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The project includes studies on medical ecology, and the zoogeography, evolution and systematics of fleas, and is primarily based upon collections made during investigations on vectors and reservoirs of rickettsial and viral investigations undertaken in New Guinea, Ethiopia, Burma, Pakistan and other areas. Fleas and other ectoparasites are important in medicine not only as vectors of diseases, but because their study contributes to our understanding of the historical biogeography, distribution, and ecology of mammals and birds, and hence with the infections harbored or transmitted by such hosts. For example, it was because of the demonstrated affinities and zoogeography of fleas, chiggers and rodents that members of this Department sought, and found, scrub typhus and tick typhus in totally unexpected geographical regions and habitats in Pakistan. This "faunal approach" led to 1) predictions, since confirmed, of the occurrence of certain infections in parts of the USSR, Pakistan, India, China and Indo-China, and 2) to the hypothesis that the infections acquired by man from other animals are actually almost all transmitted from mammals that are higher in the evolutionary scale (e.g. ruminants, rodents), instead of from those lower down (e.g. bats, marsupials and insectivores). These concepts have led to some significant generalizations about the ecology of murine typhus and Korean hemorrhagic fever. During the next year, intensive study will be continued on the distribution, host relationships, phylogeny and systematics of fleas. Papers will be prepared primarily on the systematics of the Pygiopsyllidae, the major representative of the Order in the Australian Region, and on some Ethiopian fleas, as well as on Leptopsyllids.
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{
"pile_set_name": "NIH ExPorter"
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Rotaviruses are the single most important etiologic agents of severe diarrhea of infants and young children worldwide. Because only four human rotavirus serotypes appeared to be epidemiologically important we developed a quadrivalent rotavirus vaccine containing representatives of each of the 4 serotypes: rhesus rotavirus (RRV) a VP7 serotype 3 strain (the Jennerian approach), and three human RV-RRV reassortants, each possessing ten RRV genes and a single human RV gene that encodes VP7 (a major outer shell protein) that is responsible for serotype 1, 2, or 4 specificity (the modified Jennerian approach). Four randomized, placebo-controlled, coded, efficacy trials of this orally delivered vaccine used at a dose of 4x105 PFU have been completed in 5958 infants and young children, (2 in the U.S. [783 and 695 children] and one each in Finland [2273 children], and Venezuela [2207 children]). This quadrivalent formulation, given orally in three doses, was highly effective in preventing severe rotavirus diarrhea with efficacy ranging from 69% to 91%. The vaccine was 75% to 100% effective in preventing dehydrating diarrheal illnesses. Because of the demonstrated safety (transient fever) and efficacy of the quadrivalent vaccine, the Advisory Committee on Immunization Practices in 1998 recommended its use for infants at 2, 4, and 6 months of age. Subsequently, in August 1998 the FDA granted a Biologics License to Wyeth Laboratories. However, after an estimated 1.5 million doses had been given to about 1 million infants, the CDC in July 1999 recommended postponing further vaccination because of the occurrence of 15 cases of intussusception following vaccination. Although the number of cases reported was within the expected value (considering a background rate of 51 per 100,000 [or about 1 case per 2000 infants during the first year of life in data from N.Y. State]), it was of concern that 11 of the 15 cases occurred within 1 week of receiving the first dose of vaccine. Subsequently in October 1999 the ACIP withdrew its previous recommendation because of additional data extending the initial findings of a link with intussusception. In conjunction with these events, Wyeth-Lederle withdrew the vaccine. In other studies, we evaluated collaboratively the feasibility of administering the licensed rhesus rotavirus-based vaccine to neonates to determine its reactogenicity and immunogenicity with different administration schedules. We also evaluated collaboratively several second and third generation vaccines: (i) a quadrivalent reassortant rotavirus vaccine that possesses a single VP7 gene from a human strain with serotype 1, 2, 3, or 4 specificity and the remaining ten genes from bovine RV (UK); (ii) a reassortant vaccine (Wa x UK) that contains a single gene from the human Wa strain, that encodes VP4:1A specificity, in a background of 10 bovine (UK) genes; (iii) a reassortant candidate vaccine Wa x (DS-1 x UK) that possesses the VP4 gene from the Wa strain (VP4:1A) as well as the VP7 gene from human strain DS-1 that encodes G2 specificity, in a background of 9 bovine (UK) genes; (iv) and a 30?C cold-adapted, temperature-sensitive, VP7:1 strain (D[75-82]) (see Hoshino and Ishida reports).
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{
"pile_set_name": "NIH ExPorter"
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Macrophages (MF) play a central role in atherogenesis through accumulation of lipids and production of inflammatory cytokines. Plasma phospholipid transfer protein (PLTP) is associated with MF in lesions and its' expression is up-regulated by lipid loading. We reported that PLTP produced within lesions is atheroprotective. The long-term goal of this project is to identify mechanisms whereby cell associated PLTP is anti-inflammatory and atheroprotective. The first aim will test the hypothesis that MF PLTP disrupts TLR2/1 plasma membrane dimerization and signaling, but not preformed TLR3 dimer mediated signaling in endosomes. We will assess the anti-inflammatory effects of MF PLTP on TLR2/1 and TLR3 signaling in PLTP- /-LDLr-/- mice irradiated and reconstituted with bone-marrow (BM) from PLTP+/+ or PLTP-/- mice. Cytokine production and atherosclerosis will be studied. The second aim will test the hypothesis that PLTP alters cholesterol dependent membrane domains (lipid rafts). We will determine if wild-type (WT) or mutant PLTP (which binds ABCA1 but lacks lipid transfer activity) disrupts lipid rafts and/or inhibits TLR2/1 dimerization in vivo and in vitro. The third aim will test the idea that reduced hepatic production of PLTP to reduce plasma levels of PLTP will favor the beneficial effects of cell associated PLTP. We will assess the anti-inflammatory effects of PLTP on TLR2 signaling in LDLr-/- mice with suppressed, but not deleted, hepatic PLTP. This will be accomplished with in vivo administration of mouse PLTP antisense oligonucleotides (ASO). Novel, targeted mouse models for study of chronic inflammation are used to establish the physiological significance of our hypotheses and will be complemented by in vitro studies to identify the mechanisms involved in these physiological events. Laser scanning confocal immunofluorescent microscopy is used to characterize early atherosclerosis progression. Fluorescent resonance energy transfer (FRET) is used to characterize changes in lipid rafts in response to PLTP. Finally, the proposed studies will use antisense oligonucleotides (ASO) to favor the beneficial effects of anti-inflammatory cellular PLTP while reducing the undesirable affects of circulating plasma PLTP. It is imperative that we now know whether cell-directed or plasma-directed PLTP atheroprotective therapeutics should be pursued for human treatment of cardiovascular risk factors. PUBLIC HEALTH RELEVANCE: The proposed studies will significantly alter the direction of PLTP research by providing sound evidence of its role in the regulation of intracellular inflammatory pathways. The results of our studies will lead to a new and improved understanding of MF physiology and inflammation thus providing the bases for novel treatment options for atherosclerosis, diabetes and other chronic diseases that have a strong inflammatory component.
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{
"pile_set_name": "NIH ExPorter"
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Diabetes mellitus (DM) is a prevalent and devastating chronic disease, affecting >29 million people in the United States (US). Health insurance and continued access to healthcare services are essential for optimal DM care and management; therefore, it is hypothesized that Affordable Care Act (ACA) Medicaid expansions could substantially improve access to health insurance and healthcare services for patients at risk for DM or already diagnosed with DM (`with DM risk or DM'). The ACA called for every state to significantly expand Medicaid coverage by 2014; little is yet known about the impact of ACA Medicaid expansion on the prevention, treatment, expenditures, and outcomes of patients with DM risk or DM. In a 2012 legal challenge, the US Supreme Court ruled that states were not required to implement ACA Medicaid expansions; thus (by January 1, 2015), 28 states and the District of Columbia expanded Medicaid, while 22 states did not. This `natural experiment' presents a unique opportunity to learn whether and to what extent Medicaid expansion can affect healthcare access and services for patients with DM risk or DM. We will use this unprecedented natural experiment to expand our understanding of how Medicaid expansion impacts DM prevention, treatment, expenditures, and health outcomes. We will use electronic health record (EHR) data from the ADVANCE clinical data research network, which has data from 718 community health centers (CHCs), including 470 CHCs in 12 Medicaid expansion states and 248 CHCs in 9 non-expansion states. From this dataset, we will collect detailed information on changes in health insurance, service receipt, and health outcomes, with data spanning 9 years (pre- and post-expansion), comparing states that expanded Medicaid, and those that did not. Moreover, building on our prior work, we will link EHR data from 213 Oregon ADVANCE CHCs to Oregon Medicaid claims data to assess Medicaid expenditures among patients with DM risk or DM. The specific aims are as follows: Aim 1. Compare pre-post insurance status, overall visits, and chronic disease management visits among patients with DM risk or DM, in expansion versus non-expansion states. Aim 2. Compare pre- post receipt of primary and secondary DM preventive services (e.g., screening for obesity, lipid levels, glycosylated hemoglobin) among patients with DM risk or DM, in expansion versus non-expansion states. Aim 3. Compare pre-post changes in DM-related biomarkers (e.g., body mass index, blood pressure, lipid levels) in patients with DM risk or DM among newly insured (gained Medicaid in post-period), already insured (had Medicaid coverage in pre- and post-period), and continuously uninsured (pre- and post-period) patients in states that expanded Medicaid. Aim 4. Measure pre-post changes in Oregon Medicaid expenditures among newly insured compared to already insured patients with DM risk or DM. Findings will be relevant to policy and practice, informing further improvements in the US healthcare system to ensure access for vulnerable populations and mitigate disparities in DM prevention, treatment, and health outcomes.
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{
"pile_set_name": "NIH ExPorter"
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This is the ROO activation of the funded K99 proposal Statistical Methods for Analysis of High-Dimensional Gene and Environment Data (K99ES017744) by Dr. Arnab Maity, a statistically trained Assistant Professor in the Department of Statistics at North Carolina State University (NCSU). Dr. Maity is committed to a research career in the development of statistical methodology for the analysis of high-dimensional gene and environment data. This application includes updated specific aims and research plans for the ROO phase, a description of progress during the K99 phase, evaluation reports from the K99 phase mentors, and a letter from Dr. Peter Bloomfield, Interim Head of Department of Statistics, NCSU, detailing the institutional commitment and details for Dr. Maity's career development plan. The proposed research concerns two major aims: (1) analyzing DNA methylation in the human genome and developing statistical methodology to investigate its association to environmental exposure to heavy metals and air particles, and various markers of cardiovascular disease, and (2) developing robust and efficient statistical testing procedures for genetic and environmental effects in high-dimensional genome-wide association studies (GWAS) in the presence of gene-gene and gene-environment interactions and incorporating longitudinal measures of phenotypes. The applicant has readily available data sets on genome-wide DNA methylation study in the Normative Aging Study and the genome-wide association studies of Framingham Heart Study. The proposed methods will be applied to these data sets to draw valuable conclusions regarding the interplay of DNA methylation and other genetic variants, and environmental exposures in relation to susceptibility to cardiovascular disease. Career development for Dr. Maity and accomplishment of the research aims will be facilitated by excellent research and career supporting resources available within the Statistics department at NCSU and the University, participation in various gene and environment research projects with other researchers within NCSU and outside such as UNC School of Public Health, NIEHS and SAMSI, various scientific meetings and seminars, and the rich research community in NCSU and Research Triangle Area.
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{
"pile_set_name": "NIH ExPorter"
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This proposes to etablish and operate a clinical unit in the NCI's multicenter randomized clinical trial of the efficacy of a diet low in fat (20% of calories) in the treatment of women with Stage II breast cancer. The trial's primary objective is to determine if the low-fat diet can decrease the incidence of recurrence of breast cancer. The secondary objective is to determine if the diet will decrease the incidence of second primary tumors. This application proposes the unit's participation in all three phases of the trial by the cooperative agreement mechanism; development of the protocol in two months, the feasibility trial of ten months, and the full-scale trial of 48 months. For the feasibility and full-scale trials, the unit will study two cohorts of 66 patients recruited by the Baylor Oncology Service at the Methodist Hospital, according to the protocol's criteria. The unit will collaborate with the trial's other clinical units, nutrition coordinating unit, statistical coordinating unit, and the NCI. The Baylor unit's major aims include: 1) To operate the clinic unit with economy and accuracy using a microcomputer assisted surveillance and management of clinic operations, and collection and quality control of data. 2) To gain maximum dietary adherence for this efficacy trial using a combination of standard and innovative techniques, including selection of subjects with high potential for adherence by an adherence screening examination and washout criteria, individual counseling by experienced dietitian/adherence counselers trained in counseling techniques developed by the Baylor Lipid Research Clinic aided by the 'instant' analysis of the patient's 7-day food record by the microcomputer based Nutrient Analysis System, and multimedia instruction by food models, cooking demonstrations, slide tapes, newsletters, and bulleting boards. Adherence counseling is based on the identification, diagnosis of and management of adherence problems using behavioral techniques and frequent counselor-patient contact. 3) To evaluate methods of diet recording, 24-hour diet recall, 1-7 day diet self-recording, and objective measures of diet adherence, body weight, skin-fold thickness, plasma total cholesterol, and HDL-cholesterol, and the proportion of lineleic acid in the plasma cholesterol esters. The optimum methods of clinical trial operation, adherence management and surveillance, from the feasibility trial will be applied to the full-scale trial. The project's significance is its potential for developing a dietary treatment and preventive procedure for Stage II breast cancer.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this multidisciplinary study is to determine the structural, physiological, biochemical and pharmacological changes that are produced by head injury, to recognize those changes which might be amenable to therapy and to estimate the efficiency of such therapy in the laboratory and ultimately at the clinical level. Earlier studies focused upon the ultrastructural and biochemical factors and recognized the changes in cell membranes, in energy metabolism of the cell and in blood flow. At a laboratory level we have shown in both compression and gunshot wounds that hemodilution helps reverse the deterioration in blood flow and aids survival. We have also demonstrated that dimethyl sulfoxide, a drug which belongs to both the radiation protection and cold protection series, aids survival in compression injuries. In the next year we plant to continue our studies of blood flow and membrane stabilization.
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{
"pile_set_name": "NIH ExPorter"
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Nicotine abuse leads to approximately 430,000 deaths a year in the United States. Even though most smokers want to quit, only 3% can do so without the use of other intervention. Both the psychological and physiological effects of tobacco smoke (smoking) are attributed to nicotine. The goal of this application is to discover and develop medications for the treatment of smokers. However, we expect that the compounds developed will also serve as biochemical probes useful in gaining a better understanding of the biochemical and molecular mechanisms of nicotine addition and withdrawal. Neuronal nicotinic acetylcholine receptors (nAChRs) are widely distributed throughout the central and peripheral nervous systems including several regions of the brain. Two major classes of nAChRs, alpha4beta2 and alpha7, have been identified in rat and human brains. The possibility exists that specific subtypes mediate specific functions, especially as this relates to nicotine addiction. Thus, the availability of a variety of ligands that bind with high affinity and selectivity for each subtype class is needed. It is also desirable to have both agonists and antagonists since the role of nAChRs in addiction is not known. Epibatidine is a nicotinic agonist whose biological effects appear to be mediated largely by alpha4beta2 nAChRs. Methyllycaconitine (MLA) is the most potent non-peptide competitive antagonists at U-7 neuronal nAChRs thus far reported. The high potency of epibatidine for uAP2 nAChRs and the potency and selectivity of MILA toward the brain alpha7 receptor makes these agents very useful lead compounds for the development of new ligands for studying these nicotinic subtypes. The objective of the research proposed in this application is to conduct SAR studies to develop novel epibatidine and MILA analogs for probing (alpha4beta2 and alpha7 nicotinic receptors. Pharmacological profiles of compounds identified as potent and/or selective for one of the subtypes will be examined for modulation of rodent behavior (antinociception and motor activity), for their ability to substitute for nicotine in rat discrimination, and for their ability to change nicotine self-administration in rats. Selected compounds will be studied at various alpha,beta and alpha7 combinations expressed in Xenopus oocytes to determine the nAChR subtype binding and functional profiles. Since little is known about the in vivo disposition of nAChR both before and after chronic nicotine exposure, PET and SPECT ligands will be developed to identify specific brain regions involved and to help resolve the role various nAChR subtypes play in the addiction process.
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{
"pile_set_name": "NIH ExPorter"
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Longitudinal bone growth occurs at the growth plate, a thin layer of cartilage which consists of three principal zones: the resting zone, the proliferative zone, and the hypertrophic zone. Studies in our laboratory indicate that the resting zone contains stem-like cells that are capable of generating new clones of proliferative chondrocytes. These proliferative cells undergo clonal expansion followed by cellular hypertrophy.[unreadable] [unreadable] With age, growth plate chondrocyte proliferation slows down, causing longitudinal bone growth to slow and eventually stop. Our previous studies indirectly suggest that proliferation slows because the stem-like cells of the resting zone have a finite proliferative capacity which is gradually exhausted. To test this hypothesis more directly, we measured the proliferation rate in resting zone chondrocytes using continuous bromodeoxyuridine labelling. Consistent with our hypothesis, we found that the proliferation rate decreases with age as does the number of resting zone chondrocytes per area of growth plate. [unreadable] [unreadable] Glucocorticoid excess slows growth plate senescence. To explain this effect, we hypothesized that glucocorticoid inhibits resting zone chondrocyte proliferation, thus conserving their proliferative capacity. Consistent with this hypothesis, we found that dexamethasone treatment decreases the proliferation rate of resting zone chondrocytes and slows the numerical depletion of these cells. [unreadable] [unreadable] Our findings support the hypotheses that growth plate senescence is caused by qualitative and quantitative depletion of stem-like cells in resting zone and that growth-inhibiting conditions, such as glucocorticoid excess, slow senescence by slowing resting zone chondrocyte proliferation and slowing the numerical depletion of these cells, thereby conserving the proliferative capacity of the growth plate. [unreadable] [unreadable] We have also investigated in detail one particular gene that might be involved in growth plate senescence. p27/Kip1, a cyclin-dependent kinase inhibitor, negatively regulates proliferation of multiple cell types. We therefore assessed the role of p27 in the regulation of growth plate chondrocyte proliferation. p27 mRNA expression was detected by real-time PCR in both proliferative/resting and hypertrophic zones of the mouse growth plate. To determine whether this expression is physiologically important, we studied skeletal growth in 7-wk-old mice lacking a functional p27 gene. In these mice, body length was modestly increased compared to wild-type littermates. In the proximal tibiae, proliferation of growth plate chondrocytes was increased but tibial length was not significantly greater than controls. p27 ablation had no measurable effect on growth plate morphology, including the number of proliferative or hypertrophic chondrocytes. Treatment with dexamethasone in vivo inhibited longitudinal bone growth similarly in p27-deficient mice and controls, indicating that p27 is not required for the inhibitory effects of glucocorticoid on growth plate function. p27-deficient mice had increased width of the femoral diaphysis, suggesting that p27 acts normally to inhibit periosteal bone growth. In summary, our findings suggest that p27 acts a negative regulator of growth plate chondrocyte proliferation. [unreadable] [unreadable] In clinical studies, we have investigated the relationship between skeletal development and pubertal maturation. The overall goal of these studies is to explore the primary mechanism that initiates puberty, which is currently unknown. One possible clue is that pubertal maturation often parallels skeletal maturation. Conditions that delay skeletal maturation also tend to delay the onset of puberty whereas conditions that accelerate skeletal maturation tend to hasten the onset of puberty. To examine this relationship, we previously studied boys with congenital adrenal hyperplasia and familial male-limited precocious puberty, two conditions that accelerate maturational tempo, and boys with idiopathic short stature in which maturational tempo is sometimes delayed. In all three conditions, the onset of central puberty generally occurred at an abnormal chronological age but a normal bone age. Boys with the greatest skeletal advancement began central puberty at the earliest age whereas boys with the greatest skeletal delay began puberty at the latest age. Furthermore, the magnitude of the skeletal advancement or delay matched the magnitude of the pubertal advancement or delay. This synchrony between skeletal maturation and hypothalamic-pituitary-gonadal axis maturation was observed among patients within each condition and also between conditions. In contrast, the maturation of the hypothalamic-pituitary-gonadal axis did not remain synchronous with other maturational processes including increases in weight, height, or body mass index. These data indicate that in boys with abnormal developmental tempo, maturation of the skeleton and the hypothalamic-pituitary-gonadal axis remain synchronous. [unreadable] [unreadable] This synchrony raises the possibility that skeletal maturation influences pubertal onset. To determine whether this concordance is also present in normal children, we analyzed data from 30 normal boys participating in a longitudinal study. Height, weight, and serum testosterone concentrations were assessed every 6 months and bone age every year. Pubertal onset was defined by serum testosterone greater than 30 ng/dL. The variability in bone age at onset of puberty was not less than the variability in chronological age. In addition, there was no significant correlation between skeletal advancement and pubertal advancement. Similarly, there was not a significant correlation between pubertal advancement and height age advancement, weight age advancement, or body mass index age advancement. These findings do not support the hypothesis that skeletal maturation directly influences the age of pubertal onset in normal boys. To explain both these findings in normal boys and the previous findings in boys with abnormal maturation requires a more complex model.
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{
"pile_set_name": "NIH ExPorter"
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PROJECT SUMMARY Gorham-Stout disease (GSD) and generalized lymphatic anomaly (GLA) are related diseases of the lymphatic system. GSD and GLA patients develop ectopic lymphatics in bone and gradually lose bone. Unfortunately, our understanding of the biology of GLA and GSD has not significantly advanced over the past 60 years. This gap in knowledge has made it difficult to treat GLA and GSD patients. Our long-term goal is to identify effective therapies and biomarkers for GLA and GSD. The objective of this application, which is the first step in pursuit of that goal, is to use novel mouse models developed in my laboratory to determine how lymphatic vessels form in bone and promote bone loss. PIK3CA encodes the catalytic subunit of phosphatidylinositol 3-kinase (PI3K). Our preliminary studies have revealed that mice that express an active form of PIK3CA (PIK3CALEC) in their lymphatic endothelial cells (LECs) develop irregular lymphatics in their soft tissues and bones. VEGF-C is a growth factor that stimulates PI3K signaling in LECs by activating VEGFR3. To induce excessive PI3K signaling specifically in LECs around bone, we created transgenic mice that overexpress VEGF-C in bone (VegfcOE). We have found that overexpression of VEGF-C in bone causes lymphatics to form in bone and bone loss. We have also found that rapamycin (FDA-approved mTOR inhibitor) inhibits lymphangiogenesis in PIK3CALEC mice and VegfcOE mice. This proposal will test the hypothesis that excessive PI3K/mTOR signaling in LECs stimulates the formation of lymphatic vessels in bone and osteoclast-mediated bone resorption. The specific aims of this proposal are to: 1) Characterize the development of bone lymphatics in PIK3CALEC and VegfcOE mice; 2) Determine the mechanism of bone loss in PIK3CALEC and VegfcOE mice; and 3) Determine the effect of rapamycin on lymphangiogenesis and osteolysis in PIK3CALEC and VegfcOE mice. The results from these experiments will have a significant impact on the field because they will answer fundamental questions regarding the biology of bone lymphatics. Our experiments will reveal how lymphatics form in bone and promote bone loss. This knowledge will significantly advance our understanding of processes that are relevant to the pathogenesis of GLA and GSD and could be used to inhibit lymphangiogenesis in GLA, GSD, and other human diseases.
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{
"pile_set_name": "NIH ExPorter"
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A multidisciplinary predoctoral training program in biomedical imaging and spectroscopy (BMIS) is proposed. The BMIS program will provide a unique educational experience based on a solid fundamental training in the mathematical and engineering principles of image science combined with a focus on and exposure to a broad range of biological applications. This program will draw on the existing strength of faculty and research programs in biomedical imaging and spectroscopy at the University of Arizona. Areas of emphasis include magnetic resonance imaging, magnetic resonance spectroscopy, gamma-ray imaging, x-ray imaging, optical spectroscopy, optical imaging, image processing, and image quality assessment. A specialized curriculum is proposed for students during the first two years that will involve courses in applied physiology, biology, the mathematical principles of image science, as well as the physics and engineering principles at the foundation of modern imaging and spectroscopic systems. These courses will provide students with the knowledge base necessary to carry out advanced research on the development and utilization of advanced biomedical imaging and spectroscopic technologies. In addition to the course work, students enrolled in the program will be required to complete semester-long rotations in three different research laboratories prior to selecting a laboratory and mentor for their Ph.D. dissertation work. These rotations will provide students with exposure to multiple disciplines and research environments and will help create and foster increased collaboration among researchers at the University of Arizona. Students will be recruited into BMIS through existing graduate-level education programs at the University of Arizona. The primary conduits for recruiting students will be through the graduate programs in Optical Sciences, Biomedical Engineering, and Physiological Sciences, although outstanding students from other programs such as Electrical and Computer Engineering, Physics, Chemistry, Cancer Biology, and/or Biochemistry will be eligible to participate. Program funds will be used to fully support students during their first three years. Third year students will be required to serve as student mentors to individuals entering the program. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Oxygen (O2) is not only an indispensable metabolic substrate in various enzymatic reactions including mitochondrial respiration, but also a regulatory signal that controls stability and activity of the transcription factor Hypoxia Inducibl Factor-1 (HIF-1), a key mediator of the cellular adaptation to low O2 tension (hypoxia). The fetal growth plate is a unique mesenchymal tissue because it is avascular, albeit it requires the angiogenic switch in order to be replaced by bone. Over the years, we have demonstrated that, consistent with its avascularity, the fetal growth plate has an inner hypoxic region. We have provided genetic evidence that HIF-1 is a survival factor for hypoxic chondrocytes in vivo. We have shown that mesenchymal condensations of the limb bud are also hypoxic, and lack of HIF-1 in limb bud mesenchyme delays differentiation of mesenchymal cells into chondrocytes in vivo. In this grant, we propose to identify the molecular mechanisms that mediate the role of HIF-1 as a survival and differentiation factor in cartilage in vivo. Along these lines, we have reported that viable chondrocytes at the periphery of HIF-1 null growth plates and HIF-1 null mesenchymal condensations of the limb bud are considerably more hypoxic than controls. Moreover, we have provided genetic evidence that the extreme hypoxia of HIF-1 null cells is not the consequence of reduced availability of O2 to the growth plate. Therefore, we hypothesized it had to be the consequence of increased O2 consumption. Our hypothesis is in line with the well- documented ability of HIF-1 to impair mitochondrial respiration in vitro. Based on these findings, we now propose that a key function of HIF-1 is to reduce O2 consumption in cells that are already hypoxic because of limited availability of O2, in order to prevent them from becoming virtually anoxic, a status that is not compatible with cell survival and differentiation. Specifically, we hypothesize that HIF-1 is essential for survival of hypoxic chondrocytes and for timely differentiation of hypoxic mesenchymal cells into chondrocytes by negatively regulating mitochondrial respiration, and thus mitochondrial O2 consumption. We will test our hypothesis by inhibiting mitochondrial respiration in HIF-1 null chondrocytes (Specific Aim I) and in HIF-1 null mesenchymal cells of the limb bud (Specific Aim II) in vivo and in vitro. Moreover, we will establish whether HIF-1 lowers O2 consumption in chondrocytes in vitro (Specific Aim III). Our findings may lead to a paradigm shift if we determine that, differently fro what has been reported in the context of well-oxygenated tissues, impairment of mitochondrial respiration is an indispensable requirement for survival and for early differentiation stages of hypoxic chondrocytes.
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{
"pile_set_name": "NIH ExPorter"
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Long-term potentiation (LTP) and long-term depression (LTD) are lasting, activity-dependent changes in synaptic strength that have been proposed to underlie learning and memory. These forms of plasticity are dependent on NMDA receptor activation, and recent studies have suggested that the subunit composition of these NMDA receptors (specifically whether the receptor complex contains the NR2A versus the NR2B subunit) plays an important role in whether their activation leads to LTP or LTD. However, these studies are primarily pharmacological, have provided conflicting results, and their conclusions remain somewhat controversial. The first aim of this proposal is to use pharmacological antagonists to determine whether the roles of the NR2A and NR2B in long-term plasticity change over development. The second aim is to investigate the roles of these subunits as molecular scaffolds in addition to their roles as receptor-channels. This will be done by evaluating the impact on LTP and LTD of decreasing NR2A and NR2B expression using RNAi. The third aim is to increase NR2A expression in the visual cortex of light-deprived rats to study the role of NR2A in experience-dependent plasticity. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The structure and mechanism of action of two membrane-bound ion-transporting systems, the (Na ion plus K ion)-ATPase from kidney and the Ca2 ion-ATPase from sarcoplasmic reticulum, are being characterized using nuclear magnetic resonance, electron paramagnetic resonance and kinetic methods. Nm (II) EPR studies of the (Na ion plus K ion)-ATPase and Gd (III) EPR studies of the bound metal. Water proton NMR studies provide a means of characterizing complexes of the enzymes, metals and substrates of substrate analogues, while FT-NMR methods permit determination of the conformations of enzyme-bound substrates. These structural studies will provide unique and valuable information on the active site structures of these enzymes and the mechanisms of ATP hydrolysis and ion transport.
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{
"pile_set_name": "NIH ExPorter"
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TR&D 1 Project Summary. Surface induced dissociation (SID) has emerged as an effective tool for probing the topology of protein complexes in the field of native mass spectrometry (nMS). SID has the unique advantage of efficiently cleaving the interfaces of noncovalent protein complexes to yield monomer subunits as well as small order oligomers. Additionally, SID products typically retain compact/highly structured conformations such that ion mobility measurements coupled with SID can be used to gain insights into protein subunit structure through collision cross section (CCS) measurements. This TR&D aims to improve SID technology across multiple mass analysis platforms that are used in the nMS field (namely quadrupole-time-of- flight (QTOF), OrbitrapTM, and Fourier transform ion cyclotron resonance (FTICR) platforms). Current SID devices span 3-4 cm of an instrument?s ion path with at least ten DC electrodes. These electrode voltages can be tuned to direct ions toward an off-axis surface (relative to the ion path within an instrument for the activating ion-surface collision and subsequent collection of product ions back onto the original ion trajectory axis. Lenses may also be tuned to guide ions through the device without activation, allowing for standard MS experiments to be conducted (so-called ?flythrough? mode). Aim 1 of this TR&D is to simplify the tuning of SID experiments while retaining or improving ion transmission efficiencies in both SID and ?fly-through? experiments. A new SID device containing an on-axis collision surface and ion carpet will be developed to require fewer electrodes than current SID devices, thereby simplifying tuning requirements for SID. The radial-focusing nature of the ion carpet array is expected to assist ion transmission efficiencies. The device will be implemented in a prototype higher resolution QTOF. Aim 2 will adopt advances made in Aim 1 to improve current SID performance within Orbitrap platforms for high resolution nMS experiments. There is also an SID acceleration voltage limit of roughly 70 V for current Orbitrap configurations. Collaboration with Thermo Fisher will facilitate progress by providing the schematic information necessary to incorporate independent power supplies into electronic boards on the QExactive+ EMR platform at OSU and QExactive HF UHMR platform proposed for purchase. This collaboration will also facilitate adapting technology developed from Aim 1 into the ion optics within Orbitrap Exactive platforms. Aim 3 focuses on improving SID technology in an FTICR platform. Currently, this platform has a maximum quadrupole mass isolation limit of m/z 6000, which is too low for selection of larger protein complexes. There is currently a diminished performance in the collection of collision induced dissociation fragments due to the original collision cell space being split between an SID device and truncated collision cell. Both of these shortcomings will be addressed through instrument redesign in collaboration with the vendor (Bruker) as described in Aim 3.
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{
"pile_set_name": "NIH ExPorter"
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Recent anatomic and physiologic suggest that the primate ventral thalamic nucleus (human ventrocaudal nucleus - Vc) which receives innocuous mechanoreceptive input is also involved in nociception. Studies in animal models of chronic pain suggest that this same nucleus may be involved in some types of chronic pain. We propose to examine the role of human Vc in pain by studying thalamic single unit activity recorded as part of the physiologic localization required during stereotactic procedures for the treatment of pain and movement disorders. Initial studies will attempt to define nociceptive inputs to Vc by examining neuronal responses to application of quantitative somatosensory stimuli. In pain patients, spontaneous and evoked cellular activity will be studied in regions of Vc which represent parts of the body where the patient experiences pain. These studies will attempt to identify patterns of thalamic activity associated with human chronic pain. Finally, patterns of thalamic activity and psychophysical estimates of pain will be studied simultaneously. Patterns of thalamic activity will be interpreted in light of recent advances in thalamic physiology and so may identify intracellular mechanisms involved in thalamic activity related to chronic pain. These studies may lead to the development of new therapeutic strategies for the treatment of chronic pain.
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{
"pile_set_name": "NIH ExPorter"
}
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Cardiac infarction followed by ischemic injury is a major cause of death worldwide. Digitalis, the oldest and best characterized heart failure drug, protects against ischemia-reperfusion injury, and it does so through activation of the mitochondrial ATP-sensitive K+ channel (mitoKATp). The central hypothesis of this project is that ouabain-activated signals are relayed to mitochondria, resulting in mitoKATp opening, increased production of reactive oxygen species (ROS) and activation of survival kinases that protect the heart. Our long-term goal is to uncover the mechanisms that regulate digitalis-induced cardioprotection through activation of the Na,K-ATPase / mitoKATp pathway. During this period, we will focus on the mechanism of signal transmission from the sarcolemmal Na,K-ATPase to mitochondria. Aim 1 will test the hypothesis that digitalis causes formation of functionally active caveolar microdomains (signalosomes) that interact with the mitochondrial outer membrane, leading to mitoKATp opening and inhibition of the mitochondrial permeability transition. Aim 2 will investigate signalosome recycling and the persistence of signalosome function. Aim 3 will investigate signalosome assembly and transport using electron microscopy and immunogold labeling. Aim 4 will investigate key aspects of digitalis protection and signaling in the rabbit heart model, whose cardiac function and digitalis pharmacology are closer to those in human heart. A variety of experimental approaches will be used. Signalosomes will be purified from perfused hearts and tested for functional activity on mitochondria isolated from untreated hearts. Results of these physiological studies will be supported by parallel studies of infarct size on perfused hearts. Purified signalosomes will also be subjected to biochemical and structural analyses to characterize their composition and origin.
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{
"pile_set_name": "NIH ExPorter"
}
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DESCRIPTION: (Applicant's Abstract) There is an increasingly large population receiving opiates on a chronic basis, either for treatment of opiate dependence or for treatment of chronic pain. Managing pain in opiate maintained individuals is a challenging problem. The development of opioid tolerance and dependence may result in altered pain sensitivity and subsequent response to additional opioids. Many clinicians believe addicts cannot benefit from further opiate administration because they are tolerant to the analgesic effects. Others believe that if an addict is maintained on an opiate agonist, that medication provides adequate analgesia for acute pain. Patients are reluctant to give accurate histories because clinicians often withhold opioid analgesics from patients with a history of drug abuse. Those who do complain are viewed suspiciously and their complaints ignored. Currently there are few guidelines to assist clinicians and those that do exist are based on experience and consensus rather than evidence-based research. This study proposes to examine: 1) how patients on opiate maintenance (either buprenorphine or methadone) experience pain (tolerance and threshold) and how they are different from non-opiate dependent individuals, 2) how patients maintained on methadone and buprenorphine differ from normal controls in response to added opiate and non-opiate analgesics and 3) how therapeutic plasma concentration levels of morphine for analgesia differ in methadone and buprenorphine maintained patients compared to normal controls. The results of this proposed study will help develop systematic guidelines for clinicians in the management of pain in opiate maintained individuals.
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{
"pile_set_name": "NIH ExPorter"
}
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The Veterinary Resources Program, National Center for Research Resources, has established a Watanabe hyperlipidemic (WHHL) rabbit breeding colony as a resource for investigators. Watanabe et al reported that there are two types of WHHL rabbits--those with early development and high incidence of coronary atherosclerosis (type 1) and those with late development and low incidence of coronary atherosclerosis (type 2). It has also been shown that either type of rabbit can be selectively bred to produce progeny exhibiting the phenotype of interest. Recently, it was reported that these two types of WHHL rabbits can be differentiated by their lipoprotein phenotypes. Rabbits were fasted overnight and blood was collected in EDTA vacutainers the next morning. Agarose gel electrophoresis was used to identify the lipoprotein phenotypes. Electrophoretic analysis of 14 rabbits from the NCRR WHHL rabbit colony showed that there are two lipoprotein phenotypes: one with an increased density of beta-lipoprotein and another with a beta-doublet. These phenotypes are similar to those reported for the two types of WHHL rabbits. Characterization of the NCRR WHHL rabbits as to phenotype will be of value to investigators who want to use them for their studies.
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{
"pile_set_name": "NIH ExPorter"
}
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UCI ADRC Overall Project Summary/Abstract The central theme of the University of California, Irvine Alzheimer's Disease Research Center (UCI ADRC) is to identify, quantify, and validate factors that influence the risk of Alzheimer's disease (AD) across the lifespan. Given its diverse and multifactorial nature, it is critical to understand the etiology and progression of this insidious brain disorder at multiple dimensions, including clinically and neuropathologically, and to employ novel scientific approaches so that we can advance knowledge and better diagnose, prevent and cure the disease. The UCI ADRC has a strong history of integrating basic, clinical, and translational science to gain insights into AD pathogenesis and to help meet some of the goals of the National Alzheimer's Project Act. First, our Center has a distinguished record of studying the disease in diverse patient cohorts: Uniform Data Set (UDS) Cohort, which includes cognitively normal individuals and individuals with mild cognitive impairment (prodromal AD); adults with Down syndrome, representing the largest genetically at-risk population for AD; and 90+, which show great disparity between AD pathology and cognitive loss and are among the oldest group of individuals to develop AD. Together, these cohorts enable interrogation of important novel questions related to age, susceptibility and resilience, using innovative outcomes ranging from in vivo biomarker characterization to modern pathological outcomes such as post-mortem imaging and microglial staining. Second, the UCI ADRC has a history of innovation that has helped advance the field; the development of the nation's first induced pluripotent stem cell bank for AD as part of the ADRC network and our recently established Consent-to- Contact recruitment registry are recent examples of this innovation. Third, our Center continues to evolve. Given the expertise, productivity, and resources devoted the study of special populations at UCI, we have initiated two Special Populations Cores, one focused on Down syndrome and one focused on the oldest old (90+). These Cores will interact with the other cores of the ADRC to produce novel data and resources available to ADRC investigators, including through our newly established Biomarker Core. Finally, we have established a new Research and Education Component in this proposal, which harnesses the long-standing passionate commitment to training the next generation of clinicians and scientists at UCI. In sum, the UCI ADRC brings energetic and innovative multi-dimensional and multidisciplinary approaches toward addressing the national epidemic of AD.
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{
"pile_set_name": "NIH ExPorter"
}
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Atypical antipsychotic drugs (AAPs) are indicated in the treatment for schizophrenia, bipolar disorder, psychotic depression and other psychiatric disorders. Their drawback is drug-induced metabolic derangements including weight gain, hyperlipidemia, and diabetic risks. These diabetic metabolic symptoms (DiMS) vary widely between drugs and from patient to patient. We propose to develop a novel product termed "Physiotype" to deliver personalized information for each patient on the drug- specific risks among aripiprazole, olanzapine, quetiapine, risperidone, and ziprasidone. The Physiotype consists of a multi-gene ensemble of single nucleotide polymorphisms (SNPs) that, interpreted with a biomathematical algorithm, may explain most of the inter-individual differences in DiMS among the 5 AAPs The proprietary physiogenomics technology and state-of-the-art genotyping laboratories of Genomas will be integrated with the clinical resources of the Institute of Living (Hartford CT) and of the University of Kentucky (Lexington KY), through subcontracts, respectively, to Dr. John Goethe and Dr. Jose de Leon. Our goal in Phase II is to discover SNPs predictive of differences in DiMS side effects between these 5 AAPs and to develop them into predictive diagnostic products for psychiatrists in their practice. We will recruit 200 patients treated by each of the 5 AAPs, characterize their weight and lipid profiles, and obtain their DNA for creation of a clinical registry and DNA repository. We will determine each patient's genotype at 100,000 SNPs covering all ~30,000 genes and also evolutionary conserved regions for a comprehensive, hypothesis-free search for genetic markers of DiMS. In Phase I, the collaborators have already accumulated a registry and DNA repository of 374 AAP-treated patients and their DNA. We have genotyped DNA from olanzapine- and risperidone-treated patients in the repository for an array of 384 SNPs in 222 cardiovascular, metabolic and psychiatric candidate genes and performed physiogenomic predictive modeling. We have discovered novel drug-specific DiMS markers for olanzapine and risperidone including the apolipoprotein E and leptin receptor genes, respectively. We have developed a prototype Physiotype and tested it in an independent psychiatric population. The Physiotype predicted that ~20% of patients have the most weight associated with risperidone and ~80% with olanzapine, which is consistent with known olanzapine average effects, and also pinpoints the greater risperidone-specific risk for many individuals. In Phase III, a prospective randomized trial of all 5 AAPs is planned as part of FDA review of a Physiotype device. The Physiotype will assist psychiatrists to avoid side effects by guiding drug selection for each patient according to innate characteristics unraveled and interpreted directly from the person's own DNA. The proposed program will develop DNA diagnostic products to enhance safety of atypical antipsychotic drugs (AAPs) and improve the medical management of schizophrenia and related disorders leading to better outcomes. As of now, the development of AAP side effects is unpredictable, potentially disabling to the patient, and discourages patient compliance. The products will enable DNA- guided medicine: the determination of which AAP is most suitable and the implementation of clinical safeguards, individualized to each patient, using his/her personal genome. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The Leadership and Administrative Core (LAC) is responsible for strategic planning, organization, administrative operations and evaluation of the OAIC Research and Training program. A special effort is devoted to ensure the cohesion ofthe Center and maintaining an interdisciplinary and translational research focus on the common research theme, which is sarcopenia and prevention of disability. The key LAC tasks are achieved by the Core Leader and three committees: the Executive Committee, the Independent Review Advisory Panel and the External Advisory Committee. The specific functions ofthe Leadership Core are: - To provide overall scientific leadership and direction for the OAIC research and training program. - To render administrative and budgetary support for the program - To coordinate the functions of the OAIC cores and projects in order to facilitate communication and foster translation behween basic and clinical research and ensure access of investigators to core resources - To assure the coordination of OAIC resources and functions with other research and training grants and institutional resources - To communicate with other OAlCs and the NIA, and to foster collaborations with other OAlCs - To facilitate compliance with guidelines and regulations regarding fiscal policy, human subjects, and animal care and use - To set productivity benchmarks and monitor progress of individual projects and progress of Junior Scholars (this aim is shared with the RCDC), and deal with inadequate progress - To promote quality, productivity and efficiency (timeliness) in all OAIC activities - To arrange the annual meeting of the OAIC External Advisory Committee - To maintain the OAIC web-based tracking and monitoring system to facilitate communication - To promote the use of uniform assessment batteries in OAIC supported clinical research studies to optimize the use of OAIC resources - To maintain the OAIC website and publish the OAIC newsletter Taken together, the LAC provides support for planning, organizational, evaluation, and administrative activities relating to the other Cores and to the OAIC as a whole. The LAC is responsible for monitoring, stimulating, sustaining, evaluating, and reporting progress toward the overall goals ofthe OAIC.
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{
"pile_set_name": "NIH ExPorter"
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To look at possible effects cocaine has on the brain in people of different sexes and ethnicity who are infected with HIV.
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{
"pile_set_name": "NIH ExPorter"
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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are using NCMIR to study Parkinson's disease (PD) and related neurodegenerative disorders. We hypothesize that alpha-synuclein (a-syn) causes neurodegeneration by inducing chronic mitochondrial fission through abnormal interaction with mitochondrial fission and fusion GTPases. Our current understanding of PD suggests that a-syn-mediated disruption of mitochondrial fusion/fission events may be a mechanism of neuronal toxicity. Nevertheless, this idea has never been tested. Mitochondrial fusion is thought to provide protection by facilitating the mixing mitochondrial contents, such as metabolites and mtDNA. Several observations of a-syn are in agreement with our hypothesis. First, brain tissue of PD patients and a-syn transgenic mice exhibit abnormal mitochondrial ultrastructure, respiratory complex I inhibition, and increased free radical production. In addition, overexpression of human a-syn in transgenic mice leads to dopaminergic synaptic loss. Furthermore, a-syn modulates membrane composition and forms pores similar to bacterial toxins. Intriguingly, Bax, a pro-cell death molecule of the Bcl-2 family that associates with the mitochondrial outer membrane, also has pore-forming activity and a structure similar to bacterial toxins. Recent publications show that Bax is a component of mitochondrial fission/fusion complexes in dying cells. Finally, a-syn cooperates in SNARE complex assembly that regulates exocytosis and membrane fusion. Mitofusins (Mfns) may mediate mitochondrial membrane fusion by a SNARE-like mechanism. Thus, it is conceivable that a-syn may interact with Mfns, similar to SNAREs. Dynamin-related GTPases regulate mitochondrial fission and fusion, important cellular processes that neurons must balance to maintain normal mitochondrial and synaptic activity. Dynamin-related protein 1 (Drp1) directs mitochondrial fission (division) and Mfn 1, 2 regulate mitochondrial fusion. Previous research has linked excessive mitochondrial fission to neurodegeneration and induction of mitochondrial fusion to the prevention of neuronal cell death. Bax co-localizes with Drp1 in fission complexes on mitochondria and regulates apoptotic mitochondrial fission. We believe that a-syn, like Bax, may interact with the Mfns or Drp1. Supporting this hypothesis is the observation that a-syn forms clusters on mitochondria that may constitute future or past fission sites. Mutant or abnormally folded a-syn may inhibit Mfn GTPase function and prevent mitochondrial fusion. Alternatively, a-syn may bind and activate Drp1, thereby promoting excessive fission. The consequences of such interactions might include the breakdown of long mitochondrial filaments into multiple, isolated fragments, chronic respiratory inhibition, increased free radical levels, impaired calcium buffering, energy decline, accumulation and manifestation of mtDNA mutations, and ultrastructural defects of mitochondria. Mitochondrial dysfunction, energy crisis, and oxidative stress would then cause loss of synapses, protein aggregation, and neuronal dysfunction and loss.
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{
"pile_set_name": "NIH ExPorter"
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This proposal will allow the University of Arkansas for Medical Sciences (UAMS) to obtain a state-of-the-art Orbitrap Fusion Tribrid mass spectrometer to be operated through the university's existing Proteomics Core facility. In the short term, this instrument would support seven recently funded National Institutes of Health grants with proteomic aims by providing dedicated access to the technology best suited to support them. These seven grants will advance a range of biomedical research fields, including gene regulation, development, cancer biomarker, and cancer treatment-related projects. A variety of proteomic approaches will be applied, including high-throughput protein identification, mapping of post-translational modifications, and quantification by isotopic labeling. Over the longer term, the acquisition of this new instrument will allow a strong technology facility in a traditionally underfunded state to keep pace with rapid advancements in the field of mass spectrometry.
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{
"pile_set_name": "NIH ExPorter"
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The primary objective of the activities proposed in this application is to provide optical coherence tomography (OCT) Reading Center support to the Comparison of Age Related Macular Degeneration Treatment trials (CATT). The long term goals of this project, that will be accomplished through conduct of specific CATT studies are 1) to identify by OCT those eyes that are most likely to benefit from preventative strategies or treatments designed to prevent vision loss, or improve visual acuity in eyes with AMD, 2) to determine by OCT when treatment is effective or re-treatment is required and 3) to standardize OCT protocols and grading methods in eyes with AMD. The primary objective will be accomplished through the following specific aims: Specific Aim 1: Establish the infrastructure to process OCT images that will be submitted by CATT clinical centers to the CATT OCT Reading Center. Specific Aim 2: Collaborate with the Coordinating Center to establish and implement OCT certification and training programs for CATT clinical study investigators, clinical coordinators and OCT technicians in preparation for CATT. Specific Aim 3: Provide reliable, timely, reproducible, and systematic grading of submitted OCT materials. To achieve these aims, The OCT Reading Center will use a custom, stand-alone web-based system that we have developed to establish and implement secure methods for electronic OCT data submission from the CATT clinical study sites to the CATT OCT Reading Center, and to store and back-up submitted data. We will establish secure methods to transmit data from the OCT Reading Center to the Coordinating Center. The Reading Center will also implement materials and methods for grader, photographer, clinical coordinator and ophthalmologist certification and training, and support study goals as an image resource for recruitment, Study Group meetings, presentations and publications. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Breathing is a remarkable behavior that mediates gas exchange to support metabolism and regulate pH. A reliable and robust rhythm is essential for breathing movements in mammals. Failure to maintain a normal breathing rhythm in humans suffering from sleep apnea, apnea of prematurity, congenital central hypoventilation syndrome, hyperventilation syndrome, Rett syndrome, and perhaps sudden infant death syndrome, leads to serious adverse health consequences, even death. Neurodegenerative diseases, such as Parkinson's disease, multiple systems atrophy and amyotrophic lateral sclerosis, are associated with sleep disordered breathing that we hypothesize results from the loss of neurons in brainstem areas controlling respiration. If breathing is to be understood in normal and in pathological conditions, the site(s) and mechanisms for respiratory rhythmogenesis must be revealed. A brainstem region critical for normal breathing is the preBtzinger Complex (preBtC). Studies of respiratory rhyth- mogenesis have focused primarily on preBtC neuron cellular properties or regional interactions between respiratory-related brainstem centers, largely ignoring the (micro)circuit structure within each of these essential areas, including the preBtC. While informative, these prior studies have failed to reveal validated mechanisms underlying rhythmogenesis. We break from current paradigms and propose a completely novel hypothesis: Respiratory rhythm is generated by preBtC burstlets, low level synchronous multineuronal activity, which normally trigger the high amplitude bursts necessary to produce respiratory motor output. To test this burstlet hypothesis, we will determine key cellular, synaptic and network mechanisms underlying burstlets and bursts using a novel protocol in AIM 1. To examine how the (micro)circuit structure contributes to rhythmogenesis, we take advantage of recent technological advances that allow spatiotemporally precise perturbation of neural micro(circuits) in AIM 2. We utilize an advanced optical technique, holographic photolysis, to map connections within the preBtC and determine network interactions underlying respiratory rhythmogenesis. Breathing is a rhythmic behavior of fundamental importance. By determining the mechanisms underlying the generation of respiratory rhythm, we will significantly improve our knowledge of neural control over the entire breathing cycle. These studies should make fundamental contributions to our understanding of breathing in humans in health and disease.
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{
"pile_set_name": "NIH ExPorter"
}
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In humans, apolipoprotein E (apoE) deficiency causes hyperlipidemia and vascular disease. We have recently determined that harbor seals and sea lions do not have detectable levels of plasma apoE, yet they do not develop hyperlipidemia or remnant lipoproteins typically seen in apoE deficient humans. The diet of seals and sea lions is rich in omega-3 fatty acids and may suppress VLDL synthesis, thus "masking" the impact of the apoE deficiency. The objectives of this study are 1) to determine the basis for the apoE deficiency in harbor seals and sea lions, 2) to determine the mechanism by which seals efficiently metabolize triglyceride-rich lipoproteins, and 3) to determine the role of dietary omega-3 fatty in maintaining normolipemia. First, we will determine which apoprotein(s) are responsible for receptor-mediated uptake of seal lipoproteins using both cell culture techniques and in vivo turnover studies. Second, experiments will be conducted to determine whether harbor seals and sea lions have the apoE gene and apoE mRNA using Southern and Northern blotting techniques. respectively. Third, we will assess the role of the fish-diet by feeding seals an alternative isocaloric diet high in cholesterol and saturated fat. These studies of lipoprotein metabolism are unique in that harbor seals and sea lions are animal models in which the full impact of apoE deficiency can be studied. Because their diets are composed entirely of fish, they offer a unique opportunity to determine the maximal effect of such a diet, especially in the face of apoE deficiency. Thus these studies are highly relevant to the goal of understanding the relationship of diet and lipoprotein metabolism to the causation of atherosclerosis in man.
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{
"pile_set_name": "NIH ExPorter"
}
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Early age at menarche is associated with an increased risk of breast cancer. Among determinants of age at menarche, diet appears to be of great importance because malnutrition can delay the adolescent growth spurt and menarche, overnutrition as indicated by obesity is associated with decreased age at menarche, and long-term changes in diet appear to parallel the secular trend to earlier menarche that has occurred since 1850. The objective of this 5-year study is to determine the association of specific dietary factors with age at menarche and with anthropometric measurements which indicate growth. The methodology will involve the collection of dietary data and anthropometric measurements and the determination of the age at menarche prospectively in 9-13 year old girls. The dietary data will consist of 24-hr recalls and food frequency questionnaires administered to the subjects at school at unannounced periods at six month intervals; the anthropometric measurements will be made at the same time. The occurrence of menarche will be indicated by the mother's response to a questionnaire administered at six month intervals which will also indicate the girl's food frequency usage. Dietary variation in the study population is achieved by including a school population of Seventh-Day Adventist girls to represent nonusers or light users of meat, poultry and fish and a group of girls from public school to represent the standard American diet with heavy use of meat, poultry and fish. The study will provide specific data for dietary factors which may be predictors for age at menarche such as gram estimates of the average weekday intake of specific nutrients such as total fat, animal protein, etc.; frequency of use of specific food items such as milk, eggs, etc., as reported by the subject and by her mother, as well as other data reported by the mother. The analysis of this data will permit the determination of the degree of association of specific food items and/or specific nutrients with age at menarche.
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{
"pile_set_name": "NIH ExPorter"
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The results summarized below were obtained in collaborative studies: To examine whether the recently discovered fast depolarization-induced charge movement in the M2R is responsible for M2R-mediated control of acetylcholine release, M2R KO mice were used as a tool. Inhibition of the M2R charge movement in Xenopus oocytes correlated well with inhibition of acetylcholine release at the mouse neuromuscular junction. These results suggest that, in addition to calcium influx, charge movement in GPCRs is also necessary for transmitter release control. (Kupchik YM, et al. A novel fast mechanism for GPCR-mediated signal transduction - control of neurotransmitter release. J. Cell Biol. 192, 137-151, 2011) The involvement of M4Rs in the reinforcing effects of cocaine were examined in chronic intravenous cocaine self-administration experiments. For this analysis, wild-type and M4R KO mice were used. Behavioral studies showed that M4Rs play an important role in regulating the dopaminergic reward circuitry. Thus, central M4Rs may serve as a new target for the treatment of drug addiction. (Schmidt LS, et al. Increased cocaine self-administration in M4 muscarinic acetylcholine receptor knockout mice. Psychopharmacology (Berl) 216, 367378, 2011) Different muscarinic receptor KO mice were used as tools to explore which muscarinic receptor subtype mediates vasodilation of small arteries. This analysis showed that M3Rs mediate cholinergic vasodilation in cutaneous, skeletal muscle, and renal interlobar arteries. These findings suggest that M3Rs may represent a novel target for the treatment of hypertension or local disturbances in blood flow. (Gericke A, et al.. Role of M1, M3, and M5 muscarinic acetylcholine receptors in cholinergic dilation of small arteries studied with gene-targeted mice. Am. J. Physiol. Heart Circ. Physiol. 300, H1602- H1608, 2011) To investigate the potential role of the M4R in catalepsy induced by antipsychotics (haloperidol and risperidone) as well as the anti-cataleptic effects of the non-selective anticholinergic drug scopolamine, M4R KO mice and their wild-type littermates served as tools. Behavioral studies strongly suggested that central M4Rs play a key role in mediating the motor side effects associated with the use of antipsychotic drugs. The experimental data also supported the concept that M4Rs represent an attractive target for the pharmacological treatment of antipsychotic-induced as well as idiopathic parkinsonism. (Fink-Jensen A, et al. Antipsychotic-induced catalepsy is attenuated in mice lacking the M4 muscarinic acetylcholine receptor. Eur. J. Pharmacol. 656, 39-44, 2011) Using mutant mice that lack M4Rs selectively in D1 dopamine receptor-expressing neurons (D1-M4-KO mice), we investigated the role of this neuronal population in the antipsychotic-like effects of xanomeline in amphetamine-induced hyperactivity and apomorphine-induced climbing. Interestingly, the antipsychotic-like effects of xanomeline in both models were almost completely abolished in D1-M4-KO mice, suggesting that M4Rs co-localized with D1 dopamine receptors are critically involved in mediating the antipsychotic-like effects of xanomeline. These findings support the concept that M4R agonists may prove useful for the treatment of psychosis. (Dencker D, et al. Involvement of a subpopulation of neuronal M4 muscarinic acetylcholine receptors in the antipsychotic-like effects of the M1/M4 preferring muscarinic receptor agonist xanomeline. J. Neurosci. 31, 5905-5908, 2011) Studies with wild-type and M1R KO mice demonstrated that M1R dysfunction may contribute to mood disorders and that M1Rs and the downstream ERK pathway may serve as potential therapeutic targets for alleviating manic symptoms such as psychomotor hyperactivity. (Creson TK, et al. Lithium treatment attenuates muscarinic M1 receptor dysfunction. Bipolar Disord. 13, 238-249, 2011) Previous work has demonstrated that the M3R is over-expressed in colon cancer. Studies with Apc(min/+) mice lacking M3Rs showed that the absence of M3R signaling was associated with a striking reduction in colon tumor number and volume. Similar findings were obtained with scopolamine butylbromide, a non-subtype-selective muscarinic receptor antagonist. These data indicate that M3Rs play a critical role in the pathophysiology of colon cancer and that M3R antagonists may prove useful for the treatment of intestinal neoplasia. (Raufman JP, et al. Muscarinic receptor subtype-3 gene ablation and scopolamine butylbromide treatment attenuate small intestinal neoplasia in Apcmin/+ mice. Carcinogenesis, 2011, Aug 1;Epub ahead of print)
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{
"pile_set_name": "NIH ExPorter"
}
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Regulation of mRNA export, both viral and cellular, is a complex process that is only beginning to be understood, predominantly through studies of protein import/export, and viral RNA export. During the course of HSV- 1 infection, the ICP27 protein post-transcriptionally regulates viral late gene expression and is essential for viral growth. ICP27 shuttles between the nucleus and cytoplasm, and nucleocytoplasmic shuttling correlates with the cytoplasmic accumulation of viral late mRNA. Collectively, the data are consistent with a model in which ICP27 is a viral RNA export protein; however, the precise mechanism by which ICP27 regulates cytoplasmic late mRNA accumulation remains unclear. Therefore, the objective of this study is to elucidate the details of ICP27-dependent RNA export including (1) identification of particular viral RNAs bound by the ICP27 protein, (2) identification of domains within the ICP27 protein required for interactions with RNA, and (3) determination of cellular proteins that physically associate with ICP27. A comprehensive understanding of ICP27 function may lead to the development of novel antiviral therapies, and is certain to yield important insights into the fundamental aspects of nuclear transport.
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{
"pile_set_name": "NIH ExPorter"
}
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Transient, but severe global ischemia, observed in patients during cardiac arrest and cardiac surgery or induced experimentally in animals, induces selective and delayed neurodegeneration. Pyramidal cells in CA1 are the most sensitive; CA3 and granule cells of the dentate gyrus (DG) are resistant to ischemic damage, and GABAergic interneurons in CA1 also survive. The molecular mechanisms underlying this pattern of neuronal death are not well understood. The proposed research aims to study the role of gap junctions during the several days of "maturation" of neuronal injury after global ischemia. Recent findings from this laboratory indicate that global ischemia triggers a selective upregulation of Cx36 (and Cx32) protein expression in GABAergic interneurons of the vulnerable CA1 at times prior to the onset of neuronal death, consistent with a role in the survival of these neurons. Moreover, CA1 neurons in Cx32 (Y/-) mice exhibit enhanced vulnerability to global ischemia-induced neuronal death. These data suggest that increased inhibition of pyramidal cells through synchronization of inhibitory interneurons may be neuroprotective. Gap junctions between astrocytes are also thought to have a role in post-ischemic neuronal death. Dying cells can kill resistant neighboring glial cells via glial "fratricide" (bystander death) and thereby propagate injury to neighboring regions. On the other hand, gap junctional coupling of astrocytes mediates metabolic cooperation among them and attenuates neuronal death in models of oxidative stress. The underlying hypothesis of this proposal is that gap junctions play important roles in determining neuronal death and survival following global ischemia. The research plan for the next five years focuses on changes in the abundance, distribution and molecular and biophysical properties of brain gap junctions following neurological insult. Specific Aims are 1. Characterize ischemia-induced alterations in connexin expression and gap junction properties in the vulnerable CA1 and resistant CA3 and dentate gyrus of rats and mice. Experiments will examine global ischemia-induced changes in coupling of inhibitory interneurons and expression of connexin proteins by immunocytochemistry and Western blotting and of connexin mRNAs by in situ hybridization and. Experiments will determine the effects of acute knockdown of specific connexins by antisense oligonucleotides on neuronal vulnerability and will examine neuronal vulnerability in Cx32(Y/-) mice, Cx36(-/-) mice and mice deficient in astrocyte Cx43. 2. Examine effects of oxygen/glucose deprivation on hippocampal slice cultures by immunocytochemistry, in situ hybridization and electrophysiological methods. To examine ischemia-induced changes in gap junction properties in acute slices and organotypic hippocampal slice cultures by electrophysiological methods and image analysis. The proposed research is expected to impact on the development of new treatment strategies for intervention in global ischemia, a debilitating and often fatal trauma associated with cardiac arrest in humans. Moreover, this study has important implications for research on other neurodegenerative disorders including focal ischemia, epilepsy, AIDS encephalopathy, and Alzheimer's disease.
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{
"pile_set_name": "NIH ExPorter"
}
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Each year in the U.S. approximately 220,000 infants are born to mothers who used illicit drugs during pregnancy. Others report between 4-10% of all pregnant women have used cocaine. There is growing evidence that in utero drug exposure (IUDE) to cocaine constitutes a risk leading to long-term alterations of brain functioning. Long term follow-up studies of opiate exposed children indicate that although intellectual deficits may be mild or nonexistent, behavioral abnormalities may persists into the school years. One major difficulty in studying opiate exposed children is determining which factors (environmental or direct effects or opiate exposure) result in the poor outcomes. Foster care placement for IUDE children is high. Almost half (43%) of opiate exposed children were removed from the biological parent in one longitudinal study. Cost burden of drug exposed infants are a direct result of the high incidence of prematurity, low birthweight and neonatal drug withdrawal symptoms. Neonatal drug withdrawal occurs in 42-68% in heroin addicted and 63-85% of methadone addicted newborns contributing to an increased length of stay. Other cost burdens are difficult to measure including the effects on the development and performance of the individual over a lifetime Early intervention programs including home nurse interventions are thought to enhance parenting to enrich the development in biologically and environmentally challenged infants. The goal of this continuation study is to determine if a Pediatric Nurse Specialist (PNS) home intervention will be associated with sustained effects of improved child health and demonstrate cost effectiveness in the 204 enrolled in utero drug exposed infants up to age 5 years. Additionally, the goal is to compare the intervention children to a group of non-drug-exposed control infants of comparable environment/ socioeconomic status. This ongoing study is a randomized, two-group, repeated measures controlled clinical trial to test the effectiveness of a home-based PNS intervention. Infants were randomized into the PNS intervention (N=100) or control (N=100) groups. All 204 IUDE infants were born at the Johns Hopkins Hospital and had documented IUDE based on maternal drug history by review of prenatal records and maternal urine toxicology screens at delivery and/or infant urine toxicology screens obtained within 24 hours of birth.
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{
"pile_set_name": "NIH ExPorter"
}
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This application seeks support for a project of research directed toward better understanding of the molecular mechanisms and cellular functions of ion transport across human red blood cell membranes. The overall goal of this grant project is the development and use of patch clamp techniques to answer four previously unanswered questions about the structure and functions of human red blood cell membranes. 1. Are there cryptic voltage-dependent ion channels in human red cells? If so, what are their single channel characteristics? 2. Are the kinetics of ionophore-mediated transport the same in cell membranes as in artificial planar lipid bilayers? 3. What is the net surface charge density on the cytoplasmic and external surfaces of the red cell membrane? What is the dipole potential difference that is experienced by carrier-transported ions? 4. What are the kinetic characteristics of the directly measured chloride conductance of human red cells? Can the kinetics of rapid anion exchange and low anion conductance be reconciled in a common molecular mechanism, band 3? Additionally, we will continue to investigate the mechanisms of co-transport and test the hypothesis that the ion pair between sodium and carbonate is a monovalent anion substrate for the anion exchange mechanism. Experiments are designed to answer questions relevant to these problems. They involve observations of net and tracer fluxes of inorganic ions and electrical measurements of patch clamped native and ionophore-treated human red cells in media of different ion compositions and different total ionic strengths.
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{
"pile_set_name": "NIH ExPorter"
}
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Participation in the Southwest Oncology Group (SWOG) has been a major part of the clinical investigative effort of the section of Hematology- Oncology in the Department of Medicine at the University of Oklahoma Health Sciences Center (OUHSC). This effort results in the enrollment of a consistent number of eligible and evaluable patients into SWOG trials. SWOG protocols at the University of Oklahoma provide the major opportunity for Oklahoma residents to participate in large scale, well designed clinical trials. Patients are accrued via both the parent institution and affiliated CGOPs. Most patients in the state, including a large number of Native Americans and rural Oklahomans, are treated by physicians who are part of this network. Annual accrual for the entire network has been ninety-seven patients. We have been working to increase our involvement in the scholarly aspects of clinical trials. At the member institution, we have focused our efforts in five areas including breast cancer, head and neck cancer, genitourinary cancer, lung cancer, and hematological malignancies. This effort has resulted in the establishment of multidisciplinary programs in these areas which has helped to facilitate identification and enrollment of patients on SWOG protocols. In addition, it has allowed for the development of in-house protocols which might serve as pilot studies for future SWOG trials.
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{
"pile_set_name": "NIH ExPorter"
}
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Project Summary This Phase I SBIR project is designed to develop new biodegradable hydrophilic coatings for ureteral stents. Ureteral stents are widely used to maintain drainage of the upper urinary tract and instrumental in management of stone disease, as well as in cases of other ureteral obstruction such as injury, fibrosis or malignancies. However, unlike cardiovascular stents, ureteral stents are recommended for use 3 months or less due to encrustation and biofilm formation. Over 75% of ureteral stents are encrusted at this 3 month time period, making removal difficult and causing patients pain. The best commercial coatings reduce encrustation thickness by 50% and extent of encrustion by only 22% over uncoated ureteral stents. This project focuses on novel coatings which release agents designed to minimize encrustation. In Phase I, two different agents were shown to reduce encrustation 65% over 4 weeks in vitro. In Phase II, one coating will be optimized for full scale use on ureteral stents and implanted in a 30 day GLP porcine study to prepare for FDA submission.
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{
"pile_set_name": "NIH ExPorter"
}
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Summary of Work: The study of oxidative stress has attracted considerable interest and has been the focus of much research in recent years. Cumulative oxidative damage to tissues has been implicated in a number of disease states, e.g. the aging process, cancer, and ischemia reperfusion. The study of oxidative stress in the mitochondria has shown that hydrogen peroxide is produced via the incomplete reduction of oxygen during oxidative phosphorylation. Hydrogen peroxide levels are kept relatively low under normal physiological conditions. Under certain conditions, such as inflammation, excessive amounts of hydrogen peroxide are produced. The production of excess hydrogen peroxide is thought to precede several occurrences, such as lipid peroxidation, DNA and/or protein damage, and glutathione depletion, that are characteristic of oxidative stress. We have probed the role of cytochrome C free-radicals in oxidative stress by studying the anaerobic reaction between horse heart cytochrome C (HHCC) and hydrogen peroxide using the spin-trapping reagent 3,5-dibromo-4-nitrosobenzenesulfonic acid (DBNBS). The MS of the reaction product clearly showed the presence of at least four distinct DBNBS adducts. Enzymatic digest followed by LC/ESI/MS showed an ion due to the DBNBS adduct with Tyr74 of the HHCC which is on the surface of the protein. It is thought that the heme is initially oxidized and then the radical site is transferred intramolecularly to the tyrosine. We are currently identifying other sites of radical formation based on the MS/MS analysis of the proteolytic digest of the DBNBS spin-trapped protein. In parallel experiments we are also investigating the products using MNP as a spin-trap. Investigation of cyanyl radical adducts of cytochrome c oxidase. Cyanide is a frequently used inhibitor of mitochondrial respiration. It has been widely accepted that cyanide functions as a bridge between Heme a3 and CuB. We have observed using ESR that cyanide can be oxidized to the cyanyl radical by cytochrome c oxidase [3]. It is possible that this radical can act as a suicide inhibitor by undergoing reaction with the heme to form a covalent adduct. We have investigated this reaction, and have observed that cyanyl radical does form a covalent adduct with heme. MS/MS showed that the cyanide is attached to the porphyrin ring.
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{
"pile_set_name": "NIH ExPorter"
}
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"The elucidation of molecular alterations that occur during human breast cancer development may permit the identification of preventative strategies for women at high risk. Lesions such as atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS) confer 4- and 8-10 fold increases in risk for the development of invasive carcinoma, respectively. We examined a cohort of approximately 100 human biopsy specimens using in situ hybridization to examine trends in mRNA expression levels, with the goal of identifying genes whose expression patterns correlated with increasing risk. Two genes, cyclin D and the RXR form of the retinoic acid receptor superfamily, exhibited quantitatively increased mRNA levels in DCIS specimens than in normal ductal/lobular units in the margin of the specimen. Overexpression of these two genes therefore correlates with increased risk for development of invasive breast carcinoma. Cyclin D has been transfected into the human immortal MCF-10A breast cell line. Increased anchorage-independent, but not anchorage-dependent growth was observed. Interestingly, the anchorage-independent growth of cyclin D transfectants was several fold to a log more senstive to the inhibitory effects of g-irradiation than the control transfectants. This inhibition was accompanied by increased apoptosis, and could be replicated using the apoptosis inducer Apo-2, but not using TNF-a. We conclude that cyclin D overexpression can produce colonization-competent cells in the breast, and that certain apoptosis inducers may hold promise for eliminating such cells."
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{
"pile_set_name": "NIH ExPorter"
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We propose to screen wild mice for Ia antigens which are serologically related to I-Ab, I-Ad, I-Ak, or I-Ap, and to selectively breed H-2 homozygous mouse lines with wild-derived H-2 haplotypes containing minor variants of these alleles. Preliminary studies have demonstrated that about 1 of every 5 wild mice will carry an allele whose products are serologically similar, but not identical, to one of these inbred line I-A products. A collection of such wild-derived minor variant I-A alleles will be produced by selectively breeding wild mice as semicongenic H-2 homozygous mouse lines. The structural, functional, and serologic properties of the I-A and I-E products of these lines will be thoroughly analyzed. They will be tested serologically with an extensive panel of anti-Ia alloantisera and monoclonal anti-Ia reagents. The alpha and beta subunits of the A and E molecules of serologically-related inbred and wild mice will be compared by tryptic peptide mapping procedures. The minor structural differences which distinguish the wild-derived variant and inbred line I-A products will be localized to specific fragments of the alpha or beta subunits of the A molecule. The functional differences exhibited by these minor I-A variants will be tested with the primed lymphocyte typing assay, and the lymphocyte-stimulating (LS) determinants which are unique to each minor variant allele will be described. The data will be correlated and the following questions will be answered: 1. Do all the variants of a single allele have a common evolutionary origin? 2. Are all minor structural differences in the I-A or I-E molecules equally effective at stimulating T cell proliferative responses? 3. Are structural variations occurring in certain regions or fragments of the molecules more effectively recognized by antibodies, T cell, or both?
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{
"pile_set_name": "NIH ExPorter"
}
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Although many techniques are available for detecting the occurrence of ovulation, a simple screening technique that would detect either anovulation or ovulation disorders is not readily available. It is proposed to study the feasibility of extending the use of "CUE" technology, now used for predicting and confirming ovulation, to screening for and possibly identifying such disorders as anovulation, polycystic ovarian disease, luteal phase defect, and unruptured mature follicles. It is proposed that specific salivary reading (SR) and vaginal reading (VR) signals can be related to particular steps of selection, maturation and ovum release during the ovulatory process. SR and VR data, basal body temperature, ultrasound scans, assays for estradiol, luteinizing hormone, and progesterone will be evaluated from approximately 75 cycles of subjects with ovulation disorders. The method is expected to provide for early screening of infertility patients as well as women at risk for reproductive effects of exposure to hazardous substances in the workplace or the environment.
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{
"pile_set_name": "NIH ExPorter"
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Amyotrophic lateral sclerosis (ALS), which affects about 35,000 Americans, is arguably the most pernicious of the adult-onset neurodegenerative diseases because of its relatively early onset and rapid progression between diagnosis and death. There is no effective treatment for ALS and no progress towards this goal is expected from the private sector, since confirmed "druggable targets" have not emerged and the market is too small to justify an intense research effort to identify such targets. This proposal outlines a first step towards a new class of ALS therapeutics. Aggregation of SOD1 may be pathogenic in familial ALS. Previously, we have demonstrated that aggregation of SOD is coupled to the dissociation of dimeric SOD into monomers. We believe that stabilization of the dimeric form of SOD by small drug-like molecules may be a viable strategy to develop a new class of ALS therapeutics. We propose to screen a library of 100,000 drug-like molecules for (1) Molecules that promote dimer stability in SOD (2) Prevent aggregation of SOD. In order to do this, we will develop fluorescence-based assays that will probe both dimerization and aggregation of SOD. The set of molecules identified from the screen will be subjected to secondary assays which will rely on the direct ability of these compounds to block aggregation. Our long term goal is to use a combination of screening and medicinal chemistry to develop compounds that can be tested in a mice model for ALS. Our ultimate goal is to identify candidates that justify filing for IND status.
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{
"pile_set_name": "NIH ExPorter"
}
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Mutations in the protein kinase BRAF have been found in ~70% of human melanoma. In preliminary studies, I have found that oncogenic BRAF V600E suppresses the activities ofthe tumor suppressor LKBl and its downstream kinase AMPK through indirect phosphorylation on LKBl. Moreover, this inhibition is critical for the proliferation of melanoma cells with BRAF V600E mutation. The goal of this proposal is to fiilly understand the regulation of LKBl and AMPK by BRAF signaling, examine its relevance in melanoma pathogenesis and explore its therapeutic implication. This proposal will define the molecular mechanism underlying the inhibition of LKBl- AMPK activity by BRAF V600E signaling, will investigate whether this inhibitory signaling mechanism is critical for melanoma cell proliferation, and tumor growth in mouse xenograft models, will examine the potential correlation between the active state of AMPK and ERK in human melanoma, will evaluate the effects of combined treatment of AMPK activators and MEK inhibitors on melanoma cell proliferation and xenograft tumor growth, and finally will characterize critical downstream signaling proteins of AMPK in melanoma. CANDIDATE: Bin Zheng received his Ph.D. in molecular pathology in 2002 from UC San Diego and postdoctoral trainings in the laboratory of Lewis Cantley at Harvard Medical School. His scientific advisory committee includes Cory Abate-Shen, Richard Baer, Meenhard Herlyn and Ramon Parsons, who are experts in cancer biology, cancer signaling and melanoma. The advisory committee and the vibrant scientific environment at the Columbia University Medical Center will facilitate Dr. Zheng in achieving his scientific and career goals.
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{
"pile_set_name": "NIH ExPorter"
}
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The recent emergence of antibiotic-resistant, Gram-positive bacteria presents a major challenge to today s pharmaceutical industry. Cubist Pharmaceuticals has addressed this challenge with a target-based approach to drug discovery. Cubist uses novel targets, that are essential to the pathogen s viability, in high throughput screening programs to identify potential drug candidates. Here, we present data on the acquisition and characterization of an essential target from the Gram-positive bacterium Staphylococcus aureus and propose experimentation that will lead to the identification of chemical inhibitors of this target via high throughput screening. A thorough characterization and analysis of these inhibitors to determine in vitro sand in vivo mechanisms of action is also planned. Phase II of this project will involve the optimization of these inhibitors to drug candidate status. PROPOSED COMMERCIAL APPLICATION The commercial applicability of this project lies in the recent emergence of drug-resistant Gram-positive bacteria and the ensuing need for a new chemical class of antibiotics. Cubist Pharmaceuticals has taken a target-based approach to the discovery of novel antibiotics. In this proposal we outline a research project that will allow us to identify inhibitors
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{
"pile_set_name": "NIH ExPorter"
}
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Studies were performed to investigate the regulation of carbohydrate and energy metabolism (i.e., control of adenine nucleotides) in human skeletal muscle during exercise and euglycemic hyperinsulinemia and the regulation of lactic acid production during muscle contraction. It was found that lactate production was preceded by or occurred in parallel with increases in mitochondrial nicotinamide adenine dinucleotide reduced (=NADH), suggesting, in contrast to current views that lactate production during submaximal exercise is oxygen-dependent. Glucose 1,6- bisphosphate (GP2), an important regulator of key enzymes of carbohydrate metabolism, has been shown to increase after isometric contraction to fatigue (approximately 50 seconds). To study the regulation of GP2 contents in muscle during contraction, we obtained biopsies prior to, after 20 seconds of contraction, and at fatigue. The major increase in GP2 occurred within the first 20 seconds of exercise, with no significant change there after. Preliminary results suggest that the rapid increase in GP2 is due to activation of GP2 synthase by its substrates G-1-P and G-6-P. The lack of continuous production, while the substrates increase, during the latter part of contraction, may be due to allosteric inhibition (by inorganic phosphate) of the synthase and/or activation of GP-2 phosphatase (by Ca2+ or inosine monophosphate (IMP)). A series of studies were performed to determine the in vivo regulation of AMP deaminase (deaminates AMP to IMP and ammonia), the activity of the purine nucleotide cycle (responsible for reaminating IMP back to AMP (and thereby ATP (via myokinase)). Last, it has been suggested that during euglycemic hyperinsulinemia, free glucose accumulates in the muscle of insulin.
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{
"pile_set_name": "NIH ExPorter"
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This proposal requests support for a clinical study designed to see whether lowering the hyperfiltration that occurs in early insulin dependent diabetes mellitus and inhibition of angiotensin II converting enzyme will reverse microalbuminuria and prevent the progressive decline in glomerular filtration rate that has been documented in other studies of insulin dependent diabetes. We have available 263 cases of insulin dependent diabetes presently in a longitudinal study of the natural history of diabetic retinopathy by the Department of Ophthalmology. Nearly 70 percent of the subjects do not have proteinuria detectable by routine clinical procedures. In these individuals without proteinuria, the rate of microalbuminuria will be characterized by an albumin radioimmunoassay of timed urine collections and those individuals with albumin excretion rates in excess of 15 micrograms/min will be randomized into two groups. One group will be patients with early microalbuminuria (15 to 80 micrograms/min) and the second group will contain those individuals with microalbumin excretion rates ranging between 81 and 250 micrograms of albumin per minutes. No individual with a blood pressure greater than 140/85 or who is receiving antihypertensive therapy will be included. Each of the two groups will be randomized into a treatment group and a control group and the treatment group given converting enzyme inhibitors at a level of 20 mg/day. Microalbumin excretion rates will be measured every three months and glomerular filtration rates every six months and assessment of retinal changes will be made on an annual basis. Observations will be made over a period of three years following initiation of treatment. Outcome variables will be microalbumin excretion rate, change in GFR and change in the severity of retinopathy. This study should provide a direct test of the hypothesis that the hyperprofusion of the glomeruli in early diabetes mellitus play an important pathogenetic role in the production of diabetic nephropathy.
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{
"pile_set_name": "NIH ExPorter"
}
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Enrichment Program Abstract The goal of the Michigan Center for Diabetes Translational Research (MCDTR) Enrichment Program is to: ? advance type 2 translational research in diabetes through education and training, ? promote scientific exchange among investigators, and ? encourage collaboration among trainees, diabetes researchers, and investigators from other fields locally, regionally, nationally, and internationally. To this end, the MCDTR sponsors education programs including the MCDTR Annual Symposium and partners with the Institute for Healthcare Policy and Innovation (IHPI), the Michigan Institute for Clinical and Health Research (MICHR), the Veterans Affairs Ann Arbor Healthcare System Health Services Research and Development Service (HSR&D), and the Michigan Diabetes Research Center to sponsor lectures and symposia. The MCDTR also works with the Metabolism, Endocrinology, and Diabetes T32 Training Grant and its NIDDK-funded Medical Student Training Program, the University of Michigan Medical Scientist Training Program, and the Robert Wood Johnson Foundation Clinical Scholars Program to provide training to students and fellows in T2 translational research in diabetes.
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{
"pile_set_name": "NIH ExPorter"
}
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Vascular anomalies (VAs), inborn errors in embryonic vascular development are classified into two distinct groups: hemangiomas and vascular malformations (VMs). Current therapies for VAs are limited in efficacy and have significant complications. Therefore, to improve therapy for patients afflicted with these conditions, it is critical to find new drugs or repurpose FDA-approved drugs to target VAs. Our long-term goal is to understand the underlying mechanisms that lead to pathogenesis of VAs so that better therapeutics targeting this condition can be generated. In order to pursue that goal, the objective is to identify small molecules (SMs) that will target dual-specific phosphatase-5 (Dusp-5), a member of the mitogen-activated protein kinase (MAPK) family, which is mutated in patients with hemangiomas and VMs. We have identified a serine to proline mutation at 147 AA in DUSP-5 (S147P), which results in a hypoactive phosphatase that is unable to dephosphorylate p-ERK. This results in sustained p-ERK levels, which is often associated with increased proliferation of cells such as those in VAs. Our central hypothesis is that, human S147P protein recapitulates zebrafish S148P protein function, whereby mutation perturbs the interaction with p-ERK such that DUSP5 phosphatase domain (PD) cannot be properly positioned to de-phosphorylate p-ERK. Small molecules such as SM1842, FDA-approved compounds (Suramin), and SM1842 analogs can reverse this effect, thereby permitting a switch between WT and S147P function both at the molecular level (in vitro), and in terms of cellular function. This hypothesis is formulated based on preliminary data from our group that predicts the incorrect positioning of the DUSP5 PD domain in relation to p-ERK using computational modeling studies on Dusp-5 interaction with ERK, which suggests the molecular mechanism that leads to the S147P<s hypoactivity. Further, computational docking approach with 10,500 SM compounds to the C-terminal PD of Dusp-5 enzyme identified SM1842, and other SMs that act as potent Dusp-5 antagonist in ECs. SM1842 is the most potent of the identified hits, acts as Dusp-5 antagonist in p-ERK assay, affects VEGF-stimulated p-ERK and Dusp-5 levels in endothelial cells, and restores S148P function in biochemical assays in vitro. Suramin - FDA-approved compounds (similar structure to SM1842) also affect endogenous Dusp-5 and p-ERK levels in endothelial cells. The proposed hypothesis will be tested by pursuing three specific aims: 1) Determine the structural mechanism for SM1842 in affecting Dusp-5 and S147P function; 2) Identify the optimal chemical analog for SM1842; and 3) Characterize the activity of FDA- approved compounds (suramin) similar to SM1842 in vivo and in vitro. In each of these aims, we will employ a variety of biophysical, cell biology, molecular and developmental biology approaches to unravel mechanistic basis for SM1842 and its analog to affect Dusp-5<s activity in vivo and in vitro. The approach is innovative because molecules like SM1842 that selectively affect mutant protein over WT protein, function are highly sought after by big Pharma, and this application has the potential to shift paradigm in target-based research. The proposed research is significant because benefits of this project will provide immediate clinical impact for VA patients in terms of therapy options, and importantly will translate basic science discovery into tangible clinical benefits instantaneously. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because vascular anomalies (VAs) represent an important clinical problem that has few therapeutic options. The successful development of a small molecule or FDA- approved compound that targets mutated Dusp-5 over WT Dusp-5 protein will provide the much needed therapy alternatives for patients with VAs. Thus, the proposed research is directly relevant to NIH<s mission of reducing the burden of debilitating health conditions from diseases affected by deregulated vasculature.
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{
"pile_set_name": "NIH ExPorter"
}
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The overall goal of this program project is to understand the molecular mechanisms underlying leukemia and lymphoma development and to devise novel therapeutic strategies to control lymphoid neoplasms. There are four highly interactive projects that progress from basic studies to clinically applicable therapeutic strategies with a delicate balance between in vitro and in vivo model systems. Project 1: Cell Cycle Progression of Normal and Malignant B Cells: Dr. Snow will study the role of cell cycle regulators in the ability of CD40 to synergise with BCR to induce cell cycle progression in normal B cells. They will determine the basis of the diverse response patterns of three subgroups of B cell lymphomas to CD40 signaling and the in vivo relevance of such CD40 signaling for B lymphoma growth using transgenic mice. Porject 2: Role of egr-1 gene in the growth regulation of normal B cells and B cell lymphoma: Dr. Bondada will study the basis of B cell receptor induced down regulation of the immediate early gene, egr-1 and its relation to lymphoma growth inhibition using BKS-2, a B cell lymphoma. The importance of egr-1 for B cell development and B lymphoma growth will be studied in transgenic mice that express a dominant negative form of the egr-1 protein. Project 3: Biological chaaracterization of human leukemic stem cells (LSC): Dr. Jordan will examine the novel concept that LSC are the basis of relapse of drug treated leukemias. The growth requirement of LSC will be characterized. Modulation of pro and anti-apoptotic genes will be explored to control leukemic cell growth. Project 4: Graft-versus-tumor (GVT) activity of syngeneic/allogeneic graft versus host disease (GVHD): Dr. Bryson will determine the cellular mechanisms involved in GVHD and in GVT reaction. They will test the hypothesis that cyclosporin A induced oxidative stress directly participates in the induction of SGVHD. Cellular basis of the lack of memory in GV7- responses will be studied. Support for three Cores, a transgenic and genetically defined animal facility, histopathology and administrative core, is requested to support the rsearch in this PO1 application.
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{
"pile_set_name": "NIH ExPorter"
}
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Tissue engineering of the intestine has emerged as a potential curative therapy for patients with short gut syndrome. Currently, the intestinal mucosa can be regenerated using epithelial cell clusters and polymeric scaffolds. Although the mucosa is necessary for intestinal function, it is insufficient without the muscularis of the small intestine that propels the enteric content forward. Therefore, the regeneration of the muscularis represents a critical step in realizing the clinical potential of intestinal tissue engineering. The long-term objective of the proposed research is to engineer the environment for the formation of the intestinal muscularis. Three specific aims will examine the scaffold design, signaling cues, and cellular interaction as follows. Specific Aim 1: Investigate the effects of scaffold design on muscularis formation. Sheets of aligned smooth muscle cells growing on oriented fibers will be produced. The choice of the material for the fabrication of the oriented-fiber sheets will be investigated. A spatial gradient of basic fibroblast growth factor will be created using microspheres to enhance angiogenesis in the orthogonal-layer tube. Specific Aim 2: Investigate the effect of butyrate on the phenotype of smooth muscle cells after implantation. The phenotype of the smooth muscle cells will be modulated by the delivery of butyrate in vivo. Labeled butyrate will be encapsulated in biodegradable microspheres that will release the content after a period of delay. A spatial gradient of lag time in butyrate delivery will be created to match the differential rates of smooth muscle cell proliferation in the multi- layer tube. Specific Aim 3: Investigate the effects of pacemaker cells on smooth muscle cells. To create the rhythmic contractions, we will reconstitute a network of pacemaker cells adjacent the layers of the smooth muscle cells in the contractile state. A multi-layer tube with smooth muscle cells in rolled sheets and the pacemaker cells in collagen gel surrounding the tube will form the basis of the motor unit. The effects of stem cell factor delivery, the collagen concentration, the cell density, and the arrangement of the cell types will be studied. PUBLIC HEALTH RELEVANCE: The massive loss of the small intestine results in short gut syndrome. These patients rely on intravenous nutrition that can lead to liver failure. This research will address a critical step toward the regeneration of the small intestine for patients with short gut syndrome.
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{
"pile_set_name": "NIH ExPorter"
}
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The goal of this proposal is to understand the mechanisms regulating mouse natural killer (NK) cell proliferation. By the techniques of centrifugal elutriation, sensitivity to a drug specific for dividing cells, and a single-cell cytotoxicity assay which detects lysis mediated by blast-NK cells, I have shown: 1) that interferon (IFN) and IFN inducers stimulate blastogenesis of NK cells in vivo; 2) that the primary lymphoid organ for this blastogenesis is the spleen; and 3) that this blastogenesis is accompanied by increases in turnover rates and numbers of NK cells. I recently have developed an in vitro system to support the proliferation of blast-NK cells elicited by IFN in vivo. The precursors of cells expanding, in this culture system, to mediate lysis of NK sensitive target cells are isolated in equal frequencies from both euthymic and athymic mice. This appears, therefore, to be an excellent model for the characterization of NK cell proliferation. The in vivo and in vitro systems will be combined to study NK cell expansion, the mechanisms of induction and the regulation of NK cell proliferation. Blastogenesis of NK cells will be induced in vivo by stimulation with poly I:C or with purified IFN's ( or ). Further expansion of NK cells will be carried out by multiple stimulations with IFN in vivo and by outgrowth of blast-NK cells in culture. A role for the T cell growth factor, interleukin-2 (IL-2), in the proliferation of NK cells will be determined by quantitating the expression of IL-2 receptors on in vivo elicited blast-NK cells and by growing these blast-NK cells in cloned IL-2. NK cell proliferation in the context of T cell responses will be examined during the infection of euthymic mice with lymphocytic choriomeningitis virus. The information obtained from these studies will be used to expand NK cells and induce antitumor and antiviral states in vivo.
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{
"pile_set_name": "NIH ExPorter"
}
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The longterm objective of this program is to understand the role that nerve growth factor (NGF) and its receptor (NGFR) play in forebrain cell and memory loss associated with aging and Alzheimer's disease (AD). Evidence is compelling for a dysfunction of basal forebrain cholinergic neurons in AD; there is also considerable evidence for a trophic role of endogenous NGF on basal forebrain cholinergic neurons during development and in adulthood. There are two specific aims of the present proposal, which together are designed to test the hypothesis that elevated levels of a truncated form of the NGFR (NGFRt) in urine or CSF correlated with spatial memory decline. (1) We will first confirm and extend our preliminary observations that urine levels of NGFRt in patients with AD are elevated relative to age-matched patients with no cognitive or neurological impairment, and determine if this elevation correlates positively with the degree of memory dysfunction. (2) To address the mechanism underlying the elevation of NGFRt in urine, we will study in rats the relationship of NGFRt to aging and spatial memory impairment and to basal forebrain neuron degeneration. We will accomplish this (a) by measuring NGFRt levels in urine and CSF of aged rats with poor spatial memory (water maze) ability in relation to a subpopulation of unimpaired-aged or young adult rats, and (b) by measuring the time course of changes in NGFRt levels associated with basal forebrain cell degeneration following axotomy or ibotenic acid lesions. These studies of changes in urine levels of NGFRt should lead to a better understanding of neurobiological basis of cognitive deficits associated with AD. In addition, the measurement of NGFRt in urine could be used as a simple, noninvasive method for diagnosing this disease.
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{
"pile_set_name": "NIH ExPorter"
}
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The BUSM MS Resource sponsors a weekly seminar series open to all interested persons in the Medical School community as well as investigators from other academic, research and industrial institutions throughout the Boston area. These provide an opportunity for staff members to present summaries of their ongoing research and for visiting scientists to meet and discuss their research with local scientists. Publicity for these seminars takes the form of posted notices, as well as standard mail, e-mail and telephone announcements to those outside BUSM.
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{
"pile_set_name": "NIH ExPorter"
}
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The long term objective of this research is to develop an understanding of the role of Myc in the control of cell proliferation and differentiation. One discovery that would help clarify this role is the identification and characterization of cellular proteins that interact with Myc in vivo. Thus, the specific goal of this research application is to develop the technology needed to isolate these cellular proteins. This will be accomplished by expressing in human cells a fusion protein which contains he Myc oligomerization domains fused to the bacterial lac repressor protein, followed by purification of the fusion protein complex using DNA affinity chromatography. HeLa cell lines which express high levels of the lac repressor-Myc fusion protein will be obtained by infecting cells with retroviruses which contain a lac repressor-Myc fusion gene. This research will establish that the fusion protein is capable of binding to lac operator DNA, and that it interacts specifically with cellular proteins via the Myc oligomerization domains. The fusion protein complex will be purified using a lac operator DNA affinity column, and proteins that interact with the fusion protein via the Myc oligomerization domains will be identified by SDS-polyacrylamide gel electrophoresis. The procedure will be scaled-up in order to obtain sufficient amounts of each protein for a partial amino acid sequence determination. Antibodies to each protein will be raised in rabbits, and used to screen a HeLa cell-lambda bacteriophage CDNA library to obtain cDNAs will subjected to DNA sequencing and the amino acid sequence of each protein deduced. The amino acid sequences will be compared to all sequences contained in the Protein Data Bank to identify domains that are homologous to known proteins. Northern and Western blot analysis will be used to test the hypothesis that the expression of these proteins is regulated in a growth-specific, differentiation-specific and developmental fashion. The knowledge gained from this research will lead to a better understanding of the molecular basis of cancer, and could be used to design new ways to reverse the cancer process.
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{
"pile_set_name": "NIH ExPorter"
}
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The objective of this project is to develop software for the analysis of data from large-scale genotyping and sequencing studies, building on the existing software package PLINK and its companion package PLINK/Seq. Designed to manipulate and analyze whole-genome SNP datasets, PLINK has been actively developed for over six years and has a wide base of users, with over 5000 citations in peer-reviewed journals. Over the past years, we have added considerable support for the analysis of large rare variant datasets, primarily focused on whole-exome sequencing studies in PLINK/Seq. In this renewal application, we seek to 1) provide tighter integration between PLINK and PLINK/Seq, aiming to provide a single interface for both genome-wide association and sequencing studies, particularly in the context of large statistically-imputed datasets; 2) enhance the data-integration facilities already present, across different classes of genetic variation as well as large, diverse datasets; 3) provide improved handling of family-based datasets, focused on de novo and inherited variation in (nuclear) family-based association studies; 4) to work on improving performance on very large datasets. Particular attention will be paid to ensure interoperability with other major software, file-formats and resources that are generated by the broader genetics community.
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{
"pile_set_name": "NIH ExPorter"
}
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This project is designed to assess the physiological importance of calcium ions in the regulation of energy metabolism. We have applied the fluorescent chelating-agent protocol, which was developed for the measurement cytosolic free Ca2+, to the measurement of mitochondrial matrix free Ca ((Ca2+))m. This allowed us to study directly the relationship between (Ca2+)m and the activation of the matrix enzyme pyruvate dehydrogenase for the first time in studies with isolated rat cardiac mitochondria loaded with indo-1. The KO.5 for activation was found to be 300 nM Ca2: this value was unaffected by procedures, e.g. omission of Mg2+, Na+, which change the magnitude of the Ca concentration gradient across the mitochondrial membrane. We have also studied the degree of activation of pyruvate dehydrogenase as a function of the rate of electrical stimulation of isolated guinea pig hearts: in a parallel study, mitochondria were rapidly isolated from stimulated hearts, using a protocol which prevents loss or gain of mitochondrial Ca and allows us to relate this quantity to the level of dehydrogenase activation. As the significance of dehydrogenase activation lies in the NADH/NAD ratio which can be maintained in the face of elevated work -loads, we have also measured the ratio NADH/NAD in suspensions of rat heart mitochondria with saturating and non-saturating concentrations of pyruvate or 2-oxoglutarate as respiratory substrate, in the presence or absence of Ca2+, and correlated this with the flux through oxidative phosphorylation. Finally, we have examined the ability of cardiac mitochondria from senescent dogs to oxidize substrates with emphasis on fatty acids) as a function of lifestyle, viz-sedentary versus exercised.
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{
"pile_set_name": "NIH ExPorter"
}
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Prostate cancer is now the second leading cause of cancer deaths in males, and combined with benign prostatic hyperplasia, now represents the leading neoplastic disease in man. In humans, the prostate gland has the unique function of producing and secreting extraordinarily high levels of citric acid. The major distinguishing characteristic of prostate cancer is the transformation of citrate producing prostate epithelial cells to malignant cells which are incapable of producing citrate. Our broad objectives are to elucidate and to understand the unique metabolic relationships which characterize the ability of prostate cells to achieve this function; to understand the mechanisms by which prolactin and other hormones regulate this major function; to elucidate the metabolic and hormonal alterations associated with prostate neoplasms; and to use this information for new approaches in the diagnosis, prevention, and treatment of prostate neoplasms. It has now been established that prolactin is an important regulator of prostate citrate production. The specific aims of this program is to elucidate the mechanisms by which prolactin regulates citrate synthesis of normal prostate epithelial cells. The major relationship to be addressed is the mechanism by which prolactin regulates mitochondrial aspartate aminotransferase (mAAT) and pyruvate dehydrogenase (PDH) E1alpha, the two key enzymes involved in prostate citrate synthesis. The focus of the proposed studies will be on prolactin regulation of expression of the mAAT and E1alpha genes and the role of protein kinase C (PKC). Once these relationships are established with normal prostate cells involving rat lateral prostate, the role of "prolactin-PKC-gene regulation" in human malignant prostate cells (LNCaP, PC-4 Du-145) will be established. This will represent the first concerted characterization of citrate-related metabolism in human prostate epithelial cells.
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{
"pile_set_name": "NIH ExPorter"
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Proper functioning of the vertebrate nervous system depends on individual cells with it acquiring specific identifies during embryonic development. Evidence suggest that both a cell's lineage and its environment influences its identify. However, it is unclear how different cell types arise in virtually the same position within the embryo. For instance, immediately after gastrulation, precursors of neural crest and identified spinal cord neurons are mingled in a narrow region of the neural plate, which later becomes the spinal cored. What kinds of signals cause these cells to develop different identities? Do they adopt their identities all at once, or is it a sequential process? These questions will be addressed in the context of the developing zebrafish nervous system, using powerful embryological techniques of gene mix-expression, single-cell labeling, and single-cell transplantation. This research will reveal the way multiple ell types arise in the dorsal spinal cord during development. Research into normal embryology is a prerequisite for the advance of new therapies for developmental pathologies.
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{
"pile_set_name": "NIH ExPorter"
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The effect of learning a one trial passive avoidance response and of psychotropic drugs on the turnover of acetylcholine (ACh) in discrete regions of rat brain will be studied. At various times after training in a passive avoidance paradigm, deuterated choline will be administered to rats and the turnover of ACh in various brain regions implicated in this behavior will be studied. The regions to be studied are hippocampus, striatum, and frontal cortex. Specificity will be ascertained by measuring the turnover in the posterior cortex, a region not implicated in this behavior. Some psychotropic drugs (amphetamine, haloperidol and THC) will be studied for their effects on ACh turnover in the hippocampus, striatum, hypothalamus, olfactory tubercle, frontal and posterior cortex. Where treatments (behavioral or drug) are seen to have an effect on ACh turnover, the effect of the same treatments on the levels and labelling of ACh in synaptosomes isolated from specific regions will be determined. Also, the effects of these treatments on the availability of precursors for the synthesis ACh will be determined. In these experiments, following treatment of rat, brain slices or synaptosomes will be prepared from the specific regions and the high affinity uptake and acetylation of choline and flux of labelled pyruvate and synthesis of ACh will be determined. Similarly, the proportion of pyruvate dehydrogenase in the active form will be determined following treatment. Should the results of these studies indicate, the effects of striatal lesions on behavior and turnover of hippocampal ACh will be studied. To elucidate further the role of cholinergic systems in learning and memory, the effect of an appetitive approach response on ACh turnover in discrete brain regions will be determined. The effects of cholinergic drugs on both learning and turnover of ACh will be studied in the hippocampus and striatum. Experiments to evaluate the utility of cholinergic false transmitters as probes to study cholinergic mechanisms in learning and memory will be performed. This work should show the role of the central cholinergic nervous system in behavior and a mechanism and site of action of some psychotropic drugs.
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{
"pile_set_name": "NIH ExPorter"
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Tetrahydrobiopterin (BH4) is a vital cofactor that maintains availability of amine neurotransmitters including Dopamine and Serotonin, regulates Nitric Oxide synthesis, and stimulates and modulates the Glutamatergic system. Dysregulation of neurotransmitter (NT) systems has been implicated in the pathogenesis of schizophrenia (SZ). Based on the central roles of BH4 in NT synthesis, we assayed plasma total biopterin levels (a measure of BH4) in a large sample of patients with SZ and control subjects. A highly significant mean biopterin deficit of 34% was observed for the SZ patient group when compared to controls. The observed biopterin deficit is comparable to that reported for genetic BH4 deficiency disorders, supporting its characterization as having physiological significance. Our highly significant finding, along with: a) the known roles of BH4 in NT maintenance, b) dysregulation of CNS NT synthesis observed in human and mouse BH4 deficiencies, c) evidence that plasma biopterin levels are correlated with CNS biopterin levels, and d) evidence that urinary biopterin excretion is not increased in SZ, all support our hypothesis that dysregulation of BH4 biosynthesis is involved in the etiology of SZ. SZ susceptibility has a large genetic component, and in another preliminary study, we have performed initial genetic testing of GCH1, the first gene in the BH4 biosynthesis pathway, genotyping a polymorphism for association with SZ in a sample of 132 subjects (86 SZ and 46 control subjects). The results were striking: We found that GCH1 was strongly associated with SZ: odds ratio, 5.0, p= 0.0057. We also found that GCH1 allele status predicts low biopterin in SZ patients. Based on these very positive preliminary data, and the fact that some SZ subjects without the SZ-associated GCH1 genotype also have a biopterin deficit, this exploratory study is designed to test the hypothesis that DNA variants in GCH1 and other BH4 biosynthesis-related genes, are associated with and influence biopterin levels, and are candidate SZ susceptibility loci. We will employ Re- Sequencing Arrays to screen for DNA variants that may influence gene expression and/or protein function. Sequence data generated in this study will be analyzed for association with low biopterin levels and subject diagnosis. The Specific aims of this proposed study are: Aim 1a. To design Re-Sequencing arrays that include the gene regulatory regions, intron-exon boundaries, and coding regions for the six BH4 biosynthesis pathway genes, plus 13 additional BH4 regulation-associated genes. 1b. To generate gene sequence data for 180 subjects (90 SZ subjects and 90 controls), all with assayed plasma biopterin levels. Aim 2. To analyze the sequence data using single marker association testing and haplotype analyses, and to identify DNA variants associated with SZ and biopterin deficit. Understanding the genetic basis of the SZ biopterin deficit will be primary to development of medications and may also allow pre-symptomatic and early diagnostic testing, and early treatment that may improve the clinical outcomes for persons at risk of developing SZ. 7. PROJECT NARRATIVE This study will test whether people with schizophrenia (SZ), who showed abnormally low levels of a chemical called BH4 that is very important for brain function, also have DNA abnormalities in the genes that produce BH4. If they do, we will look for ways to treat low BH4 levels, and try to improve the symptoms of patients that way. We may also be able to predict risk of SZ in people with the gene defect, and therefore may be able to start treatment as early as possible to improve the outcome for those with the disease and young people at risk of developing SZ. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The clinical investigations laboratory is a core resource laboratory for the UCLA HIV research community. In addition, because of its unique resources, extramural collaborations are also allowed. The laboratory was started in 1991 and initially focused on supporting clinical trials and application of clinical specimens to answer basic science questions. In 1993, the laboratory was refocused to support basic science investigations that needed access to specific types of clinical specimens in order to answer questions on the pathogenesis of immune destruction in HIV disease and the development of AIDS related malignancies. The laboratory houses a computerized inventory of over 15,000 serum samples, 600 plasma samples, 800 frozen viable lymphocyte preparations, as well as provides continuous fresh isolation of Kaposi sarcoma cells and AIDS lymphoma cells for basic science studies. The samples are linked to clinical status and outcome. Most patients have develop a rapid, simple serologic assay for the detection of HIV infection in the newborn, excluded Rochelimea as a causative agent for KS, examined the relationship between cytokine perturbations and the development of lymphomas, identified key molecules in the post-receptor pathway of signal translation in KS leading to proliferation, demonstrated that heavy chain utilization in lymphomas is non-random, investigated the clonality of AIDS malignancies, and isolated genetic fragments of a organism that is linked to classical and AIDS-related KS. Intramural and extramural investigators are free to propose in vitro studies using the resources of the laboratory. However, priority is given to seed grant awareness, intramural studies, and those studies which are consistent with our overall goal of understanding the immunopathogenesis of HIV infection. All studies are reviewed by the director for their scientific merit and discussed with the executive committee. Charges are levied for routine sample processing, storage, and isolation of RNA and DNA. Lymphoma and Kaposi sarcoma cell lines are provided at no charge. To date, the core has serviced over 20 investigators on a wide range of basic science questions requiring clinical materials. The unique ability to provide not only rare specimens and cell lines but the associated clinical information that places the results in context are the greatest strength of this core. It is anticipated that several of these investigations will lead to clinical trials of novel agents in 1994-95.
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{
"pile_set_name": "NIH ExPorter"
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Natural products have made enormous contributions to human health. The search for new lead compounds from natural sources continues to be a crucial element in modern anticancer drug discovery. Our long-term goal is to discover anticancer drugs that act by novel mechanisms from plant- and lichen-associated fungi of the Sonoran desert, thereby providing further support for our hypothesis that organisms living in association with one another in this unique environment elaborate a relatively unexplored repertoire of potent bioactive compounds with which to mediate their interactions and enhance their survival. The specific aims of this interdisciplinary and inter-institutional project are to: 1. expand our unique bio-resource of Sonoran desert plant- and lichen-associated fungi and optimize biosynthetic potential of 500 selected fungal strains and prepare 2,500 extracts for evaluation; 2. screen extracts for anticancer activity using cell-based biological and molecular approaches including: (a) an assay established during our previous grant period and recently modified to detect compounds capable of heat shock induction and inhibition of heat shock induction, followed by assays for direct interaction with Hsp90 including luciferase-refolding and binding assays for compounds isolated using the heat shock induction assay; (c) an assay for inhibitors of cell motility at sub-cytotoxic concentrations, and (d) a conventional dye reduction assay for inhibition of proliferation/survival based on five sentinel cancer cell lines to avoid the discovery of general cytotoxic agents; 3. dereplicate active extracts, culture on large-scale up to five promising fungal strains per year and subject their extracts to bioassay-guided fractionation to isolate and characterize bioactive compounds; and 4. define molecular mechanisms of action using genomic and proteomic approaches for the most promising compounds based on their activity, structural novelty, and potency. We expect that the most interesting compounds will serve as leads for further optimization in the development of novel anticancer agents with unique mechanisms of action. At a more basic scientific level, however, the innovative chemical biology approach we propose should help define the mechanisms by which (a) chaperones such as Hsp90 facilitate oncogenesis, and (b) small-molecules inhibit cancer cell migration involved in invasion and metastasis, leading to enhancement of our overall ability to prevent and cure cancers, and therefore have a great impact on public health.
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{
"pile_set_name": "NIH ExPorter"
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Summary: Identification of a set of cDNA clones that are differentially expressed in memory CD4+ T cells. In an attempt to determine the molecular features of memory CD4+ T cells, we have utilized cDNA microarrays to measure gene expression of freshly isolated human memory and naive CD4+ T cells from peripheral blood over 100 donors. cDNA microarray method allows us to simultaneously assess genes at genome scales, providing a powerful tool for comparative analysis of gene expression profiles and for identifying differentially expressed genes in certain cell populations. After a sequential analyses of cDNA microarray filters from commercial source (consisting of over 50,000 cDNA clones) and custom-made selected clones from the commercial source and this laboratory (~2,800 unique clones), we provide the first glimpse into gene expression patterns of memory and naive CD4+ T cells at the genome-scale. We found that freshly isolated memory and naive CD4+ T cells expressed similar numbers of genes, and that 14 cDNA clones expressed higher levels of transcripts in memory cells than in naive cells. Identification of activation induced down- and up-regulated genes in memory CD4+ T cells. We have identified 135 (130 known genes and 5 ESTs) up-regulated and 68 (42 known genes and 26 ESTs) down-regulated cDNA clones in memory CD4+ T after 16 hours of in vitro stimulation with anti-CD3 plus anti-CD28. Interestingly, the increase in mRNA levels of up-regulated genes was greater in memory than in naive CD4+ T cells after in vitro stimulation, and was higher with anti-CD3 plus anti-CD28 than with anti-CD3 alone in both memory and naive CD4+ T cells. We then confirmed the changes in expression of actin and cytokine genes identified by cDNA microarrays by Northern analysis. Furthermore, we extended mRNA analysis to protein expression and found that the levels of mRNA and protein of cytokines and actins were correlated. Together, we have identified approximately 200 cDNA clones whose expression levels changed after activation, and suggest that the level of expression of up-regulated genes is a molecular mechanism that differentiates the response of memory from naive CD4+ T cells.
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{
"pile_set_name": "NIH ExPorter"
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The objective of this project is to determine whether the major cholesteryl ester-rich lipid particle in plasma, low density lipoprotein (LDL), can transform by treatment with cholesterol esterase into a larger unesterified cholesterol-rich particle similar to those we have isolated from atherosclerotic lesions. When LDL was incubated with cholesterol esterase, the LDL cholesteryl ester was completely hydrolyzed during 4 hrs of incubation. Interestingly, only when LDL was treated with trypsin prior to incubation with cholesterol esterase was LDL cholesteryl ester hydrolyzed. When LDL was incubated in a weak buffer in which pH gradually changed from 7.2 to 6.0 during the incubation, the LDL cholesteryl ester was initially hydrolyzed from 75% to 12% (molar % of total cholesterol). However, upon prolonged incubation, a reverse reaction occurred in which re-esterification of cholesterol slowly proceeded. After 24 hrs of CEase incubation, the degraded LDL contained 55% of its cholesterol as ester. The addition of human serum albumin prevented cholesterol re-esterification indicating that removal of free fatty acids from hydrolyzed LDL attenuated the cholesterol re-esterification process probably due to lack of substrate. The structure of the LDL particle changed following hydrolysis of LDL cholesteryl ester. Small projections extended from the surface of LDL during early cholesteryl ester hydrolysis. These projections transformed into films of various sizes and shapes, some still associated with the native LDL. With complete hydrolysis, flattened vesicles of irregular shape and aggregates of flat or curled films were present. In LDL incubated with the weak buffer the cholesterol re-esterification resulted in disruption of the films and the formation of lipid droplet-like spherical particles. Our results demonstrate that LDL transforms into a larger vesicular structure during hydrolysis of its cholesteryl ester core. This newly-formed larger lipid particle may become trapped in the vessel wall and lead to vascular cholesterol accumulation.
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{
"pile_set_name": "NIH ExPorter"
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Smoking is the leading preventable cause of death in the United States. It is a major cause of cancer and other serious disease, and is by far the leading cause of lung cancer. The proposed study will test a state-of-the- art corporate smoking cessation clinic supplemented by either active or placebo nicotine replacement (transdermal nicotine system). It is hypothesized that active nicotine replacement will lead to a significant reduction in withdrawal symptoms and to improved abstinence. A minimum of 120 subjects from six companies will be randomly assigned to either active or placebo transdermal patch. All subjects will attend a 9 week smoking cessation clinic followed by several assessment sessions. Withdrawal symptom and smoking status data will be collected at each session. Self- reported abstinence will be validated by expired carbon monoxide and saliva cotinine. Twelve month outcome data will be collected independently of SBIR funding. If results are as hypothesized, a widely disseminable corporate program will have been established that for the first time systematically integrates psychological and behavioral intervention. this will lead directly to Phase II research.
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{
"pile_set_name": "NIH ExPorter"
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Enteroviruses are characterized by their high replication rates and extreme sequence plasficity, which allow them to rapidly adapt to different environments and hosts. These viruses depend entirely on the host protein homeostasis machinery, composed of molecular chaperones and quality control (QC) components such as the ubiquitin-proteasome system, for viral protein production and function. Enterovirus replication poses several challenges to the cellular protein homeostasis machinery as the need to produce high amounts of protein in a very short time places a big burden for the host protein production and folding machineries. Furthermore, enteroviral proteins tend to be large, complex and multifunctional, and thus likely to require the assistance of molecular chaperones to fold. Indeed, we have shown that the Hsp90 chaperone system is essential for capsid folding and assembly for many, perhaps most, picornaviruses, including the enterovirus polio- and coxsakie-viruses. Since other aspects of enterovirus replication Involve additional large multlprotein complexes, chaperones are likely to be broadly required for other aspects of the viral cycle. An important challenge to protein homeostasis in RNA viruses arises from their very high mutation rates, which pose a big burden to viral protein stability and are likely to produce high levels of non-functional or destabilized proteins. These mutant proteins must be either maintained in a funcfional state or eliminated from the cell to prevent dominant negative effects on viral function. We hypothesize that these functions are carried out by chaperones, which can buffer metastable proteins, as well as by the ubiquitin-proteasome system, which targets misfolded proteins for degradation. To understand the molecular and cellular mechanisms by which cellular chaperone and quality control machineries control viral protein homeostasis, and allow the virus to replicate we propose the following Aims: Aim 1: Define the chaperone components required for enterovirus replication. Aim 2: Define the role of the Quality control (QC) machinery in picornavirus replication Aim 3: Examine the plasticity and interplay of chaperone and QC pathways during viral infection
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{
"pile_set_name": "NIH ExPorter"
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Fetal exposure of man and mice to mercury (Hg) induces long-term behavioral and neuronal dysfunctions, but the mechanistic involvement of neuroendocrine immune network activities, e.g.,hypothalamo-pituitary- adrenal (HPA) axis, centrally located immune cells (microglia and mast cells), and autoantibodies to central nervous system (CNS) antigens, have not been adequately addressed. In addition, based on their genetic differences, certain mouse strains develop Hg-induced immunopathologies (autoimmune disease), which are posited to include autoantibody-induced neuroinflammation, leading to neuronal damage and dysfunction. Hg induces enhancement of type-2 immunity, which includes heightened levels of Th2 cytokines IL-4, IL-5 and IL-6 as well as IgE and elevated release of factors from mast cells, eosinophils and basophils. Some mouse strains (e.g., BALB/c mice) develop immune complexes which can localize in certain organs eliciting inflammation whereas mice with the H-2s haplotype (e.g., A.SW mice) develop autoantibodies that bind to select antigens directly initiating targeted damage. We hypothesize that based on the genetics and environmental exposures, some strains have CNS inflammation due to antibodies inducing activation of microgia and mast cells, which leads to behavioral aberrance and neuropathological outcomes that occur in adults following developmental exposures to Hg. The sequence of events that culminate in adult neuroimmune-induced pathologies are posited to be: i) altered neuroendocrine regulation, which enhances type-2 immunity; ii) increased autoimmune responses to CNS antigens; iii)modulation of immune cell (microglia and mast cells) distribution and function in the brain; and iv) elevated neuroinflammation which disrupts neuronal functions. We will first demonstrate that Hg enhances generation of autoantibodies to CNS antigens, dependent on haplotype, and then identify the autoantigens. Aim 2 will investigate the specific brain regions with autoantibodies and the phenotype of the cells present. Aim 3 will evaluate whether autoantibodies in the absence of Hg can cause similar neuroinflammation in Hg-susceptible (H-2s and Th2-prone strains) and Hg-non-susceptible strains. We suggest that certain environmental agents can initiate heightened immunity to self-antigens, including brain antigens, based on the genetics of the host. We will compare the ability of inorganic and organic Hg to initiate the described consequences.
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{
"pile_set_name": "NIH ExPorter"
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The Hedgehog (Hh) signaling pathway is integral to tissue patterning during fetal development and oncogenesis in children and adults. Genetic screens have revealed several of the signaling proteins that regulate Hh target gene expression, which in mammals include the Sonic, Indian, and Desert Hh ligands (Shh, Ihh, and Dhh), the Hh receptor Patched1 (Ptch1), the transmembrane protein Smoothened (Smo), the Gli family of transcription factors (Gli1, Gli2, and Gli3), and the Gli antagonist Suppressor of Fused (Sufu). These genes are potential targets for next-generation chemotherapies, and several compounds that inhibit Smo have demonstrated efficacy in mouse models of Hh pathway-dependent tumors, such as basal cell carcinoma, medulloblastoma, pancreatic adenocarcinoma, and prostate cancer. The Smo antagonist GDC-0449 has even caused the regression of metastatic basal cell carcinoma and medulloblastoma in human clinical trials. It has become increasingly apparent, however, that Hh pathway-dependent cancers can readily gain resistance to Smo antagonists and that certain tumors are initiated or maintained by Smo-independent Hh target gene expression. Since Smo is the most druggable target within the Hh pathway and nearly all known pathway inhibitors target this transmembrane protein, there is a clear and urgent need for small molecules that act downstream, preferably at the level of the Gli transcription factors. This application describes a high-throughput screen for small-molecule antagonists of Gli function, using a Sufu null cell line that has been stably transfected with a Gli-dependent firefly luciferase reporter (Sufu-KO- LIGHT cells). Since Sufu directly inhibits Gli function, Sufu-KO-LIGHT cells exhibit constitutive firefly luciferase expression that mediated by endogenous Gli proteins. In contrast to cell-based assays used previously to discover Hh pathway inhibitors, the Sufu-KO-LIGHT cells are unresponsive to the large number of Smo- targeting compounds typically found in chemical libraries and they do not require the overexpression of Gli1 or Gli2 to achieve Smo-independent Hh target gene expression. This novel assay will therefore rapidly identify compounds that block the function of endogenous Gli factors, which are subject to regulatory processes that can be circumvented by Gli overexpression. In addition to this new screening campaign, a comprehensive hit advancement plan is outlined, including several secondary assays for assessing the Hh pathway selectivity of lead compounds, cellular and biochemical experiments for evaluating compound action on Gli function, and studies for determining compound efficacy against Hh pathway-dependent cancer cells. Collectively these investigations will provide valuable mechanistic probes of downstream signaling events within the Hh pathway and structural leads for the development of new anti-cancer therapies. PUBLIC HEALTH RELEVANCE: Uncontrolled activation of the Hedgehog (Hh) pathway contributes to the onset and progression of several cancers and its pharmacological inhibition can induce tumor regression in mouse models and human patients. The emergence of drug-resistant tumors in these studies underscores the need for new Hh pathway antagonists, particularly compounds that inactivate the Gli family of transcription factors. The proposed research will identify novel inhibitors of Gli function through a cell-based high-throughput screen, characterize their mechanisms of action, and assess their ability to block the proliferation of Hh pathway-dependent cancers.
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{
"pile_set_name": "NIH ExPorter"
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The long-term objective of this project is to identify the fundamental mechanisms that limit the benefit a patient derives from cochlear implantation. Better treatment of profound hearing loss with a cochlear implant (improved candidacy criteria, patient prognostication, and implant designs) will depend crucially on an understanding of both the factors mediating performance, and how they can be addressed. Our first step toward this objective is to use quantitative modeling to examine the impact of different pathologies seen in the histologically-processed temporal bones harvested from users of multichannel cochlear implants. Specifically, we will use the serially-sectioned temporal bones of two subjects to build a patient-specific computer model for each that represents their unique anatomy. These models will make it possible to (1) test the model's predictions of behavioral threshold against results collected during the patient's life and (2) investigate the relative impact of specific anatomical properties on those predictions. To the extent that these patient-specific models explain features of the subjects' measured performance, they are a first step in eventually understanding the features that determine the patient benefit. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this study is to characterize the soluable and membranous forms of rat liver ap, to study the origin of the membranous and soluble forms of liver ap, to identify the molecular mechanisms by which ap synthesis is controlled in the rat liver and to identify the soluble factor in cholestatic bile which causes enhanced synthesis of menbranous ap.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this study is to demonstrate that Stractan separated young red blood cells can be transfused into autologous non-human primate donors and that these cells have an improved survival compared to whole blood. Preliminary studies have shown that young red cells from rabbits can be isolated and transfused by this method. These cells demonstrate a markedly improved survival compared to unfractionated whole blood. The eventual goal of this project will be to explore the use of this system for human transfusion, thus reducing the transfused iron load in patients with chronic anemias. BIBLIOGRAPHIC REFERENCES: Corash, L., Seaman, C., Reibman, J., Tytun, A., Piomelli, S.: Qualitatively Improved Blood Cell Transfusion: A New Approach to Therapy of Chronic Anemias. 16th Meeting, Int. Soc. Hem., Kyoto, Japan, 1976.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary. In aging, immune systems falter and the elderly are more susceptible to infection. Age related changes include decreased T-cell activation/proliferation, decreased production of IL-2 and decreased expression of IL-2R1. These are the same key immune changes that are seen in returning astronauts who have experienced altered immune function and increased vulnerability to infection during spaceflights. Loss of immune response in aging and spaceflight has been previously identified in T cells. We have previously identified microgravity induced changes in gene expression, promoter regions, transcription factors and signal transduction pathways in human CD4+ T-cells under normal and altered gravity conditions. Our preliminary data from International Space Station (ISS) described within this proposal show that in early T-cell activation at least one miRNA was upregulated during activation in normal gravity (g), while in microgravity (<g), the induction of the miRNA was muted. Moreover, three protein targets of the miRNA had increased expression in the 1.g environment suggesting a micro RNA mechanism. The changes in immune function in the elderly occur over time and are hard to evaluate. However, the same type of changes in immune response occurs microgravity, both in astronauts and in human T-cells and is easy to measure The central hypothesis is that <g regulates MiRNA in human T-cells and that weightlessness is a novel model to study the immunosuppression seen in aging. These experiments will contribute to our fundamental understanding of the molecular mechanism of immunosuppression in spaceflight and in aging. The Specific Aims are the following: (1) Identify gene expression of miRNAs during T-cell activation (using arrays and qRTPCR) under normal gravity. (2) Identify the MiRNA(s) target genes using bioinformatics and verify the changes in expression of those targets messages (qRTPCR). We will test the ability of microgravity to induce/reduce expression of MiRNAs and their targets and verify these results using miRNA overexpression viral constructs or dsRNA. (3) Analyze the protein synthesis of the upregulated/downregulated target genes (SDS PAGE gels and Western Blot proteomics) that are affected by T-cell activation under normal gravity and microgravity conditions. (4) Compare the expression of key MiRNAs candidates and genes after activation in normal gravity and microgravity, vs. that of the lymphocytes from an older population. PUBLIC HEALTH RELEVANCE: Identifying altered microRNAs and their targets in T-cells that have suppressed immune response in spaceflight but normal expression in 1g flight controls will contribute to our understanding of aging and will likely reveal important causes of immunosuppression in the elderly. In recent ISS experiments, one MiRNA is significantly downregulated in <g flown T cells when compared to onboard 1g controls;with more efficient miRNA collection, several more dysregulated MiRNAs should be identified. Because there are considerable similarities between immunosuppression in astronauts during flight and immunosuppression in the elderly, we expect that our findings will be relevant to the mission of the NIH and be broadly interesting to researchers studying molecular mechanisms of the immune response.
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{
"pile_set_name": "NIH ExPorter"
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We have funding from NIDA to complete a case-control study to evaluate if marijuana use is a risk factor for head, neck and lung cancer in adults. The GCRC is supporting the interview process and also blood collection. We will evaluate whether or not two cancer susceptibility genotypes, glutathione transferase and N-acetyl transferase, are mediators of any association between marijuana use and head, neck and lung cancer. Ten(10) cases and 35 controls have been interviewed and provided blood samples. We anticipate assessing these genotypes and other potential candidate genes after collecting samples on 30 cases and 30 controls.
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{
"pile_set_name": "NIH ExPorter"
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Recently, New Mexico was recognized as the first 'minority-as-majority' state in the contiguous 48 states, with the highest proportion of Hispanics among the states, and the highest proportion of Native Americans in the continental U.S. The University of New Mexico (UNM) today is rapidly reflecting these demographics, but more is needed to encourage some of our very talented underrepresented minority students to enter doctoral degree programs to prepare for biomedical research careers. This proposal for funding for a UNM Post-Baccalaureate Research and Education Program (PREP) is precisely what is needed. The UNM PREP program is designed with various program elements to encourage and prepare up to 60 post-baccalaureate participants in its first five years to gain acceptance into and to complete doctoral degrees in biomedical research fields. UNM PREP scholars will have a full schedule of activities that all of them participate in: conduct full-time research, be assigned to a peer mentor, attend a year long set of tours of research laboratories, participate in the graduate admission colloquium series. All PREP scholars will attend speakers' events with IMSD and MARC participants, and networking with several programs at UNM during formal activities and informal community-building events. PREP scholars will present posters at the UNM Research Fair and Research Symposium and attend professional conferences. Seminars will help prepare our PREP scholars for pursuing a research career, covering experimental research design approaches, technical writing, research ethics, and public communication. Our PREP scholars will also have the experience of teaching undergraduates about their research topic and methods. Each PREP scholar will be assessed individually to ensure later succeed in graduate studies, and a faculty advisor will work with the student to develop the study plan, and then each scholar will take up to 15 credit hours of coursework to fulfill this plan. Administrative management of the UNM PREP program will be a coordinated effort by a Program Director, a Program Coordinator, and an Executive Board and committees (Recruitment, Education & Research, Events), each responsible for a specific set of activities. Coordination with other funded programs at UNM is planned with the Program Director and Program Coordinator attending and working with the Program Officers and staff of the IMSD and MARC programs as partners in the UNM Unified Plan. Evaluations will be conducted for all activities and mentoring relations on a set schedule, and the data that is collected will be used by both the internal evaluation team and the External Steering Committee, who will assess progress in achieving goals and measurable objectives, and make recommendations for changes in the program plans. Dissemination efforts will include posting pdf documents of PREP scholar poster presentations, having PREP scholars co-author research articles with their faculty mentors, and with the Program Director and faculty mentors writing about the lessons learned in implementing PREP and presenting their results at professional conferences. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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This project will study the fundamental mechanism involved in acquired polymorphonuclear leukocyte (PMN) locomotory dysfunction in patients who sustained serious blunt trauma. This is important because PMN dysfunction noted in these patients is related to subsequent increased infection rate and death from sepsis. The probability that the mechanism involves: 1) release of PMN activating factors as a result of tissue damage from the trauma, 2) inappropriate activation of PMNs, 3) increased production of toxic oxygen species especially H2O2, 4)self-inflicted damage resulting from increased H2O2, and 5) dysfunction, will be investigated. Plasma from trauma patients will be tested for evidence of complement activation products C3a desArg and C5a desArg, and serum for spontaneous chemotactic activity. Lysosomal enzyme release, H2O2 production, intracellular glutathione and glutathione disulfide, and Conconavalin A capping assays will be used to prove or disprove steps 2, 3 and 4. PMN locomotory function will be assessed using a modified micropore filter assay. These assays will be performed on patients's plasma or serum specimens, and PMNs soon after trauma, 3 times a week during the first week after injury and twice in the second week. Associations between activating serum/plasma factors, biochemical cellular changes and dysfunction will be correlated by plotting and comparing resolution of abnormalities with time. If autooxidative damage is found responsible for this acquired PMN dysfunction, studies to determine the usefulness of antioxidants in these patients will be commenced.
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{
"pile_set_name": "NIH ExPorter"
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Summary Infantile neuronal ceroid lipofuscinosis is a devastating neurodegenerative lysosomal storage disease caused by mutations in the gene (CLN1 or PPT1) encoding palmitoyl-protein thioesterase-1 (PPT1). We have previously reported that phosphocysteamine and N-acetylcysteine mediate ceroid depletion in cultured cells from patients with this disease. We aimed to assess whether combination of oral cysteamine bitartrate and N-acetylcysteine is beneficial for patients with neuronal ceroid lipofuscinosis. 9 INCL patients (5 female and 4 males) were treated with oral cysteamine bitartrate (60 mg/kg per day) and N-acetylcysteine (60 mg/kg per day) and were assessed every 6-12 months until they had an isoelectric electroencephalogram (EEG, attesting to a vegetative state) or they were too ill to travel. Patients were evaluated by electroretinography, brain MRI and magnetic resonance spectroscopy (MRS), and electron microscopic analyses of leukocytes for granular osmiophilic deposits (GRODs). Children also underwent physical and neurodevelopmental assessments on the Denver scale. Outcomes were compared with the reported natural history of infantile neuronal ceroid lipofuscinosis and that of affected older siblings. This trial is registered with ClinicalTrials.gov, number NCT00028262. Patients were followed up for 8 to 75 months. MRI showed abnormalities similar to those in previous reports; brain volume and N-acetyl aspartic acid (NAA) decreased steadily, but no published quantitative MRI or MRS studies were available for comparison. None of the children acquired new developmental skills, and their retinal function decreased progressively. Average time to isoelectric EEG (52 months, SD 13) was longer than reported previously (36 months). At the first follow-up visit, peripheral leukocytes in all nine patients showed virtually complete depletion of GRODs. Parents and physicians reported less irritability, improved alertness, or both in seven patients. No treatment-related adverse events occurred apart from mild gastrointestinal discomfort in two patients, which disappeared when liquid cysteamine bitartrate was replaced with capsules. Our findings suggest that combination therapy with cysteamine bitartrate and N-acetylcysteine is associated with delay of isoelectric EEG, depletion of GRODs, and subjective benefits as reported by parents and physicians. Our systematic and quantitative report of the natural history of patients with infantile neuronal ceroid lipofuscinosis provides platform for future assessment of experimental treatments; (b) Since nonsense mutations account for 5-70% of all genetic disorders. In the United States, nonsense mutations in the CLN1/PPT1 gene underlie >40% of the patients with infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative lysosomal storage disease. We sought to generate a reliable mouse model of INCL carrying the most common Ppt1 nonsense mutation (c.451C>T) found in the United States patient population to provide a platform for evaluating nonsense suppressors in vivo. We knocked-in c.451C>T nonsense mutation in the Ppt1 gene in C57 embryonic stem (ES) cells using a targeting vector in which LoxP flanked the Neo cassette, which was removed from targeted ES cells by electroporating Cre. Two independently targeted ES clones were injected into blastocysts to generate syngenic C57 knock-in mice, obviating the necessity for extensive backcrossing.Generation of Ppt1-KI mice was confirmed by DNA sequencing, which showed the presence of c.451C>T mutation in the Ppt1 gene. These mice are viable and fertile, although they developed spasticity (a clasping phenotype) at a median age of 6 months. Autofluorescent storage materials accumulated throughout the brain regions and in visceral organs. Electron microscopic analysis of the brain and the spleen showed granular osmiophilic deposits. Increased neuronal apoptosis was particularly evident in cerebral cortex and abnormal histopathological and electroretinographic (ERG) analyses attested striking retinal degeneration. Progressive deterioration of motor coordination and behavioral parameters continued until eventual death. Our findings show that Ppt1-KI mice reliably recapitulate INCL phenotype providing a platform for testing the efficacy of existing and novel nonsense suppressors in vivo; (c) Neurodegeneration is a devastating manifestation in the majority of >50 lysosomal storage disorders (LSDs). Neuronal ceroid lipofuscinoses (NCLs) are the most common childhood neurodegenerative LSDs. Mutations in 13 different genes (called CLNs) underlie various types of NCLs, of which the infantile NCL (INCL) and congenital NCL (CNCL) are the most lethal. Although inactivating mutations in the CLN1 gene encoding palmitoyl-protein thioesterase-1 (PPT1) cause INCL, those in the CLN10 gene encoding cathepsin D (CD) underlie CNCL. PPT1 is a lysosomal thioesterase that cleaves the thioester linkage in S-acylated proteins required for their degradation by lysosomal hydrolases like CD. Thus, PPT1 deficiency causes lysosomal accumulation of these lipidated proteins (major constituents of ceroid) leading to INCL. We sought to determine whether there is a common pathogenic link between INCL and CNCL. Using biochemical, histological and confocal microscopic analyses of brain tissues and cells from Cln1-/- mice that mimic INCL, we uncovered that Cln10/CD is overexpressed. Although synthesized in the endoplasmic reticulum, the CD-precursor protein (pro-CD) is transported through endosome to the lysosome where it is proteolytically processed to enzymatically active-CD. We found that despite Cln10 overexpression, the maturation of pro-CD to enzymatically active-CD in lysosome was disrupted. This defect impaired lysosomal degradative function causing accumulation of undegraded cargo in lysosome leading to INCL. Notably, treatment of intact Cln1-/- mice as well as cultured brain cells derived from these animals with a thioesterase-mimetic small molecule, N-tert-butyl-hydroxylamine, ameliorated the CD-processing defect. Our findings are significant in that they define a pathway in which Cln1 mutations disrupt the maturation of a major degradative enzyme in lysosome contributing to neuropathology in INCL and suggest that lysosomal CD deficiency is a common pathogenic link between INCL and CNCL.
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{
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Core 3 - Clinical Needs Assessment and Dissemination Core 3 Overview The primary function of Core 3 is two fold. First, Core 3 is tasked with providing needs assessment in areas anticipated to advance the field of point-of-care technologies for neurologic based diseases and emergencies. This needs assessment function is two facetted covering both the need to have adequate understanding of needs assessment on proposals submitted to the Center for funding consideration and the need to have larger picture understanding of unmet clinical needs the will effect the future direction of the POC-CENT Program. The second primary function of Core 3 is to insure adequate dissemination or POC-CENT results and accomplishments to the community at large. The team members for this Core are: PI;Raj Narayan, Co-Pi;Ed Jauch, Fred Beyette, Dan Kanter, Lori Shutter, and Ton DeGrauw. The Core PI is an established clinical trialist with extensive experience in head injury research. He is the Chair of Neurosurgery and a leader on the worldwide stage for neurosurgical developments. Dr. Narayan has assembled a blue ribbon team of physicians, scientists and engineers to perform clinical needs assessment to evaluate projects that could enter the POC-CENT pathway. The team covers the breadth and depth of all the major neurological issues that could be expected. Description of the Core 3 Operations. The Team will meet quarterly or more often as needed. Clinical needs assessment will be done for two purposes. First, the team will work with the Core 2 team to provide evaluation of needs assessment for each project proposal submitted for funding consideration. Second, the team will generate an annual report that details the current clinical needs priorities for point-of-care technologies in the field of neurologic emergencies. Much of the clinical needs assessment will be done using a metrics of priorities and importance. This metrics is presented below. Are there other diagnostic options available no = 1, yes = 10 Will faster information mean better care no = 1, yes =10 Will better care improve outcome no = 1, yes = 10 Are substantial number of patients affected no = 1, yes = 10 Will specialized centers be needed no = 1, yes = 10 Will physicians embrace the technology no = 1, yes = 10 Will nursing or paramedical staff embrace the technology no = 1, yes = 10 Will there be an opportunity to impact other disciplines no = 1, yes =10 Will it require extensive in servicing and training no = 1, yes = 10 Will it save time for the clinical personnel no = 1, yes = 10 Will it save the hospital/practice money no = 1, yes =10 TOTALS: 10 to 100 Table 5: Clinical needs assessment criteria While it is sometimes difficult to assign a numerical score to subjective items as perceived by our experts, it is important to be able to compare projects based on the clinical need. The expertise and subjective input of the experts will be encouraged with making final decisions and recommendations for all projects.
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{
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The goal of this proposal is to identify biosynthetic enzymes catalyzing reactions leading to the formation of gossypol in the cotton plant. Information of this kind can be useful in the genetic removal of gossypol from Gossypium hirsutum through the selection of mutants lacking these enzymes. The proposal calls for the purification and structural analysis of a cotton root prenyltransferase which catalyzes the formation of farnesylpyrophosphate and a cotton root cyclase which utilizes the farnesylpyrophosphate in the formation of a cyclic sesquiterpenoid, a precursor of gossypol. In addition, experiments are proposed which can establish if gossypol is a phytoalexin, and if this phytoalexin property is due to a de novo synthesis of gossypol or simply a formation of a "free"-active form of gossypol from a "bound"-inactive form.
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{
"pile_set_name": "NIH ExPorter"
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The overall objective of the proposed research is to elucidate the molecular mechanism of sensory processing in E. coli and S. typhimurium chemotaxis. Sensory-motor regulation in bacteria represents a useful model system for the study of receptor- mediated control of cell behavior. Receptor activity is controlled by 2 enzymes, an S-adenosylmethionine:glutamate methytransferase and a glutamyl methylesterase. Genetic studies have identified 4 additional cytoplasmic proteins required for chemotaxis, the products of the cheA, cheW, cheY, and cheZ genes. The specific goals in this grant period include determining the structure-function relationships which underlie the activities of the receptor modification enzymes. What is the role of the regulatory domain of the esterase? Is a thioester adduct between the receptors and the esterase important in chemotaxis? What is the physiological significance of the methylation reactions in chemosensing? The biochemistry of the auxiliary signalling proteins will also be investigated. Specific projects include determining the role of CheA in the regulation of the methylesterase, determining the possible role of nucleotides in CheW function, and elucidating the mechanism and significance of CheZ methylation. The coordinate functioning of these components will be investigated using a variety of genetic approaches, including the selection of second site mutations which compensate defined lesions in critical structural features of the Che proteins. Finally, attempts will be made to extend results obtained with chemotaxis to cognate regulatory systems in bacteria including the apparatus in B. subtilis which triggers the developmental switch from vegetative growth to endospore formation, and the mechanism in E. coli which regulates porin expression.
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{
"pile_set_name": "NIH ExPorter"
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Age-associated changes in immune function in humans and animals are quite important with regard not only to the general health of aged persons but also to the general features of the immune system itself. Elderly subjects have been shown to be more susceptible to viral and bacterial infections and are believed to be more susceptible to cancer. There have been a number of hypotheses for the diminished immune responses observed in elderly subjects including involution of the thymus, active immunosuppression, replication senescence of immune cells, cellular signaling defects, and alterations in cytokine expression profiles. A series of clinical studies has revealed that elderly subjects, in contrast to their younger counterparts, exhibit poor cellular and humoral immune responses to vaccines even in the presence of standard adjuvants. Currently, many laboratories are focusing their research efforts into developing more effective stimulants for use with known vaccines to be tested with elderly populations. However, the poor description of alterations in innate and acquired immune function during the aging process has limited therapeutic intervention. The current project utilizes peripheral white blood cells obtained from normal healthy volunteers of different ages to gain insight into the biological, biochemical, and molecular mechanisms underlying age-associated changes in human immune function. In comparison with immune cells obtained from younger individuals, aged leukocytes also display distinctive patterns of protein phosphorylation, cytokine synthesis and gene expression, effects on cell migration and trafficking, and cell-cycle progression. As several immune subpopulations (e.g., CD28-, CD25+) have been shown to be dramatically increased in the circulation during various disease states (including arthritis, AIDS, and aging), we believe that more detailed molecular and biochemical analysis of these subsets will not only yield valuable information about the immune deficits associated with aging and disease but may also lead to possible immunotherapeutic interventions to boost immune responses.
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{
"pile_set_name": "NIH ExPorter"
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PROJECT SUMMARY/ABSTRACT Improving the long-term survival of kidney transplants (KT) is a national priority in the US. The prevalence of end-stage renal disease is increasing, and the organ shortage is growing. Unfortunately, long-term graft and patient survival has not improved since the 1990s. The half-life of a deceased donor kidney is currently only fifteen years. The most common cause of kidney transplant failure is patient death. In order to address this health crisis, the transplant community needs to focus on modifiable risk factors for patient death and other adverse long-term outcomes after kidney transplantation, such as frailty. Frailty is a clinical syndrome characterized by decreased physiologic reserve and is common in patients with chronic kidney disease (CKD). Frailty prior to KT has been associated with increased post-transplant mortality and has been shown to be modifiable in non-transplant patients. However, there is a significant knowledge gap regarding frailty after KT, including risk factors for its development, biomarkers with which to identify it, and interventions with which to improve it. Cellular senescence is an exciting new area of frailty research that directly applies to these deficits. During the process of senescence, metabolic stressors cause cells to enter a state of permanent growth arrest. Senescent cells accumulate throughout the body and secrete factors collectively called the senescence- associated secretory phenotype (SASP) which induce formation of other senescent cells and cause surrounding tissue damage. Cellular senescence is a mechanism of aging and age-related diseases such as frailty. Components of the SASP serve as biomarkers of frailty in non-transplant populations and may help us identify KT recipients at high risk of functional decline and premature death. In addition, frailty biomarkers could ultimately serve as surrogate endpoints in clinical trials designed to improve frailty. The overall objective of this application is to 1) identify frailty trajectories after kidney transplantation, 2) identify biomarkers of frailty after kidney transplantation, and 3) conduct a phase II clinical trial examining the preliminary efficacy, feasibility and acceptability of an exercise intervention on post-transplant frailty. The proposed K23 application involves the use of innovative biomarkers and behavioral interventions to improve frailty and long-term outcomes after KT, a priority for the NIDDK which focuses on bridging translational research gaps to improve the health and quality of life of patients with CKD. The candidate has exceptional resources available to her: a multidisciplinary team of expert mentors; access to a large volume of transplant patients; and excellent career development activities, i.e., formal courses and workshops in statistical methods, biomarker development, and behavioral clinical trials. Together, these resources will allow the candidate to achieve her long-term goal of becoming an independent investigator and nationally recognized expert on the use of biomarker technology and behavioral interventions to improve frailty and long-term outcomes after KT.
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{
"pile_set_name": "NIH ExPorter"
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OTHER PROJECT INFORMATION - Project Summary/Abstract This proposal seeks support to add detailed measures of health and economic security to a long-term follow-up study of low-income middle and older aged adults participating in a unique randomized neighborhood mobility experiment know as Moving to Opportunity (MTO). The MTO demonstration used a randomized lottery to offer some public housing residents but not others the chance to relocate to less disadvantaged neighborhoods using a rental housing subsidy. MTO experimentally generated large difference in neighborhood characteristics for otherwise comparable groups of low-income, disproportionately minority families. At the time of our proposed survey fieldwork (10 to 12 years after random assignment) 31% of the adult sample will be age 50 or older, with a mean age of 46.8. An interim study found that compared to the control group, adults in the treatment groups had substantially lower estimated prevalence of serious mental illness 4-7 years after randomization, were 20% less likely to be obese and showed signs of improved diet and exercise, but had no differences in labor market outcomes. However environmental toxicity, dietary quality, physical activity and the burdens of psychosocial stress are difficult to capture through surveys. The effects of MTO on economic security could potentially increase over time due to health improvements or if adults become more socially integrated into their new areas, but the possibility of convergence across groups in neighborhood attributes over time suggests labor market impacts could instead attenuate over time. This proposal has four main aims: " To expand our long-term data collection to administer to all MTO adults a set of detailed assessments of health (including blood samples to measure disease precursors such as cholesterol, C-reactive protein, and glycosylated hemoglobin)and DSM-IV mental health disorders, and indicators of economic security such as earnings, savings, assets, and participation in programs such as TANF or SSI. The interim MTO impacts on diet, exercise and obesity are about as large as those found with public health lifestyle interventions, which have been shown to impact biomarkers of the sort we propose to collect for MTO. " To estimate long-term effects of the MTO experiment on these outcomes in the total sample and sub- samples defined by age, gender, and other baseline risk factors. " To decompose the total effects of the intervention on these outcomes and narrow down the the set of behavioral mechanisms through which the aggregate effects occurred. " To develop a public MTO data archive that is made available for secondary analysis. The overarching goal is to better understand the effects of MTO and neighborhood mobility on adult long-term health and economic security as well as the processes by which these outcomes are affected. This study will help shed light on larger disparities in health and economic outcomes across race and class lines. NIA funding is essential to administer the full range of outcomes described here to all MTO adults.
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{
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Free-running and light-induced circadian activity rhythms of wolf spiders will be measured using digital circuitry and punched paper tapes for computer analysis. The point will be to determine intensities of a number of different wavelengths of light which will have the same phase-shifting effect upon free-running (dark) activity, that is, which will appear equally bright. Such intensities will allow determination of behavioral spectral sensitivities for comparison with electrophysiologically measured spectral sensitivities of these animals' eyes.
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{
"pile_set_name": "NIH ExPorter"
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Our long-term goal is to understand the molecular and biochemical mechanisms of RNA interference (RNAi), a conserved post-transcriptional gene-silencing mechanism mediated by microRNA (miRNA) and small interfering RNA (siRNA). These tiny regulatory RNAs play critical roles in many fundamental biological processes. In particular, misregulation of miRNAs has recently been linked to human diseases, such as cancer. Moreover, RNAi has been widely used as a powerful gene-silencing tool to perform functional genomic studies in multiple model systems. The specificity and potency of RNAi highlight its potential for developing novel and efficient therapeutic avenues to treat human diseases. We have previously identified two Dicer complexes, DCR-1/R3D1 and DCR-2/R2D2, as the respective miRNA- and siRNA-generating enzymes in Drosophila. In addition, the DCR-2/R2D2 complex binds nascent siRNA and facilitates its incorporation into the siRNA-induced silencing complex (siRISC). Based on these studies, the goal of this proposal is to address several outstanding questions in the Drosophila RNAi pathway with an emphasis on the miRNA pathway. We will investigate the inherent specificity and mechanism of miRNA biogenesis (Aim 1). We will test our hypothesis that the DCR-1/R3D1 complex senses asymmetry of miRNA and facilitates miRNA loading onto its effector complex, miRISC (Aim 2). Finally, we will develop a reconstitution assay to explore the mechanism of miRISC assembly and to identify and characterize new components of miRISC assembly (Aim 3). These studies will significantly advance our understanding of the Drosophila RNAi pathway. The origins of human diseases, such as cancer, can be generally attributed to loss-of-function of important genes (tumor suppressor genes) and/or gain-of-function of pathological genes (oncogenes). For unknown reasons, RNAi in human cells is less efficient than RNAi in Drosophila cells. Thus, comparison of the Drosophila and human systems will allow us to identify the underlying differences and, hopefully, to optimize RNAi in human cells. This could lead to the development of novel and efficient small RNAs-based gene-silencing methods to cure human diseases by specifically shutting down pathological genes. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Inflammatory bowel disease (IBD) encompasses Crohn's disease and Ulcerative colitis, diseases the cause of which is currently unknown. In recent years, tremendous progress has been made towards understanding the pathogenesis of IBD. The evidence strongly points to a role for both a microbial and an immune component. Several species of pathogenic bacteria residing within the gut lumen have been implicated, and these include invasive Escherichia coli, Helicobacter, Clostridium and Enterococcus species. Furthermore, increased levels of the inflammatory cytokines interieukin (IL)-17 and IL-23 have been reported within the intestinal mucosa. These two cytokines are critical for the function and maintenance of T helper type 17 (THI 7) cells, a recentiy identified subset of helper T cells that play important roles in host defense and organspecific autoimmune inflammation. Genome wide array analyses have identified prominent IBD susceptibility genes involved in the IL-23-TH17 immune response pathway, immune suppression, and innate immunity to microbial pathogens. Collectively, these studies point to an imbalance in the symbiotic relationship between host and microbe, such that loss in tolerance to the intestinal microbiota and increased TH17 related inflammation are now considered to be responsible for the pathogenesis of IBD. The exact mechanisms for why and how this occurs in patients with IBD is unclear. Our recent identification of one physiological trigger that drives THI 7 responses within the intestinal lamina propria provides a clue. We found that innate immune recognition of apoptotic intestinal epithelial cells dying during an infection induces differentiation of naive CD4 T cells into effector THI 7 cells. Here we aim to test our hypothesis that an inability to protect the intestinal epithelium during infections with damaging and invasive enteric pathogens serves as an instigator of IBD. We will characterize the antigen specificities of infection-induced THI 7 response to self and non-self antigens, and define the regulatory mechanisms that control self-specific populations. We will ask whether an absence or a dysfunction of these regulatory populations within a genetic susceptibility background for IBD, leads to failure in protecting and repairing the epithelium damaged from infection, progressive loss of barrier functions against luminal bacteria, and an inability to suppress the aggravated inflammatory response.
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{
"pile_set_name": "NIH ExPorter"
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Acute myeloid leukemia (AML) is a heterogeneous group of genetically diverse hematopoietic malignancies with variable responses to treatment. Around 10% of AMLs are involved in chromosomal rearrangements of the mixed lineage leukemia (MLL) gene with over 60 fusion partners. The critical feature of MLL-rearrangements is the generation of a chimeric transcript consisting of 5' MLL and 3' sequences of a partner gene (80% involving AF9, AF6, AF10, ELL or ENL in AML). The prognosis of MLL-associated leukemia is poor. A group of important oncogenes, including homeobox A (HOXA) genes, MEIS1, FLT3, MYB, and MYC, are frequently up-regulated in MLL-associated leukemias, and play a key role in the self-renewal of leukemia stem cells (LSCs) carrying MLL-rearrangements. However, clinically significant therapies have not been developed to effectively target these genes yet. Thus, better understanding of the molecular mechanisms underlying the pathogenesis of MLL-associated leukemia, and the development of effective therapeutic strategies based on such understanding, are urgently needed. MicroRNAs (miRNA) are a class of small, non- coding RNAs that play important roles in post-transcriptional gene regulation. Very recently, we reported that miR-150 is significantly down-regulated in most AML cases, and its repression is critical for MLL-AF9-mediated cell transformation and leukemogenesis; miR-150 functions as a pivotal tumor-suppressor gatekeeper in the MLL-fusion/MYC/LIN28?miR-150?FLT3/MYB/HOXA9/MEIS1 signaling circuit, through targeting FLT3/MYB directly and MYC/LIN28/HOXA9/MEIS1 indirectly (Jiang X., et al. Cancer Cell. 2012). Hypothesis: miR-150 is required for both development and maintenance of MLL-rearranged AMLs and for the self-renewal of the relevant LSCs. Therefore, the restoration of miR-150 expression/function holds significant potential to be clinically applicable to treat this type of presently therapy-resistant disease. Specific Aims: 1) To determine whether repression of miR-150 is required for both development and maintenance of MLL-rearranged AMLs; 2) To determine whether repression of miR-150 is required for the self- renewal of LSCs of MLL-rearranged AMLs; and 3) To determine whether restoration of the expression/function of miR-150 (delivered by nanoparticles) is an effective new strategy for treating MLL-rearranged AMLs. Study Design: 1) We will use mouse bone marrow transplantation (BMT) models to determine whether ectopic expression of miR-150 can significantly inhibit both development and maintenance of all five major sub- types of MLL-rearranged AMLs (i.e., MLL-AF9, -AF6, -AF10, -ELL and -ENL). 2) We will conduct both competitive repopulation and limiting dilution assays to determine whether ectopic expression of miR-150 can significantly inhibit the self-renewal of relevant LSCs. 3) We will develop novel targeted nanoparticles based on FLT3L (FLT3 ligand)-directed dendrimers complexed with miR-150 oligos, followed by assessment of their specificity and efficacy in targeting/treating MLL-rearranged AMLs both in vitro and in vivo.
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{
"pile_set_name": "NIH ExPorter"
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PROJECT SUMMARY/ABSTRACT Cleavage of amyloid precursor protein (APP) by b-site APP cleaving enzyme-1 (BACE-1) is the rate- limiting step in production of Ab, whose deposition is the pathological hallmark of Alzheimer?s disease (AD). Despite a rapidly growing burden of health care for the aging United States, there is a fundamental gap in understanding how trafficking and mutations of APP influence neuronal function and contribute to AD pathogenesis. Continued existence of this gap represents a critical problem because, until it is filled, AD prevention and treatment based on molecular understanding of the disease progression remains inaccessible. Since neurons can grow axons that are up to a meter long, continuous imaging of APP trafficking and processing in live neurons at the single-molecule level requires extremely photostable fluorophores. Our lab has recently developed a new class of upconversion nanoparticles (UCNPs) that are immensely photostable over months. The overall objective of this project is to use our novel photostable UCNPs to perform single-molecule imaging of the trafficking and processing of APP in live human induced neurons (iNs) ? an excellent model system as many human diseases are not fully recapitulated in mouse neurons. The central hypothesis is that mutations of APP lead to impaired axonal transport and render APP more vulnerable for b-cleavage by BACE-1. This hypothesis has been formulated on the basis of previous work on culture mouse neurons and transgenic mouse models. The rationale for the proposed research is that ultralong-term single molecule imaging of WT and mutant APP in human iNs will reveal axonal transport defects caused by AD-associated mutations, providing important insights into their relationship to AD. Guided by strong preliminary data on the novel experimental platform, the hypothesis will be tested by pursuing the three specific aims: 1) Measure the trafficking dynamics of endocytosed APP in human iNs; 2) Determine how axonal transport is impaired by mutations of APP in human iNs; and 3) Visualize the association dynamics of APP and BACE-1 in human iNs. A battery of techniques including single-molecule imaging, nanotechnology, biochemistry and stem cell technology will be used to interrogate APP trafficking. The approach is innovative because it departs from the status quo by utilizing extremely photostable UCNPs to perform long-term single- molecule tracking, novel analysis for non-invasive determination of motor number, and the use of human induced neurons. The proposed research is significant because it is expected to characterize in depth the trafficking and association dynamics of APP and BACE-1 with unprecedented spatiotemporal resolution, as well as uncover the extent to which APP mutations impair axonal transport, thereby shedding light on future prevention and treatment of AD.
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{
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PROJECT SUMMARY Project II. MicroRNAs in Anesthetic Cardioprotection (PI: Mingyu Liang) The goal of Project II is to test the hypothesis that miR-21 contributes to cardioprotection conferred by anesthetics in animal models and in human cardiomyocytes. MicroRNAs are endogenous small RNA molecules that regulate a wide range of cellular functions primarily through reducing the abundance of target proteins. MicroRNAs have been shown to be powerful regulators of cardiac development, injury, and remodeling. However, the role of microRNAs in anesthetic cardioprotection and its impairment by diabetic conditions remains unknown. We and other investigators have reported that microRNA miR-21 contributes to ischemic preconditioning of the heart and the kidney and to xenon-induced protection of the kidney. Importantly, we have obtained very exciting preliminary data strongly supporting an important role of miR-21 in isoflurane-induced cardioprotection and its impairment by diabetic conditions in animal models and in human cardiomyocytes generated from non-diabetic individual-derived induced pluripotent stem cells (iPSCs) (N-CM) or from a type 2 diabetic patient-derived iPSCs (T2-CM). Based on these novel findings, we propose in Aim 1 to use gene knockout or transgenesis to further examine the role of miR-21 in anesthetic cardioprotection in animal models and in possible restoration of anesthetic cardioprotection in models of type 2 diabetes. In Aim 2, we will translate the findings to human using patient-specific cardiomyocytes including N-CM and T2-CM. Finally, in Aim 3, we will investigate the molecular mechanisms involved by examining miR-21 target genes and their downstream pathways linked to mitochondrial function and mitochondrial permeability transition pore dysregulation as well as additional mechanistic pathways. Project II shares the central theme of the PPG, which is to investigate cellular mechanisms underlying anesthetic cardioprotection and its impairment by diabetic conditions. Project II will interact extensively with Projects I and III by studying shared mechanistic pathways leading to mitochondrial dysfunction and cell death. Results of Project II will be used to test, validate, and extend the mathematical models developed in Project III.
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{
"pile_set_name": "NIH ExPorter"
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Primary Objective To determine, in a population of 5-9 year old children with asthma, if regular use of either of two classes of anti-inflammatory medications (inhaled corticosteroids or cromolyn sodium), compared to regular bronchodilator medication and to each other, results in greater lung function and less bronchial hyperresponsiveness over a five year period. Secondary Objectives To determine if either of the anti-inflammatory medications, compared to bronchodilator medication and to each other, results in less patient morbidity, measured by frequency of asthma symptoms, days of limited activity, and days lost from school. To determine if either of the anti-inflammatory medications compared to regular bronchodilator medication and to each other results in less use of health care resources such as emergency room visits or hospitalizations. To compare the long term safety and side effects of the two classes of anti-inflammatory medications, compared to bronchodilator medication and to each other. To compare the effects of the two classes of anti-inflammatory medications, compared to bronchodilator medication and to each other, on physical and psychosocial development, and on quality of life.
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{
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Many biologically important substances contain the imidazole ring with varying degrees of sustitution. We are looking at the reaction of cyanothioformamide as a possible starting material for the synthesis of uniquely substituted imidazoles. 1) The reduction of 1,2-dithiones with H2S is expected to give new 1,3-disubstituted-4-thiohydontoins. 2) The reaction of isocyanates with cyanothioformamides gives a dimeric disulfide by reduction. The scope and pathway for this reaction will be studied.
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{
"pile_set_name": "NIH ExPorter"
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Genomic stability is vital to the health of the individual and the preservation of the species. This stability can be threatened by DNA damage from endogenous and exogenous sources. However, mutations, which are heritable sequence changes in the DNA, can also arise as a result of errors made during replication of undamaged DNA. Recently an entirely new family of error- prone DNA polymerases has been discovered. Found in all three domains of life, these polymerases are beneficial because they can replicate past DNA lesions, but they are also potentially detrimental because they make frequent errors, even on undamaged DNA. If the activities of these polymerases are not controlled, they could be a potent source of the mutations that lead to genetic disorders such as cancer. The hypothesis underlying the proposed research is that cells can and must control the activities of their error-prone DNA polymerases. To test this hypothesis, pathways that control the activity of E. coil's error-prone DNA polymerase IV (Pol IV) will be found and characterized. The specific aims are: (1) to identify new regulatory factors that affect the mutagenic activity of Pol IV; (2) to characterize the proteins and pathways regulating Pol IV levels or activity; and, (3) to further characterize the regulatory factors that are already identified. Because Pol IV is a close homologue of eukaryotic error-prone DNA polymerases, higher organisms, including humans, may use similar control mechanisms.
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{
"pile_set_name": "NIH ExPorter"
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[unreadable] Now in its seventh year, the Regenerate international conference and exposition is a collaborative effort that unites leaders within the tissue engineering and regenerative medicine communities, offering a premier educational program for basic scientists, clinicians, students, business leaders, entrepreneurs, and representatives of government funding agencies. This annual endeavor is organized in an effort to foster interactions that will accelerate development of new technologies to benefit patients worldwide. This [unreadable] meeting is intended to unite leaders within these multidisciplinary fields and offer a premier educational program. It is co-sponsored by the University of Toronto, the institutional host of Regenerate 2007. The purpose of this proposal is to provide travel and registration support for graduate students and postdoctoral fellows. An important consideration is the diversification and participation of all students. Toward this end, funds are allocated for disadvantaged and under-represented students to attend the [unreadable] meeting. Graduate students and post-docs need to be exposed to the broader tissue engineering and regenerative medicine research advances; Regenerate 2007 will provide the opportunity to be exposed to cutting-edge research and to interact with the scientists and clinicians involved in the field. In Regenerate 2007, the multi-disciplinary leaders have been invited to give a series of plenary addresses throughout the conference and include: stem cell biology, imaging, transplantation and translation. [unreadable] The primary purpose of this proposal is to provide support for graduate students and post-doctoral fellows to attend Regenerate 2007. We plan to provide travel awards for approximately 50 graduate students and post-doctoral fellows to attend and actively participate in Regenerate 2007 through oral and poster presentations, networking events and co-chairing of sessions. Meeting participation provides a significant networking opportunity for students as they prepare for post-graduate careers and the career center will enhance this experience as employers interview potential employees on site, at the meeting. We have designed this series of events, in collaboration with student leaders, to achieve these goals. [unreadable] Regenerative Medicine promises to change the way we approach medicine, where symptoms are not merely treated but diseases are reversed. Regenerative Medicine will ultimately obviate the need for autografting or transplantation from a donor. The successful translation of Regenerative Medicine to clinical application has already been achieved for some tissues, providing the landscape for its translation for other tissues and organs in the future. To achieve these goals, the stakeholders in government, industry and academia must come together, as they will at Regenerate 2007. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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This component will continue elucidating the molecular mechanisms underlying "ontogenesis" of kappa opioid receptor (KOR), i.e. production of KOR protein during developmental stages and differentiating neurons. Studies in the previous funding cycles have delineated several regulatory pathways for the control of KOR mRNA production during developmental stages, principally transcriptional control. This concludes the first phase of studies focusing on the regulation of KOR mRNA synthesis, a hall markd of KOR neurogenesis involving signaling pathways of retinoic acid (vitamin A) and nitric oxide and requires chromatin remodeling of KOR gene regulatory regions, as well as more recent findings in epigenetic control. Importantly, ontogenesis of KOR appears to be controlled, most crucially, by post-transcriptional mechanisms such as mRNA stability, transport and translation. This renewal component will focus on translational mechanism by extending from our preliminary studies that have identified Netrin-1 as a translational stimulator for KOR, and Grb7 as a translational represser of KOR. Two specfiic aims are: 1. To elucidate the mechanism that activates KOR translation via Netrin-1/Grb7 pathway. We will focus on i) regulation of translational initiation of KOR by Grb7, including studies of its molecular and biochemical features and funcitonal domains, and KOR RNA sequences bound by Grb7 which can be activated by Netrin-1, and ii) specific translational initiation step that is targeted by Grb7 including initiation factors and subcellular distribution and possible circularization of KOR mRNA. 2. Pharmacological and physiological relevance of Grb7/Netrin-1 signaling to KOR ontogenesis. We will i) perform gain- and loss-of-function studies to validate the relevance of Netrin-1/Grb7 in KOR ontogenesis using stem cells and primary neurons, and ii) examine the physiological relevance of KOR synthesis in neuronal activity such as in a specific pain circiitry and during nerve injury or as a stress response.
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{
"pile_set_name": "NIH ExPorter"
}
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Targeting gene product in tumors is very important for systemic gene delivery because only a portion of genes are successfully delivered into tumors by using the most successful systemic tumor-targeted gene delivery technology. More importantly, some gene products such as IL-12 are more effective when co-localized with tumors, compared to staying in blood or other organs One simple approach to targeting gene product into tumors is to generate and use fusion genes encoding IL-12 and tumor-targeted mini-peptide. Unexpectedly, we found that none of the known tumor-targeted peptides boost IL-12-induced anti-tumor efficacy. In the current grant period, however, we discovered the novel peptide VNTANST, which boosts accumulation of IL-12 protein into tumors when a fusion gene encoding IL-12-VNTANST, was administered into muscles distant from the tumors. Using a gene therapy approach for DNA delivery and using a fusion gene product IL-12-VNTANST for tumor-targeting (tumor-targeted IL-12 gene therapy has shown great potential in inhibiting tumors in multiple tumor models. We also revealed that this tumor-targeting IL-12 (IL-12-VNTANST) binds to vimentin, a protein known as an intracellular protein and increased in metastatic tumors. An intriguing question is how the extracellular IL-12-VNTANST binds to an intracellular protein? Significantly, our preliminary study showed that vimentin is found on the cell membrane of malignant epithelial cells. Our objective in this renewal application is to determine whether membrane vimentin localization is increased in some of the metastatic tumor cells and thus has a significant role in binding to IL-12-VNTANST and boosting IL-12-anti-tumor efficacy. We will also examine how to effectively using this targeted IL-12 gene therapy for treating membrane positive metastatic tumors in various organs using tumor-targeted IL-12 gene therapy. We hypothesize that tumor-targeted IL-12- VNTANST gene therapy has a great potential for treating membrane vimentin positive metastatic tumors. The proposed study is innovative because it may definitively confirm that the membrane vimentin localization plays a key role for tumor metastasis and for boosting the proposed targeted IL-12 gene therapy. Confirmation of membrane vimentin dependence would also allow us selecting proper tumor-bearing patient in the future to perform this therapy, which is so called personalized immune therapy, an important direction of novel therapeutics and a long term of goal of our effort.
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{
"pile_set_name": "NIH ExPorter"
}
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Natural products that interfere in a specific and potent manner with cellular processes have proven to be useful probes in cell biology research. In particular, agents that exhibit specific cellular effects have served as powerful biochemical tools for dissecting molecular mechanisms of signal transduction pathways involved in various cellular functions. This proposal seeks to further exploit the rich potential of marine natural products in this regard building on our previous success with the marine alkaloid pateamine A, a novel eukaryotic protein synthesis inhibitor, in collaboration with Prof. Jun Liu (Johns Hopkins). Principal aims include a highly unified, enantioselective strategy to the complex, bioactive marine alkaloids palau'amine, axinellamine, and styloguanidine from a common cyclopentane intermediate bearing either cyclic urea-hydantoin or bis-cyclic guanidine heterocycles. This strategy is suggestive of one possible biogenesis of these natural products and as a means to study the various hypotheses posited in the literature, we propose the synthesis of singly- and doubly-15N-labelled oroidin and sceptrin for biosynthetic feeding experiments in collaboration with Prof. Ted Molinski (UCSD). A novel and concise cascade process will be studied for the synthesis of the antimetastatic agent agelastatin A enabling cellular receptor isolation studies in collaboration with Prof. Coran Watanabe (TAMU) building on known SAR data. Employing our synthesis of gymnodimine, we propose synthesis of haptens for eventual development of an ELISA assay for this marine toxin in collaboration with Prof. Chris Elliott (UK). The synthetic strategies proposed and completed (in the case of gymnodimine and phakellstatin) will be "put to work" to enable synthesis of biotin and fluorophore conjugates of palau'amine, phakellstatin, and agelastatin A to enable a molecular level understanding of the immunosuppressive and antitumor activities exhibited by these agents and also to begin unraveling the biosynthesis of the oroidin alkaloids. The biomechanistic and biosynthetic studies are enabled by continued, productive collaborations with Prof. Liu and new collaborations with Profs. Watanabe and Molinski. Innovative aspects of this proposal include: (1) Proposed controlled, oxidative cyclizations from the common cyclopentane intermediate providing a highly unified strategy to several complex, oroidin dimers. (2) A novel N-acyliminium cascade process for a concise and versatile synthesis of agelastatin A and congeners further demonstrating the nucleophilic character of imidazolones. (3) Synthesis of `committed,'complex 15N-labelled precursors to provide the first direct data regarding intermediates in the proposed biogenetic pathways to more complex oroidin-alkaloids. (3) Hypothesis-driven biomechanistic studies of spirocyclic imine marine toxins building on our total synthesis of gymnodimine. The end results of these studies will be identification of further useful biochemical tools and potential leads for drug development as demonstrated with pateamine A. PUBLIC HEALTH RELEVANCE: The natural products targeted in this proposal all possess potent biological effects including antibacterial, immunosuppressive, and antitumor effects thus having potential for the treatment of human disease including bacterial infection, inflammation, and cancer. We propose unique, concise synthetic strategies to prepare these natural products and derivatives to address questions regarding the molecular details of their interactions in cells. Subsequent mode of action studies of these compounds including cellular target elucidation will contribute to fundamental studies in cell biology and define the potential of these natural products as drug leads. In addition, the studies proposed will begin to shed light on the biosynthetic pathways used by the producing marine sponges that produce these complex natural products.
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{
"pile_set_name": "NIH ExPorter"
}
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About 2-3% of the world population is infected with hepatitis C virus (HCV). Majority of HCV infections lead to chronic hepatitis. Hepatic steatosis (fatty liver) is recognized as a common histologic feature of chronic hepatitis C and occurs at an average rate of 50%. At molecular level, we and others have observed that HCV gene expression leads to alteration of cellular lipid metabolism. These changes manifest in elevated expression of genes involved in cholesterol/fatty acid biosynthetic pathways. In the current application, we propose investigate the role of HCV-induced oxidative stress in the activation of transcription factors, which include sterol regulatory element binding proteins (SREBP), liver X receptor, (LXR), PPAR1 and PCG-12. Attempts will be made to identify individual HCV gene product(s) playing a key role in steatosis. We further propose to investigate the mechanisms by which HCV gene expression impairs the lipidation of apolipoprotein B-100 (apoB). ApoB represents the major structural component of VLDL. Our recent studies identified the possible ternary complex composed of HCV NS5a protein, apoB and microsomal triglyceride transfer protein (MTP). Whether these protein-protein interactions lead to intervention of apoB lipidation will be investigated. Other possible mechanisms leading to the block in the secretion of apoB will be investigated. The results of these studies will provide unique insight into the ability of HCV gene expression in affecting lipid homeostasis and likely guide the design of future antiviral strategies in the treatment of liver disease associated with HCV infection.
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{
"pile_set_name": "NIH ExPorter"
}
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The Administrative and Outreach Core is responsible for managing the SPORE resources and facilitating communications between the SPORE components, other collaborators, and with other SPOREs in the NCI. This is accomplished through a series of SPORE Seminar Series, oversight committees, organized administrative and scientific meetings of SPORE investigators, institutional representatives and external reviewers. In addition, the outreach goals of the Breast Cancer SPORE will be coordinated through this core.
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{
"pile_set_name": "NIH ExPorter"
}
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Our research into the pathophysiology of presbyopia, the age-related loss of the eye's ability to focus on near objects, has found evidence in a monkey model that the aging ciliary muscle is restricted in accommodative movement by its posterior elastic attachments. In the Parent R21 grant application, our goal is to determine the role of the posterior zonule in accommodation and presbyopia. In this supplement application (Notice number: NOT-OD-09-058) we wish to expand our investigation by determining the role of the choroid, another of the ciliary muscle's posterior elastic attachments, in restricting the accommodative movement of the ciliary muscle with age. Human accommodative amplitude (the ability of the eye to focus on near objects) declines progressively with age, beginning in the second decade of life and perhaps earlier, and is completely gone by age 50-55 years.[1] No individual appears exempt, making presbyopia (literally, "old eye") the most common ocular affliction in the world. Although certainly not a blinding condition, and correctable by various optical means, presbyopia's cost in devices and lost productivity is substantial.[2] Although much useful and relevant information has been garnered from studies in living and postmortem human eyes, the invasive techniques required to answer some of the most critical questions cannot be employed in the living human. While the eyes of subprimate species either do not accommodate or accommodate by mechanisms very different from that of the human,[3] the accommodative apparatus of the rhesus monkey eye is very similar to that of the human eye.[4] [5] [6] Rhesus accommodation declines on a relative time scale that is essentially identical to that of the human.[5] Our group has utilized the rhesus monkey to contribute significant new information relevant to presbyopia pathophysiology. In this model, we have demonstrated that the ciliary body excursion during accommodation diminishes with age, probably due to an age-related decrease in elasticity of the posterior attachments.[7, 8] This restricts the degree of accommodative amplitude. We have also shown that the movement of the lens equator decreases with age, again resulting in reduced accommodative amplitude.[7, 8] Further, although histological data from excised postmortem human eyes shows the older human ciliary body at rest in an anterior/inward position, we have preliminary imaging data that suggest this may not be the case in vivo (see Preliminary Studies). Classical teaching attributes presbyopia to "lenticular sclerosis," or "lens hardening,"[2, 9-15] so that the lens cannot change shape, but the definitive mechanism that results in presbyopia remains elusive. In search of a way to restore some degree of accommodative amplitude, we hypothesize that age-related immobility of the muscle is due to posterior restriction. Further, if these posterior restrictions are eliminated, mobility of the muscle can be restored and facilitate the function of accommodating intraocular lenses (IOLs). PUBLIC HEALTH RELEVANCE: Our goal is to determine what role the extralenticular tissues play in the pathophysiology of age- related ciliary muscle immobility in the non-human primate, and to determine whether the resulting model is relevant to human presbyopia. This may be crucial in enabling the function of next- generation intraocular lenses (IOLs).
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{
"pile_set_name": "NIH ExPorter"
}
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This is a resubmission of an application for a K23 award for Dr. Christine Dehlendorf, whose research focuses on how to improve women's use of contraception, with specific emphasis on communication with health care providers about family planning and on racial/ethnic disparities in reproductive health outcomes. She has conducted two primary data collection projects in these areas and has an observational cohort study that includes audiotaping of contraceptive counseling visits currently underway. Her research training includes completion of a fellowship in Family Planning as well as a Masters in Clinical Research. This award will allow her to develop skills in patient-provider communication, decision science, health information technology, implementation and dissemination science, and health care intervention research. Following completion of this award, she will be in a position to establish an independent research program, including applying for an R01 award to fund a randomized controlled trial of a decision support intervention for contraception. UCSF provides an ideal environment for the development of Dr. Dehlendorf's research career. She has assembled a team of mentors from Family and Community Medicine, Obstetrics and Gynecology, Epidemiology and Biostatistics, and Internal Medicine, and she will have ample logistical and practical support from the Department of Family and Community Medicine, where she is already a member of the faculty. Women in the United States have extraordinarily high rates of unintended pregnancy, with poor and minority women disproportionately experiencing this adverse reproductive health outcome. This high rate of unintended pregnancy is caused in part by the under-use and misuse of effective contraceptive methods. As in the United States all non-barrier methods of contraception require consultation with a health care provider, clinicians who provide family planning have an opportunity to positively impact contraceptive use during contraceptive counseling. Little research has investigated this interaction, however, and there have been few attempts to use decision support in the family planning context. Dr. Dehlendorf's specific aims consist of analysis of her observational cohort study of contraceptive counseling to determine which aspects of counseling are associated with continuation of contraception and use of effective contraceptive methods (Aim 1) as well as to study the extent to which there are racial/ethnic disparities in contraceptive counseling (Aim 2);development of an interactive computerized contraceptive decision support tool for use in waiting rooms of clinics providing family planning services (Aim 3);and pilot testing of this tool in four clinics (Aim 4). The development and testing of this computerized contraceptive decision support tool will be informed by the field of implementation and dissemination science, so that if this intervention is found to be successful, it will be appropriate for widespread application. Through completion of these aims, Dr. Dehlendorf will be ready to submit an application for an R01 grant to fund a cluster randomized trial of this intervention. PUBLIC HEALTH RELEVANCE: Approximately 50% of pregnancies in the United States are unintended, which results in poor maternal and child outcomes and contributes to the cycle of disadvantage experienced by racial and ethnic minorities. Improving the use of contraception has the potential to improve this important public health outcome and decrease reproductive disparities. This resubmission proposes the development and pilot testing of an interactive computerized decision support tool for contraception that is targeted towards the needs of diverse populations and is designed to increase the use of effective contraceptive methods, as well as decrease discontinuation of and improve adherence to these methods.
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{
"pile_set_name": "NIH ExPorter"
}
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The synthesis and testing of several new highly lipophilic agents with potential multiphasic antitumor activity will be undertaken. The prototype compounds are designed to probe the physicochemical requisites for chemotherapeutic access to central nervous system tumors. This series may also test a hypothetical link between the mechanisms of action of the nitrogen mustards and the nitrosoureas.
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{
"pile_set_name": "NIH ExPorter"
}
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Schizophrenia is a devastating psychiatric condition that affects up to 1% of the US population. During the previous funding period, we reported that a loss of hippocampal interneuron function, recapitulates the hyperdopaminergic state thought to underlie psychosis in patients. In addition, we have recently provided one of the first reports describing the feasibility of interneuron transplants (derived from fetal medial ganglionic eminence) as a novel therapeutic approach for the treatment of schizophrenia. In these initial studies we focused primarily on the hyperdopaminergic state thought to underlie psychosis. In the present proposal, we will now address the issues of negative and cognitive deficits that are not only severely debilitating, but are not well treated by antipsychotic medications. Towards this end, we plan to examine the role of two separate interneuron populations, namely parvalbumin and somatostatin, in discrete but interconnected neuronal systems (the vHipp and mPFC) that likely underlie these symptoms clusters in schizophrenia. More broadly, this concept is central to understanding dimensions of behavior and neurobiological measures as detailed in the Research Domain Criteria project (RDoC). Taken together, the studies proposed will provide an understanding into the role of discrete interneuron populations in the pathophysiology of schizophrenia and will examine the utility of stem cell transplants as a potential treatment for this disease.
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{
"pile_set_name": "NIH ExPorter"
}
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