have a major impact on public health, as it may facilitate the development of mechanism-based treatments in autoimmunity, such as CD40 blockade. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The overarching goal of this amended proposal entitled Long-term effects of depression care on services use and mortality in late life is to assess the long-term effect on use of health care services and mortality of a primary care practice-based intervention for depression. Our work has shown that after 5 years of follow-up, older adults with major depression in practices randomized to the practice-based intervention condition were less likely to die over the follow-up interval. A goal of this competing renewal proposal to MH065539 is to extend our 5 year mortality follow-up to 8 years and to add claims data, supplementing available data on depression and depression interventions over 2 years with dates and causes of death from the National Center for Health Statistics (National Death Index Plus) and with health services claims data from the Center for Medicaid and Medicare Services. Specific aims of this study are: (1) to assess how the effectiveness of a practice-based intervention (intent-to-treat analysis) and increasing intensity of different types of depression treatment (as-treated analysis) in reducing all-cause mortality over the course of an 8-year follow-up interval differs between older adults with major depression, clinically significant minor depression, or suicidal ideation versus older adults without significant depression or suicidal ideation; and (2) to assess how the effectiveness of a practice-based intervention (intent-to-treat analysis) and increasing intensity of depression treatment (as- treated analysis) in reducing use of health care services over the course of an 8-year follow-up interval. To accomplish these aims, we will capitalize on screening, clinical assessment, treatment, and follow-up assessments of 1,226 persons which occurred for up to 2 years in PROSPECT, building on our work with 5- year mortality follow-up data. We propose to study the relationship of major depression, other depressive syndromes, and suicidal ideation with long-term outcomes (use of health services and mortality) in the context of an intervention directed at the practice level that increased individual-level exposure of older patients to antidepressant medication and psychotherapy	{ "pile_set_name": "NIH ExPorter" }
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. Identification and Characterization of Key Proteases Necessary for Ovulation	{ "pile_set_name": "NIH ExPorter" }
Recent developments in technology and the expanding power of computers have made it possible for large numbers of people to have direct access to an increasingly large array of information and services. One task that is fundamental to most-technology-based systems is information search and retrieval. Although this topic has received attention within the human- computer interaction literature, the available data on older people is limited. The data which is available suggests that older people have more difficult than young people using technology to perform these types of tasks. The goals of this research project are to investigate the ability of older people to use technology to perform information search and retrieval tasks so that interface guidelines can be developed which accommodate this population. The project will investigate a range of information search and retrieval tasks that cut across living domains (e.g. home, work) and involve a variety of technologies. The initial study will focus on telephone-based voice menus and will investigate design parameters associate with these types of systems such as speech rate, prosodic stress, and menu structure. This study will also investigate the potential benefits of screen phones, which combine voice and text menus, for older people. The second set of studies will involve using the World Wide Web to search for information. Data will be gathered on age differences in search performance and navigational efficiency. Strategies for enhance performance such as improve user mental models of Web systems and modifications in interface design will be evaluated. The third set of studies will focus on database inquiry tasks which are information search tasks commonly performed in work settings. The goals of this study are to evaluate if extended practice can minimize age performance differences for these types of tasks, and toe examine if changes to the design of the interface have an impact on performance. Consistent with the other projects in the Center the studies will examine the relationships between cognitive abilities and task performance. The data generated from the research will be used as input to design guidelines.	{ "pile_set_name": "NIH ExPorter" }
Functions of the Core This Core builds on the nanotechnology strengths at the University of Washington (UW), one of the oldest state-supported institutions of higher education on the west coast, and one of the preeminent research universities in the worid. The UW Center for Nanotechnology (CNT), directed by Professor Francois Baneyx, was established in 1997, and has more than seventy-five faculty members from the Departments of Chemistry, Physics, Bioengineering (e.g.. Prof. Gao), Chemical Engineering {e.g., Prof. Baneyx), Electrical Engineering, Materials Science and Engineering, Biochemistry, Genome Sciences, Physiology and Biophysics, and Microbiology. Its Ph.D. Program in Nanotechnology established through a National Science Foundation's Integrative Graduate Education and Research Traineeship (NSF-IGERT) award was the first of its kind in the nation;the program provides graduate students with excellent interdisciplinary education experiences in nanoscale science and technology and emphasizes career path development. CNT is also home to the NanoTech User Facility (NTUF), which provides the Pacific Northwest nanotechnology community with access to advanced characterization and nanofabrication equipment. In 2004, NTUF expanded its role to the national level by becoming one of 14 nodes in the National Nanotechnology Infrastructure Network (NNIN), an NSF sponsored network of 13 universities, whose mission is to establish the infrastructure for current and future research and education needs in nanoscience and nanotechnology. NTUF houses leading-edge instruments, that will be critical for the characterization of quantum dots (Qdots) discussed below. Imaging tools include a Leica inverted fluorescence microscope, a Renishaw inVia Confocal Raman Microscope, an FEI field emission SEM with electron beam lithography capability, a Vecco Nanoscope scanning probe microscope with scanning tunneling microscopy and liquid mode force-distance measurement capabilities, a Zeiss LSM510 laser scanning confocal microscope interfaced with an atomic force microscope, a Woollam imaging ellipsometer, and a recently acquired Tecnai 200 kV scanning transmission electron microscope (S/TEM) with tomography capability. Fabrication tools include soft lithography, a Nabity e-Beam lithography system in the SEM, an Oxford OpAL atomic layer deposition system and a Heidelberg maskless pattern writer. NTUF also performs in-house nanotechnology tool development by drawing on faculty research and expertise. In addition. Prof. Baneyx, Gao, and Yost also have state-of-the-art instruments in their own laboratories for nanoparticle fabrication, purification, and characterization (see investigator equipment lists). Within this framework, the proposed Core has the following specific functions: (1) Preparation of well-characterized and purified nanoparticles in sufficient quantities for the U19 projects; (2) Construction of an aerosol generation and nanoparticle carrier system for exposing air-liquid interface cultures (Project 1) and mice (Project 2) to Qdots.	{ "pile_set_name": "NIH ExPorter" }
DESCRIPTION (Applicant's Abstract): This competing continuation application requests funds to extend a longitudinal project examining the role of family and mental health factors in HIV risk exposure during the transition to adolescence to mid-late adolescence -- the time when most youth are likely to be involved in sexual and other risk behaviors (e.g., IV drug use) that can lead to exposure to HIV, other sexually transmitted diseases, early pregnancy or fatherhood, and other negative health consequences. A sample of 315 urban African-American families living in high rise housing neighborhoods in Chicago were first contacted when their children were pre-adolescent (10-12 years of age) and followed through early adolescence (12-14 years of age). Continued funding is requested to follow the sample to mid-late adolescence (16-18 years of age), in order to better understand the influence of pre and early adolescent experiences upon HIV risk exposure in later adolescence. At the initial two data waves, extensive videotape interaction and interview data were collected on individual (mental health, cognitive level, pubertal development), familial support, supervision/monitoring, control/decision-making, conflict, affect, problem solving, and communication) and friendship (support, values) factors, as well as upon HIV/AIDS knowledge and attitudes, and HIV risk behavior. The proposed follow-up wave would focus on these same factors, adapting as needed to new developmental levels. For example, in addition to videotaping the family, same sex friends would be requested to be involved in data collection, as such relationships may be powerful in communication of risk and behavioral decision making. In addition to assessment of friendship relationships, qualities of romantic relationships (with the possibility of both same and other sex relationships) will be assessed. An extensive strategy for recruitment and tracking and for adapting of assessments is proposed; included in this strategy is development of a youth collaborative board to advise on these issues. Results will be used to further develop existing programs aimed at primary and secondary prevention of HIV risk exposure for urban African-American youth and families, as well as to explore new possibilities for the incorporation of friendship and romantic partner relationships into these programs.	{ "pile_set_name": "NIH ExPorter" }
Natural Killer T (iNKT) cells have evolved to recognize glycolipid antigens presented by CD1d molecules. Following stimulation through their T cell receptor (TCR), iNKT cells respond very rapidly, as is characteristic of innate rather than adaptive responses. iNKT cells have been implicated in the regulation of immune responses associated with a broad range of diseases, including autoimmunity, allergy, infectious diseases and cancer. iNKT cells can be activated through two major pathways. Microbe-specific CD1d-restricted lipids can stimulate iNKT cells directly. Alternatively, iNKT cells can be activated via recognition of CD1d-restricted self-antigens, in combination with inflammatory cytokines. This suggests that iNKT cells might play a role in the response to nearly all infectious agents. Furthermore, recognition of "self" by iNKT cells might play an important role in cancer immunity as iNKT cells have been shown to play a critical role in the immune surveillance of carcinoma, sarcoma and hematopoietic malignancies. Similarly, the inflammatory cascade triggered during ischemia-reperfusion injury and sickle cell disease appears to be a direct consequence of iNKT cells activation by self-derived lipids. Importantly, self-antigens that can trigger an iNKT cell response may be responsible for the positive selection of these cells during their development. The nature of the self-lipid(s) that are involved in these processes is currently a subject of controversy. We have engineered iNKT T cell receptors that conserve the same specificity than "regular iNKT TCR" but have a higher affinity for the antigen-CD1d complex. Using these unique tools, we have started to identify self-antigens that can bind the TCR when presented by CD1d molecules. Through distinctive and multi-pronged approaches we propose to examine what is/are the self-ligands of iNKT cells and how they are recognized. Finally, we will assess, using a highly innovative approach, how these newly identified self-ligands affect iNKT cell development. These studies will define the guidelines to optimize iNKT cell ligands and ultimately regulate iNKT cell function, with important implications for glycolipid-based vaccine development. PUBLIC HEALTH RELEVANCE: Natural Killer T cells (NKT cells) represent a unique lymphocyte population that has evolved to recognize glycolipid antigens presented by a non-polymorphic MHC class I molecule, CD1d. These cells have been implicated in the regulation of immune responses associated with a broad range of diseases, including autoimmunity, allergy, infectious diseases and cancer. NKT cells can be activated by two major ways. They can either recognize foreign lipid antigens expressed by microbes or they can be activated by self-lipids under certain inflammatory conditions. This recognition of "self" is a trademark of NKT cells that appears important to many of their ascribed functions, from immune surveillance of tumors to ischemia-reperfusion injury. The overall goal of this grant is to understand more fully the nature of the self-lipids responsible for this activation and to better define how NKT cells recognize them. An understanding of iNKT cell receptor-antigen recognition is of fundamental importance to understand the function of these cells and could potentially be exploited for therapeutic usage to improve human health.	{ "pile_set_name": "NIH ExPorter" }
This proposal focuses on an important but unexplored problem - howsteroid hormones, which are well known risk factors, interact with p53 mutations to produce aneuploidy and malignancy, and how the chromosomal segregation protein Separase is involved? Our sex-steroid dependent p53-mice preneoplastic breast cancer model allows a unique approach to this problem. In this model, steroidal induction in p53 mutant mammary glands results in chromosomal instability, aneuploidy and tumor formation analogous to that seen in majority of human breast cancers. We propose a paradigm that there is a set of proteins whose deregulation promotes aneuploidy (termed PRAN; Promoter of Aneuploidy) including chromosomal instability which results in loss or gain of whole or parts of chromosomes, and that PRAN proteins are interactively regulated by steroid hormones and the tumor suppressor p53. Our published and new preliminary data provide the first evidence that steroid hormones play a role in the regulation of mitotic proteins involved in sister chromatid cohesion and separation. We propose that the combined effect of mutation of the tumor suppressor p53 and signaling by steroid hormones produces aneuploidv in breast cancer by affecting expression of key proteins involved in chromosomal separation. We focuses on the elements that regulate chromosomal segregation, particularly sister chromatid cohesion/separation proteins, as candidate PRAN proteins, since chromosome missegregation during mitosis can lead to aneuploidy. A key gene in this analysis is ESPL1, which encodes an endopeptidase called Separase that separates joined sister chromatids by cleaving cohesin Rad21/SCC1/MCD1 during the metaphase to anaphase transition. The hypothesis is that hormonal stimulation of p53 null mouse mammary glands results in misexpression of the ESPL1 gene, thus promoting aneuploidy and breast cancer formation. This proposal applies in vivo transplantaion of p53 mutant and wild type (WT)mammary cells that are stably transfeeted with ESPL1, and an ESPL1 transgenic mice model to test the PRAN paradigm following hormone treatment. Steroid and p53 regulation of ESPL1 at the transcriptional level is studied by characterizing the ESPL1 promoter region. These objectives will be accomplished by pursuing two specific aims: 1) Functional role of Separaseoverexpressionin aneuploidy, and 2) Transcriptional regulation of ESPL1 gene expression. The proposed study not only elucidate underlying mechanisms of hormone-induced aneuploidy, a fundamental unresolved question in cancer biology, but also likely to identify a new class of proteins that are responsible for chromosomal instability and breast cancer progression.	{ "pile_set_name": "NIH ExPorter" }
This project will examine the hypothesis that insulin is critical for exercise-induced increases in rates of protein synthesis and skeletal muscle hypertrophy. Insulin Dependent Diabetes Mellitus causes an accelerated loss of skeletal muscle mass. This loss may be counteracted by resistance exercise which results in net protein accretion, however the role of insulin during such accretion has not been systematically studied. Specific Aims: We will determine whether elevations in rates of protein synthesis after acute and chronic resistance exercise are due to insulin- dependent increases in peptide-chain initiation. Insulin regulates specific eukaryotic initiation factors in peptide-chain initiation (the first steps in the translation of mRNA into protein), and that regulation is more pronounced in type II skeletal muscle fibers. Insulin availability during and after exercise may be critical for increasing rates of protein synthesis after resistance exercise. We will then determine whether the ability of muscle to hypertrophy is compromised in rats rendered insulin- deficient and whether such a decrement is due to altered regulation of specific steps in peptide chain initiation. Experiments and Methods: Rats are conditioned to rise up on their hindlimbs and push a switch in response to a light cue (to avoid electric shock). Resistance is added using weight packs on a Velcro vest (weights over the scapula). Acute resistance exercise will be weight lifting sessions on 4 separate days. Chronic resistance exercise will last 3, 6 or 9 wks with progressively greater weights being added to the backpacks The effect of prevailing hypoinsulinemia on the ability to increase rates of protein synthesis after acute exercise and the ability of muscle to hypertrophy due to chronic exercise will be elucidated using 70 and 90% partial pancreatectomy of mature (250g) rats. Rates of protein synthesis will be measured by incorporation of 3H-Phenylalanine using either an in vivo tritiated phenylalanine flooding dose technique or in situ bilateral hindlimb perfusion 16 hr after acute or chronic exercise. Because the effects of insulin on peptide-chain initiation are more pronounced in fast-twitch fibers, we will compare and contrast the affects of graded insulinemia on gastrocnemius (predominantly Type II), soleus (predominantly Type I), and extensor digitorum longus (EDL, predominantly Type II, but not extensively recruited during the exercise). Changes in ribosomal aggregation and the ability to form the 435 preinitiation complex in individual muscles will be used as markers of alterations in peptide-chain initiation. Mechanisms of the proposed involvement of peptide chain initiation will be evaluated by measuring the expression of eukaryotic initiation factor 2B (eIF-2B) and eIF-4E, the activity of eIF- 2B and phosphorylation state of eIF-4E. Significance: The proposed studies will be the first to systematically assess a role for insulin in protein synthesis during periods of net protein accretion. We predict a critical and fiber-specific role for insulin in mediating exercise-induced hypertrophy.	{ "pile_set_name": "NIH ExPorter" }
A clinical protocol was written to study cerebral glucose utilization in man in relation to aging, which employs positron emission tomography and 18F-2-deoxy-D-glucose as a tracer of cerebral glucose utilization. The protocol was approved and technical aspects of the emission tomography method were defined. Subjects were screened for the absence of neurological disease and for the study, which will commence in the forthcoming year.	{ "pile_set_name": "NIH ExPorter" }
Schistosomiasis is a neglected tropical disease that affects more than 200 million people in the developing world. Currently only a single drug (praziquantel) is available to treat this disease, highlighting the importance of studies aimed at understanding basic biological processes in these organisms. One remarkable characteristic of schistosomiasis is that the parasites responsible for this disease (i.e., schistosomes) can live in the blood stream of their human hosts for decades. How these parasites thrive in this immunologically hostile environment is not known. To address the longevity of these parasites in their mammalian host we previously demonstrated that schistosomes possess a population of somatic stem cells, which we refer to as neoblasts, which are capable of generating new intestine and muscle cells. Our preliminary studies for this proposal led us to a striking observation: a large fraction of these neoblasts differentiate to produce parts of the parasite's surface coat, a syncytial tissue known as the tegument. Our data suggest that tegumental cells are rapidly turned over, relying on neoblasts for their continuous renewal. As a result, many tegument-associated molecules are drastically down regulated within days following stem cell depletion. Since the tegument is the primary interface between the parasite and its host, understanding the stem cell-tegument relationship will provide important clues about how schistosomes defend themselves from immune attack. Based on these observations, we hypothesize that an important function of the neoblasts is to rapidly regenerate the parasite's tegument, thereby ensuring a functional host-parasite interface and evasion of host immunity. To address this hypothesis we propose the following two specific aims: (1) discover regulators of the neoblast-to-tegument transition and (2) understand the role for neoblasts in maintaining tegumental function in vivo. In the first aim, we will use transcriptional profiling and in situ hybridization to identify genes expressed in schistosome stem cells. We will then use RNA interference to determine which of these genes are required for the differentiation of the neoblasts into tegumental cells. In the second aim, we will use a combination of electron microscopy and immunological approaches to determine the effects that neoblast ablation has on tegumental morphology and function. Concurrently, we will surgically-transplant parasites incapable of making new tegumental cells into mice and examine the effects on worm survival. These studies are innovative as they are the first to explore the function of the neoblasts in the biology of this devastating parasite and bring a number of new methodologies to the table for studying these organisms. Collectively, these studies will determine how neoblasts contribute to parasite survival in the context of the host immune system, potentially leading to new therapeutic approaches targeting these important pathogens.	{ "pile_set_name": "NIH ExPorter" }
Over the last decade, studies have revealed that diminishing mutant huntingtin levels in mouse models of Huntington's disease (HD) leads to the amelioration of pre-existing symptoms, raising the tantalizing possibility for a cure. One of the primary events that accompany symptomatic reversal is the concomitant clearance of the aggregated mutant protein. Despite the intense debate that surrounds the role of protein aggregation in the pathogenesis of HD, a great amount of effort has been put forward to inhibit or disaggregate these proteinaceous intracellular deposits, with limited success. More recently, efforts to drive the turnover of these structures have been proposed, with some promise. One difficulty with these studies has been the inability to target protein degradation pathways in such a way to either enhance or impede the selective elimination of the aggregates, often leading to unwanted, nonspecific consequences that obscure the interpretation of the studies outcome. Recent studies have emerged demonstrating that the protein degradation pathway macroautophagy is capable of the selective degradation of various cargo including ubiquitinated protein aggregates. We have identified the Autophagy linked FYVE domain protein (Alfy) as essential to this process: Importantly not only does depletion of Alfy inhibit the macroautophagic clearance of aggregated mutant htt, but it does so without inhibiting basal and starvation-mediated macroautophagy. Moreover, over-expression of Alfy in neurons led to fewer mutant huntingtin inclusions. In response to the PAS-10-183 Validation of Novel Therapeutic Targets for Huntington's disease, we propose to use Alfy to genetically determine whether the clearance of aggregated mutant huntingtin represents a valid therapeutic approach in HD. PUBLIC HEALTH RELEVANCE: Huntington's disease is a devastating, inherited neurodegenerative disorder that affects 5 to 10 persons per every 100,000. By understanding the cellular events that affect the disease may not only help cure HD, but may also shed insight into sporadic disorders such as Parkinson's disease, Alzheimer's disease and Lou Gehrig's disease. Our goal is to determine if the elimination of protein aggregates, a common feature across most adult onset neurodegenerative diseases, represents a viable therapeutic strategy.	{ "pile_set_name": "NIH ExPorter" }
The broad objective of the research proposed in this competing continuation application is to identify predictors of risky and safer sexual behavior and sexually transmitted disease (STD) among South African adolescents. This is a collaborative effort by a multidisciplinary team of researchers from the United States and the Republic of South Africa to conduct a long-term follow-up of an ongoing clustered randomized controlled trial of an HIV/STD risk-reduction behavioral intervention. The Specific Aims are to (1) determine whether the effects of the HIV/STD risk-reduction intervention on self-reported sexual behavior are evident at 36-month and 48-month follow-up; (2) determine whether the intervention reduced the rate of clinically documented STDs; (3) identify participants' characteristics and theoretical mediator variables assessed at baseline that predict sexual-risk behavior at long-term follow-up; and (4) identify participants' characteristics and theoretical mediator variables assessed at baseline that predict STD incidence. The participants will be the 1,057 Xhosa-speaking grade 6 learners from the 18 schools in Mdantsane and Berlin in the Eastern Cape Province, South Africa who were randomized to Let Us Protect Our Future a 12-hour HIV/STD Risk- Reduction Intervention or a 12-hour health-promotion control intervention on health issues unrelated to sexual behavior. Male and female Xhosa-speaking co-facilitators from the community led both interventions. The theoretical basis of the interventions was social cognitive theory and the theory of planned behavior. Extensive formative research, pilot studies, and input from a Community Advisory Board were employed to ensure the contextual appropriateness of the interventions and other protocols. The participants will be about 16 years of age at the time of the proposed 48-month follow-up, an age at which many South African adolescents have their sexual debut. The proposed research, then, provides a unique opportunity to examine predictors of sexual risk behavior at a critical period in the development of South African adolescents. Confidential self-report questionnaires will be employed to assess sexual risk behaviors and theoretical variables hypothesized to mediate the intervention's effect on such behavior 36 months and 48 months postintervention. Urine specimens will be collected and tested for Chlamydia trachomatis, Neisseria gonorrhea, and Trichomonas vaginalis using DNA-amplification technology. To address the Specific Aims, we will analyze the data with generalized estimating equations (GEE) and random effects models. The results of this study will contribute to scientific knowledge regarding the long-term effects of HIV/STD risk- reduction interventions for South African adolescents and the predictors of sexual risk behavior and STD rates as the participants approach middle adolescence. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
a. Functions of the module. This proposed module has evolved from the Histology Module to meet the expanding imaging needs of CVS Core users. During the past five years many exciting developments have taken place in vision science at Rochester and with those developments, the needs for imaging have increased substantially and have broadened to new technologies. Some of the major developments include the establishment of the Rochester Center for Brain Imaging (RCBI), with its state-of-the-art 3T magnet, the joint acquisition by CVS and the University of Rochester Eye Institute of the Confocal Imaging Facility, and a number of newly hired vision scientists, working in areas ranging from cell and molecular biology, to cortical circuitry, physiology and psychophysics. The proposed module will consist of three components: Histology Facility, Confocal Imaging Facility and Functional/Structural MRI Facility. A highly qualified full-time technician will staff each of these three components.	{ "pile_set_name": "NIH ExPorter" }
The primary purpose of the proposed research program is to improve the understanding of the effects of normal aging on language processing in bilinguals. In the proposed studies, the investigator will extend new methods and concepts from the literature on aging in monolinguals to the special problems faced by multilingual adults, with emphasis on the potential for between-language interference in fluent comprehension and production. The proposed experiments focus on the interplay between semantic and/or linguistic inhibition, and their contribution to lexical access in and out of a discourse context. Participants will be Spanish-English bilinguals (elderly bilinguals, and college-age controls under normal and stressed conditions), tested in a series of "on-line" (real time) methods for the assessment of word comprehension and production. All experiments are administered in both Spanish and English. Individuals will be screened to assess language history and language dominance in "off-line" (untimed) and on-line (timed) situations. The proposed on-line tasks will include word repetition (i.e. "auditory naming"), picture naming, picture-word priming, and picture-word Stroop tasks. The investigator's results to date with a subset of these methods suggest that the ability to process words from Language A while listening to Language B is markedly slowed in elderly bilinguals, and in young bilinguals processing the same stimuli under adverse conditions. These laboratory findings have implications for the language switching commonly experienced by bilinguals in daily life. Results will also provide information relevant to competing accounts of cognitive declines in normal aging (based primarily on studies of monolinguals), including slowed or reduced activation, reductions in working memory, and/or a reduced ability to inhibit competing responses that are not relevant to the task.	{ "pile_set_name": "NIH ExPorter" }
Our goal is to develop a viral vector to study the molecular interactions between arthropods and the pathogens they transmit causing diseases such as malaria, lymphatic filariasis, dengue fever, African sleeping sickness, Chaga's disease and Lyme disease. A viral vector for gene expression and gene silencing in arthropods is a powerful tool for studying the function of the genes involved in disease transmission. Vectors derived from Sindbis virus (SIN) have proven to be very useful in this type of research and few other viral vectors are under development. Use of SIN, however, is constrained by the narrow host range, limited tissue dissemination, inability to express larger genes and human pathogenicity of the virus. During the previous period of support, we initiated studies to develop a highly efficient viral vector system based on Flock House Virus (FHV), a simple RNA virus and a member of the Nodaviridae family of insect origin. FHV has a small genome (4.5 kb) composed of two separate positive sense single stranded RNAs packaged into a coat protein with no envelope. It is not a human pathogen and has the ability to adapt to a wide range of insect hosts as well as hosts from different kingdoms such as plants and yeast. During the initial period of support we have extended the host range of FHV to four mosquito genera (Aedes, Anopheles, Culex and Armigeres), a species of tsetse (Glossina morsitans) and a reduviid (Rhodnius prolixus). These insects showed no adverse effect when infected with appropriate doses of FHV. We have made several vector constructs and demonstrated their abilities to express forein genes in mosquito and tsetse cells. In this continuation application we propose to (1) assess infection potential of FHV to two species of ticks (Ixodes scapularis and Ornithodoros turacata) that transmit Lyme disease and Relapsing Fever (2) continue to improve existing FHV RNA-based vectors for transient gene expression in above insect cells, and develop DNA-based vectors for prolonged expression, and (3) use FHV vectors to study the effects of expressing and/or silencing genes involved in immunity and pathogen transmissibility of Anopheles, tsetse and ticks. We expect to establish FHV vectors as a powerful new system to study gene function in a wide variety of medically important arthropods.	{ "pile_set_name": "NIH ExPorter" }
A retrospective analysis of 54 patients undergoing concomitant aortic, mitral and tricuspid valve replacement at the NIH was performed. Pre- and postoperative catheterization data was subjected to statistical analysis as well as patient age, patient sex, history of prior cardiac surgery and date of operation. Patients were grouped by valve type to assess differences in long-term morbidity and mortality. Computer generated cumulative survival and event-free survival curves were compared. Findings included the following: 1) Operative survivors had a significantly lower preoperative mean pulmonary artery pressure, 2) decrease in pulmonary artery systolic pressure following operation had a positive correlation to long term survival, 3) advanced age had a significant influence on operative mortality but patient sex and history of prior cardiac surgery did not, 4) patients with all porcine valves had an improved raw survival and event-free survival compared with other combinations of prosthetic valves.	{ "pile_set_name": "NIH ExPorter" }
A major problem in neurobiology is how the central nervous system is able to coordinate the movements of different parts of the body during a behavior. The long-term objective of the proposed project is to help elucidate the mechanisms by which such coordination is achieved. Experiments will be carried out on the American cockroach. Insects will be pinned out in such a way that some of their legs will be free to move while intracellular recordings are taken from neurons impaled in the connectives which join two adjacent thoracic ganglia, or in the ganglia themselves. Detailed study will be made of neurons which respond preferentially to natural or imposed movement of one or more legs. This study will include morphological work with neurons filled with cobalt chloride or Lucifer yellow to determine the extent and location of dendritic and axonal arborizations. The morphological information will be used to help locate areas of synaptic interaction between the interneurons and sensory or motor neurons, so that the way sensory input influences motor output can be determined.	{ "pile_set_name": "NIH ExPorter" }
The proposed research will challenge the standard approach to lumbar interbody fusions (LIFs) by utilizing medical imaging, finite-element (FE) models and emerging materials to guide novel device design. These fusions are performed annually on 500,000 patients who suffer from degenerative disc disease, instability, and scoliosis. This proposal offers an innovative, multi-disciplinary approach to studying LIFs. Conventional CT will be correlated and utilized to predict local strength and modulus values across the endplates. These data will also be used to create patient-specific, FE models with the matching material properties. FE simulations will be used to evaluate the complex biomechanics of the spine for a wide variety of cages/rod designs, materials, and configurations. Furthermore, simulations will yield valuable information regarding the stress distribution across the endplates as well as overall construct stiffness as a function of bone formation and fusion. Lastly, poly(para-phenylenes) (PPPs) are a new class of aromatic polymers with strength and modulus values higher than poly(ether-ether-ketone) (PEEK). PPPs have superior manufacturability compared to current cage materials and will be made into porous devices. The porosity of the device will be spatially tailored to match the properties of the implant across the endplates, while also promoting osteointegration of bone. Our hypothesis is that a porous interbody fusion cage would reduce complications (subsidence and adjacent-level disease) and improve clinical outcomes by (1) spatially tailoring the modulus of the implant to the endplate, (2) more evenly distributing stresses across the entire endplate, (3) lowering the overall construct stiffness value, and (4) allowing for osteointegration of bone into the implant. If successful, the proposed research would shift the paradigm for how LIF cages and constructs are designed as well as lower rates of subsidence, implant failures, and adjacent-level disease. The proposed team consists of an inter- disciplinary group with backgrounds in clinical surgery, mechanical and biomedical engineering, and materials science.	{ "pile_set_name": "NIH ExPorter" }
It is proposed to use synthetic antigens to investigate a number of parameters of the immune response. As an extension of findings that spacers between the haptenic and immunogenic determinants of bifunctional antigens modulate the anti-hapten response, the upper limit of spacer size will be determined by synthesizing bifunctional molecules with large rigid spacers. Monofunctional molecules which differ only slightly in structure but profoundly in immunogenicity will be compared with respect to macrophage processing, tolerogenicity and induced changes in levels of prostaglandins and cyclic nucleotides in lymphoid cells. The object is to elucidate at what stage of antigen-immune system interaction discrimination between immunogens and non-immunogens takes place. Monofunctional immunogens which differ in the specificity of their induced delayed hypersensitivity responses will be compared with respect to the specificity of tolerance and helper activity they induce, in order to assess the cellular relationships of these functional activities. It will be determined if the unresponsiveness induced by monofunctional antigens is mediated by suppressor T-cells. Finally, it will be determined if T-cells in A/J mice specific for the azobenzenearsonate epitope bear surface molecules which share the cross-reactive idiotype found on antibodies, and, if so, whether the molecules are biosynthetic products of T-cells.	{ "pile_set_name": "NIH ExPorter" }
The sulfinic acid reductase, sulfiredoxin (Srx), was recently identified as the enzyme in yeast responsible for the reduction of the sulfinic acid moiety (Cys-SO2-) within several oxidatively-inactivated peroxiredoxins (Prxs). This discovery shattered the dogma of the irreversibility of overoxidation for the Prx enzyme family. Moreover, the ATP- and Mg2+-dependent repair or "retroreduction" of the overoxidized Prxs may modulate the role of these enzymes as regulators of hydrogen peroxide-mediated intracellular signaling. Based on the analysis of the in vivo oxidation state for a variety of human Prxs, repair of different Prx isoforms by Srx appears to proceed at different rates, in spite of their high overall degree of sequence identity. Our recent structure determinations of human Srx in complex with either phosphate or ADP have revealed a new protein fold and a novel nucleotide binding motif. Other preliminary data have confirmed some aspects of the proposed scheme for yeast Srx action, although several surprises in our results (the isolation of a disulfide-bonded rather than thiosulfinate-linked intermediate, and the reduction of the Srx:Prx complex by glutathione rather than thioredoxin) have suggested that further investigation is required. The goals of this proposal are: to determine the crystal structures of human Srx in complex with cofactors and human Prxs (Aim 1); and to carry out steady-state and partial turnover experiments coupled with site-directed mutagenesis to elucidate the nature and reaction rates of intermediates, and to characterize residues critical to catalysis (Aims 2 & 3). These investigations will contribute significantly to our understanding of the molecular origins of sulfinic acid reductase action and the novel sulfur chemistry involved in this process.	{ "pile_set_name": "NIH ExPorter" }

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