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Extracellular matrix (ECM)-degrading proteinases of the matrix metalloproteinase (MMP) gene family have been implicated in the pathogenesis of tumor progression, yet also play a role in normal tissue remodeling. This project will examine at the level of molecular regulation how MMPs alter the microenvironment of mammary gland during development and remodeling. The hypothesis being tested is that the molecular targets of MMPs are essential in the interaction between mammary epithelial cells and their mesenchymal stroma. In the previous grant period, using transgenic mice and mammary cultures, the investigators showed that a critical balance exists between MMPs expressed by stromal cells and mammary epithelial cells to regulate lobuloalveolar development, lactational differentiation and involution, and that misregulation of an MMP results in inappropopriate development, and in neoplastic and malignant alterations. The present proposal addresses the mechanisms by which these MMPs orchestrate the cross-talk between mesenchymal and epithelial compartments. The approach uses parallel in vivo and culture experiments to analyze mammary gland development and dysfunction. Ductal and alveolar mammary epithelial cells and mammary fibroblasts with genetically altered MMP or inhibitor levels will be studied in culture, and in normal or transgenic mice, and analyzed for growth, morphogenesis, differentiation, and invasive behavior. The mechanisms by which MMPs stimulate ductal proliferation, branching and lobuloalveolar development will be compared and contrasted with the mechanisms by which MMPs induce involution and apoptosis in mammary alveolar epithelia cells. In both cases the contribution of mammary stromal cells will be evaluated. The MMPs that mediate these two distinct sets of processes and their molecular targets in the extracellular matrix or on the cell surface will be elucidated. The direct effects of MMPs on the phenotype of epithelial cells and fibroblasts will be studied to evaluate their roles in altering proliferation, inducing fibrosis, invasion and tumorigenesis. These approaches will elucidate how MMPs alter the cellular microenvironment, and what the critical responses are in normal and abnormal mammary gland epithelium and stroma. These studies address an important aspect of women s health, that of how MMPs regulate normal and neoplastic mammary gland function. These studies may form the basis of intervention and therapy in breast cancer, not only in the late malignant stages, but potentially in the premalignant lesions.
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{
"pile_set_name": "NIH ExPorter"
}
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Observational follow-up of the landmark United Kingdom Prospective Diabetes Study revealed that early tight glycemic control had persistent benefits among adults with newly diagnosed diabetes even a decade after the trial had ended. This phenomenon has been dubbed the legacy effect of glycemic control and has not been observed for blood pressure control. The existence of the legacy effect implies that the timing of glycemic control, for example, early vs. late in the disease course, may have clinical consequences. This raises new fundamental diabetes translational research questions. In clinical practice, how soon after diagnosis do patients achieve hemoglobin A1C (A1C) and blood pressure (BP) control and for how long? How do different long-term patterns of control affect future health outcomes? And what do patients believe regarding how and when the timing of therapies to achieve A1C and BP control affects their health outcomes? I am an internist who is committed to exploring whether the legacy effect exists in clinical practice and how to translate this novel observation from clinical trials into improved clinical care. Using a large, observationl cohort of adults from Kaiser Permanente Northern California with newly diagnosed diabetes (1995-2005), I will (Aim 1) characterize common patterns of a) A1C and b) BP control over e10 years of follow-up and (Aim 2) compare complications/mortality rates over e15 years of follow-up among patients with various 10-year patterns of a) A1C and b) BP control. Then in order to develop a theoretical model for how patients perceive the concept of time in their diabetes-related health decisions, I will conduct semi-structured interviews with patients with Type 2 diabetes to (Aim 3) explore their beliefs regarding the timing of A1C and BP control. The results of these studies will be used to inform a future R01 behavioral intervention focused on improving patient outcomes through effective risk communication and appropriately timed efforts to intensify A1C and BP control. During this career development award, I will work closely with my mentor, Dr. Elbert Huang, who is a nationally- recognized investigator in medical decision making for older adults with diabetes and Director of the Quantitative Analysis Core and Enrichment Programs of the Chicago Center for Diabetes Translation Research (P30) at the University of Chicago, and my co-mentor, Dr. Andrew Karter, an accomplished diabetes epidemiologist at Kaiser Permanente Northern California Division of Research, in addition to my advisory panel of leaders in longitudinal data analysis, endocrinology, health economics and behavioral psychology. This proposed research, along with additional training in advanced epidemiology, biostatistics and behavioral psychology, and the expertise of my mentorship team will help me achieve my long-term goal of becoming an independent clinical investigator with expertise in the optimal timing of Type 2 diabetes treatments and its risk communication.
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{
"pile_set_name": "NIH ExPorter"
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PROJECT SUMMARY Stress disturbs many homeostatic functions, including reproduction. While previous studies have extensively examined the role of physical, psychosocial, and immunological stressors on reproduction, to date, the mechanisms remain unknown. Stress impacts multiple levels of the hypothalamic-pituitary-gonadal (HPG) axis, decreasing Gnrh expression and basal gonadotropin (luteinizing hormone) secretion. Recent advances in the field have allowed for an ultra-sensitive LH assay that will be used in the proposed Aims to examine changes in LH pulses induced by stress. The proposed Aims will examine if upstream neuroendocrine peptides regulate the reproductive axis during stress. Two neuropeptides of interest are RFRP-3 and kisspeptin. RFRP-3 is an inhibitory neuropeptide that inhibits production of the HPG axis via a central inhibition of GnRH neurons and thus decreased LH levels. By contrast, kisspeptin is a stimulatory neuropeptide that functions as the pulse generator to increase GnRH output. Together, RFRP-3 and kisspeptin are important regulators of hypothalamic GnRH neurons and may be potential targets during stress. To determine if RFRP-3 and kisspeptin regulate reproduction during stress, it is necessary to examine both males and females. The stress response is highly sexually dimorphic, with females being more sensitive than males. To date, there are no studies that examine sex differences in stress and reproduction. Therefore, the objective of this project is to examine the role of restraint stress in mediating reproduction, namely through RFRP-3 and kisspeptin neurons and LH pulsatility, in both males and females. Aim 1 examines if there is a sex difference in the effects of restraint stress on RFRP-3 and kisspeptin neurons and LH pulses. Aim 2 explores if RFRP-3 neurons are necessary to inhibit reproduction during restraint stress. As previously stated, RFRP-3 neurons are inhibitory to the HPG axis. Thus, this Aim will determine if ablation of an inhibitory signal will be important in mediating stress inhibition of reproduction as well. Aim 3 will identify if stress functions through glucocorticoid receptor (GR) localized on RFRP-3 or kisspeptin neurons. This Aim will provide a potential mechanism if selective ablation of GR on RFRP-3 or kisspeptin neurons abolishes the inhibition of reproduction by stress. The crosstalk of stress and reproduction is critical in identifying mechanisms of reproductive problems associated with stress including metabolic disease (obesity, anorexia nervosa), infertility during in vitro fertilization treatment, and additional reproductive problems such as hypogonadotropic hypogonadism and polycystic ovarian syndrome. Furthermore, identifying if sex differences exist in stress-induced inhibition of reproduction is critical to future experiments as well as treatment options in humans. Overall, this work is critical to understand and improving our understanding of reproductive physiology and its disruption by stress.
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{
"pile_set_name": "NIH ExPorter"
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The long-term goal of this project is to understand what mechanisms are involved in prostate cancer, which may help to discriminate aggressive, fatal types from indolent, less aggressive ones and develop an improved framework for choosing new drug targets. Interrogation of disease mechanisms and risk stratification could be accomplished by leveraging knowledge about inherited risk factors such as germline genetics. The ability to use the majority of inherited genetic information in the clinical setting has been limited because many popular analytic approaches are agnostic to gene structure and the role of gene products in biological mechanisms. This important context is not commonly integrated into studies of genome-wide genetics and often only referenced post-analysis to interpret significant findings. In answer to this problem, the proposed study will utilize pathway analysis, recent innovative approach for the study of germline genetics. This method is adapted from gene expression studies and investigates the effects of genetic variants from an entire set of genes that are related to one another through biological mechanisms or cellular functions, termed pathways. Pathways are treated as the unit of analysis rather than individual genetic variants, thus aggregating association signals and improving statistical power to detect true phenotype associations. The aims of this proposal are to identify the germline genetic pathways involved in aggressive prostate cancer development and disease progression, and determine whether any genes or pathways are shared between the two phenotypes. This will be accomplished by first estimating the combined effects of rare and common variants for each gene (Aim 1a), followed by the organization of these genes into pathways (Aim 1b), which will be tested against each other to identify those involved in aggressive prostate cancer development (Aim 1c). The process will also be conducted for a time to disease progression phenotype (Aim 2a); then genes and pathways involved in the development of both disease outcomes will be identified (Aim 2b). Shared genes and pathways may be promising targets for understanding what differentiates between latent and aggressive prostate cancer. Information about unshared genes and pathways can also highlight mechanisms that are important at only certain phases of cancer development. The interdisciplinary training environment for this proposal will provide the applicant an opportunity to gain technical skills, to build research competency, and to develop content area expertise for her career as a future independent researcher.
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{
"pile_set_name": "NIH ExPorter"
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It is important to characterize hepatosteatosis, the first stage of alcoholic liver disease (ALD), in order to identify therapies to reverse hepatosteatosis and/or prevent progression to more advanced disease stages. Lipid metabolism in hepatocytes is regulated by sterol regulatory element-binding protein-la (SREBP-1a) and peroxisome proliferator-activated receptor-alpha (PPARalpha), The lipid aldehydes 4-hydroxy-2-nonenal (4-HNE) and malondialdehyde (MDA) are generated in response to chronic ethanol consumption, and are capable of covalently modifying proteins. The experiments proposed by this application are designed to test the general hypothesis that covalent modification of the transcription factors SREBP-1alpha and PPARalpha by 4-HNE and MDA alters binding affinity to their respective promoter sequences, as well as their rates of ubiquitin-dependent degradation, both of which may result in lipid accumulation in hepatocytes. Adducts will be identified by tandem mass spectrometry (MS/MS), and promoter binding activity of modified SREBP-1alpha and PPARalpha will be compared to native protein binding by electrophoretic mobility shift assays (EMSA). Ubiquitination and proteasomal degradation will be measured in a rabbit reticulocyte lysate (RRL) system.
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{
"pile_set_name": "NIH ExPorter"
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This grant focuses on one of the most critical yet most poorly understood mechanistic events in urolithiasis, the attachment of urinary crystals by the epithelium of the kidney papillary tip which then serves as maturation sites for urinary stone growth. Stone research literature supports a relationship between tissue injury and stone disease. Their studies on crystal attachment to inner medullary collecting duct cells (IMCD) have demonstrated that calcium oxalate monohydrate (COM), hydroxyapatite (HA), and uric acid (UA) crystals attach to IMCD cells cultures in a structurally specific, saturable and inhibitable manner. Crystal attachment appears to be based on specific atomic interactions between molecular arrays on the surfaces of stone crystals and on the surfaces of lipid rich domains on the surfaces of IMCD cells. They hypothesize that renal epithelial cells will only bind kidney stone crystals if there is a specific perturbation in outer plasma membrane leaflet phospholipid composition and/or structure resulting in the formation of phospholipid domains which are structurally complementary to the molecular arrays on crystal surfaces. Such changes might be associated with renal epithelial injury. Specific Aim I. They propose to correlate the attachment of COM, calcium oxalate dihydrate (COD), HA, and UA crystals to IMCD cells with specific phospholipid composition and structure in the plasma membrane of these cells. They also propose to define the changes in the composition and structure of the phospholipid domains as a function of crystal attachment following PS and PI phospholipid enrichment. Specific Aim II. They propose to study crystal attachment to IMCD cells as a function of I) prior exposure of IMCD cells to ionic conditions at or near supersaturation conditions as would be expected in stone forming patients demonstrating recurrent episodes of crystalluria, ii) exposure of IMCD cells to crystals coated with selective lipids or urinary protein macromolecules identified as potential stone growth inhibitors or promoters, and iii) exposure of IMCD cells to one stone crystal type followed by a second exposure of a second crystal type to assay for lipid domain composition or structure perturbations as primers for more effective crystal attachment. Specific Aim III. They propose to crystallographically model the molecular structures on the surfaces of all five crystals as well as the structures of an assembly of phospholipid headgroups both as single as well as admixed phospholipid structures. They also propose to epitaxially analyze the dimensional basis as well as the specific molecular bonding matches that would support long range interactions between the various stone crystals and molecular structure in lipid-rich domains in the outer leaflet of IMCD cell plasma membranes.
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{
"pile_set_name": "NIH ExPorter"
}
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The overall objective of this proposal is to develop hepatitis C virus (HCV)-specific immunogens that may be useful as prophylactic and/or therapeutic vaccines for the prevention or treatment of chronic HCV infection. Two general nonexclusive approaches are proposed. In the first approach (Specific Aim 1) we propose to develop a prophylactic HCV vaccine designed to elicit neutralizing antibodies to the HCV E2 protein. For this purpose we have developed the woodchuck hepatitis core protein (WHcAg) as a particulate vaccine carrier platform. The WHcAg platform is capable of accommodating a variety of inserted B cell and CD4 + T cell epitopes and elicits extremely high levels of antibodies to the inserted B cell epitopes and primes insert-specific CD4 v T cells. Three categories of E2-specific neutralizing B cell epitopes will be inserted into the WHcAg vaccine platform: (a) highly conserved, non-HVR1 E2 epitopes that we have identified; (b) consensus sequences derived from the highly variable HVR1 region of E2, which will address the problem of genetic variability of HCV; and (c) conserved "framework motifs" present within the HVR1 region. The E2- WHcAg hybrid particles will be optimized for protein expression, assembly competence, yield in the E. coil expression system, antigenicity and immunogenicity. Because strong T cell responses (both CD4 vand CD8 v) against HCV antigens and especially the nonstructural 3 (NS3) protein have been linked to viral clearance in acute and chronic HCV infection, our second approach will be aimed at developing a NS3/4A-specific DNA vaccine candidate (Specific Aim 2). The NS3 protein is highly conserved and an advantage of a DNA vaccine is the ability to elicit CD4 v Th cells and CD8 v CTL as well as antibody. We have found that a NS3/4A gene elicits significantly more efficient immune responses than the widely used NS3 gene. It is anticipated that a NS3/4A DNA vaccine may be used for prophylactic or therapeutic applications either alone or in combination with E2-WHcAg hybrid particles.
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{
"pile_set_name": "NIH ExPorter"
}
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Anterior cruciate ligament (ACL) injury is prevalent and often leads to instability, quadriceps muscle weakness and osteoarthritis. (OA) Many of those with chronic ACL deficiency have evidence of knee OA years after ACL rupture. Weakness and instability experienced by those who do not compensate well for ACL deficiency (non-copers) independently lead to compensation strategies that could precipitate or worsen knee OA. These poor compensation strategies do not uniformly resolve after surgical reconstruction and those who undergo reconstruction also have an increased risk of developing knee OA. The overall goal of this work is to determine whether effective rehabilitation programs to dynamically stabilize the knee reduce the adaptations that lead to osteoarthritis in a population at great risk for the development of knee OA. Eighty individuals with ACL rupture who are scheduled for surgery will be randomly assigned to a group that includes a form of neuromuscular training called perturbation training, or a standard group. They will be evaluated before and after surgery using motion analysis and radiography. In vivo measures of kinematics, kinetics, tibial translation and EMG based models of joint compression will be used for comparison. This randomized trial is designed to demonstrate that, after preoperative rehabilitation that includes perturbation training, movement patterns adopted by non-copers: 1) demonstrate improve joint stability and reduced muscle cocontraction, 2) persist after reconstruction and 3) result in better functional outcomes after reconstruction and 4) lead to the development of less knee pain and OA over time than standard preoperative treatment strategies. The information derived from this project will provide valuable insight into the management of the approximately 100,000 Americans who rupture their ACLs each year and undergo reconstructive surgery. More importantly, if the perturbation training program's ability to induce dynamic knee stability actually results in joint protection, its application to others at risk for the development knee OA may help reduce the incidence of this disabling clinical condition.
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{
"pile_set_name": "NIH ExPorter"
}
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Proper salivary gland function is critical for the maintenance of oral health and physiological well-being. Saliva functions to moisten the oral tissues as an aid for speech, as a solvent important for taste, and as a masticatory wetting agent/lubricant important in swallowing food. Saliva also plays an important role in host defense and is a major contributor of the innate host defense of the upper gastrointestinal tract. The salivary glands are a major component in the mucosal immune system of the oral cavity that confers antigen- specific immunity to oral and mucosally-acquired pathogens. We demonstrated that the salivary gland can act as a mucosal inductive site in addition to an effector site and tha antigenic stimulation of the salivary gland is sufficient to induce immunity at proximal and distal mucosal sites as well as systemic sites. Understanding the immunobiology of the salivary glands is also of particular interest because of rapidly developing progress in gene therapy and tissue engineering as it relates to the salivary gland. The ability to re-engineer the salivary gland via gene transfer in vivo with the resultant in situ restoration of fluid secretion and to utlize salivary endocrine secretory pathways for systemic gene therapeutics provides additional reason for gaining a better understanding of how salivary gland immunity interacts with the expression of transferred gene(s), especially because the transferred gene(s) are frequently expressed utilizing recombinant viral vectors. Murine cytomegalovirus (MCMV) infections have served as a tool for investigating salivary gland immunity. In addition, MCMV infections have already been used for some time as models for human CMV infection. Human CMV infections are common but do not lead to significant disease in immune-competent healthy adults. Nevertheless, in immune-compromised patients (transplant recipients or AIDS patients) CMV infection becomes a serious complication and source of mortality. Therefore, understanding the immune responses to MCMV infections can lead to a better understanding of the pathogenesis of CMV infections in humans. In addition, human CMV can be spread through oral and respiratory secretions and through sexual transmission. This makes investigating the mucosal responses to MCMV, in particular, relevant to infection in humans. In this proposal we will utilize the MCMV model to extend our proof of concept that salivary gland vaccination in particular cannulation of the parotid salivary gland, could be an important new clinical route for mucosal vaccines. This project is ideally suited for a short-term summer exposure to research of interest to the dental student that can easily be expanded over the course of the academic year. We expect that the outcomes of these studies will greatly add to our understanding of salivary gland induced mucosal immunity. The award of this AREA grant will meet the goals of student research at the School of Dental Medicine by fostering scholarship and critical thinking skills, by adding to the body of scientific knowledge, and by facilitating recruitment of students into academic, dental research careers.
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{
"pile_set_name": "NIH ExPorter"
}
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Osteoporosis, both post-menopausal and senile, is a major health care problem of the elderly, increasing in importance as our population ages. It results from an imbalance has, as a long-term goal, the understanding of the role that osteoblastic protein secretion plays in differentiation, from the standpoint both of secreted, soluble effector molecules, and secreted proteins which help to shape and form the extracellular matrix (ECM). Currently, there is little information on the number of different proteins that are secreted by osteoblasts, their identities and characteristics, and the roles they play in inducing or maintaining osteoblastic phenotype. There is virtually no information concerning the role of the ECM in these same processes. This proposal addresses these areas, using high resolution electrophoretic techniques as well as Northern blot analysis. Specific aims 1 and 2 are to identify and characterize the secreted proteins of human osteoblastic cells in culture and correlate their expression with the expression of known marker proteins such as osteonectin or osteocalcin. Specific aim 3 will examine whether or not bone cells obtained from individuals of different ages express the same proteins, to the same extent. Specific aim 4 will correlate the secreted proteins of bone cells with the proteins found in human bone (Project 5), while specific aim 5 will explore the effects of various soluble effector molecules (e.g., PTH, vitamin D3) on protein secretion in these cells. In the above studies, we will emphasize a new antigen from Saos-2 cells, P80. Specific aims 6-9 will examine the effect of bone ECM on osteoblastic phenotype in several ways. Specific aim 6 will examine in detail a new finding that MG-63 cells alter shape and osteonectin production when grown on type I collagen; under this specific aim we will also explore other purified components of ECM for phenotypic effects. Specific aim 7 will examine natural ECM as a growth substrate for osteoblastic cells, while specific aim 8 asks whether the age of the natural ECM (e.g., age of donor) alters its effects. Finally, in specific aim 9 we will study the relationship between the ECM effects and effects of hormones and vitamins on expression of osteoblastic phenotype.
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{
"pile_set_name": "NIH ExPorter"
}
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Oxidative stress has been suggested to play an important role in a wide variety of disease processes including hyperoxiainduced lung and retinal injury and the normal aging process. We have reported the discovery of novel bioactive prostaglandin F,-like compounds, termed FZ isoprostanes (F,-IsoPs) that are produced in vivo non-enzymatically by free radical induced peroxidation of arachidonic acid (AA) and that similar compounds are also produced in the brain from oxidation of docosahexaenoic acid (DHA). Measurement of these compounds has proven to be a major advance in our ability to assess oxidative stress status in vivo. We have also shown that compounds that have E-, D-, A-, and J-type rings and highly reactive isolevuglandins (IsoLGs) are products of IsoP pathway. We have recently discovered a series of novel lipid peroxidation products that we speculate contain a furan ring (IsoFs). These compounds will be structurally characterized and their mechanism(s) of formation will be determined using a number of analytical approaches utilizing mass spectrometry. In the presence of high oxygen tension, we have found that the formation of IsoFs is highly favored over the IsoPs. Thus, studies are proposed to utilize measurements of IsoFs to investigate the role of free radicals in hyperoxia-induced pathologies. New IsoPs and lsoFs will also be synthesized and tested for bioactivity. We have obtained evidence that the formation of IsoPs is increased dramatically in aged rats. This will be further characterized and correlated with other products of oxidation and antioxidant defenses in rats. We will also explore the hypothesis that this phenomenon maybe the cause of multi-organ failure in the elderly. Studies are also planned to further characterize the spectrum of lsoP-like and related compounds that can be formed from oxidation of DHA. Studies are also planned to explore the biochemistry and pathobiology of IsoLGs, focusing on their preferential adduction to membrane proteins with attendant alteration in the function of receptors and ion channels. We will also explore whether these compounds, which are initially formed esterified in phospholipids, are substrates for platelet activating factor acetyl hydrolase. Finally, we will test the hypothesis that free radicals and IsoPs play an important role in the renal failure that ensues in glactosamine treated rats, an animal model of the hepatorenal syndrome.
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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. Antenatal Screening and Consent in a Research Network Model Study Aims: 1) To obtain consistently performed, timed and interpreted neuroimaging studies in extremely preterm infants enrolled in SUPPORT: a. cranial US at 4-14 days of age b. cranial US at 35-42 weeks PMA c. MRI at 35-42 weeks PMA 2) To compare early and late US and MRI findings between Low and High SpO2 groups within each randomized ventilation strata, and between Early CPAP and Control ventilation groups within each randomized oxygenation strata. 3) To utilize the NICHD Neonatal Research Network follow-up programs to assess neurodevelopmental outcomes at 18-22 months corrected age, as described in SUPPORT. 4) To examine the independent associations of neuroimaging findings with neurodevelopmental outcomes through predictive modeling. a. Regression models will assess the absolute and relative value of early and late cranial US, and brain MRI, alone and in combination with traditional risk factors, to predict abnormal and normal neurodevelopmental outcome at 18-22 months. b. Through stepwise regression modeling, assess the value of neuroimaging findings, alone and in combination, in predicting neurodevelopmental outcomes over and above the value of early risk factors or early and in-hospital risk factors alone.
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{
"pile_set_name": "NIH ExPorter"
}
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Diphtheria, which re-emerged as a major epidemic disease in Russia andthe Newly Independent States of the former Soviet Union during the 1990s, is a paradigm for toxin-mediated bacterial disease. Tuberculosis, which causes more deaths worldwide than any other bacterial infection, is a paradigm for intracellular bacterial infection. Very similar iron-activated regulatory proteins, called the diphtheria toxin repressor (DbcR) and the iron-dependent regulator (IdeR), control virulence in C. diphtheriae and M. tuberculosis, respectively. Db<R and IdeR are prototypes for a family of metal-dependent global regulators predominantly found in Gram-positive and acid-fast bacteria. During the current project period, we used genetic and biochemcial methods to identify iron-regulated genes and gene products in C. diphtheriae, and we developed lead compounds that promote activation of DbcR or IdeR at lower concentrations of ferrous ions than are needed to activate DtxR or IdeR by itself. During the next project period we will extend these studies in several important ways. In Aim 1, using C. diphtheriae, we will characterize the structure and function of several important iron-regulated genes, determine the molecular basis for DtxR-dependent and Db<R-independent regulation of gene expression by iron, investigate the roles of specific sigma factors within the iron regulon, and investigate oxidative stress pathways and their interactions with iron-dependent gene regulation. In Aim 2, we will characterize the multiple siderophore-dependent iron-uptake pathways in C. diphtheriae that participate in maintenance of iron homeostasis. We will characterize interaction of diphtheriabactin (the siderophore produced by C. diphtheriae) with its dip0582/ciuA receptor, and we will identify the genetic systems that enable C. diphtheriae to use other siderophores (e.g., desferrioxamine, ferrichrome, rhizoferrin and the recently discovered siderophores from C. pseudotuberculosis). In Aim 3, we will apply structure- based methods to develop "second-generation" peptides and other molecules with high potency for stimulating or inhibiting Db<R or IdeR activity. We will test these novel compounds for biological activity, first in pound coli or C. glutamicum systems in which recombinant IdeR controls expression of reporter genes, and then in M. tuberculosis. Because ideR is essential in M. tuberculosis, and because expression of a "hyper- repressor" variant of DtxR attenuates the virulence of M. tuberculosis, we will determine whether the novel activators and inhibitors of IdeR that we are developing have antimicrobial activity against M. tuberculosis and potential value as chemotherapeutic agents for treatment of tuberculosis. RELEVANCE (See instrucUons); Two Closely related iron-dependent regulatory proteins, Db<R and IdeR, control virulence of the bacteria that cause diphtheria and tuberculosis, respectively. Our studies will show how DtxR functions as a master regulator controlling the effects of iron on gene expression and virulence in the diphtheria bacillus. We will use structure-based methods to design novel compounds to activate or inhibit DbcR/ldeR-type regulatory proteins. We will evaluate these compounds for potential value as novel drugs for treatment of tuberculosis.
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{
"pile_set_name": "NIH ExPorter"
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Beta-adrenergic receptor (AR) is a pivotal regulator of cardiac function. However, enhanced beta1-AR signaling promotes apoptotic heart cell death, which is implicated in myocardial remodeling and heart failure. The goal of this study is to determine whether the classic Gs-adenylyl cyclase-cAMP-PKA pathway is essential to beta1-AR apoptotic effect. To avoid complicated interactions between beta-AR subtypes, we created a genetically "pure" beta1-AR experimental setting by expressing the mouse beta1-AR in the null background of beta1/beta2-AR double knockout adult mouse cardiomyocytes using adenoviral gene transfer. Here we show that beta1-AR stimulation is markedly increases myocyte apoptosis, as evidenced by increased TUNEL staining positive cells, DNA fragmentation (cell death ELSIA and DNA laddering). Beat1-AR-induced apoptosis is abolished by a beta-AR antagonist, propranolol (1 uM). To our surprise, specific PKA inhibitors, including Rp-CPT-cAMP (100 uM), H89 (5 uM) and a peptide inhibitor (PKI, 5uM), while blocking forskolin (0.1 uM)-evoked myocyte apoptosis, cannot prevent beta1-AR-induced apoptotic cell death. Ironically, blocking Ca2+ influx by a L-type Ca2+ channel inhibitor nifidipine (1 uM) or buffering intracellular Ca2+ with EGTA-AM (1 uM) fully protects heart cells against beta1-AR-mediatd apoptosis, suggesting that a PKA-independent increase in intracellular Ca2+ is obligatory to beta1-AR apoptotic effect. To delineate the downstream events of beta1-AR/Ca2+ apoptotic signaling, we first evaluated the potential role of a Ca2+/ calmodulin-dependent phosphatase, calcineurin, since this phosphatase has been implicated in beta-AR-induced apoptosis in cardiomyocytes. However, inhibition of calcineurin with cyclosporin A (5 uM) or FK506 (10 uM) does not affect b1-AR-mediated apoptosis. In sharp contrast, inhibition of Ca2+/calmodulin kinase II (CaMKII) with KN-93 (0.5 uM) or a peptide inhibitor (AIP, 10uM) fully abolishes b1-AR-promoted apoptotic cell death. This is consistent with the fact that beta1-AR induces a robust increase in CaMKII activity in a PKA-independent manner. Furthermore, overexpression of CaMKII-dC, a predominant cardiac CaMKII isoform, in cardiac myocytes using adenoviral gene transfer markedly enhances beta1-AR mediated myocyte apoptosis. Thus, beta1-AR apoptotic effect is mediated by a CaMKII-, rather then PKA-, dependent mechanism. These findings shed light on our understanding of beta1-AR cardiac detrimental effects, also underscore novel therapeutic strategies for the treatment of heart failure.
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{
"pile_set_name": "NIH ExPorter"
}
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Among individuals with chronic kidney disease (CKD) receiving maintenance dialysis therapy the proportion of African American patients is significantly higher compared to their non-Hispanic white counterparts, and traditional risk factors of cardiovascular disease such as hypercholesterolemia, hypertension and obesity show seemingly anomalous, inverse associations with adverse outcomes. The exceptionally high prevalence of end- stage renal disease (ESRD) among African Americans is likely a result of the complex interaction of their higher rates of CKD incidence and/or faster CKD progression, lower mortality, and lower likelihood of kidney transplantation. The anomalous cardiovascular risk factor profile in ESRD could be the result of short-term competing risks related to malnutrition and inflammation, with the seemingly unfavorable traditional cardiovascular risk factors associating with a better nutritional state, which could differentially affect African Americans an hence provide one explanation for their better survival in ESRD. Whether similar paradoxical differences in mortality rates in non-dialysis dependent CKD stages exist is not clear; their presence, extent, the CKD stage of its occurrence and their mechanisms of action all need to be clarified in sufficient detail to allow for the design of proper diagnostic and interventional strategies towards cardiovascular risk reduction and towards alleviating racial disparities in outcomes. In the spirit of PA-09-196 we will utilize data obtained from the national VA research database which is the only large administrative database with detailed socio-demographic and clinical information on very large numbers of individuals (over 4 million individuals including ove 0.5 million with CKD) across all parts of the US. We will examine the population-wide dynamic effects of incident CKD and mortality on racial composition and on changes in cardiovascular risk factor profiles by examining longitudinally a cohort of patients with normal estimated glomerular filtration rate. We will explore the effects of various socio-demographic characteristics, co-morbidities, biochemical measurements and medication use on mortality and progressive CKD using complex epidemiologic methods including joint modeling to assess the effect of longitudinal changes in risk factor parameters on mortality and marginal structural models in order to adjust for both baseline and time-dependent confounders. This three-year project will generate a wealth of information to more reliably examine the above hypotheses related to racial and cardiovascular discrepancies in the outcomes of patients with all levels of kidney function that could have significant public health implications.
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{
"pile_set_name": "NIH ExPorter"
}
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We propose to carry out a series of experiments all bearing on the general question of the physiological mechanisms of color vision. In single cell recording studies, we will examine the responses of cells in various cortical areas to equiluminant color stimuli, as well as luminance-varying stimuli, to examine the extent to which color information is processed in various cortical areas and the transformation, if any, which takes place with respect to the chromatic characteristics of the light. In addition, in both psychophysical and recording experiments we will examine the role played by color information in spatial vision. Specifically we will compare the way in which spatial variations defined by color differences and those defined by luminance differences are analyzed by the visual system.
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{
"pile_set_name": "NIH ExPorter"
}
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To extend studies concerning serum and spinal fluid complement components in patients with central nervous system manifestations of S.L.E. including an analysis of participation of the alternate pathway of complement activation by measurement of serum levels of the C3 proactivator. Further, to demonstrate conclusively in vivo activation of complement in the CSF by documenting the presence of reaction products of complement components (e.g., alpha2D, C3b, or C4i) or activated components (e.g. Cl) in CSF. Another means of documenting complement activation will be to measure other complement properties within the CSF and to compare such complement profiles with serum profiles indicative of activation of the classic or alternate pathway. Secondly, to investigate abnormalities of spinal fluid complement in neuropsychiatric disorders other than S.L.E. Assays of complement components and an attempt to demonstrate reaction products or activated components will be carried out. Third, to extend previous studies from our laboratory concerning the in vivo metabolism of C3 in human disorders. Fourth, to study the quantitative immunology of red cell sensitization with complement components in acquired hemolytic anemias in order to correlate such quantitation with clinical findings and with serologic reactions to further clarify the role of complement in immune red cell destruction. Methods used will be the use of hemolytic assays and immunochemical techniques; immunodiffusion, electrophoresis in antibody containing gels and radioimmunoassay. The in vivo kinetics of radiolabelled, purified complement proteins will be studied to determine catabolic and synthetic rates. Finally, a sensitive new immunochemical method based on antigen inhibition of lysis of complement sensitized sheep cells will be used to study the significance of in vivo red cell bound complement components.
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{
"pile_set_name": "NIH ExPorter"
}
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Many receptors are composed of multiple proteins joined in a complex and are associated with ?stabilizing? proteins. Androgen receptors are transcription factors that are associated with a stabilizing protein called Sigma1. If you effectively inhibit Sigma 1 (or otherwise block its function) one consequence is that androgen receptor signaling is lost in its entirety. Androgen drive is particularly important in prostate cancer. For this reason, prostate patients who fail to adequately respond to chemo often undergo chemical or surgical castration. Castration is helpful to varying degrees; however, many patients continue to exhibit castration resistant prostate cancer (CRPC). CRPC occurs because androgen receptors are always signaling ? even in the absence of testosterone drive (i.e., there is a baseline level of signaling that takes place in the absence of any androgen agonist activity). This is called ?constitutive? signaling. Context's drug destabilizes the androgen receptor. This destabilization effectively eliminates all signaling, including baseline/constitutive. A key driver of androgen receptor (AR) signaling in CRPC is the presence of AR variants, including splice variants and inhibitor-binding mutants, which renders abiraterone and enzalutamide, the market leading drugs ineffective. Targeting Sigma1 is a novel strategy that inhibits AR signaling regardless of AR variant expression. This vastly expands the landscape of men likely to respond to therapy, including those with resistance to abiraterone or enzalutamide due to AR variants, and provides the opportunity for novel combination therapies. Moreover, we have already developed and characterized a series of drug-like small molecule Sigma1 modulators that are poised for optimization, allowing rapid translation of our preclinical studies to Phase I trials. Specific aims of this proposal are: Aim 1: To provide an improved drug-like Sigma1 inhibitor suitable for advancement as a Preclinical Candidate. The lead compound, CT-110, is an orally bioavailable small molecule that potently binds Sigma1 and induces tumor regression in vivo; however, CT-110 is not a Preclinical Candidate (PCC) due to hERG channel inhibition and modest plasma exposure. We seek to optimize CT-110 using medicinal chemistry approaches to maintain potency while improving selectivity and optimizing in vitro pharmacokinetic and ADMET properties. The resulting optimized lead from this aim will be evaluated for in vivo pharmacokinetics and 7-Day toxicology in mice. Aim 2: To establish the in vivo efficacy of an improved Sigma1 inhibitor in a model of CRPC. We will evaluate the improved Sigma1 modulator in CRPC cell lines (C4-2, 22Rv1) that reflect key hallmarks of CRPC, including AR overexpression and constitutively active AR variants. The ability of our improved Sigma1 modulator to inhibit tumor progression will be evaluated, in addition to markers of target engagement and disruption of the AR axis. These studies directly respond to the overarching challenge of developing effective treatments and addressing mechanisms of resistance.
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{
"pile_set_name": "NIH ExPorter"
}
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The inactivation of tumor suppressor genes or the activation of oncogenes have been revealed as underlying causes for many tumors. The challenge is now to define their presumed roles in normal growth regulation, and to determine how their deregulation leads to tumor formation. It is the focus of this project to elucidate the function of one such tumor suppressor gene, NF2. Germline deletions in NF2 cause Neurofibromatosis II, a disease predisposing for certain nervous system tumors. These tumors have lost/inactivated the NF2 protein due to a further mutation in the second NF2 allele. Moreover, the inactivation of NF2 is also seen in sporadic brain tumors. NF2 is structurally related to the ezrin/radixin/moesin (ERM) family of membrane-cytoskeleton linker proteins. This relation places NF2 into a new class of cytoskeletal tumor suppressors. The hypothesis to be tested is whether cytoskeletal proteins like NF2 and the ERM family are involved in growth inhibiting pathways, either directly or indirectly by chanelling these signals from the membrane to the nucleus. To this end, we have cloned the homologs of NF2 and the ERM family in C. elegans, hoping to take advantage of the genetic power of this invertebrate system. In C. elegans it will be possible (1) to observe the impact of these proteins on normal growth and development, (2) to genetically manipulate these proteins in vivo and observe the effects in the living organism and (3) to determine pathways using genetic screens discovering interacting partners. The preliminary results on this project have been obtained by the candidate at the MGH Cancer Center directed by Dr. K. Isselbacher (Sponsor). This environment is dedicated to the investigation of tumor suppressor genes, oncOgenes, cell signalling and the cell cycle; and further dedicated to use invertebrate genetics for this purpose. Dr. F. Solomon, MIT, as an expert on cytoskeletal proteins, has been involved in the designing phase of this project and, as a ce-Sponsor, will have dominant scientific impact on its further development. The C. elegans expertise is provided by the candidate's coworker Dr. J. Fleming and by Dr. Horvitz who will serve as a consultant. The candidate is a Pediatric Oncologist with completed clinical training plus research training, with a major interest in basic research. Dr. Medearis, Chief of the Department of Pediatrics at MGH, concurs in the candidate's wish to maintain her clinical work on a restricted schedule in order to support her future development with the longer term perspective of an academic career in Pediatric Oncology. This award will allow the candidate to receive additional training in the areas of cytoskeletal proteins and C. elegans, which will be indispensable for the proposed project and which will expand the candidate's current understanding of oncogenesis. With this further period of training the candidate is expected to assume the role of an independent investigator with her own research program.
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{
"pile_set_name": "NIH ExPorter"
}
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Resulting from organ retrieval, cold preservation and a period of warm ischemia during the surgery, ischemia-reperfusion injury (IRI) often leads to primary graft non-function, may predispose to late chronic rejection, and contributes to the shortage of donor organs available for transplantation. At present, there is no specific treatment available to prevent IRI in transplant recipients. Moreover, mechanisms of IR-induced tissue damage, and how clinically-relevant innate immune responses contribute to graft rejection represents one of the most challenging yet understudied problems in clinical liver transplantation. We have identified the role of intrinsic PACAP neuropeptide in maintaining hepatic homeostasis in a non-transplant model of hepatic warm IRI. We have shown that treatment with PACAP neuropeptide ameliorated liver IRI by depressing macrophage function in cAMP-PKA dependent manner. Our pilot data from an ongoing clinical study show that PACAP and its receptors are preferentially induced in human IRI-resistant liver transplants, as compared to those suffering from severe IR-damage. Here, we propose to analyze the function of distinct PACAP signaling pathways in a clinically relevant mouse model of prolonged hepatic cold ischemia followed by orthotopic liver transplantation (OLT). Hypothesis: PACAP-mediated cAMP-PKA signaling at the macrophage (innate immune arm) - hepatocyte (defensive arm) interface regulates pro-inflammatory (pathogenic) and cytoprotective (homeostatic) functions in OLTs subjected to IRI. Aim 1: To define the role of PACAP neuropeptide in innate immune function in liver IRI. Hypothesis: PACAP inhibits TLR4 macrophage activation and promotes immune homeostasis in IR-stressed OLTs. Aim 2: To evaluate the role of PACAP neuropeptide in hepatoprotection in liver IRI. Hypothesis: PACAP enhances parenchyma cell autophagy and promotes hepatocyte survival. By identifying a novel PACAP mediated-neural regulation in the milieu of IR-stressed OLTs, this proposal puts forth the advances in hepatic immune modulation in a completely new context of innate immunity and parenchyma cytoprotection. Based on our preliminary experimental data, supported by clinical observations, findings from this exploratory proposal may have far reaching basic and practical ramifications, as taming of IR-triggered innate inflammation at the graft site is now considered critical for improving both short and long-term transplantation outcomes.
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{
"pile_set_name": "NIH ExPorter"
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We have previously shown that a wide variety of virally and spontaneously transformed mammalian fibroblasts secrete a major transformation-dependent phosphoprotein with a molecular weight of about 62,000. Elevated secretion of the phosphoprotein occurs regardless of the transforming agent and secretion is temperature sensitive in a cell line that is temperature sensitive for the transformed phenotype. Elevated secretion of this major phosphoprotein also correlates with tumorigenicity of cells of epithelial origin and distinguishes neoplastic from hyperplastic epithelium. The 62K phosphoprotein is apparently not related to viral or tumor-specific phosphoproteins which have been described by others. Recently, I have determined that the 62K secreted phosphoprotein binds avidly to fibrin clots, binds selectively to lysine and benzamidine-Sepharose, and shares antigenic determinants with a component of normal plasma and serum. Because fibrin deposition commonly accompanies tumor growth in vivo, it may be that the 62K phosphoprotein associates with tumor-fibrin in vivo. The goals of this proposal are to purify the 62K phosphoprotein extensively, to characterize it biochemically, to determine its biological functions, and to relate these properties to the biology of tumor growth. The affinity of the phosphoprotein for fibrin clots will be analyzed further, and it will be determined if the phosphoprotein is related to any of the proteins with known affinities for fibrin. Purified preparations of the phosphoprotein will be analyzed for proteolytic activity, and the effect of dephosphorylation on this activity (if found) as well as the phosphoprotein's affinity for fibrin will be examined. Antibody to the 62K phosphoprotein will be used to purify the antigenically related component(s) of plasma and serum and they subsequently will be characterized. Metabolic labeling of tumor-bearing animals and immunofluorescent staining of tumor sections will be employed to study the expression and localization of the phosphoprotein in vivo. Finally, additional tumorigenic and nontumorigenic cells (including human) will be compared for secretion of the phosphoprotein to further test its apparent correlation with tumorigenicity.
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{
"pile_set_name": "NIH ExPorter"
}
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A microsomal galactose oxidizing system (M.G.O.S.), identified in this laboratory, will be further purified, characterized, and classified according to its reaction mechanism. The product of the reaction, galactonic acid or its lactone precursor, will be tested as a potential inhibitory metabolite of lysosomal, microsomal, and glycolytic enzymes. The hypothesis that lysosomes participate in the process of hormone-mediated enzyme induction will be further investigated utilizing the starved-refed rat, diabetic rats, the genetically obese mouse, and the beige mutant mouse. In vitro studies are planned with cultured primary rat hepatocytes and the mouse fibroblast subline, 3T3-L1, which differentiates into adipocytes. Nuclear RNA synthesis studies will be conducted to augment the previous probes of the basic hypothesis. Finally, an isozyme of nucleoside diphosphate kinase has been discovered to be a microtubule-associated protein. During the grant period the kinase will be purified, and its properties and possible regulatory role in microtubule assembly mechanism will be assessed.
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{
"pile_set_name": "NIH ExPorter"
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Abstract: Subcortical pathology is a common feature in aging, Alzheimer's disease and vascular dementia but has been challenging to study with micron resolution in vivo. Optical methods such as two-photon microscopy image the superficial cortex at the micron-scale, but the resolution of these conventional microscopic methods degrades rapidly beyond 600 microns imaging depth. Standard whole-brain magnetic resonance imaging (MRI) methods do not yet provide cellular-level resolution and are expensive. Thus, there is a pressing need for methods to assess deep cortical and subcortical perfusion and cellular injury at the microscopic level, thus bridging the gap between existing superficial optical microscopy and macroscopic imaging. This proposal will develop, validate, and demonstrate advanced optical microscopy methods for longitudinal imaging of subcortical structures in the mouse brain using 2200 nm Optical Coherence Microscopy. 2200 nm imaging, in which tissue scattering is reduced by 2.5 and 1.5 compared to 1300 nm and 1700 nm, respectively, will enhance the delivery of ballistic (as opposed to multiply-scattered) photons to the focal spot, and enhance the proportion of photons backscattered from the focus that are detected without further scattering. Both of these benefits will substantially improve the signal localization, spatial resolution and signal-to background ratio when imaging deep in the brain. These methods will push penetration depths further into the living mouse brain, imaging subcortical structures (i.e. hippocampal proper and dentate gyrus) and pathology at higher resolutions than were previously possible.
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{
"pile_set_name": "NIH ExPorter"
}
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Emotional and cognitive characteristics were studied in epileptic patients following unilateral left or right temporal lobe resection. The integrity of attentional and perceptual (visual, auditory, and tactile) systems were evaluated using standard and experimental procedures. Physiological events (skin conductance) were monitored and recorded during test performance. The research examined the role of the temporal lobe in establishing specific limbic associations between left and right hemispheres in regulating cognitive functions and emotional experiences in man. Tachistoscopic studies identified a critical perceputal role for right temporal mechanisms especially during the early stages of visual processing. The left and right temporal lobes contribute differentially to specifying the identify of a stimulus, but not to its position or orientation in space. Left temporal mechanisms encode verbal information during initial learning. Modest compensation for memory defects following temporal lobectomy may be achieved with strategy which combines overt and covert imagery with praxic encoding. In affective sectors, left temporal patients tend to neutralize reactions to nuances with emotional coloration; right temporal patients, in contrast, rate these materials similar to normal individuals. Unike normal individuals, however, the left and right temporal lobectomy patients were hyporesponsive to affective material as indexed by skin conductant measures. Moreover, both lobectomy groups failed to take advantge of the emotional characteristics of information to facilitate memory. These data suggest that unilateral temporal lobectomy disrupts the normal linkage of cognitive-affective associations mediated by temporal limbic interaction. There were, however, beneficial effects to surgical treatment in that patients, following temporal lobe surgry, were less deviant from normal subjects in emotional behavior.
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{
"pile_set_name": "NIH ExPorter"
}
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Anxiety can have a great effect upon the behavior of individuals exposed to stressful situations. In many instances this anxiety may prevent the emission of behaviors that are necessary to alleviate the stressful situation (e.g., making it impossible to study for an upcoming examination that one is anxious about). In many instances minor tranquilizers are prescribed by physicians to help decrease anxiety in their anxious patients. Such individuals can become dependent on these drugs and thus may persistently self-administer and abuse the compound. This research proposal is for investigations of the neurochemical changes that occur in the brains of animals exposed to a stressful situation resulting in anxiety. The objectives of the research are a) to study changes in neurotransmitter systems during the acquisition and extinction of an animal model for anxiety; and b) to concurrently study the acute and chronic effects of minor tranquilizers on the behavior and neurotransmitter systems. The animal model consists of inducing the conditioned emotional response (conditioned suppression of positively reinfoced responding) in rats and measuring changes in the turnover of biogenic amines (serotonin, dopamine, norepinephrine and acetylcholine) in brain areas which interact to process emotion. Since benzodiazepines, barbituates, meprobamate and alcohol are known to act on neurotransmitter systems in naive animals, it is important to establish their role in reversing conditioned suppression. Evidence exists to suggest that neurochemical mechanisms differ in naive versus behaving animals. These drugs have a tranquilizing effect due to a complex interaction between neurochemistry and individual components of the anxiety-producing environment. Following an in-depth analysis of these multiple interactions, one can characterize the neurochemical changes that occur as a result of anxiety and the alleviation of these changes by minor tranquilizers. A better understanding of the brain mechanisms associated with anxiety and how they are affected by minor tranquilizers may permit more effective treatment of this condition and the development of more specific anti-anxiety agents to minimize the abuse of these compounds by humans.
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{
"pile_set_name": "NIH ExPorter"
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The Biostatistics Core Component 2 will provide data processing, data management, data analysis and[unreadable] statistical support for each of the other cores, components and projects within the proposed[unreadable] Methamphetamine Abuse Research Center (MARC). This single biostatistical unit serves as an important[unreadable] interface for the transmission of diverse types of data and processed information among core and project[unreadable] investigators. It assures that data and information from different cores and projects will be processed as[unreadable] uniformly as possible to facilitate integration of results and of effort. The environment provided by a single[unreadable] biostatistics unit also facilitates the effective and timely return of processed data and information to the core[unreadable] and project investigators in support of their objectives. In addition, this Core will assist in the use of public[unreadable] databases and computer software, and provide the computer infrastructure for the microarray and gene[unreadable] mapping (QTL) studies. As the Center evolves, we will work toward development of a central database for[unreadable] the MARC patterned after the successful database we have developed for the NIH-NIA Alzheimer's Center[unreadable] at OHSU.
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{
"pile_set_name": "NIH ExPorter"
}
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The overall objective of this project is to test the hypothesis that the effects of saturated fat (SF) on lipoprotein markers of cardiovascular disease (CVD) risk are influenced by food sources of dietary protein. There is growing epidemiological evidence that consumption of red meat is associated with greater incidence of CVD than either white meat or non-meat foods. Pathophysiological support for the validity of this association is provided by preliminary evidence from our group that a high beef diet has a more deleterious effect on lipoprotein measures of CVD risk than we have observed for mixed protein diets. Specifically, we have found that a high protein, high SF diet with a moderate red meat content selectively induces increases in intermediate density lipoproteins (IDL) and larger LDL particles that have been found to be much more weakly associated with CVD risk than smaller LDL. In contrast, a more recent study from our group has found that, with a similar intake of SF, high beef consumption results in a preferential increase in levels of small and medium sized LDL particles, both of which are strongly related to incident CVD. To date however, no studies have directly compared the lipoprotein effects of red meats with other food sources of protein in the context of both high and low saturated fat intake. We specifically hypothesize that increases in plasma levels of LDL cholesterol (C), and apolipoprotein (apo) B, induced by SF are greater when the major food source of protein is red meat rather than either white meat (poultry) or non-meat foods, and that this is due to increased levels of small and medium sized LDL particles. We therefore propose a clinical trial in which 180 healthy men and women will be randomized to high SF (15%) or low SF (7%) diet groups, and within each group, consume diets with equivalent amounts of protein derived from red meat, white meat, and non-meat sources for 4 wk each in random order. Our Specific Aims will test whether: (1) with high SF, the red meat diet, compared to the other food sources of protein, will result in higher levels of LDL-C, apoB, small and medium sized LDL particles, and total/HDL-C; (2) with low SF, dietary protein source will not be related to any of these measurements; (3) with both the white meat and non-meat diets, increased LDL-C with high vs. low SF will be due primarily to increases in IDL and/or large LDL, whereas with red meat the additional increase in small and medium LDL will result in greater increases in apoB. In addition to these aims we will test for possible metabolic determinants of dietary effects on apoB-containing lipoprotein subclasses, including post-heparin plasma hepatic lipase activity, which is critical for production of smaller LDL, and LDL receptor activity as assessed in peripheral blood mononuclear cells, a system demonstrated to reflect physiologically relevant LDL receptor regulation. Finally, we will examine potential dietary influences on other metabolic biomarkers of CVD risk, including HDL subclasses and apoproteins, insulin sensitivity as assessed by HOMA-IR, measures of inflammation including CRP and multiple cytokines, and endothelial function using a non-invasive fingertip method.
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{
"pile_set_name": "NIH ExPorter"
}
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The objective of this study is to assess the safety, tolerability, and effectiveness of electrical stimulation of the subthalamic nucleus and the globus pallidus for the treatment of patients with advanced Parkinson's disease (PD). The proposed study is an open-label study with a double-masked randomized assessment of PD function at 3 months.
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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. Damage to medial temporal lobe structures in monkeys impairs performance on a task of visual recognition memory, the Visual Preferential Looking Task (VPLT). This project seeks to identify the neuronal mechanisms that support memory performance on the VPLT in awake, behaving monkeys. In the past year, we finished data collection and analyses which demonstrated that increased gamma-frequency (40-100 Hz) synchronization among hippocampal neurons is associated with enhanced recognition memory. We showed, on a trial-by-trial basis, that Increased synchrony during encoding predicts successful subsequent recognition memory performance. We have also completed analyses that demonstrate that individual hippocampal neurons modulate their firing rates based on stimulus novelty. Furthermore, the magnitude of this modulation is correlated with recognition memory performance. Together, these studies suggest that hippocampal mechanisms, both at the single neuron and network level, underlie successful recognition memory. Finally, we have recently collected data that demonstrate the existence of theta-band oscillations in the primate hippocampus, and that the phase of these oscillations is reset by eye-movements during visual exploration. In addition, our recent data showed that theta-band phase modulates the power of gamma-band activity. Together, these data suggest that phase resetting of the theta-band oscillation to an 'ideal phase'upon fixation onset may align hippocampal activity to an optimal state with respect to gamma-band synchronization and memory formation.
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{
"pile_set_name": "NIH ExPorter"
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[unreadable] [unreadable] Polyglutamine (polyQ) disease is a major cause of inherited neurodegeneration in the United States. The nine known polyQ diseases are caused by CAG repeat expansions that encode glutamine repeats in otherwise unrelated disease proteins. Spinocerebellar ataxia type 3 (SCA3), the most common dominantly inherited ataxia, is caused by an expansion of a glutamine repeat in the C-terminus of ataxin-3. One major question in polyQ disease research is the identity of the putative toxic intermediate(s) that triggers neurodegeneration. This grant aims to identify the toxic species that underlie SCA3. In aim 1, two transgenic SCA3 mouse models will be used to investigate the molecular and cellular pathological features of SCA3 over time in an effort to identify the toxic intermediates that lead to neuronal dysfunction and cell death. Aim 2 addresses the link between mutant (expanded) ataxin-3 and protein quality control mechanisms in disease pathogenesis. Specifically, both cell culture and mouse models will be used to investigate the interactions of ataxin-3 with a known modifier of polyQ disease, the co-chaperone and ubiquitin ligase C-terminus of Hsp70 interacting protein (CHIP). Neurodegeneration in many diseases, including Alzheimer's, Parkinson's and the polyglutamine diseases, is thought to be due to misfolding of proteins in neurons. This project will investigate protein misfolding in polyglutamine disease, which currently affects about 100,000 Americans; however, the findings will be broadly relevant to protein misfolding diseases in general. The results may also identify new therapeutic targets for polyglutamine disease. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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Mitogen-activated protein kinases (MAPKs) are serine-threonine protein kinases that are activated by diverse stimuli. MAPKs contribute to the control of cell and tissue homeostasis, cell division and cell death. The MAPK regulatory module includes three kinases that establish a sequential activation pathway comprising a MAPK kinase kinase (MKKK), MAPK kinase (MKK) and MAPK. Currently, mammalian MAPKs comprise five families: ERKI/2, p38 alpha,beta,gamma,delta, JNK1,2,3, ERK3/4 and ERK5. Each family of MAPKs has distinct, but sometimes overlapping, functions in terms of substrate recognition and control of cellular pathways. The hypothesis of this proposal is that two related MAPK kinase kinases, MEKK2 and MEKK3, regulate the MEK5-ERK5 module. In addition, MEKK2 regulates JNK and MEKK3 regulates p38. MEKK2, by binding the adaptor protein LAD, selectively couples to receptor tyrosine kinases and Src. MEKK3 binds a structurally unique adapter protein we have named osmosensing scaffold for MEKK3 (OSM). OSM in response to osmolarity changes translocates MEKK3 from the cytoplasm to plasma membrane ruffles and sites of membrane actin polymerization. Recruitment of OSM/MEKK3 to the plasma membrane is associated with MEKK3 activation. OSM is the first identified adaptor protein in mammalian cells for the activation of MAPK pathways in response to osmolarity changes. Targeted gene disruption of MEKK3, that binds OSM, is embryonic lethal, demonstrating the importance of this pathway in cell physiology. The aims of this proposal include: 1) Define the function of OSM in osmosensing for the regulation MAPK pathways 2) Define the function of the PB 1 domain for interaction of MEKK2 and MEKK3 with MEK5. 3) Generate a conditional MEKK3 knockout so the role of OSM/MEKK3 signaling in adult mice and differentiated cell types can be defined. These studies will define the mechanism for OSM/MEKK3 regulation of MAPK activation in osmoregulation, a critical regulatory response for homeostasis.
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{
"pile_set_name": "NIH ExPorter"
}
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State legislators throughout this country are making difficult policy decisions about complex issues involving drug abuse and addiction without the scientific expertise to critically evaluate these issues. These legislators typically do not have the background or opportunity to learn the science pertinent to abuse and addiction. Indeed, many are unaware that there is a scientific basis for preventing or treating drug abuse. Scientists have generally not helped the situation. Scientific findings are presented in journals that are not easily accessible or understandable, and scientists are often not able to present their knowledge in ways lay audiences can grasp. As a result, instead of policy debates proceeding from a scientific base, they tend to be political in nature, and much susceptible to special interest influences as well. We believe this situation is correctable. During the past four years the Addiction Studies Program has brought carefully selected drug abuse scientists from basic and applied areas together to present accurate scientific information about abuse and addiction to audiences that now total over 100 journalists from the electronic and print media. In the course of a two-day workshop, lectures are paired with ample interpersonal interaction and detailed discussion of the available scientific information on specific issues of public concern. The journalists have been very receptive to the information. Attendees have produced a book, numerous newspaper and magazine articles, radio and television shows about addiction, and a movie script. Now, in partnership with the Treatment Research Institute and the National Council of State Legislators, we propose to extend and tailor this workshop approach to educate at least 100 state legislators over the next five years. These workshops will match available scientific information to the important policy questions facing legislators and will include an evaluation of the impact of those efforts.
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{
"pile_set_name": "NIH ExPorter"
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Abstract In the United States, skin and soft tissue infections (SSTIs) are one of the most common indications for antibiotic treatment, representing 10% of hospital admissions and 6 million doctor visits a year. SSTIs range from self- limiting furuncles and cellulitis to life-threatening sepsis and tissue necrosis. Recent studies suggest more than 50% of all SSTIs were caused by staphylococcal or streptococcal species, in particular, methicillin-resistant staphylococcus aureus (MRSA). We have engineered a novel class of cell-penetrating antimicrobial therapeutics, called CAHPs, which demonstrate broad-spectrum activity against clinically relevant Gram+ and Gram- pathogens, while exhibiting minimal toxicity in human cells. These compounds have been shown to penetrate infected target cells, breakdown established bacterial biofilms of Category B and C pathogens, and to act as anti-inflammatory agents. Identifying a CAPH lead candidate as a novel therapeutic agent for the treatment of, initially, cutaneous infections addresses a highly unmet medical need. Ultimately, its use can be expanded into a broad spectrum antimicrobial therapeutics. The initial product will be an IV formulation that can be administered in community and nosocomial settings. An IV formulation is preferable for treating systemic cSSTIs, especially if hospital-acquired, and could later also be supplemented with a topical product for use in both communal and hospital settings. The Phase I portion of this proposal will involve characterization of the current prelead candidates for their antimicrobial potency and in vitro toxicity as well as for their pharmacokinetic characteristics. The most promising candidate will be tested in in vivo efficacy models of systemic and topical infections and assessed for its exploratory safety profile. The end result of this work will be a novel, first-in-class antibacterial drug candidate for treatment if SSTIs ready for preclinical development activities leading to the opening of an IND. We have assembled a team of expert advisors and collaborators to ensure successful completion of this research plan.
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{
"pile_set_name": "NIH ExPorter"
}
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The purpose of this study is to test the efficacy of atorvastin for the treatment of elevated cholesterol and triglycerides.
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{
"pile_set_name": "NIH ExPorter"
}
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(Applicant's Abstract) Patients with asthma can express clinically significant airway responses to challenge with very low doses of relevant allergen. We hypothesize that mast cells, through their ability to amplify the local expression of allergen-induced inflammatory responses and to produce many mediators, cytokines, chemokines and growth factors, can contribute importantly to the development of the airway hyperreactivity (AHR), chronic inflammation, and tissue remodeling associated with asthma. In experiments using genetically mast cell-deficient WBB6Fl-Kitw/Kitw-v mice and the congenic +/+ mice, we already have found that, in mice sensitized to ovalbumin (OVA) without adjuvant and then challenged via the airways, mast cells have a critical role in significantly enhancing allergen-induced AHR to aerosolized methacholine, eosinophil infiltration, and numbers of proliferating cells within the airway epithelium. However, the specific mechanisms by which mast cells promote the inflammatory, structural and functional changes associated with this asthma model are very incompletely understood. We therefore will develop further and characterize in detail "mast cell-dependent" models of asthma, and compare them to a "mast cell-independent" model of the disease, focusing on the patterns of allergen-induced inflammation, structural tissue changes, and AHR, and alterations in tissue- and cell-specific gene expression, that are induced in these settings. To define the specific mast cell-dependent mechanisms that contribute to these changes, we will investigate their expression in mast cell-deficient Kitw/Kitw-v mice, congenic wild type mice, and Kitw/Kitw-v mice that have been selectively reconstituted with in vitro-derived wild type mast cells or mast cells that express genetically-determined alterations in activation and/or mediator/cytokine growth factor production. The specific aims will test 3 hypotheses: 1. Mast cells can significantly amplify the expression of multiple phenotypic characteristics of asthma; 2. Mast cells can express such function via distinct mechanisms, including the IgE-dependent production of diverse mediators, cytokines, chemokines and growth factors; 3. Mast cells can influence the expression of multiple phenotypic characteristics of asthma by inducing distinct patterns of qualitative and/or quantitative changes in gene expression in the surrounding target cells and tissues.
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{
"pile_set_name": "NIH ExPorter"
}
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The Computational Biophysics Section studies problems of biological significance using several theoretical techniques: molecular dynamics, molecular mechanics, modeling, ab initio analysis of small molecule structure, and molecular graphics. These techniques are applied to a wide variety of macromolecular systems.[unreadable] [unreadable] Dr. Larkin's research involves the application of Quantum Mechanical/Molecular Modeling(QM/MM) and Molecular Dynamics simulation techniques to enzymatic reaction pathways. Specifically, Dr. Larkin is investigating reaction mechanisms of boronic acid inhibitors of Beta Lactamase enzymes that are responsible for increased resistance to penicillin antibiotics.[unreadable] [unreadable] Dr. Woodcocks research, in collaboration with Membrane Biophysics Section, involves high level ab initio computations in vacuum and with the IEFPCM implicit solvent modelon 5-hydroxy-methyl-tetrahydropyran to investigate the effects of water on the exocyclic torsional surface. Rotamer populations evaluated from the omega (C-C-C-O), theta (C-C-C-O) solvent surface agree almost quantitatively with experimental values for the closely related methyl-alpha-D-4-deoxyglucopyranoside. Potentials of mean force obtained from the two surfaces show substantial solvent stabilization of the TG (omega = 180 plus or minus 60 degrees) rotamer and the barriers at omega = 120 and 240 degrees, but solvent destabilization at the cis barrier (omega = 0 degrees). Natural bond orbital analyses indicate that energetics of these effects are largely explained by overstabilization of the vacuum GT (omega = 60 plus or minus 60 degrees) and GG (omega = 300 plus or minus 60 degrees) rotomers. Solvent stabilization of theta conformations provides entropic stabilization.[unreadable] [unreadable] Dr. Woodcock and Edward O'Brien have been working on a comprehensive study to identify the strengths and weaknesses with advanced sampling techniques for simulations. More specifically, they are using standard Replica Exchange (REX) methodology in combination with multiplexed REX (MREX) to examine the effectiveness of self-guided Langevin dynamics (SGLD). This work is being carried out on both all-atom systems (i.e. a model beta-hairpin peptide) and a coarse grain representation of the ACBP protein. Preliminary results are very encouraging and should allow us to define a new paradigm for performing enhanced sampling simulations. [unreadable] [unreadable] Dr. Damjanovic is studying the structural consequences of ionization of internal groups in proteins. The work is performed in collaboration with experimental group of Prof. Bertrand Garcia-Moreno (JHU). Through experiments it is known that some variants of staphylococcal nuclease (SN) in which an ionizable group is buried in the protein interior exhibit conformational rearrangements triggered by pH induced ionization of internal groups. Through a relatively novel technique for advanced sampling, i.e., Self-Guided Langevin Dynamics (SGLD) simulation method, changes in secondary structure that are consistent with experimental findings are found. The simulations suggest that ionization of internal groups enhances water penentration which, in certain cases, can lead to unfolding of the protein core.[unreadable] [unreadable] Dr. Damjanovic is also studying the regulatory interactions in nitrogen regulatory protein C (NtrC). NtrC can exist in two conformations: inactive and active. Binding of a phosphate group stabilizes the active form. SGLD simulations of this protein indicate large conformational heterogeneity in the region involved in structural transition, and suggest that the transition pathway involves partial unfolding. Continuum electrostatics calculations and SGLD simulations suggest that phosphorylation may result in charging of a nearby His residue, which in turn may further modulate the conformation of a key helix.[unreadable] [unreadable] Dr. Okurs research involves examining the structural and energetic basis for diabetes. In collaboration with Professor Michael Weiss of Case Western Reserve University School of Medicine, NMR experiments on a single chain insulin analog (SCI-57, wild type insulin chains A and B linked with a 6-residue connecting peptide, PDB_ID: 2JZQ) were investigated and we observed a significant increase in stability with respect to wild type insulin. We speculate that the connecting peptide has a dampening effect on the fluctuations between the two chains. We have been investigating this stabilizing effect via molecular dynamics simulations of wild type and single chain insulin using CHARMM. Initial results are consistent with the hypothesis that the single chain analog has more defined structure than the wild type insulin. Detailed analysis of the fluctuations as a probable cause for increased stability is still ongoing.[unreadable] [unreadable] Mr. O'Brien's research has focused on the development and application of the Molecular Transfer Model (MTM). The MTM is a statistical mechanical method for accurately predicting osmolyte, denaturant, and pH effects on a protein's thermodynamic properties by combining ergodic molecular simulations with experimentally measured, or theoretically computed, transfer free energies of individual amino acids. The MTM has been shown to be in quantitative agreement with a number of published experiments. The MTM is the first, and only example in which molecular simulations have been capable of studying equilibrium folding and unfolding as a function of osmolyte type, osmolyte concentration, and pH. All previous computational studies used temperature to examine folding/unfolding. For this reason the MTM is useful because it can predict quantities that can be directly compared to experiment and it offers a molecular level interpretation of these phenomena based on the simulation structures it utilizes.[unreadable] [unreadable] We have applied the MTM to understand protein denatured state properties under varying solution conditions. We found that denatured state collapse occurs as urea concentration decreases and that residual secondary structure persists even at high denaturant concentrations. We also applied the MTM to examine how accurate FRET (Forestor resonance energy transfer) inferred denatured state properties are. Using measurements of the FRET efficiency as a function of denaturant concentration, in conjunction with simple polymer models (such as the Gaussian chain model), a number of experimental research groups have inferred denatured properties such as the radius-of-gyration and persistance length. Using the MTM, we found that error associated with these inferred properties can be as large as 25%, suggesting that this approach results only in qualitative estimates of denatured state properties.[unreadable] [unreadable] Another area of research Mr. O'Brien is focused on is confinement effects on protein folding, due to its relevance to a number of problems in biology and biotechnology. We have examined the stability of different helix-forming sequences upon confinement to a carbon nanotube. We showed that the interplay of several factors that include sequence, solvent conditions, the strength of nanotube-peptide interactions, and the nanotube diameter determines confinement-induced stability of helicies. Our results provide a framework for interpreting a number of experiments involving the structure formation of peptides in the ribosome tunnel as well as transport of biopolymers across nanotubes.
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{
"pile_set_name": "NIH ExPorter"
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To investigate how human liver cells prevent or retard the malignant effects of chronic exposure to carcinogens present in the environment, the enzymology of the excision repair of DNA damaged by model environmental carcinogens will be investigated. In these initial studies a key DNA repair enzyme, human liver DNA endonuclease specific for apurinic/apyrimidinic sites (apurinic DNAse), will be studied. Preliminary studies have demonstrated that autopsy liver is a rich source of apurinic DNAse and have also suggested that the activity of apurinic DNAse in autopsy livers is fairly stable. In initial experiments, phosphocellulose chromotography has revealed multiple species of human liver apurinic DNAse. Each species of apurinic DNAse will be purified further by column chromotography on DNA cellulose and hydroxyapatite. The specificity of endonucleolytic incision by each enzyme will be examined by assaying incision with apurinic DNA, with apyrimidinic DNA, and with DNA damaged methyl methane sulfonate, by nitrogen mustard, by N-acetoxy-2-acetylaminofluorene, and by ultraviolet irradiation. Each purified apurinic DNAse will be tested for associated non-endonucleolytic activities including DNA exonuclease, DNA phosphatase, and DNA glycosidase. Taken together these studies should begin to define how human liver cells repair DNA damaged by environmental carcinogens.
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{
"pile_set_name": "NIH ExPorter"
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Maintaining optimal health, both physical and cognitive, throughout the aging process is critical to minimizing healthcare costs and morbidity and mortality associated with diseases of aging. The integrity of the vascular system is essential for healthy aging. Aging-related structural and functional disturbances in the macro- or microcirculation contribute the development of cognitive dysfunction and declining physical performance. Early life factors, from birth through childhood and adolescence, may play an important role in successful cognitive and physical aging via the aging of the vascular system. In the proposed study we will examine the role of vascular aging in maintenance of cognitive and physical performance by recruiting 1,257 participants in the Bogalusa Heart Study cohort who participated in cardiovascular risk factors examinations at least twice in childhood and twice again in adulthood. Participants will undergo cognitive function testing, physical function assessments and vascular risk factor examination with noninvasive studies of vascular structure and function. Birth weight and childhood socioeconomic and risk factor data is available for all individuals. Longitudinal analysis will be used to examine the relationship of early life risk factrs to subclinical vascular disease markers, while linear models will be used to examine the role of vascular risk factors and subclinical markers in maintenance of cognitive and physical function. This study represents a unique opportunity because all vascular disease risk factors have been collected prospectively from early life to middle-age in this bi-racial (black/white), rural community, allowing for exploration of race and gender relationships with cognitive and physical function from mid-life. The proposed research will link vascular risk factors across the life span and subclinical vascular markers in early middle age with cognitive and physical performance in later middle age. In doing so, we will identify risk factors, timing and subpopulations for intervention that could reduce the incidence of cognitive and physical decline in old age and improve the rate of successful aging for persons across the nation.
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{
"pile_set_name": "NIH ExPorter"
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Sexually violent themes have become increasingly prevalent in both pornographic and popular literature over the last several years. Exposure to violent pornography has been associated with a number of rape-related attitudes and behaviors. Alcohol consumption has been implicated in the commission of rape, but very little research has addressed the relationship between alcohol consumption and responses to sexually violent materials. The only two studies that have examined this connection indicate that expectancies associated with alcohol consumption are responsible for increased arousal to and interest in deviant sexual material. However, a systematic examination of the interaction between alcohol and differing levels of violence portrayed in these materials has yet to be undertaken. Furthermore, the impact of these materials on women has also been neglected. Social influence theory may provide a valuable framework to examine these relationships. Alcohol may serve as a cue, or discriminative stimulus, to engage a set of norms associated with sexual responding that would otherwise be considered unacceptable. To study this proposition, four experiments are proposed to examine the interrelationships among the pharmacological versus expectancy effects of alcohol consumption, normative perceptions, and responses to sexually violent material in both men and women. The contribution of individual difference traits will also be examined.
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{
"pile_set_name": "NIH ExPorter"
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The confocal Core Family will provide equipment and technical expertise for co-investigators in cystic fibrosis research at the Mayo Foundation. This unique core facility will allow investigators to examine subcellular localization of proteins and interacting proteins, to determine levels and time course or changes in intracellular ions and other fluorescently- labeled second messengers, to use caged compounds to examine the physiologic mechanism of rapid changes in intracellular second messengers, to routinely make three-dimensional reconstructions of living and fixed cells at high resolution, and to make use of optical tweezer technology to manipulate intracellular organelles. This confocal core facility is a state-of-the-art facility with several unique capabilities, including the ability to use two-photon excitation for imaging and high resolution photolysis of caged compounds. In the near future, technical development of a new spectral imaging system will be completed, allowing up to eight fluorescent dyes of various wavelengths to be imaged simultaneously. In addition, the confocal facility has implemented infrared optical tweezers, and has extensive three-dimensional reconstruction software capabilities. In particular, the core facility will provide assistance in the imaging of improperly folded and mutant CFTR gene products, proteins associated with pI/Cln and their intracellular distribution, and the co-localization of mucin with other proteins and its expression within tissues and cells.
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{
"pile_set_name": "NIH ExPorter"
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Intestinal tract disease is responsible for a substantial amount of the morbidity and mortality among people with the genetic disease cystic fibrosis (CF). The manifestations of CF intestinal disease frequently occur during infancy and result from pancreatic exocrine insufficiency (PI), resulting in profound nutrient malabsorption and malnutrition, and consequently in growth failure and intestinal obstruction. Despite aggressive, early nutritional intervention with pancreatic enzyme replacement and other therapies, early growth failure remains common among infants with CF, suggesting that characteristics other than PI contribute to growth and nutritional failure in infants with CF. The identification of thes growth and nutritional determinants in children with CF therefore remains a key research goal in CF care. Recently, a clue to a potentially critical contributor to nutrient and energy uptake in children has emerged from the study of the microbes present in the gastrointestinal tract- the gut microbiota. Research has identified a complex relationship between gut microbiota, nutritional intake, nutrient absorption, and other health measures. GI tract microbes are known to contribute significantly to human nutrient metabolism and energy harvest, and they are altered in many disease states. Therefore, infant GI tract microbes may represent important determinants of CF nutritional outcomes, which in turn can significantly impact severity of lung disease and overall longevity. We recently showed that infants and children with CF had GI microbiota that differ markedly from those of children without CF, and that this dysbiosis correlated with severity of GI dysfunction. However, the relationships between CF GI microbiota with growth and other clinically important outcomes have not been studied. We hypothesize that the intestinal microbiomes among children with CF correlate with severity of malabsorption, intestinal inflammation, gastrointestinal signs and symptoms, vitamin and mineral deficiency, and nutritional status. To test these hypotheses, this ancillary study will define the fecal microbiomes of 250 infants with CF being enrolled in an ongoing multicenter study of infant nutrition (the Baby Observational NUtrition Study (BONUS), funded by the Cystic Fibrosis Foundation and the National Institutes of Health) using ultra high-throughput, culture-independent sequencing methods. With these resources, we will (1) define the fecal microbiomes, composed of the microbiota (the species of microbes present) and metagenomes (the microbial genes present, which reflects the metabolic capacity of the microbiota) of at least four stool samples per subject over the first year of life, and (2) determine the relationship of fecal microbiomes, and their evolution over the year of study, with nutritional and clinical parameters (including weight, height, body mass index, growth rate of each of these parameters among subjects, serum vitamin levels, fecal nutrient and metabolite content, GI symptoms, and measures of inflammation). We will use massively parallel next- generation DNA sequencing methods, combined with advanced bioinformatic analytical methods, with which we have extensive experience. Our goal is to define the role of the infant CF GI microbiome in growth and nutrition, with the hope of identifying interventions that will improve early growth, and thus long-term outcomes, in CF.
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{
"pile_set_name": "NIH ExPorter"
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Data analysis in aging research poses distinctive challenges that require careful selection and tailoring of statistical methods, because of older adults'multiple comorbidities / medications, competing rtsks, and high drop out rates due to death or disability. In addition. In the current context of limited health care resources in the face of an Impending surge in the number of older Americans, consideration has to be given to the costeffectiveness of interventions to restore or maintain the independence of older adults. ACEC will provide important guidance to OAIC researchers and projects, as well as other UCLA aging researchers, in the selection and conduct of appropriate statistical and economic analyses. The Analysis/Cost-Effectiveness Core will provide analytic support in research study design (focusing on power considerations), statistical data analysis, and interpretation and accurate description of findings, for: [unreadable] OAIC Career Development Awardees, [unreadable] OAIC supported pilots and development projects, [unreadable] UCLA junior researchers conducting research that meet the UCLA OAIC mission, theme, and goals, [unreadable] Externally funded UCLA research projects that meet the UCLA OAIC mission, theme, and goals The core will also provide training workshops / tutorial seminars on state-of-the-art methods for statistical data analysis, comparative effectiveness studies, and cost effectiveness analysis. It will also assess the cost-effectiveness of successful clinical interventions studied in the OAIC Finally, the ACEC will disseminate new analytic approaches to comparative effectiveness studies, as they become available. In collaboration with the other OAIC resource cores, ACEC will support externally funded aging research projects, promote aging research aimed at maintaining the independence of older adults, ensure the quality of its products, hasten its translation into clinical practice, and nurture a new cohort of aging researchers and future leaders.
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{
"pile_set_name": "NIH ExPorter"
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Gene-environment interactions may be defined as a genotype's phenotypic expression being altered by the environment, e.g., the weaker effect of FTO polymorphisms on body mass index (BMI) in exercisers compared to non-exercisers. However, our preliminary analyses suggest that the phenotypic expressions of FTO polymorphisms are diminished in lean vis-a-vis overweight individuals. This suggests an alternative interpretation (Figure 1). Based on the fact that exercisers are leaner than non-exercisers, we hypothesize that physical activity affects BMI, which in turn affects FTO gene expression, rather than exercise affecting FTO gene expression directly. Our preliminary analyses of lipoproteins and BMI in 1800 subjects suggest that a genotype's phenotypic expression often increases with the quantile of the phenotype, i.e., when the value of the phenotype is high relative to its distribution in the population. We refer to this dependence as quantile- dependent penetrance. This dependence differs from the standard regression model, which assumes that the same relationship between the dependent and independent variables (e.g., phenotype vs. genotype) applies to all quantiles of the dependent variable. We propose to apply quantile regression to data available through the NHLBI Candidate-Gene Association Resource (CARe), DBGaP, and other studies to assess whether quantile-dependent penetrance applies to most other genotype-phenotype relationships. Although our preliminary analyses lacked the statistical power to assess this phenomenon for individual SNPs, its demonstration in genetic risk scores suggests that the majority of SNP effects must also be quantile dependent. We will also test whether prior assertions of gene- environment interactions are attributable to quantile-dependent penetrance, whether allowing the genotypic expression to vary with the percentile of the trait distribution significantly increases the phenotypic variances explained, and whether quantile-dependent penetrance can be extended to SNP-SNP interactions. This proposal is hypothesis driven; i.e., we hypothesize that most genotype-phenotype associations increase substantially with the percentile of the phenotype. This hypothesis is based upon the premise that the most important gene-environment interaction involves an individual's own physiological environment within which the genes are expressed. The lowest to highest percentiles of a trait's distribution represent a range of physiologic parameters, genetic make-ups, and gene-gene interactions whose presence may be essential for the genetic variant to be expressed. To our knowledge, quantile-dependent penetrance has not been proposed as a primary basis for genotype-phenotype relationships, or as an alternative to gene-environment interactions.
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{
"pile_set_name": "NIH ExPorter"
}
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Understanding of the spectrum of the HIV and HCV epidemics in injection drug users (IDU) has been advanced by a number of outstanding prospective studies conducted in various part of the world, including, North America, Europe, and Australia. Each of these studies has furthered our understanding of disease prevalence and incidence, risk behaviors, and natural history, including clinical, immunological and virological factors. This application seeks funding for the first global collaboration of prospective studies of HIV and HCV in IDU. The "International Collaboration of Incident HIV and HCV in Injecting Cohorts" (InC3), is merged international multi-cohort project of pooled biological and behavioral data from nine prospective cohorts of IDU. The InC3 collaboration will facilitate new in-depth studies of HIV and HCV infection not possible from each individual study. This collaboration will allow us to study 4,091 IDU who have been followed for a collective 9,016 person-years of observation, 859 incident HCV infections with longitudinal follow up, and 575 HIV infections. This proposal includes six research questions that represent some, but not all, of the potential breadth and depth of the proposed InC3 research collaboration, including: assessment of temporal trends in HIV and HCV incidence, examination of HCV incidence by HIV status and sexual behavior, estimates of rates and determinants of HCV viral clearance and reinfection, examination of the effects of HCV infection disclosure are on risk behaviors, and an evaluation of outcomes related to clinical treatment of acute HCV in IDU. Many of these questions remain unanswered due to the relatively small number of small numbers of behavioral, viral, or clinical events in each study. However, since many have been meticulously characterized in each of these rich longitudinal cohorts, merging data will offer the statistical power needed to make firm conclusions. The significant and innovative scientific agenda and the InC3 leadership will be supported by an administrative core and Data Coordinating Center (DCC). InC3 will promote and support the development of new study questions and research opportunities through supporting research exchange and training for participating junior scientists. Lastly, some gaps in biological data will be addressed with support for laboratory measures from stored specimens. This proposed InC3 study, will accomplish the following: (1) creation of a rich new data source that will fill scientific knowledge gaps that heretofore have not been possible in single studies alone;(2) enable new research relevant to understanding, preventing, and treating HIV and HCV in IDU;(3) the creation of a multilateral collaborative of researchers and scientists committed to long term cooperation and partnership in areas including: epidemiology, behavioral studies, clinical research, and laboratory science. Many of the investigators of this proposed multicenter project are experienced and successful in pooling and analyzing large epidemiological data in both a national, and international, context. Despite the obvious benefits to public health, such an organized effort has never been attempted with cohorts that have studied both HIV and HCV. InC3 is significant, innovative, will advance research relevant to the global burden, consequences, prevention, and treatment of HIV and HCV infections among IDU. PUBLIC HEALTH RELEVANCE: This international collaboration: the InC3, will create a merged international multi-cohort project of data pooled from nine prospective studies of IDU conducted in four countries over 22 years. The collaboration will result in the largest dataset available for analyses and research on HIV and HCV infections in IDU, including acute HCV infection and HIV/HCV co-infections. The combined expertise and research data will advance research relevant to the global burden, consequences, prevention, and treatment of HIV and HCV infections among IDU.
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{
"pile_set_name": "NIH ExPorter"
}
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It is clear that repeated exposure to stress increases the risk of developing and expressing symptoms of human disorders of fear and anxiety;however, little is known about the mechanisms by which this occurs. Many studies have shown that numerous brain areas, including the hippocampus, are adversely impacted by chronic stress. For example, chronic stress has been directly linked to atrophy of neuronal processes, reduction of neural plasticity, and deficits in behavioral performance on mnemonic tasks. However, recent data indicates that one brain region, the amygdala, exhibits a relatively unique response to chronic stress. That is, chronic stress causes growth of dendritic processes in amygdala neurons and increases in anxiety. These findings suggest the intriguing hypothesis that chronic stress produces fundamentally opposing effects in the hippocampus and amygdala. We have behavioral data supporting this hypothesis: Chronic stress enhances amygdala-dependent delay fear conditioning but impairs hippocampus-dependent trace fear conditioning. In this proposal, we will combine sophisticated multi-site tetrode recording techniques and cutting-edge molecular methods (viral-mediated gene transfer) with immunohistochemical and behavioral techniques in rats to further examine this hypothesis and explore two mechanisms by which opposing effects may occur. First, we will characterize chronic stress-induced changes in both spontaneous and associative spike firing in the hippocampus and amygdala. Second, we will determine whether glucocorticoids, making up one component of the neuroendocrine response to stress, produce opposing effects on electrophysiological, behavioral, and gene expression measures in the amygdala and hippocampus. Third, we will determine the contribution of growth hormone, a gene that is oppositely regulated in the hippocampus and amygdala after chronic stress, to stress- induced changes in behavioral measures of fear memory. Because of the important role that the amygdala plays in regulating anxiety and fear, experiments assessing the unique effects of chronic stress on the amygdala may shed light on the specific mechanisms by which chronic stress exacerbates mental illnesses such as post-traumatic stress disorder. Identifying mechanisms by which stress exacerbates fear may lead to new therapeutic targets for the treatment of mental illness. PUBLIC HEALTH RELEVANCE: This research will compare the impact of chronic and intense stress on the hippocampus and amygdala and determine the contribution of glucocorticoids (a hormone secreted in response to stress) and growth hormone (a hormone that is uniquely upregulated in the amygdala after chronic stress) to these effects. This work will shed light on the multiple ways in which chronic stress can enhance fear and anxiety, and it will identify novel therapeutic strategies by which stress-enhancement of fear memory may be reduced. This is particularly important as chronic stress is a potent trigger for affective mental illnesses such as post-traumatic stress disorder (PTSD), but the mechanisms by which stress triggers this class of mental illnesses is unknown.
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{
"pile_set_name": "NIH ExPorter"
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A symposium on developmental reproductive biology and sexual differentiation will be held in conjunction with the 12th annual meeting of the Society for the Study of Reproduction at the University of Laval, Quebec, Canada. Three scientists will discuss the current "states of the art" with respect to the immunogenetics, endocrine-genetics and epithelial-stromal interactions in sexual differentiation, while another three will consider sex refersal in lower forms, the role of hormones in brain development, and in the psychosexual differentiation of the human. Together, these six presentations will evaluate the fundamental aspects of developmental reproductive biology and also consider how comparable are the processes of the differentiation of the peripheral reproductive organs and the brain. The proceedings of the symposium will be published in the February, 1980 issue of the Biology of Reproduction.
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{
"pile_set_name": "NIH ExPorter"
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Arterial thrombotic diseases such as myocardial infarction and stroke are the leading cause of death in the U.S. Their socioeconomic and psychological impacts are immense. Drugs that can efficaciously reduce or eliminate these diseases are of great clinical importance and should have major commercial applications. Unfortunately efficacious drugs are limited. We consider it important to develop new anti-thrombotic drugs. In our pilot work we have identified anti-platelet properties in the ethanol-soluble fraction of a Chinese fungus, Cordyceps sinensis. The fraction exhibited inhibitory activity against platelet aggregation and secretion. Its inhibitory effect appears to be independent of thromboxane A2 and may be related to cAMP elevation. We propose in this SBIR application to purify the active component, evaluate its biological activity, elucidate its mechanism of action and assess its anti- thrombotic efficacy. The Phase I study will determine the feasibility of the purifying procedures and testing anti-platelet properties. We plan to carry out a sequence of solvent-solvent extractions which have been developed in our laboratory and is effective in segregating active compounds on the basis of their polarity into one or two highly enriched fractions. The inhibitory effect of the concentrated fractions on platelet aggregation and secretion will be evaluated in a lumiaggregometer. The fractions will be then processed by step-by-step procedures to isolate the active component. The Phase I study will lay the foundation essential for the full development of the active component into anti-thrombotic drugs in Phase II and subsequent clinical studies.
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{
"pile_set_name": "NIH ExPorter"
}
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The Yale Mouse Metabolic Phenotyping Center (MMPC) was established in the spring of 2000 with the mission to advance research in the area of diabetes by providing novel experimental tools to the scientific community for phenotyping mouse transgenic models of diabetes and related disorders using state-of-the-art methodology that is not widely available. The goals of this program are to: 1) broaden the scope of techniques available to investigators; 2) standardize key methodologies; 3) expedite the completion of research; and 4) compile and make accessible phenotyping data to the scientific community upon its publication or two years after the completion of phenotyping services, and 5) develop novel methodology for phenotyping transgenic mice through an national administered Pilot & Feasibility Program. The cornerstone of the Yale MMPC is its two research cores, the Integrative Physiology Core and the Metabolomics Core. The Integrative Physiology Core performs glucose-insulin clamps using both radioactive and stable isotope technology to assess insulin action in liver, skeletal muscle and fat of awake mice. This core also performs hyperglycemic clamps to assess in vivo B-cell function as well as hypoglycemic clamps to assess complications relating to defective glucose counter-regulation. This core also performs 24-hour, automated, non-invasive assessment of activity, food/water consumption and energy expenditure in an environmentally controlled room. The Integrative Physiology Core also contains a Mouse Imaging Sub-Core, which performs state-of-the-art noninvasive measurements of intracellular metabolites in muscle, liver and brain of transgenic mice using MRI methods which in turn is used to assess in vivo metabolic flux through critical biochemical pathways as well as whole body measurement of fat and muscle tissue with MRI. The Metabolomics Core provides automated high throughput clinical chemistry analyses of microliter samples of mouse plasma and urine. In addition this core provides state-of-the-art GC-MS and LC/MS/MS analysis of plasma, urine and tissue samples obtained from transgenic mice. The Animal Care Core provides a centralized facility for coordinating the import, initial quarantine, screening and monitoring of mice. This Core provides stable, biocontainment housing, husbandry and health care for mice submitted to the Center and determines the microbiological profile of each imported cohort. The Administrative Core oversees the operation of the Center, coordinates the importation of all mice to Yale, performs all of the material transfer agreements, coordinates and schedules meetings and teleconferences between investigators and MMPC Core Directors, coordinates the Pilot & Feasibility Program and administers the Enrichment Program. Overall the Yale MMPC serves as a national research center that provides investigators both inside and outside Yale access to unique, state-of-the-art, standardized methods to further characterize their novel transgenic mouse models of complex metabolic diseases in a cost efficient manner.
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{
"pile_set_name": "NIH ExPorter"
}
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The long-term goal of this project is the chromosomal mapping and characterization of mutations causing four skeletal dysplasias: nail-patella syndrome (NPS), Langer-Giedeon syndrome (LGS), Ellis van-Creveld syndrome (EvC), and cartilage-hair hypoplasia (CHH). The skeletal dysplasias are a diverse group of over 100 clinical disorders whose underlying biochemical causes are, for the most part, unknown. Delineation of the genes responsible for these varied phenotypes will undoubtedly provide insight into the mechan- isms of normal chondrogenesis and growth. This project will focus on four conditions which are particularly amenable to new approaches for human genomic analysis. The approach to all four conditions will be similar: (1) establish a chromosomal localization by traditional genetic linkage analysis, homozygosity mapping, or identification of gross chromosomal deletions (2) use of yeast artificial chromosome (YAC) cloning and pulsed field gel electrophoresis technology to define DNA fragments closer to, and eventually encroaching upon, the actual mutant locus and (3) establish by sequence analysis the specific mutation causing the disorder. The first step in this process has already been accomplished for NPS and LGS, both of which have known chromosomal locations. NPS is an autosomal dominant condition localized to human chromosome 9q34 and closely linked to the adenylate kinase and ABO blood group loci. LGS, which may be a "contiguous gene syndrome" involving loci for both multiple exostoses and trichorhinopharyngeal syndrome type 1, has been localized to 8q24 on the basis of patients with microscopically detectable deletions involving this band. EvC and CHH are autosomal recessive skeletal dysplasias whose biochemical bases are unknown. Both are unprecedentedly prevalent in the Old Order Amish. This fact in combination with the structure of the Amish "founder populations" mean that these disorders are well-suited to the use of "homozygosity mapping" as well as the traditional genetic linkage approach. Restriction fragment length polymorphisms associated with candidate genes encoding known structural proteins of the cartilage matrix and with anonymous DNA fragments spanning the human genome will be used. Fifty-five nuclear families with EvC, and 72 families with CHH be available for these studies.
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{
"pile_set_name": "NIH ExPorter"
}
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The primary goal of this project is aimed to improve the understanding and control of diastereofacial selectivity in asymmetric organic reactions. The conformational preferences of chiral alkenes will be determined through the J averaging method with the variable temperature NMR technique. The free energy differences for the two diastereomeric transition states will be estimated through the isomeric products ratio. The stereochemistry of the products and the conformational preferences of the starting materials should form a basis for the understanding of the important factors in the transition states, therefore the control of the pi-facial selectivity. Based on our preliminary results, the modern high field NMR technique is well suited to study the relative conformational stabilities of chiral alkenes. Although the significance of the conformational profiles of the chiral alkenes has been recognized for sometime, presently experimental techniques are either difficult to interpret or cannot be applied to a wide range of reagents. The information obtained through the VT NMR study should prove to be of great value and interest to the chemical community. Previous studies aimed at the understanding of electronic effects have concentrated on comparing C-C bonds with slightly different polarities. This project employs substrates which have either a C-H bond or a C-0 bond anti to the attacking nucleophile. Thus the properties of the implied orbital interaction in the transition state are different enough to allow little ambiguity in the final analysis of the results.
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{
"pile_set_name": "NIH ExPorter"
}
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This investigation will include the construction of physical and deletion maps of the human chromosomal region 11p11.p12. Overlapping YACS and cosmids will be identified through the construction of a physical map. The deletion map will allow definition of a minimal region of chromosomal loss in patients with multiple exostoses and possibly with parietal foramina and/or mental retardation. Cosmids identified through the physical mapping effort will be used to perform exon trapping and identify candidate genes potentially involved in a contiguous gene deletion syndrome that includes multiple exostoses. All candidate genes will be analyzed for a potential role in multiple exostoses. It is also possible that this work will result in the identification of genes involved in other associated, but unrelated, traits such as parietal foramina or mental retardation.
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{
"pile_set_name": "NIH ExPorter"
}
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The purpose of this study is to determine the phenotype of multiple extracellular matrix molecules in adult mouse glomeruli from different strains of mice. Our postulate is that mice prone to develop glomerulosclerosis have a higher level of expression of genes coding for extracellular matrix at baseline prior to the appearance of lesions. We used the increased sensitivity afforded by the polymerase-chain reaction to assess alpha-1 type I and several alpha chains of type IV collagen mRNA as well as laminin subchains and tenascin in freshly microdissected normal adult mouse glomeruli. RT-PCR reactions for mRNA encoding these components were also performed using mesangial cell lines previously isolated from the same strain of mice. Type IV collagen mRNA was easily detectable in normal adult mouse glomeruli as well as in the cell lines. On the other hand, type I collagen mRNA was not detected in normal glomeruli, despite increasing the number of PCR cycles from 25-45 (roughly a 1000 fold increase in sensitivity). Assays using competitive PCR were developed. The current study provides evidence that the expression of types I and IV collagen in normal glomeruli is regulated at the pretranslational level in vivo. They also provide evidence for a continuous turnover of basement membrane in the adult glomeruli. We have applied this method to a mouse strain which develops spontaneous glomerulosclerosis, the ROP-OS mouse. This mouse is originally derived from a radiation-induced mutation. Our preliminary data indicate that the mutation induces a rapid glomerulosclerosis on a ROP background but has little effect on a C a C57B background. The ROP-OS mouse has a 3 fold increase in mRNAs coding for type IV collagens and laminin early in life at a time when morphologic evidence of glomerulosclerosis is quite discrete.
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{
"pile_set_name": "NIH ExPorter"
}
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We have recently observed that the deleterious effect of systolic overload induced by hypertension on cardiac performance and contractile proteins in rats can be partially or completely prevented or reversed by exposing the hypertensive rats to a chronic swimming program. The purpose of the present proposal will be to further elicit the types of physical training that might protect or reverse the effects of the pathologic overload on the heart. Experiments will be conducted to determine whether physical training by running is effective; whether pre-training the animals can prevent the deleterious effect of subsequent systolic overload; and whether physical training can reverse the effects after deleterious effects of systolic overload have been well established. Several others models of pathologic overload besides renal hypertension will be investigated. These will include: Dahl hypertensive rats, spontaneous hypertensive rats, rats with mild and with severe hypertension due to coarctation of the aorta, rats with overload induced by arterial venous fistula and rats with compensatory hypertrophy associated with myocardial infarction. Additionally, myopathic hamsters will be investigated to determine the prevention or reversibility of that lesion. Experiments will include heart perfusions, studies of myocardial vascular responses, analysis of contractile protein enzymology and isoemzymes, and the role of thyroid hormones.
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{
"pile_set_name": "NIH ExPorter"
}
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Summary of Work: During the past year, studies on HIV protease were aimed primarily at determining the conformational selectivity of proteolytic cleavage, and its consequences. In particular, three of the eight cleavage sites on the Gag-Pol polyprotein which are the target sequences for HIV protease involve Araa-Pro bonds, where Araa corresponds to the aromatic amino acids tyrosine or phenylalanine. Since imide bonds formed with proline exhibit significant cis/trans isomerism, this leads to a question concerning the specificity of cleavage of the Araa-Pro bonds. This specificity can be determined under conditions in which the rate of cleavage greatly exceeds the isomerization rate - i.e., high enzyme concentrations and low temperature. In order to study the reaction, we have utilized two approaches: 1. A fluorogenic substrate peptide analog of the p17/p24 cleavage site of the gag polyprotein with the sequence: Arg-Glu(EDANS)-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(DABCYL)- Arg was used for HIV protease assays. Hydrolysis of the Tyr-Pro bond results in increased separation of the DABCYL fluorophore form the EDANS quencher, leading to an increase in fluorescence. 2. A fluorinated substrate: Ser-Gln-Asn-FPhe-Pro-Ile-Val-Gln, where FPhe = L-4- fluorophenylalanine, was used for NMR studies. The fluorine nucleus acts as a useful reporter group for both cis/trans isomerization and for cleavage, and in fact provides distinct signals depending on where the peptide is cleaved. As of this date, the 19F NMR studies were limited to the model aspartyl protease, pepsin, since HIV protease proved too unstable for use in the NMR experiments. Specificity for cleavage of the trans imide bond conformation was demonstrated in all cases. The conformational selectivity of proteolytic cleavage of Araa-Pro imide bonds by HIV protease provides a possible explanation for the accumulation of the cellular protein cyclophilin by HIV. In particular, the cyclophilin could act as an auxiliary enzyme for the protease, converting inactive, cis imide bonds into trans bonds which are substrates for the protease.
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{
"pile_set_name": "NIH ExPorter"
}
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Frontotemporal dementia (FTD) is a neurodegenerative disorder characterized clinically by changes in personality, social behavior, executive function, and/or language dysfunction, often in association with a movement disorder. The clinical manifestations of FTD are correlated with degeneration of the frontal and anterior temporal lobes. FTD can present either sporadically or as a familial disorder. Some of these kindred demonstrate an autosomal dominant pattem of inheritance. Elucidation of linkage to chromosome 17 and the subsequent identification of mutations in the tau gene (MAPT) in FTD cases provided direct evidence that tau protein dysfunction can lead to neurodegeneration. More than 30 different MAPT mutations have been identified. Studying familial forms of FTD can help elucidate the etiology and pathophysiology of FTD. Since FTDs are clinically heterogeneous, it is imperative to collect and study a large number of kindred to identify new mutations in MAPT and discover novel disease-associated genes. We began to collect DNA samples from individuals and families with FTD or related neurodegenerative disorders with the goal of enabling research genetic studies of such conditions. We now propose the formal establishment of a Genetics Core as a part of the Program Project Grant (PPG), Frontotemporal Dementias: Genotypes and Phenotypes to continue and expand this effort. The Genetics Core will provide genetic counseling to individuals and families with FTD and support the on-going collection and storage of DNA from these families. Limited genetic analysis of known genes associated with FTD will be performed to identify mutations. Finally clinically-relevant genetic tests will be translated to the clinical (CLIA-approved) laboratory. The collection of well-defined FTD cohorts and/or identification of new mutations will support genetic discoveries from Project 2, help advance the clinical characterization of FTD in Project 1, enable detailed biochemical and immunohistochemical analyses of FTD (Project 3), lead to the generation of new animal models of disease (Projects 2 & 4), and serve as a resource for genetic analysis of pathologically diagnosed cases of FTD in the Neuropathology Core to facilitate correlations between genotype and the clinical, neuropathological, and biochemical phenotypes.
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{
"pile_set_name": "NIH ExPorter"
}
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The proposed research study seeks to use and expand an existing prospective longitudinal adoption study, the Early Growth and Development Study (EGDS: R01HD42608) to better understand how genetic factors, prenatal drug exposure, and postnatal rearing environment operate together to influence toddler development, and ultimately, an individual's risk of later drug use. We propose to recruit 200 yoked adoption units (birth parents and adoptive families linked through the adopted child - approximately 866 individuals) and to capitalize on the existing 350 yokes recruited by the EGDS study. The 550 total yokes (approximately 2,382 individuals) will provide sufficient power to examine possible interactions among genes, prenatal drug exposure and postnatal rearing environment that EGDS was not designed to study. Using the same assessment strategy and measurement as EGDS, the new 200 yokes will be followed longitudinally and DNA will be collected from all participants in the full sample of 550 yokes. This expanded sample and longitudinal assessment, combined with the collection of DNA, enables the examination of the following specific aims: (SA1) Understanding how genetic risk for later drug use is expressed behaviorally in the toddler period;(SA2) Estimating prenatal exposure effects independent of genetic risk;(SA3) Estimating postnatal family environmental effects independent of genetic risk and prenatal exposure;and (SA4) Clarifying how genetic risk and prenatal exposure to drugs are moderated by postnatal family environment. Integrating the three possible mechanisms of the transmission of drug abuse to children - genetic, prenatal exposure and postnatal environment - has important potential for informing public health. Specifically, the proposed study is designed to disentangle the effects of genetic, prenatal and postnatal environmental factors on early child development and to examine interactions and correlations among these factors. This will help guide intervention efforts aimed at the prevention of drug abuse in children of drug-dependent or drug-abusing parents by identifying which postnatal family environmental factors have the most impact on ameliorating risk from genetic and prenatal exposure factors.
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{
"pile_set_name": "NIH ExPorter"
}
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The overall objective of the research in this laboratory has been to attempt to identify non-histocompatibility antigens which may play a role in the immune response to allo- and xenografts.
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{
"pile_set_name": "NIH ExPorter"
}
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This case-control study is designed to investigate factors which may be related to the decreased risk if IDDM experienced by Hispanics compared with Anglos in Colorado.
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{
"pile_set_name": "NIH ExPorter"
}
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The benzodiazepine-GABA-chloride ionophore receptor complex has been demonstrated to be involved in the physiologic and psychologic effects of ethanol. Diazepam, a benzodiazepine, binds to this receptor complex, and demonstrates a cross-tolerance to ethanol. Recent studies have shown that diazepam-induced alterations in eye movements offer a useful measure of benzodiazepine receptor sensitivity in humans. Preliminary findings at the NIMH and NIAAA suggest an increased sensitivity to the effects of diazepam in alcoholics as measured by saccadic eye movements, in alcoholics. In this study subjects will be administered diazepam and sub- sequently evaluated for changes in EEG, ERP (event-related potentials). body sway, vigilance tracking, memory, mood assessment and expectancy, ACTH, cortisol, prolactin, and growth hormone. This study has been terminated. The results are being analyzed and will be submitted for publication.
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{
"pile_set_name": "NIH ExPorter"
}
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Gene-environment interactions may be defined as a genotype's phenotypic expression being altered by the environment, e.g., the weaker effect of FTO polymorphisms on body mass index (BMI) in exercisers compared to non-exercisers. However, our preliminary analyses suggest that the phenotypic expressions of FTO polymorphisms are diminished in lean vis-a-vis overweight individuals. This suggests an alternative interpretation (Figure 1). Based on the fact that exercisers are leaner than non-exercisers, we hypothesize that physical activity affects BMI, which in turn affects FTO gene expression, rather than exercise affecting FTO gene expression directly. Our preliminary analyses of lipoproteins and BMI in 1800 subjects suggest that a genotype's phenotypic expression often increases with the quantile of the phenotype, i.e., when the value of the phenotype is high relative to its distribution in the population. We refer to this dependence as quantile- dependent penetrance. This dependence differs from the standard regression model, which assumes that the same relationship between the dependent and independent variables (e.g., phenotype vs. genotype) applies to all quantiles of the dependent variable. We propose to apply quantile regression to data available through the NHLBI Candidate-Gene Association Resource (CARe), DBGaP, and other studies to assess whether quantile-dependent penetrance applies to most other genotype-phenotype relationships. Although our preliminary analyses lacked the statistical power to assess this phenomenon for individual SNPs, its demonstration in genetic risk scores suggests that the majority of SNP effects must also be quantile dependent. We will also test whether prior assertions of gene- environment interactions are attributable to quantile-dependent penetrance, whether allowing the genotypic expression to vary with the percentile of the trait distribution significantly increases the phenotypic variances explained, and whether quantile-dependent penetrance can be extended to SNP-SNP interactions. This proposal is hypothesis driven; i.e., we hypothesize that most genotype-phenotype associations increase substantially with the percentile of the phenotype. This hypothesis is based upon the premise that the most important gene-environment interaction involves an individual's own physiological environment within which the genes are expressed. The lowest to highest percentiles of a trait's distribution represent a range of physiologic parameters, genetic make-ups, and gene-gene interactions whose presence may be essential for the genetic variant to be expressed. To our knowledge, quantile-dependent penetrance has not been proposed as a primary basis for genotype-phenotype relationships, or as an alternative to gene-environment interactions. PUBLIC HEALTH RELEVANCE: We have previously shown that the effect of the genotype on a phenotype increases with the percentile of the trait distribution in genetic risk scores for total cholesterol, triglycerides, high-density lipoprotein cholesterol, and body mass index. This phenomenon was demonstrated using baseline data for a study of 1800 subjects. The purpose of this proposal is to demonstrate this phenomenon in much larger, more diverse data sets and to extend the findings to other variables. We believe that this phenomenon could be a general principal of phenotype-genotype relationships. The results could more sharply define gene-environment interactions into: a) effects of the environment on the genotype, which affects its phenotype expression, and b) effects of the environment on the phenotype which affects the penetrance of the genotype.
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{
"pile_set_name": "NIH ExPorter"
}
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The University of Alabama (UAB) Multipurpose Arthritis and Musculoskeletal Diseases Center (MAMDC) is a multidisciplinary program which fosters and promotes research related to the causes, diagnoses, treatments, and improved care of patients with arthritis and musculoskeletal diseases. In order to ensure operational effectiveness, the Administrative Unit of the MAMDC is specifically designed to initiate, promote, and maximize interactions among the Center Director and principal staff; relevant institutional academic and administrative units; staff at the National Institute of Arthritis, Musculoskeletal and Skin Diseases (NIAMS); regional, state, and local health agencies and organizations; and individuals and families served by the UAB MAMDC. The Administrative Unit is designed to enable proper governance of the various MAMDC Components through committees and regularly scheduled meetings and seminars serving to enhance communication among the various investigators and administrative staff. To assist the Center Director in his effort to ensure the highest quality in the operation of the UAB MAMDC, five committees with appropriate charges have been established: the Executive Committee; the Research and Development Committee; the Institutional Advisory Committee; the External Advisory Committee; and the Clinical Record System Utilization Committee. The Administrative Unit which has evolved at the UAB MAMDC during the past 14 years has proven to be effective in assisting the Director to ensure that the goals of the MAMDC are fulfilled.
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{
"pile_set_name": "NIH ExPorter"
}
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Chronic kidney disease (CKD) is a growing public health problem that affects 10% of Americans, increases morbidity and mortality, and imposes a huge economic burden. Persistent renal tubular injury is an important component of tubulointerstitial fibrosis (TIF), the common feature of progressive CKD of any etiology. Growth factors such as transforming growth factor-? (TGF??) are important determinants of how tubula epithelia respond to injury. Although excessive TGF-? signaling promotes TIF, blocking TGF-??using genetic tools results in increased tubular damage after mouse models of CKD, suggesting that TGF-??signaling has protective effects as well. TGF-??mediates many differet cellular effects including G1 arrest of cell cycle progression and epithelial de-differentiation. Tis proposal tests the hypothesis that TGF???signaling in the proximal tubule promotes survival ater chronic renal injury by inducing cell cycle arrest and epithelial de- differentiation. The first ai investigates how TGF-? alters cell cycle progression and how these changes affect epithelial survival using murine models of injury and pharmacologic inducers of G1 arrest. To test this, we will use genetically modified mice that lack the TGF?? receptor (T?RII) specifically in the proximal tubule and induce chronic kidney injury by either angiotensin/ uninephrectomy or aristolochic acid. We anticipate that mice lacking T?RII have impaired G1 arrest and reduced epithelial survival after injury and that this improves with administration of palbociclib, an FDA-approved inducer of G0/G1 cell cycle arrest. Proximal tubule cells with and without T?RII in vitro will be used to understand the signaling pathways through which TGF-? induces cell cycle arrest. The second aim explores mechanisms whereby TGF????lters epithelial differentiation nd how these changes impact survival. The role of epithelial differentiation in chronic kidney injury will be defined using mice with epithelial-specific deletion of Snail, a key transcription factor downstream of TGF-?? Proximal tubule cells with and without T?RII will be transfected with siRNA and constructs in vitro to test the hypothesis that Snail mediates TGF-?-dependent de-differentiation. The third aim defines how the Wnt/?-catenin pathway, important in renal injury, interacts with TGF-? to affect epithelial survival. Genetic models and cell culture techniques wil be used to test the hypothesis that TGF-? increases canonical Wnt/??catenin signaling to improve epithelial survival through increased de-differentiation and G0/G1 cell cycle arrest. This proposal uses innovative genetic approaches and a new strategy for analyzing injury in a quantitative and location-specific way. These studies will provide valuable insights into the roles of cell cycle and epithelial plasticity in the epithelial response to CKD, focusing on the proximal tubule which is the target of acute kidney injury and a mediator of CKD. However, information about cell cycle, epithelial plasticity and response to chronic injury has implications beyond the kidney as most organ dysfunction results from chronic epithelial injury.
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{
"pile_set_name": "NIH ExPorter"
}
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The general goal of this project is to delineate the mechanisms involved in the regulation of the activation of thymus-dependent (T) and thymus-independent (B) lymphocytes. To this end, studies of the genetic regulation of T lymphocyte activation directed at an analysis of the role of molecules encoded in the major histocompatibility complex (MHC) have been carried out. These experiments strongly suggest that T lymphocytes possess antigen-binding receptors which recognize complex antigenic determinants involving both conventional antigen and an MHC structure expressed on an auxilliary cell. The other principal approach has been to analyze the requirements for B lymphocyte activation by studying the responses and surface markers of B lymphocytes derived from a strain of mice with an X-linked defect in B lymphocyte maturation or function.
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{
"pile_set_name": "NIH ExPorter"
}
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Massachusetts Alzheimer?s Disease Research Center: Biomarker Core The Biomarker Core of the Massachusetts Alzheimer?s Disease Research Center (MADRC) supports and performs biofluid biomarker research to contribute to our understanding heterogeneity in Alzheimer?s disease and Alzheimer?s disease related disorders (AD/ADRD) and to accelerate progress towards a cure. We carry out core functions, marshaling resources to enhance the research mission through Aim 1's collection, curation and distribution activities for blood and cerebrospinal fluids (CSF) for MADRC, local and national research and Aim 2's standardized performance of essential molecular diagnostic assays of amyloid- b42 and tau to contribute to biological classification of AD/ADRDs in the MADRC Research Cohort and other samples. We develop and implement new strategies and accelerate progress towards a cure in Aim 3 and 4's refinement and validation of novel ELISA, Simoa, and MS assays targeting multiple pathophysiological pathways in blood and CSF, and in Aim 5's express focus on understanding those assays' utility for clinical and clinical trials settings. These novel approaches are piloted in the MADRC collections of blood and CSF spanning diverse AD/ADRD diagnoses, stages of disease, prognosis, demography (including sex as a biological variable), risk factors, heterogeneous pathophysiological drivers, and treatment. Successful pilot work and biofluid sample resources are then leveraged to facilitate larger focused research projects in and outside of the MADRC community. We build the future in Aim 6 by providing expertise, education, training and laboratory resources to faculty, trainees and students for best practices in biofluids biomarker research, and in Aim 7 by providing education to researchers, clinicians and the public on the advances, utility and responsible interpretation of biomarker data in ADRD research and ultimately how biomarkers can be used in caring for people with AD/ADRD.
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{
"pile_set_name": "NIH ExPorter"
}
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Interaction of prokaryotes with their viruses (phages) and plasmids accounts for horizontal gene transfer (HGT) that underlies the spread of antibiotic resistance and emergence of human pathogens. Bacteria evolved numerous systems to limit HGT. A novel prokaryotic defense system against foreign DNA is based on CRISPR (clustered regularly interspaced short palindromic repeats) cassettes and cas genes. A CRISPR cassette consists of direct repeats interspersed with spacers of highly variable sequence. Small CRISPR RNAs (crRNAs) bound to a large Cas proteins complex recognize foreign DNA, matching the spacer sequence present in crRNA, and destroy it. This process is referred to as CRISPR interference. Spacers in CRISPR cassettes are excluded from interference. Viral or plasmid-derived DNA is acquired by CRISPR cassette, becoming a spacer, in a process called CRISPR adaptation. Acquisition of host-derived spacers must be avoided, for it will lead to self-interference. Neither stage of CRISPR response is fully understood. We propose to study CRISPR function in Escherichia coli, the best-studied prokaryote. CRISPR/cas loci of laboratory E. coli are dormant. We developed genetic systems to study both stages of E. coli CRISPR response. We will use these systems and genetic, biochemical, crosslinking, laboratory evolution, and modeling approaches to: Aim 1. Analyze CRISPR interference and identify rules that govern self versus non-self DNA recognition by CRISPR interference machinery; characterize in vitro Cas protein-crRNA complexes formed with foreign DNA targeted for degradation, and localize the sites of crRNA-mediated target cleavage. Experiments will be performed with existing systems targeting the M13 phage and with new systems interfering with lytic T-odd phages and RNA phages of E. coli. Aim 2. Analyze CRISPR adaptation and determine i) rules that govern self versus non-self DNA discrimination by CRISPR adaptation machinery; ii) sequences outside CRISPR cassette that affect spacer acquisition; and iii) molecular details of the process that leads to appearance of extra spacer-repeat units in CRISPR cassette. Cas protein complexes formed with foreign DNA targeted for adaptation will be characterized in vitro and in vivo by trapping them at protein roadblocks. To better understand CRISPR-mediated viral-host dynamics and co-evolution we will monitor spacer acquisition in CRISPR cassettes of the host and viral mutations that render CRISPR interference ineffective in continuously infected cultures and develop a mathematical model of this process in collaboration with a group of bioinformaticians. As a result of proposed work novel molecular mechanisms operational during CRISPR response will be revealed and new ways for strain engineering and gene silencing in prokaryotes will be developed. The significance of proposed work will not be limited to E. coli, since CRISPR loci are found in more than 40% eubacteria and in 95% of archaea.
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{
"pile_set_name": "NIH ExPorter"
}
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To establish that one night's sleep deprivation will produce a rapid, sustained response to antidepressant drugs initiated at the same time in about 50% of depressed subjects; and to determine whether or not baseline thyroid function will predict response to antidepressants and SD.
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{
"pile_set_name": "NIH ExPorter"
}
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Arthropod-borne flaviviruses, and especially dengue viruses, cause a wide range of important human diseases for which there are no specific therapies. To address this critical shortfall in preparedness to confront these emerging and re-emerging viruses we have established a program to investigate host factors as targets of anti-dengue therapy. We have discovered many novel drug targets using functional genomics and en masse biochemical approaches. Among these are the 3'-5' exonucleases of the DnaQ/DEDDh superfamily of enzymes: EXD2, WRN and ERI3 (PRNPIP)). These enzymes are highly related to virally encoded exonucleases in SARS coronavirus and Lassa fever virus suggesting that the DnaQ/DEDDh superfamily of enzymes is widely used by pathogenic viruses and thus inhibitors of these enzymes could have broad spectrum of activity. We propose to characterize these enzymes in detail and to identify compounds that inhibit their activity and dengue infection. This will be achieved by 1) Developing in vitro assays for EXD2, WRN and ERI3 and 2) developing in vivo (yeast-based) assays to screen inhibitors of EXD2, WRN and ERI3. Significance to public health. Flaviviruses, and especially dengue virus, are an emerging threat to public health in the US, a current risk to our armed forces and other citizens deployed around the world, and a major problem globally. At this time there is little that can be done to prevent or treat the majority of flaviviral infections and therefore development of anti-flaviviral drugs is of crucial importance.
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{
"pile_set_name": "NIH ExPorter"
}
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In the adult heart, cell death following myocardial infarction (MI) initiates an inflammatory reaction that removes dead cells and contributes to scar formation and cardiac repair. Since the regenerative capacity of the adult mammalian heart is limited, induction of this innate immune response could be maladaptive and compromises cardiac contractile function. In neonatal mice, the heart can regenerate fully without scarring following MI; however, this regenerative capacity is largely lost rapidly after birth. A proactive role of the immune system and its response to injury has been proposed to be a central mediator of neonatal heart regeneration. However, the exact mechanisms by which neonatal adaptive immunity modulates heart regeneration are largely unknown. We found that induction of MI in post-natal day 1 (P1) mice induced an inflammatory response that failed to activate key inflammatory serine proteases (ISPs), enzymes released upon leukocyte activation and are the primary reason for tissue damage at the sites of inflammation. In contrast, activation of ISPs was observed when MI was performed at P7 or later, a time when the regenerative capability of the heart is very low. Because activation of ISPs occurs early after myocardial injury, is an important regulator of the inflammatory response and functionally modulates a number of protein substrates that regulate cell growth and function, we hypothesize that activation of ISPs plays an active role in regulating cardiac regeneration and repair. Pilot study shows that inhibition of ISPs in vivo using mice deficient in DiPeptidyl Peptidase I (DPPI), a key enzyme necessary for the cleavage and activation of major ISPs, enhanced cardiomyocyte proliferation during post-natal development and in adult hearts subjected to MI compared to wild-type, along with an improvement in cardiac remodeling and function. Interestingly, DPPI deletion also enhanced the survival of stem cells in the infarcted area, suggesting that ISPs modulate post-MI repair by affecting not only cardiomyocyte growth and proliferation, but also by modulating stem cell survival and growth. These data support the hypothesis that activation of ISPs impairs endogenous cardiac repair and leads to cardiac dysfunction post-MI. Here, we will determine whether inhibition of ISPs enhances endogenous cardiac repair and regeneration in neonatal and adult heart after MI injury. We will also define the mechanisms by which ISPs modulate cardiac regeneration of neonatal and adult hearts. The long term goal is to develop novel strategies targeting DPPI to enhance cardiac repair after MI, for which not a single drug is currently available.
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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. Neurometabolism and Outcome in Traumatic Brain Injury [unreadable]a magnetic resonance imaging and spectroscopy study The overall aim is to determine the usefulness of neurochemical and structural markers of brain injury determined by non-invasive magnetic resonance spectroscopy and imaging to predict cognitive outcome and monitor recovery following traumatic brain injury. Approximately 100 survivors of TBI have been studied longitudinally (~1, 3, and 6 months post-injury) with MRI, MRS and neuropsychological testing as outcome measures. They will be compared to a similar number of matched uninjured controls. This component of the project is to quantify altered brain volumes associated with TBI.
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{
"pile_set_name": "NIH ExPorter"
}
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This project attempts to elucidate the relationship between membrane antigens and an Fc receptor which appear in Herpes simplex type II-infected cultured human retinoblastoma cells. Further, both of these will be related to the increased Con A lectin binding ability of HSV II-transformed cells previously noted in the literature. The appearance of an Fc receptor and increased lectin binding of a following infection time when viral-directed membrane antigens are appearing suggests a relationship between these three membrane events. We suspect the Fc receptor is a viral-directed membrane-associated protein, as are the increased Con A binding sites. We are attempting to prove or disprove this assumption using ferritin and hemocyanin-labelled antibodies and lectins. The ability to bind host IgG may confer survival advantages to HSV-infected cells, enhancing both viral replication and latency.
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{
"pile_set_name": "NIH ExPorter"
}
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This research project will develop an engaging game for high school students using simulation of immune cells in the human body fighting infection. The approach combines visually stunning 3D graphics and advanced training technologies including automated case scenario generation, intelligent tutoring, automated debriefing, live conferencing, and performance assessment to provide a constructive environment for both male and female students to build and deploy elements of the immune system in response to pathogens and infectious agents. The game will be designed to include both "virtual world building" and "first-person action" elements. In addition to understanding the cellular basis of the immune response, students will be presented with case vignettes teaching the importance of proper hygiene and lifestyle choices to prevent infection and disease. The proposed STTR project involves a collaboration between SimQuest and the Federation of American Scientists, and the Phase I tasks include the formation and management of an Advisory Board for content and curriculum development, creation of an instructional plan, and development of a Design Document for the game. It is anticipated that the proposed combination of attractive 3D game play, validated content and constructive instructional strategies will produce an effective game for teaching immunological concepts to young adults.
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{
"pile_set_name": "NIH ExPorter"
}
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The Imaging Core is an integral component of this Program Project and will conduct all of the in vivo imaging studies in experimental animal models. The main objective of using non-invasive molecular-genetic imaging in this Program Project is to assess the spatial and temporal dynamics of stem cell trafficking and engraftment, as well as T cell trafficking and lymphoid and myeloid tumor cell targeting. The Imaging Core will utilize several dedicated pre-clinical and clinical imaging instruments, including: bioluminescence imaging (BLI) system, pre-clinical micro-PET/CT for imaging in mice, clinical PET/CT system for imaging non-human primates, Radiochemistry Facilitiy, and Reporter Vecotr Laboraotury of the Dept. of Experimental Diagnostic Imaging (EDI), UT-MDACC. Specifically, for Project 1, the Core will perform repetitive imaging studies to compare spatial-temporal dynamics of engraftment: a) in NOG/SCID mice of two different human cord blood-derived CD34+ hematopoietic stem cell populations (CB-HSC) differentially labeled with FLuc and RLuc reporter genes for BLI: 1) CB-HSC from two unmanipulated cords; 2) unmanipulated vs in vitro expanded CB-HSC, and 3) CB-HSC unmanipulated and expanded with/without fucosylation to improve homing to the bone marrow. For Project III, the Core will perform: a) BLI and 18F-FEAU microPET/CT in NOG/SCID mice to assess spatial-temporal dynamics of trafficking of HSV1-tk/Luc-labeled CB-derived T cells expressing CD19-specific chimeric antigen receptor (CAR+) and compare to that of the CB-derived tri-virus-specific (CD19)CAR+ T cells; b) repetitive PET/CT with 18F-FEAU in rhesus macaques to non-invasively monitor the trafficking and homing of adoptively transferred autologous T lymphocytes expressing HSV1-tk reporter gene and CD19-specific or CD20-specific CARs. For Project IV, the Core will use BLI and microPET/CT to assess the distribution, expansion, and persistence in mice of (1) human AML labeled with RLuc/RFP reporter, (2) high affinity PR1-CTL, and (3) low affinity PR1-CTL labeled with HSVI-tk/FLuc reporter gene. In summary, BLI will enable cost-effective high-throughput studies in mice, while 18F-FEAU PET/CT in mice and nonhuman primates will facilitate transition of the proposed cellular therapies into the clinic.
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{
"pile_set_name": "NIH ExPorter"
}
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The actions of the intracellular messengers inositol trisphosphate (InsP3) and calcium ions will be investigated in invertebrate photoreceptors. Illumination of invertebrate photoreceptors produces InsP3 via the phosphoinositide (PI) pathway of signal transduction. Molecular components of the PI pathway have been identified within invertebrate photoreceptors, but a complete explanation of their role in transduction has not yet emerged. Although the PI pathway is unlikely to play a role in excitation of vertebrate photoreceptors, a complete understanding of invertebrate phototransduction can be applied to other cells that respond to hormones or neurotransmitters which activate the PI pathway, including iris smooth muscle cells and retinal neurons. In Limulus (horseshoe crab) photoreceptors, InsP3 releases calcium ions from endoplasmic reticulum. The released calcium ions "mimic" the effect of light by opening sodium channels in the plasma membrane. Novel InsP3 isomers will be used to demonstrate that the response to InsP3 is mediated by stereospecific receptor site. We will continue to examine feedback inhibition by calcium ions of InsP3-induced calcium release, in order to demonstrate its rapid reversibility. Feedback control of calcium release may play a role in lowering intracellular calcium ion concentration (Cai) during stimulation of mammalian cells, preventing cell injury and death during ischemia. Metabolism of InsP3 will also be investigated using novel InsP3 analogues. Prior injection of a novel inhibitor of the enzymes that destroy InsP3, l-chiro 1,4,6,InsP3, causes single injections of InsP3 to produce sustained oscillatory bursts of calcium release. These bursts are not normal components of the light response. The inhibitor will be used to determine whether rapid destruction of InsP3 after a light flash prevents their occurrence. Inhibition of InsP3 metabolism by analogues should have general applications, since there are few drugs presently targeted against the PI pathway. Lithium's anti-manic effects may result from inhibition of inositol phosphate metabolism. A second set of experiments will determine whether the InsP3-induced elevation of (Cai) is necessary for visual excitation or whether another pathway for excitation must exist. We will use fluorescent calcium indicators to determine whether excitation of Limulus photoreceptors by light can occur in the absence of detectable elevation of Cai. We will measure Cai during the collapse of the light response of mutant fly photoreceptors (nss Calliphora), which are unable to sustain a response to bright illumination.
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{
"pile_set_name": "NIH ExPorter"
}
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The overall goal of this project, Cumulative Environmental Effects: Expanding Research with the Hopi Tribe, is to use a community-based participatory research (CBPR) approach to collaborate with the Hopi Tribe investigating household exposures to inform policy decisions. Household exposures are major sources of environmental hazards encountered by many American Indian and Alaska Native (AI/AN) communities. Household exposures include combustion by-products from heating and cooking, particulate from nearby mining and other land uses, and water and food contamination. These exposures, and co-exposures such as unemployment and poor access to preventive programs and health services, impact respiratory health and obesity among children and adults, and warrant evaluation of AI/AN household exposures within a social ecological framework. The Hopi Tribe in northern Arizona has identified several areas of concern, including the impact of burning coal and biomass in homes for heating and its potential impact on respiratory health. Other issues include concerns about arsenic and uranium species in drinking and surface water. Arsenic exposure is linked to compromised lung function and an increase in body mass index. This application seeks to expand existing relationships to include Hopi officials in the Hopi Environmental Protection Office and university environmental scientists and health promotion experts. The project proposes to: 1) Characterize magnitude of environmental exposures to particulate matter (PM), arsenic species, uranium and other contaminants from air, water, and food in selected households on Hopi tribal lands; 2) Evaluate how exposures are moderated by social determinants of health, social capital, community resilience and other cultural assets; and 3) Expand Hopi capacity to address areas of environmental concern. The effort will build additional capacity within the Hopi Tribe to evaluate and mitigate environmental hazards of concern to the Tribe. The proposed joint project provides an opportunity to develop and strengthen a relationship built on trust between the Hopi Tribe and university researchers and to increase the capacity of the Hopi Environmental Protection Office to monitor its air and water quality. Anticipated results include modeling of cumulative exposures to arsenic species and particulate among Hopi residents and addressing environmental concerns of the Tribe in terms of health inequities. The project will build the Hopi Tribe's capacity to conduct research on adverse exposures and develop programs that inform tribal environmental and health policies for a sustainable future.
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{
"pile_set_name": "NIH ExPorter"
}
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Description: The role of Core D is to perform dietary assessments, anthropometric measurements, body composition/energy metabolism measurements, and nutrition counseling and support. A complete list of the study parameters is presented in Table 1. Project 1 conducts the continuing surveillance of the "Nutrition for Life" (NFL) cohort that has currently been followed from one to three years and involves comprehensive longitudinal data on each subject. This Core will continue to set the protocols, quality control, training, and analyses on the cohort data. This cohort data includes dietary assessments (3-day food record (3DFR) and food frequency questionnaire (FFQ)), anthropometric measurements (weight, height, skin folds, hand grip, and circumferences), body composition measurements (bioelectrical impedance (BIA) and dual energy x-ray absorptiometry (DEXA)), and energy expenditure and metabolism measurements. This core will also provide nutrition counseling and support for the cohort. Project 1 will now include use of DEXA for determinations of body composition (yearly). Project 2 will utilize most of the services and measurements listed in this Core. This project has a population study group of 100 with each participant enrolled for one year. An average of 20 persons per year will be enrolled in the study. Individual nutrition counseling will be used for this study due to the specific nature of the nutrition interventions and the staggering of enrollment. Use of this Core?s services by Project 2 is delineated in the table on Page 381. Project 3 will contain three studies and will utilize many of the services and measurements offered in this Core (table on page 381). Study 1 will include only NFL participants and will not require any additional measurements for the NFL participants. Study 2, cohort sub-study with intensive metabolic measurements, will have 50 participants with each participant enrolled for one year. Study 3, Metformin for abdominal adiposity, will have 30 participants and will last for six months. Individual nutrition counseling will be utilized for this project because of the more detailed and instructional nature of the nutrition intervention and the importance of tracking. Staggered recruitment also necessitates individual counseling. Project 4 will be carried out at two sites (Children?s Hospital, Boston, MA; and Rochester Medical Center, Rochester, NY) by Tracie Miller, M.D. Procedures and protocols for the methods cited in this Core have already been adapted for this special population of infants and children.
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{
"pile_set_name": "NIH ExPorter"
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The aim of this research is to identify long QT syndrome families not linked to one of the previously described loci and to identify mutations in families linked to the LQT1, LQT2 or LQT3 loci. We intend to obtain concurrent electrocardiograms (ECG) and blood samples for leukocyte DNA analysis to examine for linkage between the LQT1, 2, 3 and 4loci on chromosomes 11, 7, 3 and 4 respectively and a long QT interval on the ECG. Identification of families not linked to one of these loci will be important in identifying other genes that can cause this life- threatening disorder. Also, identifying mutations in families linked to one of the previously-described genes will further our understanding of the disease process.
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{
"pile_set_name": "NIH ExPorter"
}
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The objective of this project is to implement and evaluate a nursing practice model to assist older, rural women in managing severe urinary incontinence (UI) while living at home. The nursing practice model is designed to structure the efforts of a nurse, an older, and a primary caregiver (where applicable) to establish measurable goals for continence and to develop a plan for managing UI that is sensitive to both the individual needs of the older and the sociocultural environment within which she resides. The specific aims are to: (1) describe and evaluate processes and outcomes associated with a nursing practice model - behavioral management for continence (BMC) - that includes specific techniques for managing UI; (2) evaluate the impact of model-specific factors as well as individual and social context characteristics on the attainment of behavioral goals; (3) evaluate the impact of the BMC intervention, individual, and social context characteristics on urine loss comparing treatment and control groups; and (4) conduct a cost-effectiveness analysis comparing BMC to formal community based and institutional services. The quasi-experimental design includes random assignment to treatment and control groups, repeated measures over time, and the use of both survey and clinical data. The sample will consist of 320 women 65 years of age or older who live in a nine county rural area in north-central Florida and experience severe stress, urge, or mixed UI. The principal outcome measure will be change in UI; however, additional bladder control variables, process measures, individual and social context characteristics, and cost-effectiveness data also will be analyzed. The significance of the project lies in: (a) delineating a nursing practice model (BMC) to enhance continence in a vulnerable, rural population; (b) obtaining a better understanding of factors that impact the attainment of continence goals; (c) evaluating the impact of BMC on UI for elders living at home; and (d) assessing the cost of BMC versus other community based and institutional alternatives.
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{
"pile_set_name": "NIH ExPorter"
}
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This study will examine the relationship of drug side effects to the daily functioning of elderly ambulatory patients receiving care in the primary care practice of a teaching hospital. The study will test two hypotheses: (a) that drug side effects lead to diminished functioning, particularly in extended mobility and social activities and (b) that a strong patient/provider relationship characterized by high levels of communication and close monitoring of drug regimens will lessen the number and degree of side effects and thus lessen decrements in functioning. A telephone interview will be conducted with 800 patients sampled randomly from persons aged 65 and over who attend the practice; data will be obtained on use of prescribed and non-prescribed drugs, adherence to regimens, side effects experienced, interactions with clinicians, and functional capacity, as well as socio-demographic characteristics and use of health services. These data will be supplemented by medical record abstracts. In-person interviews will be carried out with the clinicians staffing the practice (residents, nurse practitioners, attending physicians) with regard to their prescribing, patient education and drug regimen monitoring procedures. A protocol for assessing the relationship between specific drugs and symptoms will be used to differentiate between side effects and signs/symptoms of disease processes. The functional decrements most likely to be affected by drug side effects are expected to be mobility within the home and community, and interactions with other people. There is evidence that maintenance of social activity levels is significant in resistance to disease, recovery following acute illness, and adjustment to limitations due to chronic diseases. The study will provide important information on drug use and prescribing patterns, and on the factors which influence the effects of drugs on functioning. Further, the findings will provide an understanding of an area increasingly important for the expanding elderly population, and a basis for possible targetted intervention studies.
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{
"pile_set_name": "NIH ExPorter"
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DESCRIPTION (provided by investigator): While we have developed a rich knowledge base on family caregiving to frail elders, as well as on interventions designed to support caregivers, most research to date has focused on the primary, geographically proximate caregiver. Yet, it is estimated that as many as 11% of family caregivers in the U.S, and 9% of those providing care to persons with dementia, live more than two hours away from the care receiver. We have little in-depth knowledge about this growing phenomenon, especially as it applies to caregivers across diverse SES and racial/ethnic groups. This R21 application seeks to extend the scope of prior discoveries regarding family care to impaired elders by specifically focusing on the long-distance caregiver (LDC). The long-term goal of the proposed research program is to develop a knowledge base that will be used to adapt existing evidence-based caregiver interventions (e.g., REACH models) to the needs of LDCs and, ultimately, to test adapted models in the field. This R21 application is the first step in this process; with the project goal of examining LCD experiences in-depth to understand how LDCs, deal with, and are affected by, the structural constraint of distance and how the experience differs by key characteristics. The specific objectives are: (1) To examine the long-distance caregiving consequences (i.e., caregiver well-being, service needs) and identify subgroup differences related to ethnic and socioeconomic status, gender, distance to the care recipient (travel hours), the cultural value of familism, and cognitive status of the elder. (2) To identify the primary stressors (e.g., caregiving involvement and burden) and secondary stressors (e.g., work/family conflict) experienced by LDCs, the strategies employed to deal with these stressors and to identify subgroup differences. (3) To determine the internal and external resources (e.g., coping skills and social support) which mediate the association between primary and secondary stressors and caregiving outcomes, including models with travel distance, ethnic/socioeconomic status, gender, familism, and elder cognitive status as important exogenous factors. (4) In order to determine how, if at all, recruitment strategy may influence findings, to explore differences in caregiver well-being and service use between our primary sample of LDCs to elders linked to formal services and a secondary sample of LDCs recruited from a representative community panel. The innovation of this research lies in its unique sample (diverse ethnic and SES LDCs with primary/shared responsibility) and focus on issues most relevant to LDCs (e.g., communication, technology) rather than measures/methods. In-depth telephone interviews will be conducted with 300 LDCs (2+ hrs. travel) who are primary/co-primary caregivers to an ethnically/racially diverse sample of elders who receive formal residential or home-based long-term care services. A secondary sample (n=50) will be drawn from a representative community-based sample of middle- age and older adults. A mixed method design is used, with open and structured parts of the interview and corresponding qualitative (thematic) and quantitative (SEM) approaches to data coding and analyses.
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{
"pile_set_name": "NIH ExPorter"
}
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Immunological memory is a defining feature of the adaptive immune system. Ag-experienced cells respond differently from nave cells. Understanding how such responses differ is of fundamental importance. Yet, there is relatively little knowledge about intrinsic differences between nave and memory cells. Our goal has been to elucidate properties that distinguish memory B cells from their nave precursors. We are approaching this task using a combination of gene expression analysis, functional and biochemical characterization of memory cells and genetic strategies to test the roles of memory-specific molecules in vivo. In the previous period, we overcame a major hurdle to the study of memory B cells by developing a system that creates large numbers of such cells. In this system, the definition of a memory cell is one that had seen and divided in response to Ag and remained as part of an expanded population in a resting state. We first characterized memory cells in this system with phenotypic and functional analysis, then moved to gene expression profiling, identifying multiple genes and proteins that were differentially expressed. These genes highlighted several novel themes with respect to memory B cell function. In addition, we found that memory B cells were comprised of clear phenotypic subsets. We propose first to investigate the ontogeny and functions of these memory B cell subsets. In Aim 1 we will test several hypotheses about the origins of different subtypes of memory B cell. In Aim 2 we will better characterize certain memory B cell subsets and test hypotheses about how they function differently from each other in vitro and in vivo. One theme we discovered in the last period is that B7-family members with inhibitory function are overexpressed on some or all memory B cells. In Aim 3 we use genetic approaches to determine the functions of these molecules on memory B cells, again using both in vitro and in vivo methods. It is quite intriguing that two of the most overexpressed genes we found in memory cells are receptors that are intimately associated with stem cell homeostasis. Though increased in expression by 81- and 15-fold by RT-qPCR, the functions of these genes in the immune system in general and memory B cells in particular are virtually unknown. We hypothesize that these genes play roles in regulating memory cell homeostasis and differentiation, much as they would in other long-lived stem cell populations. In Aim 4 we will used floxed alleles of these genes, along with CD19-Cre mice, and novel mice we recently produced that allow inducible Cre activity specifically in B cells, to determine the functions of these genes in B cell development, responses, and memory formation, as well as in memory cells that were established normally and then induced to delete these genes. Together, these experiments should provide significant advances in an important but relatively understudied area: the origins and function of murine memory B cells.
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{
"pile_set_name": "NIH ExPorter"
}
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DESCRIPTION: (Applicant's Abstract) The adoptive transfer of tumor-reactive T lymphocytes mediates regression of established malignancies. In animal models, tumor-draining lymph nodes (LN) are a rich source of sensitized T cells that can be activated ex vivo to acquire effector function. In this application the generation of tumor-sensitized T lymphocytes in LN, enrichment of this population, and analysis of effector cell function will be investigated. Well defined murine tumor models of adoptive immunotherapy will be utilized to test the hypothesis that antigen presenting cells transfer tumor antigens to draining LN. Several adjuvants including GM-CSF will be injected paratumorally to study augmentation of LN T cell sensitization. In the draining LN, antigen primed T cells display altered levels of several membrane markers, among them CD62L (L-selectin), that will be used to enrich tumor-sensitized cells from irrelevant T cells. The production of cytokines upon contact with tumor cells and therapeutic efficacy will be analyzed in the segregated T cell subsets. To mediate tumor regression T cells must contact tumor and transferred cells infiltrate tumors even in immune privileged sites such as the central nervous system. The homing of transferred T cells to tumor and effector mechanisms will be analyzed in mice bearing intracranial tumors. The cell adhesion molecules that mediate adherence of T cells to the tumor vasculature will be defined by their expression in situ, on T cells isolated from tumors, and by interference with tumor infiltration by blocking mAb. T cell mediated tumor regression is highly specific and the contribution of selective trafficking or retention of tumor-reactive cells to this specificity will be determined. Although ex vivo activated T cells are not cytolytic in the standard 51Cr release assay, they do produce cytokines upon contact with tumor. The production of cytokines in vivo, and the participation of accessory cells in mediation of tumor regression will be investigated.
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{
"pile_set_name": "NIH ExPorter"
}
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The purpose of the proposed studies is to define precisely the processes involved in iron uptake, utilization or storage, and eventually release by cells in key tissues throughout the body. Emphasis will be placed on studies of physiologic systems and iron compounds normally present in health or disease. Attention will be directed initially to iron metabolism by isolated rat liver cells, the placenta, and the Fletcher-Huehns phenomenon of selective release of iron from the two binding sites on transferrin. Liver cells isolated by the use of chelates, enzymes or mechanical means will be used to measure binding of radioiron from rat transferrin with determination of the number and avidity of iron binding sites, competition by other metals and proteins, the formation of intracellular ferritin, and the release of iron under various stresses to iron metabolism. Transferrin-bound radioiron uptake by placental structures of pregnant rats will be analyzed to identify placenta iron receptor sites and small molecular weight carrier substances involved in this unidirectional iron transfer. Functional heterogeneity of iron atoms on transferrin will be explored in vivo to determine whether selective release of iron to erythroid precursors, liver cells, placenta and other tissues occurs.
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{
"pile_set_name": "NIH ExPorter"
}
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Herpes simplex virus type-1 (HSV-1) is highly prevalent among adult human population. The candidate's long-term career goal is to maintain an outstanding research program in studying early events that allow the virus to enter into cells and establish a productive infection for spread. To date, the candidate's research has made a significant contribution to the understanding of HSV-1 entry receptors and uptake mechanisms. Most recently the candidate's laboratory has made new advances in understanding the interactions of HSV-1 with host cells via live cell fluorescence microscopy. The salary support provided by an Independent Scientist Award (K.02) will significantly reduce teaching and administrative responsibilities for the candidate to pursue a new line of research and be more available for the training of graduate students and fellows in the candidate's laboratory, which will enhance their overall productivity. The present proposal extends the research being conducted by the candidate on HSV-1 entry and spread mechanisms. It explores the significance of heparan sulfate (HS) in viral transmission and spread processes. It is based on a novel observation that initial virus-host interactions occur at membrane processes such as filopodia that express HS. This interaction is then exploited by the virus for an efficient and targeted delivery to the cell body for entry and spread to neighboring cells. Although this phenomenon seems to exist for all the herpesviruses examined so far by the candidate and mimic surfing phenomenon reported with retroviruses, the exact molecular mechanism for the transport process remain unknown. It is also unclear whether the viruses travel exogenously or intracellularly on the membrane processes. The candidate hypothesizes that the virions travel exogenously until they reach the cell body. Depending on accessibility to entry receptor, the virions either enter into a cell or travel again via membrane processes to reach a neighboring cell body for entry. Experiments will be performed to determine the exogenous nature and the molecular basis of the herpesvirus surfing phenomenon. Significance of the surfing in virus transmission will be addressed using an animal model. The proposed work in this application will provide a new dimension to the current work being done in the candidate's laboratory and it will also open up a new and biomedically significant area for future research. RELEVANCE: HSV-1 infections are highly prevalent among humans. About 80% of world's adult population may be seropositive for the virus. The disease manifestations range from more common fever blisters to rare cases of encephalitis. The virus via its envelope proteins interacts with receptors on our cells and this interaction is crucial for the initiation of the infection. Our research may suggest new strategies to stop virus transmission and spread among humans.
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{
"pile_set_name": "NIH ExPorter"
}
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Development in mammals involves the modulation of gene expression during cell differentiation. We are investigating two sets of genes active during mouse pre-implantation development. One set contains homeobox sequences which have been shown to control spatial relations in Drosophilia development. We have recently indentified clones from a mouse cDNA overian library which cross-hybridize with Drosophilia homeobox probes and are determining if these mouse genes contain homologous homeobox sequences. The second set of genes we study code for the three sulfated glycoproteins of the mouse zona pellucida (ZP-1, ZP-2 and ZP-3). The products of this gene family are germ-line specific and active only during meiosis where their coordinate regulation occurs in the absence of cell division. We have developed monoclonal antibodies which are specific for ZP-2 and ZP-3. Using these immunological probes we have isolated cDNA clones coding for ZP-3 from a ovarian cDNA library in the Lambdagtll expression vector. ZP-3 message is expressed as a 1.7 kb polyadenylated mRNA which is found exclusively in ovarian tissue. Southern blot analyses of restriction enzyme digests of DNA from brain, liver and ovary reveal similar genomic organization of ZP-3, although there is hypomethylation of the ZP-3 locus in ovarian tissue. Mouse ZP-3 cDNA cross-hybridizes with DNA from a wide variety of mammals including rat, rabbbit, dog, pig, cow and human. Parenteral administration of anti-zona monoclonal anitbody results in antibody localization to the ovary where it coats growing oocytes. Although ZP-3 is thought to be the species-specific sperm receptor, monoclonal antibodies to either ZP-3 or ZP-2 appear equally effective in inhibiting in vitro and in vivo fertilization. The contraceptive effect is long-term but totally reversible and the presence of the monoclonal antibodies had no adverse affect on early development. These observations couples with our ability to express recombinant ZP-3 as fusion proteins will allow us to test the efficacy of a contraceptive vaccine based on the zona proteins.
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{
"pile_set_name": "NIH ExPorter"
}
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The origin of the central nervous system during embryogenesis in Drosophila melanogaster depends upon the proper segregation of neuroblasts from the primative ectoderm and the subsequent differentiation of neuroblast-derived neurons into the segmentally organized nervous system. We have recently been able to isolate embryonic neuroblasts from gastrula-stage Drosophila embryos insufficient quantities to use as an antigen for preparing monoclonal antibodies and to examine neural specific genes and gene products. In addition, we have identified two female sterility mutations which affect the early embryonic determination step at which ectoderm cells become either neuroblasts or epithelium. We will prepare monoclonal antibodies against freshly isolated neuroblasts and from neuroblasts as they differentiate In Vitro into recognizable neural nets. We will also prepare cDNA probes to poly (A) containing RNA of differentiating neurons In Vitro to screen genome libraries of neural specific genes. Finally, we will clone the genes for the two female sterility mutations affecting neurogenesis. With these probes we will analyze the genetic organization of the major genes involved in neurogenesis and we will use the monoclonal antibodies as tools to investigate the development and organization of the embryonic nervous system. These investigations will provide a model for the understanding of early developmental switches and for the early differentiation of complex nervous systems.
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{
"pile_set_name": "NIH ExPorter"
}
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Overactive bladder (OAB) and interstitial cystitis/bladder pain syndrome (IC/BPS) are diseases with unknown pathology and etiology. OAB and IC/BPS have devastating psychological and social impacts on quality of life, but treatments for these diseases are clinically challenging. Sacral neuromodulation is a FDA approved treatment for OAB. Currently it is only offered to OAB patients after pharmacotherapy has failed. Since its FDA approval for OAB, sacral neuromodulation has been shown to be effective in treating IC/BPS in many clinical trials and is currently listed in the AUA guideline o IC/BPS as one of the suggested treatment options. In addition clinical studies have showed that pudendal neuromodulation is superior to sacral neuromodulation. It successfully treated OAB or IC/BPS patients who have failed sacral neuromodulation. Furthermore, recent multicenter clinical trials have indicated that tibial neuromodulation is effective for treatment of OAB. Although neuromodulation is an effective treatment for OAB or IC/BPS, the mechanism underlying neuromodulation (sacral, pudendal, or tibial) is still unknown. More surprisingly there is very limited effort in basic science research aimed at revealing the mechanism of action underlying neuromdulation. In this project we propose to determine the central sites of action for neuromodulation by answering the following questions: 1. whether neuromodulation acts in the spinal cord or in the brain? 2. Whether it acts on the ascending or descending limbs of the spinobulbospinal micturition reflex pathway activated by bladder A?-fiber afferents or on the spinal micturition reflex pathway activated by noxious C-fiber bladder afferents? 3. What neurotransmitters are involved in neuromodulation? 4. How brain sensory and motor activity associated with bladder nociception/overactivity is altered by neuromodulation? Different neuromodulation methods (sacral, pudendal, or tibial) will be investigated and compared to answer these questions. Identifying the sites of action for different neuromodulation therapies will surely guide clinicians and patients in selection of the optimal treatment strategies and thereby significantly improve the clinical outcomes in treating OAB or IC/BPS. Information about the sites of action and the neurotransmitter mechanisms underlying neuromodulation could also be useful in developing new drugs to treat OAB or IC/BPS. Our studies will significantly benefit millions of Americans suffering from OAB or IC/BPS.
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{
"pile_set_name": "NIH ExPorter"
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The MBRS Program at Fisk University is designed to strengthen biomedical research at this institution and contribute to the solution of basic health problems through two projects that involve two faculty members, three graduate and three undergraduate student trainees. The specific projects are (1) organic chemical syntheses of stilbene derivative as potential anti-HIV and anti-cancer agents; (2) study optically the processes of nucleation in the temperature controlled growth of single crystals, suitable for crystallography, of proteins and other biomolecules.
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{
"pile_set_name": "NIH ExPorter"
}
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The clinical research mission of the CCR is as follows: * Conduct hypothesis-based translational clinical trials that evaluate new therapeutic approaches * Develop novel approaches to early cancer detection and prevention * Develop and deliver novel technologies for molecular characterization, detection, diagnosis, and monitoring of cancer * Emphasize understudied diseases and cancers with increasing incidence, poor prognosis, or involving special populations * Develop better preclinical models and methods to expedite development of novel interventions for cancer To accomplish these goals we must have participants on our clinical trials and the NCI has a national referral based to serve all US citizens. To enter participants we must travel them to Bethesda, lodge and feed them while here.
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{
"pile_set_name": "NIH ExPorter"
}
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In order to study the effect of ethanol and other drugs on folate metabolism, a quantitative assay for hepatic conversion of folic acid to N-5-methyltetrahydrofolic acid (MTHF) will be perfected in normal subjects. This assay will involve an intravenous injection of tritiated folic acid followed by an intravenous flushing dose of unlabeled folic acid, with separation of tritiated folic acid from tritiated MTHF by anion exchange column chromatography of plasma and urine. In this way, a normal ratio for hepatic conversion of folic acid to MTHF will be established. Normal volunteers will then be studied on oral ethanol regimens and control regimens. The ratio of conversion of folic acid to MTHF for the ethanol study of each subject will be compared to the control ratio for that subject and to the established normal ratio. In another protocol, the effect of oral ethanol on urinary excretion of folate metabolites will be investigated. It is anticipated that these studies will show that ethanol does not enhance urinary folate loss and will establish whether or not ethanol impairs conversion of folic acid to MTHF. The effect of short-term ingestion of diphenylhydantoin on the serum folate level as determined by the L. casei assay and the radioassay will also be investigated. If diphenylhydantoin is shown to lower the serum folate level, the causal mechanism will be sought by the prcedures described above for the study of the effect of ethanol on folate metabolism. Finally, the frequency, degree, and hematologic consequences of folate deficiency in women taking oral contraceptives will be explored. It is hoped that the mechanisms by which ethanol and diphenylhydantoin impair folate metabolism will be established, and that the knowledge gained about folate metabolism will benefit many fields of scientific endeavor.
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{
"pile_set_name": "NIH ExPorter"
}
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Acientobacter baumannii is a gram-negative bacillus, which was initially known for causing health-care associated infections. Most recently the experience in Iraq/Kuwait/Afghanistan has reaffirmed the importance of A. baumannii in causing infections in wounded combatants. Further the prevalence of multi-drug and pan- resistant isolates of A. baumannii is increasing, placing us at risk for entering a post-antibiotic era. As a result, the incidence of Acinetobacter infection in all venues is increasing worldwide. The changing epidemiology and incidence of infections due to Acinetobacter establishes it as a pathogen of increasing medical importance. Needless to say, treatment of infections due to Acinetobacter has become challenging. Improved outcomes will require the development of new preventative and/or therapeutic strategies. Unfortunately, significant knowledge gaps exist that need to be filled to accomplish these goals. Vaccine candidates need to by identified and assessed. Virtually nothing is known about capsular polysaccharides in A. baumannii. Surface polysaccharides such as capsule have formed the basis for several successful vaccines directed against extracellular pathogens such as H. influenzae type b, S. pneumoniae, and certain serotypes of N. meningitidis. This approach may also be practical for Acinetobacter. Therefore, the first goal of this proposal, to determine the number and relative prevalence of A. baumannii capsule serotypes, will be accomplished in aim 1. These data will lay the foundation for immunization and protection studies designed to assess capsule as a vaccine candidate, which will be completed in aim 2. Also in aim 2 studies will be performed to determine the structures of 3-5 of the most prevalent capsular serotypes. This information will be used to optimize the method and design of capsule- protein conjugates as needed to optimize immunogenicity. These data will also be used to predict possible immune reactivity to human epitopes. Lastly, structural data will be an additional means to assess the conservation of capsular epitopes. This proposal will generate new and important data and fill knowledge gaps necessary for the development of new preventive approaches to combat A. baumannii, a pathogen of increasing medical importance in the civilian, veteran, and active military populations.
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{
"pile_set_name": "NIH ExPorter"
}
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This proposal seeks continued funding for the Pancreatic Cancer Genetic Epidemiology (PACGENE) Consortium, a multicenter, multidisciplinary team which has the goal to identify susceptibility genes for :amilial pancreatic cancer (FPC). The PACGENE Consortium has shown that coordinated case finding, :amily recruitment, and linkage analysis are productive: we have identified novel putative regions on at least 4 chromosomes, and are positioned to use the pedigree material to further gene discovery. The PACGENE onsortium comprises 7 data collection/analysis centers in the U.S. and Canada, 2 Cores (Data Management/Analysis and Pathology/Tissue Genotyping), and is guided by Steering and External Advisory ommittees. Our specific aims are: AIM 1. To validate linkage findings for gene discovery with a new, final sample of FPC families. We will (a) generate a validation sample of 85 new FPC pedigrees and follow up our ixisting cohort of FPC pedigrees to increase their informativeness; (b) perform a genomewide 5800 single nucleotide polymorphism (SNP) linkage analysis on this validation sample (genotyping will be sought at the Center for Inherited Disease Research at no cost to this grant); AIM 2. To perform a familial case-control association study to validate SNP associations identified in other consortia studies. We will genotype 500 FPC probands and 500 unrelated family controls using a custom SNP marker panel identified by the genomewide association studies of the Cohort Consortium, the Pancreatic Cancer Case-Control Consortium, and our own linkage analyses; and AIM 3. To identify the genes in candidate regions identified by linkage and association. We will (a) fine map regions identified through linkage analyses (including chromosomes 2p, 9p,17p, and 22q); (b) perform genomewide loss of heterozygosity allelotyping of microdissected early stage tumors (PanlNs and IPMNs) from FPC probands (comparing tumor to normal), with a focus on regions of interest; and (c) resequence the best candidate genes in the regions identified. Public Health Relevance: Over 32,000 new cases of pancreatic cancer occur in the U.S. each year, almost all rapidly fatal. Our research will help identify the genes for FPC, furthering knowledge about the etiology of pancreatic cancer that should accelerate translation to care. Risk assessment will be improved, and identified FPC genes will lead to development of more effective strategies for early detection, prevention, and therapy.
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{
"pile_set_name": "NIH ExPorter"
}
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The proposed research addresses the gap in knowledge between nutritional status of an organism and the response of its genetic material. Nutritional stimuli influence the structure as well as the function of liver chromatin. I have previously studied how rat liver chromatin responds to varying intakes of carbohydrates, protein, and fat and have demonstrated (J. Nutr. 110:105, 1980) that the relationship of dietary components alters the highly ordered structure of chromatin. The specific objectives of this proposal are (1) to determine by what manner nutritional stimuli alter the structure of liver chromatin, (2) to determine whether differences exist in the complexity and abundance classes of messenger RNA (mRNA) populations under various nutritional states, and (3) to determine whether there are modifications in chromosomal protein constituents of chromatin. To determine how nutritional intake alters chromatin structure, chromatin from purified nuclei will be mildly digested with micrococcal nuclease (E.C.3.1.4.7). The fragments generated will be analyzed as to size by polyacrylamide gel electrophoresis and as to sequence organization by DNA-DNA reassociation. The complexity of mRNA populations will be determined by RNA-DNA hybridization. Chromosomal proteins associated with chromatin fragments generated from nuclease incubation will be purified and analyzed by two-dimensional gel electrophoresis.
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{
"pile_set_name": "NIH ExPorter"
}
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In the current project, genetic determinants of diabetic nephropathy and related traits are being sought using techniques of genetic linkage and association analysis. Lymphoblast cell lines have been established from informative pedigrees. DNA is available from other families in nuclear pellets extracted from blood specimens obtained in the epidemiologic studies. An autosomal genome-wide linkage study identified suggestive evidence for linkage to diabetic nephropathy on chromosome 3q and chromosome 7q. Efforts to identify potential causative variants in both of these regions are currently underway using both a systematic analysis of linkage disequilibrium and analyses of candidate genes. Through collaboration with multicenter Family Investigation of Nephropathy (FIND) consortium, additional informative families are being analyzed for linkage. Genome-wide association strategies are also being used.[unreadable] [unreadable] In the last year, a genome-wide linkage study in the first 378 families recruited from the multiethnic FIND consortium identified potential diabetic nephropathy susceptibility loci on chromosomes 7q, 10p, 14q and 18q. Several regions linked with quantitative measures of renal function were also identified. Markers were genotyped for the full collection of families from the FIND consortium and linkage analyses are currently underway. A genome-wide association study in these families was planned. In addition, a genome-wide association study comparing allele frequencies in pooled DNA from Pimas with end-stage renal disease with pools from individuals with long duration of diabetes without evidence of nephropathy identified the PVT1 gene on chromosome 8q as a strong candidate for susceptibility to diabetic nephropathy.[unreadable] [unreadable] Current efforts are focused on analysis of linkage and genome-wide association in the full collection families from the FIND consortium. With collaborators, dense linkage disequilibrium maps are being generated in the candidate regions on chromosomes 3q and 7q. Replication studies of PVT1 and other candidate genes are also being pursued. Additional American Indian families informative for linkage and association studies of diabetic nephropathy continue to be recruited. In conjunction with collaborators, further recruitment of families for these studies has been initiated in Micronesia.
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{
"pile_set_name": "NIH ExPorter"
}
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Previous work has demonstrated survival of peptidergic, diaphorase, and pyramidal neurons in fetal cortex transplants which connect to host cortex and subcortical structures. This proposal addresses the question of whether these transplants are functional, and if so, whether this is due to appropriate connections or to trophic interactions between host and transplant. To answer whether transplants are functional, rats are first trained to perform a forelimb motor task. One week after lesioning forelimb motor cortex, one group receives fetal frontal cortex, one receives fetal cerebellar, and one receives gelfoam implants. The motor behavior of each transplanted group is compared to the gelfoam implanted controls. Following this, transplants are removed. It is hypothesized that forelimb motor function will deteriorate following removal of functional transplants. Secondly, it will be determined whether adult host brain thalamic connections to fetal parietal cortex transplants are functional. The whisker sensory barrelfield of adult rats is removed, and fetal parietal cortex transplanted into the cavity. Later, the vibrissae contralateral to the transplants are stimulated. 2-deoxyglucose is injected and the resting and stimulated transplant glucose metabolic rates determined using a double label autoradiographic method. Transplants with functional host connections should be metabolically activated during the whisker sensory stimulation. Lastly, trophic effects of fetal cortical transplants on neonatal host thalamus will be examined. One week after removal of the frontal cortex of newborn rats, either fetal frontal cortex, fetal cerebellum, fetal parietal cortex, minced fetal frontal cortex, or gelfoam is transplanted into lesion cavities. Since marked host thalamic atrophy normally occurs ipsilateral to fetal frontal cortex lesions, this will determine which transplants ameliorate the atrophy. Host-transplant connections are examined in the above experiments to determine which effects correlate with formation of connections. Cortex injury due to strokes, head trauma, and other diseases can lead to devastating, permanent motor and sensory impairments in humans. The data from this proposal will help determine whether fetal transplants might offer some hope for treating such deficits.
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{
"pile_set_name": "NIH ExPorter"
}
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Meeting national needs in the public health sector and particularly in research areas within the biomedical and behavioral science will depend on the growth and diversification of the workforce entering this sector. Community colleges educate large numbers of students, especially underrepresented minorities (racial, ethnic, first-generation college, and economically disadvantaged). With specific attention to their unique needs, a greater percentage of this population has the potential to complete four-year degrees and graduate programs and become researchers in the health sciences. The Virginia Bridges Program aims to increase the number of underrepresented (UR) Virginia residents pursuing degrees and research career paths in the biomedical and behavioral sciences. A bridge between New River (NR) and Northern Virginia (NOVA) Community Colleges and Virginia Tech (VT) will generate interest and skills in the biomedical/behavioral sciences among UR students, beginning their first year at NR and NOVA through engaging research experiences. Additional research opportunities combined with academic support programs and mentoring will promote persistence, achievement, and timely progress through their degrees. Through reflective practices that span all four undergraduate years, Bridges students will define their own academic path.
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{
"pile_set_name": "NIH ExPorter"
}
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This project represents a program of coordinated studies designed to study behavioral and familial precursors of schizophrenia and schizophrenia-like disorders. A recently completed five-year follow-up of index cases seen originally in adolescence revealed that two parental attributes, communication deviance and negative affective style, predict the occurrence of extended schizophrenia spectrum disorders at follow-up. Currently, a longer term follow-up is proposed to estimate more stable diagnostic outcomes for the index cases.
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{
"pile_set_name": "NIH ExPorter"
}
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The major objective of the proposed research is the characterization of a single-locus genetic effect that influences behavioral performance in a learning situation. The proposed analysis should determine whether the locus being characterized influences performance by affecting stimulus sensitivity, activity level, responses to initial conditioning trials, or perhaps learning ability. The generality of the effect among various genotypes and upon other behaviors will also be assessed. The primary genetic methods will be Mendelian analysis and a new technique, the recombinant-inbred strain technique. Behavior will be assessed in active and passive avoidance situations, activity wheels, the open-field, water mazes, and operant- conditioning avoidance situations.
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{
"pile_set_name": "NIH ExPorter"
}
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During the last several years, great advances have been made in our understanding of angiogenesis, the study of blood vessel growth and proliferation. Several of the agents which promote this process have been well characterized. One promising agent, endothelial cell growth factor (ECGF), has been synthesized through recombinant techniques. The purpose of this investigation is to utilize ECGF in a canine model of effect myocardial angiogenesis, and to direct this process to ameliorate myocardial ischemia. In our model, the left anterior descending coronary artery (LAD) of dogs is occluded gradually over a 2 to 3 week period by an ameroid device applied to the proximal vessel. The internal mammary artery (IMA) is implanted into the region normally supplied by the LAD. It is known that collateral vessels develop from the IMA and other sources to supply the territory normally perfused by the LAD. Myocardial blood flow is generally sufficient under rest conditions, but is inadequate under conditions of stress. We have developed a means to quantitate the maximal conductance between the IMA and the LAD vascular bed, assessing myocardial perfusion, the physiologic result of angiogenesis. Ameroids will be applied to the LAD of 16 dogs. The IMA will be implanted in the LAD area. Dogs will be randomly assigned to receive infusions of ECGF of normal saline. After 8 weeks, resting and maximal myocardial blood flow be quantitated using radiolabeled microspheres. The maximal IMA to LAD bed conductance will be calculated, and comparisons made between groups. In addition, vessels will be examined morphometrically and various hematologic, biochemical, and immunologic parameters will be assessed in the 3 groups, to determine potential adverse effects of ECGF.
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{
"pile_set_name": "NIH ExPorter"
}
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In recent years, considerable progress has been made in understanding the origins and clinical management of large congenital melanocytic nevi (LCMN). New results from genetics, cell biology, epidemiology, and therapeutic research are flanked by enormous efforts made by patient initiatives around the world, spearheaded by groups in the U.S. and European countries. There is enormous potential for synergy between all of these players. The 2011 International Expert Meeting in Tbingen, Germany enabled the exchange of the most up-to-date, sometimes unpublished information from basic biology and clinical medicine among outstanding researchers in the field. This 2013 International Expert Meeting in Marseille, France will foster and enable the coordination of novel, interdisciplinary and international collaborations among participants. The leadership of major U.S. and European LCMN advocacy groups will attend the conference, to further refine the international initiatives established in 2011. These outcomes will continue making important contributions to the improvement of diagnosis and treatment for patients with this disfiguring and potentially life-threatening disease.
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{
"pile_set_name": "NIH ExPorter"
}
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It has become abundantly clear that aging takes a toll on all three chemical senses: taste, smell, and common chemical sense (CCS). Of these, taste seems the most resilient; although old people, on the average, lose the ability to detect faint levels of tastes, the perception of supra-threshold taste magnitudes remains relatively intact. In contrast, losses of smell and CCS frequently pervade the entire sensory continua from low to high stimulus levels. These are accompanied by loss of ability to identify and discriminate odors. The present proposal concerns the relation of olfactory and CCS losses to other aspects of chemosensory function about which much is known from youthful persons, and concerning which there is reason to hypothesize that aging might matter. These include (a) Olfactory affect, or the perception of the pleasant-unpleasant dimension of odors, the main question being to what extent affective narrowing takes place in old age and to what extent it pertains to the loss of suprathreshold magnitude; (b) Olfactory memory, or the ability to recognize odors shortly after inspection and then at intervals of a day and a week. Questions of interest include: the relation of odor recognition to olfactory identification and discrimination, to visual recognition, and to gender. (c) Adaptation, or loss of sensitivity over time with ongoing stimulation, especially as regards its role in the detection of a gas warning agent, ethyl mercaptan; (d) Olfactory--CCS mutual inhibition, or the power of nasal irritants to mute smells, and vice versa; (e) Detection of danger-related substances, in the present study the ability to detect smoke and ethyl mercaptan; and (f) Age-related dementia (especially Alzheimer's disease) as it affects three basic functions of olfaction: detection, odor identification, and perception of the affective dimension, i.e., the capacity to discriminate between good and bad odors and tastes.
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{
"pile_set_name": "NIH ExPorter"
}
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