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Controlling gene expression in primary cultured cells, such as human keratinocytes is difficult due to the stratum corneum, which ranges from 10-50 urn in thickness and serves as a barrier to the intracellular environment. Methods to introduce gene regulation agents such as oligonucleotides have met with limited success, as both the penetration depth and effectiveness of the regulation strategies vary. The proposed work will develop oligonucleotide-nanoparticle antisense agents as methods for controlling gene expression in keratinocytes. This work will take advantage of newly discovered conjugate properties of the nano-materials including enhanced stability, high penetration ability, and non-toxicity. The ability of these agent to work on genes of relevence in skin biology models will be tested.
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{
"pile_set_name": "NIH ExPorter"
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Since auditory function can be assessed objectively by recording Auditory Evoked Potentials from the brainstem, we propose to study in term and premature neonates affected by hyperbilirubinemia. Such factors as development, hypoxia and therapy will be examined for their influences. Serum Bilirubin levels will be correlated with auditory function in an attempt to ascertain a more precise understanding of the mechanism of deafness in these infants. It is expected that a more precise index of Bilirubin toxicity will be found for purpose of treatment and prognosis. From those infants who succumb in the course of study, neuropathologic confirmation of the location and extent of CNS damage will be obtained.
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{
"pile_set_name": "NIH ExPorter"
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This application seeks continuing support for the on-going Chromosome Metabolism and Cancer Training Program that trains researchers exploring the links between chromosome metabolism andcancer. Advances in the studies of genes and genomes have greatly facilitated the understanding of chromosome metabolism (gene expression, chromatin assembly, segregation, telomeres and centromeres, DNA repair, etc.) and illustrated its importance in cancer initiation and progression. The 25 training faculty are experts in this area from the Basic Sciences Human Biology Divisions of the Fred Hutchinson Cancer Research Center. Many are leading scientists in their own fields, and together they form a cohesive group that approaches the issue from different but complementing angles. They include structural biologists in addition to molecular and cellular biologists, and utilize various model organisms such as nematode, fly and yeast in addition to vertebrates and cultured cells in their studies. The breadth and depth of this program should help generate excellent young scientists with creative approaches to understanding cancer mechanisms. Two predoctoral positions are requested to maintain the current level of support of this successful program that was initiated ten years ago following the formation of the Molecular and Cell Biology (MCB) Graduate Program. The postdoctoral positions will also be maintained at the current level of five. Predoctoral trainees will be recruited among the MCB students, who enter the PhD program through vigorous competition and are among the best in the nation. Postdoctoral trainees are drawn from an outstanding pool of fellows working with CMTP Faculty. Predoctoral students are appointed to the training grant after they have chosen a permanent laboratory and completed lab rotations and a portion of their required coursework. Postdoctoral training consists of independent research, supplemented by rigorous quarterly presentations in front of training grant faculty and experience in organizing CMTP-sponsored seminars. In addition, all trainees will benefit from the rich activities available for researchers at this Center, including the strong interdisciplinary programs that expose basic scientists to clinical and public health aspects of cancer. It is hoped that this program will create young scientists with in-depth understanding of chromosome metabolism and commitment to explore its various links to cancer cause, cure and prevention.
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{
"pile_set_name": "NIH ExPorter"
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The proposed research investigates automated, programming techniques for the objective evaluation and education, including socialization, of severely retarded children. The procedures permit full control over stimulus and reinforcing events, including social variables, and constitute a comprehensive intervention in the lives of children whose peculiar behavior problems make them difficult or impossible to teach by more traditional methods. The research will investigate the educational setting and conditions sufficient, and perhaps necessary, to achieve unambiguous and continuous stimulus control of the children's learning. The setting is a highly specialized teachning room for the individual child, where the interactions between the child and the teaching conditions are intensively examined and modified constructively.
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{
"pile_set_name": "NIH ExPorter"
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Overexpression of P-glycoprotein (P-gp) and/or multidrug resistance-associated protein (MRP1) is associated with resistance of tumors to a wide range of chemotherapeutic drugs. Many cancers initially respond well to chemotherapeutic treatment, but these tumors eventually become resistant to those cytotoxic drugs. Some other cancers are resistant to anticancer drugs, even at the beginning of treatment. Therefore, no matter whether the resistance is "acquired" or "intrinsic," multidrug-resistance is one of the major obstacles to the successful treatment of many types of cancers. The mechanism of multidrug resistance conferred by either P-gp or MRP1 involves extrusion of the drugs out of the tumor cells. Both P-gp and MRP1 are members of the ABC superfamily of transport proteins, typically containing two membrane-spanning domains and two nucleotide binding domains. However, MRP1 differs from P-gp in that it contains an extra membrane-spanning domain at the N-terminus. ATP (binding and hydrolysis) is required in both cases. However, the transport steps differ in that P-gp extrudes the unmodified drugs directly, while the drugs transported by MRP1 protein require the presence of a hydrophilic compound, e.g., glutathione- or glucuronide-conjugates. Since the mechanism of multidrug resistance conferred by P-gp or MRP1 is to reduce the level of drug accumulation inside of the cell, inhibiting of the function of P-gp or MRP1 may reverse the drug resistance. Therefore, much effort is being devoted to discover specific inhibitors of these pumps. Present candidates as modulators of the process include verapamil, cyclosporin A, Cremophor, ardeemins, PSC833, rifamycins, RU486, MS-209, non-steroidal inflammatory drugs, acrolein, pyridine analogues, ONO-1078, chloroacetaldehyde, imidazothiazole derivatives, and even some protein kinase inhibitors. However, in order to develop well-designed specific modulators, it is important to understand the mechanisms of drug transport carried out by each individual protein. The first specific aim is to further characterize the ATP finding/hydrolysis and substrate transport by MRP1 protein. The second aim is to determine whether MRP1 protein phosphorylates itself or is a substrate for other protein kinases. This is based on our hypotheses that the phosphorylated MRP1 protein is an intermediate (i.e., there is an autophosphorylation) during ATP hydrolysis and substrate transport. The third aim is to define the substrate binding site(s). New insights gained from these aims should provide the basis for novel means of combating multidrug resistance.
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{
"pile_set_name": "NIH ExPorter"
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Project summary: The major goal of this research proposal is to elucidate the function of serotonin (5-HT) in feeding and feeding-related behaviors in Drosophila melanogaster. 5-HT affects the feeding behavior of both vertebrates and invertebrates, however the precise molecular mechanisms by which this occurs are largely unknown. 5-HT also influences numerous other behaviors and physiological processes that could affect feeding such as locomotion, circadian rhythm and digestion. Using the genetically tractable fruit fly, I will address the following questions: How does 5-HT affect both feeding and other feeding-related behaviors? Are there specific subpopulations of 5-HT neurons that affect feeding directly? What is the role of the serotonin receptors in these behaviors? I will address these questions with the following specific aims: Aim 1. Loss-of-function and gain-of-function manipulations of 5-HT signaling. Aim 2. Identification and characterization of 5-HT neuronal subsets. Aim 3. Analysis of 5-HT receptors in feeding behavior. Relevance to public health: Obesity is a serious health concern that confronts millions of people worldwide. In the U.S., more than 65% of adults and 15% of children are considered overweight. Exploration of feeding behavior in model organisms will promote our understanding of how complex behavioral states, such as hunger and satiety, are controlled in the nervous system. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Potassium channels of the cardiac cell membrane play a major role in initiating and controlling the heart beat. Thus, for example, cardiac pacemaker activity is slowed by the neurotransmitter acetylcholine by way of its activation of an inwardly-rectifying potassium channel in the cll membrane. The mechanism of this activation is complex. It is thought to proceed through the stimulation of membrane-bound muscarinic receptors which activate a signal-transducing guanine nucleotide binding protein. Interaction of this activated transducer molecule with the potassium channel, either directly or indirectly by way of intermediate steps, results in channel opening which produces the electrophysiological effect. The objective is to elucidate the molecular mechanisms by which the function of cardiac potassium channels is controlled by neuroendocrine, pharmacological and biochemical factors. Initially, the project will combine electrophysiological and biochemical techniques to establish the molecular mechanisms of the control of K+ channels in living heart cells produced by enzymatic disruption of the adult heart. Specifically, the ability of purified quanine nucleotide binding proteins to open K+ channels will be examined to establish which of the signal transducing G-proteins activate K+ channels and whether or not activation occurs by direct interaction of the G-protein with the channel protein. These studies will be followed by efforts to incorporate, from vesicles of heart sarcolemma into lipid bilayer membranes, functional K+ channels that respond to modulation by the appropriate G-protein. Reconstitution of the signal transducing complex in a simple, chemically well-defined system is necessary for a molecular understanding of the mechanism by which the system functions. It allows one to make molecular changes, for example by altering the G-protein or the membrane lipid composition, specifically designed to test hypotheses concerning the function of the K+ channel and its interaction with G-protein. Later stages of the project will also attempt to isolate and biochemically characterize the K+ channel protein by taking advantage of its specific interaction with G-protein. If successful, this procedure should permit the sequencing and eventual cloning of the K+ channel.
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{
"pile_set_name": "NIH ExPorter"
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ABSTRACT: The goal of this proposal is to provide technically-advanced support for the specific aims of NCI- funded projects for medical imaging affiliated with the University of Washington, the NCI Quantitative Imaging Network (QIN), and the NCTN ECOG-ACRIN cooperative group. My medical imaging career began at UW in 1985, where I was fortunate to work with Drs. Michael Graham and Alex Spence, who laid the foundation for quantitative analysis and clinical oncology, respectively. My early work in PET imaging revolved around managing animal imaging projects, a full biochemistry lab and a nascent human PET glioma imaging program. As PET imaging developed under the PET Program Project (P01-CA042045, KA Krohn, PI), we transitioned to human imaging and clinical trials research, probing different biochemical pathways using numerous PET radiotracers. I developed an avidity for compartmental modeling analysis of kinetic image data, initially with guidance from Drs. James Bassingwaithe, Michael Graham and Finbarr O'Sullivan, and later in collaboration with Drs. Tony Shields, David Mankoff, Janet Eary, Hannah Linden and Paul Kinahan, all of them funded by NCI grants. I am currently Director of the Imaging Core lab and a member of Paul Kinahan's U01 grant. I have responsibility for participating in the image analysis working groups of the QIN, extended planning of PET imaging projects, imaging protocols and designing data analysis methods for new experiments. I am a senior contributor to our overall program as well as a provider of novel approaches to data management, quantitative image analysis and data simulations for numerous cancer projects in Radiology and Oncology. Due to the cost of imaging, the radiation dose and other issues, my personal ambition is to extract as much quantitative information as possible from well-designed and executed cancer imaging studies using PET radiotracers and advanced MR methods. My career goal is to provide independent, but collaborative, support for the specific aims of NCI grants for medical imaging projects at with UW, the QIN and ECOG-ACRIN. Additionally, I have national recognition as manager of the ACRIN-UW PET Core Lab, member of the ECOG-ACRIN Head and Neck, Experimental Imaging Sciences and Brain Tumor Committees, and a key member of the NIH-Cancer Imaging Program's QIN Working Groups. The innovative protocols I developed for dynamic imaging are a substantial improvement for extracting quantitative information from dynamic PET, and are now a national clinical trials standard. I routinely present my advanced PET imaging results at national and international scientific meetings (EORTC, SNMMI, WMIC, ACRIN and QIN/NCI) and publish/review manuscripts annually in medical journals. In summary, I have developed a nationally recognized advanced lab for PET image analysis, funded entirely through NCI grants for 30 years. The overall benefit NCI receives for sharing my labor across NCI funded projects will have an impact on current and new investigators, providing them with quantitative methods of analysis for PET/CT and MRI image data using robust, repeatable and validated methods.
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{
"pile_set_name": "NIH ExPorter"
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Myeloid leukemias are characterized by aberrant blockade of differentiation due to expression of leukemic oncoproteins such as PML/RAR-alpha. Elucidation of the mechanisms responsible for normal myeloid differentiation will further our understanding of differentiation blockades in leukemias, and foster the development of novel differentiation therapies. Thus, our long-term objective is to define the intracellular signaling pathways that lead to myeloid differentiation, and the role these pathways play in leukemogenesis. In preliminary studies we have observed rapid and prolonged activation of the MEK/ERK signal transduction pathway during myeloid differentiation. Furthermore, MEK/ERK activation was required for differentiation in myeloid cell lines. This contrasts with known roles for MEK/ERK activation in proliferation and survival, and constitutive MEK/ERK hyperactivation in myeloid leukemias. Together, these studies suggest a dichotomy of roles for MEK/ERK activation: promoting differentiation under normal conditions, and transformation under conditions where differentiation is blocked. Therefore, we hypothesize that prolonged activation of the MEK/ERK pathway is critically important for cytokine-induced myeloid differentiation. We further hypothesize that activation of the MEK/ERK pathway cooperates with differentiation-inhibiting leukemic oncoproteins to promote leukemogenesis. To test this hypothesis, we will: 1) elucidate the importance of MEK/ERK activation during cytokine-induced myeloid differentiation of myeloid cell lines and primary cultures of normal murine myeloid progenitors;2) determine the functional consequences of MEK/ERK activation during cytokine-induced myeloid differentiation;3) examine whether MEK/ERK activation promotes myeloid differentiation in vivo through the generation of transgenic mice that inducibly express constitutively active MEK enzyme in myeloid lineage cells;and 4) determine whether MEK/ERK activation cooperates with expression of PML/RAR-alpha to promote myeloid leukemogenesis.
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{
"pile_set_name": "NIH ExPorter"
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Otitis media (OM) is the most common cause of acquired pediatric hearing loss and may delay language and cognitive development. It is the leading diagnosis in the U.S. for children and is the primary reason for prescribing antibiotics and performing surgery. The etiology and pathogenesis of OM are poorly understood. This has hampered the development of rational therapeutic approaches. The primary goal of this proposal is to gain a better understanding of the role of bacteria and viruses in OM. Elucidation of the pathogenic cascade in OM has been impeded by the inherent limitations of conventional cultural methods for these microbes. These obstacles can be overcome by use of the polymerase chain reaction (PCR) which has revolutionized the study of infectious disease. Chronic OM effusions are being obtained from a well- characterized pediatric population during myringotomy and tube placement. These specimens are then subjected to comparative analysis for microbial infection using PCR-based assays and culture methods. Using this molecular method, Hemophilus influenzae and Streptococcus pneumonia DNA have been detected in the majority of culturally-sterile middle-ear effusions of children. To determine if these findings are indicative of low-grade active infections the molecular environment of the middle-ear will be characterized. The fate of nucleic acids. and DNase activity, will be ascertained using a well-characterized animal model (Chinchilla laniger). A multiplex-PCR-based assay will be developed to detect and differentiate upper respiratory tract viruses in pediatric middle-ear effusions. The results of this analysis will be compared with culture. The chinchilla model will be joined with one of the most recent advances in molecular medicine to gain a fundamental understanding of viral pathogenesis in OM. In situ PCR is the molecular equivalent of immunohistochemistry an can demonstrate the location of amplified nucleic acids in histologic sections. PCR and in situ PCR will be used in the chinchilla model to determine if OM is a consequence of direct viral invasion of the middle- ear mucoperiosteum following nasopharyngeal infection, or rather a sequelae of viral-induced Eustachian tube (ET) dysfunction. In future studies this molecular diagnostic system will be used in children to establish whether acute OM is the result of a direct viral infection of the middle ear, or rather the result of virally-induced ET dysfunction with concomitant bacterial infection.
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{
"pile_set_name": "NIH ExPorter"
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The long-range objective of the proposed research is to elucidate basic principles and mechanisms governing the prenatal and postnatal development and plasticity of prefronatal association cortex in primates. The research is composed of four integral parts: (1) analysis of the outgrowth, synaptogenesis and organization of certain classes of prefrontal cortial connections during the course of prenatal and postnatal development; (2) assessment of the functional development of prefrontal cortex and related subcortical structures; (3) investigations of the structural and (4) functional plasticity which follows early injury to prefrontal cortex. The studies are conducted on rhesus monkeys because the expanse of association cortex in this species as well as its neuronal organization, protracted development and capacity for mediating complex behavior makes it an excellent animal model for the study of normal and pathological development of human neocortex. The project combines behavioral studies with a battery of advanced neurobiological methods (light and electron microscopic autoradiography, horseradish peroxidase and acetylcholinesterase histochemistry and (14C)-2-deoxyglucose mapping of the central nervous system). These techniques are used in conjunction with recently refined procedures that permit experimental manipulation of the primate fetal brain with subsequent return of the fetus to the uterus and its survival to and after term. Information from these studies will increase understanding of the development and modifiability of complex regions of the human brain and may lead to the discovery of means of prevention or treatment of cognitive disorders resulting from congential and neonatal brain injury.
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{
"pile_set_name": "NIH ExPorter"
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The long-range goal of the proposed research is to gain a detailed understanding of the cellular and molecular mechanisms underlying long- term potentiation (LTP) of synaptic transmission. Changes in the efficiency of transmission at synapses, such at LTP, are thought to be an important mechanism by which the function of the brain is modulated and by which memories might be stored. One of the most important and controversial issues surrounding LTP is whether its expression involves the presynaptic terminal. The proposed experiments provide critical specific and direct tests of the role of the presynaptic neuron in LTP, with an emphasis on determining the biochemical mechanisms involved. Some of the experiments are geared towards testing the role of the diffusible messenger nitric oxide (NO), which may act as a "retrograde messenger" from the postsynaptic neuron to the presynaptic neuron in LTP. LTP will be studied using electrophysiological recordings from pairs of synaptically coupled rat hippocampal neurons in organotypic culture. Pharmacological agents will be injected into presynaptic neurons so as to test their effects on LTP. The specific targets of the experiments include, but are not limited to, signal transduction pathways which might be activated by NO in presynaptic terminals.
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{
"pile_set_name": "NIH ExPorter"
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This project involves evaluating improved man-machine interface methods and identifying ways that modern microprocessor technology might benefit physicians and the practice of medicine. Interface methods that offer promise are voice input/output, bar codes (such as used with point of sale equipment), and touch screen selection systems for use with video displays. Potential applications include medical record keeping, medical education, and computer-aided diagnosis.
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{
"pile_set_name": "NIH ExPorter"
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The ultimate objective of this research plan is to establish a multi-disciplinary, multi-institutional Network for Translational Research to develop, standardize, validate, and optimize a targeted, multi-modal optical/nuclear imaging platform that uses labeled probes to identify dysplastic mucosa in the digestive tract for the early detection of cancer in patients at increased risk. The goal of the Primary Project is to establish network infrastructure, standardize imaging protocols and validate performance measures among three clinical centers for detecting preferential binding of fluorescent-labeled peptides to sessile colonic adenomas > 5 mm in diameter used as a model for dysplasia. Wide area endoscopy is used to localize regions suggestive of peptide adherence and confocal microscopy provides confirmation of binding to dysplastic colonocytes rather than being trapped in mucus or debris for increased detection specificity. Peptides will be topically applied to the local mucosa via a spray catheter during routine screening colonoscopy to demonstrate the proof of concept of this novel imaging methodology for future applications that provide delivery via an enema. We have combined the strengths and resources from academia and industry to establish a world class team of investigators from the University of Michigan, Stanford University, Mayo Clinic, Olympus Medical Systems Corp, and STI Medical Systems Inc to pursue these aims. Pilot clinical studies will be performed to prepare this Center for a future multi-center clinical trial by the end of the funding period. In addition, four Task-Specific Projects have been proposed to support the progress of the Primary Project. They include investigating the use of radio-labeled peptides to localize dysplasia on imaging with SPECT/CT for ultimate use in high risk individuals to determine the screening interval for colonoscopy. Furthermore, novel optical instrumentation will be developed using a dual axes confocal architecture to image into the submucosa for future use in assessing tumor invasion and micrometastases. In addition, new peptides will be discovered that bind to flat dysplasia that cannot be appreciated on standard white light endoscopy using techniques of phage display and gene expression profiles to provide functional targets. Peptides have been chosen for use as molecular probes because of their clonal diversity, small size, and minimal immunogenicity, and are well-suited for clinical use because of their rapid binding kinetics, deep tissue penetration and lack of toxicity. At the end of this funding period, we will be prepared to validate this integrated strategy in a multi-center clinical trial. Public Health: The proposed studies will result in the standardization and validation of a novel, multi-modal imaging platform that uses labeled peptides to target the presence of pre-malignant mucosa in the digestive tract of individuals at increased risk for the early detection and prevention of cancer. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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This is a multi-group research study which plans to evaluate the overall progression-free survival of children with average-risk medulloblastoma treated with craniospinal radiation and local boost radiotherapy plus one of two adjuvant chemotherapy regimens differing in the replacement of CCNU by cyclophosphamide. The long-term neurocognitive, endocrinologic and cardiopulmonary sequelae of radiotherapy with adjuvant chemotherapy will be determined, as well as the feasibility of routine surveillance scans to detect sub-clinical recurrent disease. Another objective of this study will be to evaluate the sensitivity of molecular and biochemical techniques to predict progression-free survival and disease relapse. Following surgery, patients will be randomized to receive Regimen A or B of treatment. Both regimens will include 2340 cGy of craniospinal radiation and 3240 cGy of boost radiation directly to the primary tumor with weekly vincristine doses. Six weeks following the completion of radiotherapy, patients will begin eight cycles of maintenance chemotherapy for Regimen A; CCNU, cisplatin, and vincristine or Regimen B; cyclophosphamide, vincristine and cisplatin.
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{
"pile_set_name": "NIH ExPorter"
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Cells require ordered movement of proteins and lipids from one membrane-bound compartment to another, while maintaining the organization, function and heterogeneity of the donor and acceptor membranes;vesicular membrane traffic carries out these functions. Clathrin coated vesicles are the most prominent form of traffic from the plasma membrane to endosomes (endocytosis), a pathway by which various ligands and their receptors enter cells. Clathrin coated vesicles are also important for traffic between endosomes and the trans-Golgi network (TGN). We propose to continue our studies of molecular mechanisms in clathrin-mediated membrane traffic, using direct observation by live-cell imaging with single-molecular sensitivity and high spatial and temporal resolution, to answer the following questions. (1) What are the molecular events required to initiate formation of a clathrin coated pit? (2) What are the molecular components of coats trapped at various stages of growth and closure: early and late abortive coated pits, and pits stalled at a relatively late stage, by imposing tension of the membrane and by inhibiting actin dynamics? (3) What is the mechanism of clathrin uncoating mediated by Hsc70 and its co-chaperone auxilin and how is the uncoating coordinated with assembly so that the two processes do not compete? PUBLIC HEALTH RELEVANCE: Vesicular membrane traffic is the principal mechanism for moving proteins and lipids among membrane-bound compartments in a cell. Clathrin coated vesicles are the most prominent form of traffic from the plasma membrane to endosomes (endocytosis), a pathway by which ligands such as hormones, transferrin, immunoglobulins, LDL, viruses and their receptors enter cells.
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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. MultiSeq [1] (http://www.scs.uiuc.edu/[unreadable]schulten/multiseq/) is a unified bioinformatics analysis environment within VMD that allows one to organize, display, and analyze both sequence and structure data for proteins and nucleic acids. Special emphasis is placed on analyzing the data within the framework of evolutionary biology.
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{
"pile_set_name": "NIH ExPorter"
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Thermal injury represents a common form of trauma associated with significant mortality and morbidity. Despite the remarkable improvements in critical care and wound management, end-organ effects of systemic inflammatory reactions characteristic of multiple organ edema and dysfunction remain life-threatening problems in patients with major burns. Of great concern is the development of microvascular barrier dysfunction, a fundamental cellular process underlying inflammatory edema that has yet to be understood at the molecular level. The overall goal of this project is to identify key signaling molecules and structural components responsible for the microvascular hyperpermeability response to thermal trauma. Within this context, a major hypothesis is developed stating that Src tyrosine kinase-signaled endothelial cytoskeletal contraction and junctional disorganization induce microvascular barrier dysfunction during thermal injury. Correspondingly, four specific aims will be achieved: 1) to characterize burn-induced microvascular leakage; 2) to examine the mechanism by which endothelial cytoskeletal contraction causes microvascular hyperpermeability; 3) to elucidate the role of VE-cadherin junctional disorganization in mediating endothelial barrier dysfunction; and 4) to verify the common signaling role of Src in the regulation of cytoskeletal and junctional structure and function. The study will test the hypotheses using a clinically relevant rat model of scald burn in combination with pharmacological approaches and molecular biology techniques. The study will provide new insights into the cellular and molecular control of endothelial barrier function. Information derived from this study will enhance our knowledge on microvascular pathobiology of not only burns but also other types of injuries associated with inflammatory responses. Furthermore, identification of signaling or structural molecules directly responsible for microvascular hyperpermeability should lead to the development of new therapeutic targets specific to the injurious process.
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{
"pile_set_name": "NIH ExPorter"
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Summary of work: Most studies of age differences in personality have been conducted in the United States using self-report measures. For this project, two studies were conducted across cultures using observer ratings of personality. In the first, we obtained both self-reports and informant ratings of the same individuals in Russia (N = 800) and the Czech Republic (N = 705). In both cultures, the same pattern of age differences was found: Neuroticism, Extraversion, and Openness declined across the lifespan, whereas Agreeableness and Conscientiousness increased. Informant data generally replicated self-report findings, although the effects were weaker. In the second study we gathered college students' ratings of an anonymous target, either 18-21 or 40+ years old, in 50 cultures, from Iceland to Ethiopia. Adults were rated as higher than college-age targets on Conscientiousness and lower on Extraversion and Openness in most cultures. Age differences on Neuroticism and Agreeableness were much weaker. Although there are still some unexplained differences between self-report and informant rating methods of assessing personality, the data overall demonstrate that age differences are universal, supporting the hypothesis that personality change is part of the intrinsic maturation of the human species.
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{
"pile_set_name": "NIH ExPorter"
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Hedgehog (Hh) signaling is employed in controlling cell fates in most developing tissues and organs, as well as during many regeneration events. Defects in Hh signaling lead to birth defects and cancer. Many mechanistic mysteries remain regarding how an Hh signal is transduced. Using high-throughput RNAi screening, we identified Neuropilins (Nrp) 1 and 2 as novel, specific regulators of vertebrate Hh signaling. Nrps are single-pass transmembrane proteins implicated in the reception of a diverse set of secreted ligands, including Semaphorins and VEGF165 and in cell adhesion and cell migration. In fibroblasts the inhibition of Hh signal transduction resulting from blocking Nrps is as strong as the effect of blocking cilia formation or blocking Smoothened function. Conversely, over-production of either Nrp sensitizes cells to Hh signals. New components of the Hh pathway are uncovered infrequently; the Nrps were probably missed due to their partial redundancy. Our discovery of two proteins whose functions are required by this important morphogenic pathway has the potential to bring fundamental changes to current models of Hh signaling and to enlarge the understanding of Nrp functions in other signaling pathways. Aim 1. Determine the mechanism by which Nrps regulate Hh signal transduction. Nrps could influence Hh signal transduction by directly associating with known Hh pathway components, or mediating other signals that converge with Hh transduction, or by altering cell properties or processes that are required for Hh transduction. We will investigate each of these possibilities by determining which steps in Hh signaling are affected, whether cell adhesion or migration changes are involved in the effect of Nrps upon Hh signaling, and what proteins interact directly with Nrps. Aim 2. Determine which domains of Nrp are needed to support Hh signaling, and whether known Nrp co-receptors, ligands, or effector molecules are capable of Hh pathway cross- regulation. We will investigate which domains contribute to Hh signal transduction in two ways: engineered domain deletions, and a high-throughput screen for point mutations that interfere with Nrp support of Hh signal transduction. Nrps transduce VEGF and Semaphorin signals, acting as co-receptors for VEGF receptors and Plexins, respectively. We will test whether VEGF, VEGF receptor, Plexin receptors, or Semaphorins are involved in the effect of Nrps upon Hh signal transduction. Aim 3. Investigate how Nrps influence Hh- dependent development and tumorigenesis. Using mice that carry mutations in both Nrp genes, we will control temporal and tissue-specific removal of Nrp functions to test their involvement in several Hh-dependent developmental processes in vivo. Using newly created lentiviruses, which encode specific inhibiting RNAs that block the nrp genes, we will infect primary cultures of Hh-responsive cells and monitor effects on Hh target gene expression.
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{
"pile_set_name": "NIH ExPorter"
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The objectives of this proposal are to obtain an understanding of the selective inhibition of fat intake by enterostatin, the aminoterminal pentapeptide of pancreatic procolipase. The hypothesis to be investigated is that enterostatin is secreted in response to the intake of dietary fat and acts as a feed-back regulator of fat intake. Excessive intake of dietary fat is thought to be a major factor in the development of obesity and the high incidence of cardiovascular disease but little is known of the physiological control of fat intake. We shall investigate the amino acid sequences and structural requirements for maximal biological activity of the peptide in the inhibition of fat intake, the regulation of synthesis and secretion of enterostatin, the site(s) of action of the peptide, its mechanism of action and its activity in a number of animal species in order to explore the physiological role of enterostatin. Enterostatin analogues will be synthesized and used to investigate the structural requirements necessary for the inhibition of fat intake after central and peripheral administration. The site of production of the peptide will be studied by Northern blot analysis using cDNA probes to parent molecules and an ELISA assay will be used to study serum and intestinal levels of enterostatin. With these techniques the regulation of synthesis and secretion of the peptide in response to dietary and endocrine manipulations that alter fat intake will be studied. As enterostatin is effective in reducing fat intake after both central and peripheral administration the possibility that enterostatin has a peripheral site of action will be investigated by studying the effects of enterostatin infused intravenously or into the gastrointestinal tract, and the possibility that this peripheral action is mediated through control of gastric emptying or at intestinal or hepatic sites via afferent vagus nerves will be studied. Central sites of action will be investigated by infusions of enterostatin into specific brain nuclei in rats stereotaxically implanted with guide cannulas. The interrelationship of the enterostatin inhibition of fat intake with the galanin stimulation of fat intake will be studied and the possibility that either or both of these systems are modulated through mu-opioid receptors will be explored in two ways. Firstly, the interactions of centrally injected enterostatin, galanin, and opioid agonists and antagonists in regulating fat intake will be studied; secondly, ligand binding assays will be used to identity specific receptors for enterostatin and investigate the possibility that enterostatin interacts directly with mu-opioid receptor systems. Finally we shall investigate the ability of enterostatin to suppress fat intake in other animal species (mice, dogs and baboons). As a result of these studies, we will gain important insights into the regulation of fat intake and the possible role that an endogenous peptide has in this process. This new information may be the impetus for a new therapeutic approach to treating obesity.
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{
"pile_set_name": "NIH ExPorter"
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This COBRE application builds on the foundation laid by the initial COBRE (RR15640). The Neuroscience COBRE has as its programmatic objective the establishment of a sustainable Center for Neuroscience. Significant progress has been made. Neuroscience was identified as one of four Areas of Distinction in the University Academic Plan. A critical mass of Neuroscience Center faculty is developing by the augmentation of existing faculty with the addition of three recent hires and the approval for 3-4 more Neuroscience faculty to be recruited during the next support period. Core research facilities have been established that have enhanced the biomedical research capability of investigators and which are necessary for the continued growth of the Neuroscience Center. Innovative research programs have developed as a result of COBRE established Core facilities for imaging the brain, and analysis of the genetic and protein structure of the nervous system. Multidisciplinary teams have formed that are comprised of electrical engineers, electro physiologists, and behavioral neuroscientists. Collectively these teams have as their thematic focus the study of activity dependent changes in neuronal function which are essential for the survival of the animal and normal brain function. The Neuroscience COBRE investigators will work on interrelated projects that seek to understand how central nervous system circuitry and neural pathways adapt to changes in sensory information. The model systems used by investigators will explore mechanisms underlying activity dependent changes in neural systems due to alterations in visual (including photoperiod), somatosensory, and nociceptive input. Approaches to these functional questions will be multidisciplinary with the investigators sharing electrophysiological, pharmacological, molecular, and neuroanatomical techniques. Through collaborative efforts, computational processing algorithms may be developed to model activity dependent changes in mature, functional systems. Collectively, the projects will show the degree to which inhibitory circuits shape the neuronal architecture of incoming sensory inputs at multiple levels of the nervous system and in multiple sensory systems. The overall scientific objective is to advance our understanding of neural systems controlling sensory processing, neurochemical and neuroendocrine signaling and behavior, so as to facilitate prevention and treatment of: pathological states resulting from sensory restriction, neuroendocrine dysfunction, and chronic pain.
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{
"pile_set_name": "NIH ExPorter"
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The goal of this proposal is to obtain mechanistic insights into the role of cardiac mitochondria in defining the substrate for atrial fibrillation (AF), the most common arrhythmia encountered in clinical practice. With a projected 6-fold increase in the prevalence and a cost exceeding $15 billion per year, AF remains a major national health problem. Despite the recognition that aging increases susceptibility of the atria to fibrillation, with a 100 fold higher prevalence in the older-elderly compared to young adults, the molecular bases for this remains unknown. Changes in hemodynamic, vascular, and metabolic factors that accompany aging or associated disease contribute to functional and structural atrial remodeling promoting cardiomyocytes loss and fibrosis that increases susceptibility to fibrillation, however, the molecular bases for such alterations contributing to the progression of atrial dysfunction are not well defined. In our preliminary studies, using human atrial tissue, a distinct transcriptional downregulation of genes regulating mitochondrial energetics and signaling pathways involved in energy production and utilization, cell loss and fibrosis was demonstrated with aging and AF. Additionally, functional defects with impaired capacity to maintain cellular energetics and ionic homeostasis under stress were demonstrated in senescent mitochondria that can be ameliorated by modulating mitochondrial membrane permeability. Based on these, we hypothesize that susceptibility to AF in the elderly results from diminished mitochondrial functional reserves in the atria that promote cardiomyocyte loss and fibrosis due to enhanced sensitivity of the myocardium to energetic failure, calcium overload and oxidative injury during stress, thus facilitating development and progression of the substrate for AF. We propose 1) to identify differences in atrial structure and function, energetics and mitochondrial susceptibility to stress in patients with low or high risk for the development of AF and those with paroxysmal, persistent or permanent AF;2) to identify mechanisms underlying atrial energetic deficits and mitochondrial dysfunction predisposing to enhanced cell loss and fibrosis;and 3) to determine the protective role of mitochondrial modulation against mitochondrial and cellular injury during metabolic stress in patients at risk for or with AF. These aims will be achieved using atrial tissue obtained from patients undergoing coronary artery bypass surgery without or with risk factors for AF (heart failure, hypertension, or mitral regurgitation) or a history of paroxysmal, persistent or permanent AF. An integrative approach combining clinical information with in vivo and in vitro atrial structural and functional data obtained by imaging with comprehensive cellular and mitochondrial studies assessing differences in ultrastructural, functional, molecular, genetic and proteomic changes in atrial tissue between those at risk of AF, who develops AF following surgery and those with AF, not only is highly innovative but also of high clinical significance. The results will provide new insights into the role of mitochondria priming the substrate for AF and identify novel targets for the development of therapeutics toward prevention of AF. (End of Abstract)
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{
"pile_set_name": "NIH ExPorter"
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Intramuscular injections of plasmid DNA has been shown to express transgenes at relatively efficient levels. Experiments are currently underway to define the mechanism of DNA uptake by muscle cells. We would like to use the confocal microscope to examine the efficiency of nuclear uptake of DNA under various conditions after cytoplasmic microinjection.
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{
"pile_set_name": "NIH ExPorter"
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Determine if the regional cerebral blood flow (rCBF) of patients with isolated amnestic syndromes differs from that of healthy persons, particularly during performance of memory tasks.
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"pile_set_name": "NIH ExPorter"
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The National Network of Libraries of Medicine program is intended to provide health science practitioners, investigators, educators, and administrators with timely, convenient access to information. The program is coordinated by the National Library of Medicine and carried out through a nation-wide network of more than 3,000 health science libraries and information centers. It incorporates the individual health practitioner within the institutional network by involving all of the libraries in the Network in establishing direct contact with health professionals. The Network also includes eight Regional Medical Libraries, or RMLs. These are major institutions designated by NLM to administer and provide backup services in each of eight geographical regions. Objectives for the RMLs are: to promote awareness of and access to biomedical information resources for health professionals through expanded and targeted outreach programs, to develop and improve the biomedical information resources in the regions, and to support the sharing of these resources within the regions and throughout the nation. Region 1 provides services to the states of Delaware, New Jersey, New York, and Pennsylvania.
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{
"pile_set_name": "NIH ExPorter"
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Models of HBsAg specific cell mediated lympholysis have been developed and utilized to examine the cytotoxic T lymphocytes (CTL) in hepatitis B patients. Procedures are being developed to establish antigen specific T lymphocyte cell lines. One T cell clone has been isolated from HBV vaccine recipient by repeated alternated stimulation with antigen and interleukin 2. Its antigenic specificity and HLA restriction have been established. The HLA restriction has been mapped and the T cell specific antigenic site at primary structural level is being sought after with synthetic polypeptides.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary Synthetic inhibitors of protein-protein interactions (PPIs) are emerging as a promising approach for discovery of new therapeutics. However, despite recent progress, PPIs remain challenging targets and the in vivo potential of small molecule PPI inhibitors remains insufficiently characterized. We will develop rationally designed small molecule inhibitors of an interaction that is critical for HPV-associated head and neck squamous cell carcinoma (HNSCC). Strong preliminary results, including in vivo animal data, support our strategy. HNSCC is the sixth most common cancer with an annual incidence of approximately 600,000 cases worldwide. Recent evidence supports the recognition that HPV infection is a major risk factor for HNSCC, in particular oropharyngeal SCC. High-risk HPV16 is by far the most frequent, ~90%, HPV type detected in HNSCC. Epidemiological data indicate that the prevalence of HPV-positive HNSCC has rapidly increased by about 3-fold in the past three decades in the United States and Europe. Based on these alarming numbers, it has been suggested that an epidemic of HPV-positive HNSCC will emerge in the near future. Therefore, there is a clinical need to identify alternative therapeutic strategies to manage HPV-positive HNSCC patients. In HPV16-associated HNSCC, the tumor suppressors, p53 and Rb, are expressed as wild type but inactivated by the HPV16E6 and E7 proteins. Simultaneous reactivation of p53 and Rb offers an appealing strategy for developing anti-cancer leads for HPV16-associated HNSCC. p53 and Rb function are modulated through post-translational acetylation by the transcriptional co-activator p300. HPV16E6 and HPV16E7 bind the CH1 and CH3 protein-protein interaction domains of p300. Since p300 is known to be a limited resource, we hypothesized that HPV16E6 and HPV16E7 hijack p300 activity in HPV16-positive HNSCC. We will leverage our preliminary results and advance prototype HPV16E6-p300 small molecule modulators as rationally designed molecularly targeted therapeutics for HPV16-positive HNSCC. The project will have broad impact on discovery of PPI inhibitors because little is known about the in vivo potential and selectivity of small molecule PPI modulators. We have assembled a multi-disciplinary team of experts in head and neck cancer biology and signal transduction (Pan) and chemistry (Arora) to successfully complete this proposal.
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{
"pile_set_name": "NIH ExPorter"
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Androgen deprivation therapy (ADT) is being increasingly used as both a primary and neoadjuvant therapy for patients with localized prostate cancer (PCa). Our recent studies have demonstrated that the metabolic information provided by 3D MR spectroscopic imaging (MRSI) can improve the specificity of magnetic resonance imaging (MRI) in identifying prostate cancer prior to and after therapy. However, our preliminary data also indicate that the anatomic and metabolic atrophy induced by ADT is duration dependent and the ability of combined MRI/MRSI to assess the presence of residual/recurrent cancer can be reduced at certain times after therapy. Therefore, the time-course of anatomic and metabolic atrophy following hormone therapy needs to be established. Preliminary proton magic angle spinning nuclear magnetic resonance (1H MAS-NMR) spectroscopy studies of surgical specimens have also identified several metabolic changes that may further increase the specificity of MRSI for the identification of cancer after ADT. In this fellowship, serial in vivo MRI/MRSI data and 1H MAS-NMR data obtained from biopsy samples will be combined with clinical data to examine the anatomic and metabolic changes that occur in patients undergoing ADT. The goal of this work is to improve the MRI/MRSI assessment of cancer after hormone ablation and to better understand the biochemical consequences of ADT. The clinical importance of this work combined with Dr Swanson's strong interest and background in the application of NMR to biological questions and the extensive experience in MRI/MRSI of prostate that we have at UCSF makes this an ideal topic for his post-doctoral fellowship.
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"pile_set_name": "NIH ExPorter"
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Unique opportunity to study 12 year old boy with pseudohypoaldosteronism, to determine 1) his sodium and potassium replacement requirements, 2) the effect of variations in intake of these ions on muscle function and endurance, 3) the effect of an angiotensin converting enzyme inhibitor on electrolyte balance, and 4) the effect of oral K-exelate on the control of hyperkalemia. It is hoped this information will lead to improved care for this patient and better understanding of the treatment of this rare disorder.
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{
"pile_set_name": "NIH ExPorter"
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Leydig cells within the testis are the source of androgens that promote virility at both fetal and adult stages, but Leydig cell populations are distinct at each age. As such, adult and fetal Leydig cells function to synthesize testosterone in distinct cellular and endocrine environments. The AIMs of this proposal are focused on regulation of Star, the known gatekeeper in controlling access of cholesterol to enzymatic activity of the series of steroidogenic enzymes required for conversion to testosterone. Here we focus on novel regulatory events that exert dynamic interactions with Star chromosomal loci that will explain how fluctuating Star transcript accumulation can relate to changes in androgen synthesis. Previously, we used high-resolution fluorescent in situ hybridization (HR-FISH) to localize and quantify a unique pattern for primary, spliced, and mRNA species accumulation for Star compared to other steroidogenic genes within single Leydig cells. We will use this and other innovative techniques to compare results from studies that investigate individual adult and fetal Leydig cells in vitro, within MA10 cells and primary cultures, and in vivo, within whole testes. Pulsatile LH stimulates primarily cAMP/PKA signals to promote testosterone synthesis in adult Leydig cells. While we have substantial means to explain how Star transcription is turned ON by LH/PKA, we understand little about what happens when the pulse is removed, and even less about the interpulse interval. In AIM 1, we will test the hypothesis that the events occurring at Star loci during this interval are just as critical to controlling testosterone output as the initial stimulus. Meanwhile, the external stimuli that maintain androgen synthesis in fetal Leydig cells are less clear, but evidence points to paracrine signals, with PKA activity playing a role. Another paracrine factor, Sertoli cell-derived Desert Hedgehog (Hh) is known to initiate fetal Leydig cell differentiation, but its role in their maintenance has not been tested. Once differentiated, fetal Leydig cells produce androgens at a steadily increasing rate until late gestation. Therefore, in AIM 2, we will test the hypothesis that regulatory events on Star loci facilitate a controlled increase in androgens within the fetal Leydig cell that compare to those that occur during the interpulse interval in adult Leydig cells. Our findings have the potential to explain fundamental biology underlying steroidogenic control and will have a profound impact on our ability to explore mechanisms by which disturbances in testosterone synthesis, as in endocrine disruption, cause significant clinical ramifications in males from all stages of life.
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{
"pile_set_name": "NIH ExPorter"
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SUMMARY There is an urgent need to understand the diverse mechanisms of cardiac arrhythmias that lead to sudden death and stroke, which remain major health concerns and place more than 10 million Americans at risk. The 2017 Gordon Research Conference (GRC) on Cardiac Arrhythmia Mechanisms, ?The New Basics: Model Systems, Emerging Technologies and Precision Medicine? addresses this need by emphasizing the importance of basic science discovery, the power of model systems and the new technologies that ultimately enable patient-specific, mechanistic approaches to disease diagnosis and treatment. The long-term objective of this meeting is to provide a forum for investigators ? junior, mid-level and senior ? to engage in focused discussion on the latest advances in the field of cardiac arrhythmia mechanisms. The meeting fosters the development of trainees by supporting an associated Gordon Research Seminar on Cardiac Arrhythmia Mechanisms at which trainees present their research findings and engage in panel discussions with mentors on matters of career pathways and professional development. The specific aims of the conference are: ? To bring together investigators who study the mechanisms of cardiac arrhythmias from different disciplinary perspectives using state-of-the-art methodologies. In the intense and informal atmosphere of the GRC, such interactions can be transformative to participants? research programs. ? To bridge the gap between basic science and clinical applications by highlighting new discoveries and innovations that are translational while preserving the rigor of basic science and exploiting its potential for creating paradigm shifts. ? To provide the opportunity for lively networking interactions that foster the development of junior investigators as they develop lifelong relationships with each other and with more senior investigators. The GRC provides an unparalleled environment for the development and emergence of the next generation of leaders in our field. ? To highlight neglected areas in our understanding of cardiac arrhythmia mechanisms and to identify and prioritize novel research directions. These goals are closely aligned with the mission of NHLBI. They will be achieved during a five-day intensive meeting in which investigators of different professional levels, disciplines and occupations will present their work, engage in group and individual discussions, and establish relationships and collaborations that will transform current practices and advance our understanding of cardiac arrhythmia mechanisms.
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{
"pile_set_name": "NIH ExPorter"
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Our objective is to examine the pattern(s) of tissue injury resulting from the activation of the efferent limb of the intestinal (or systemic) immune response by antigen challenge administered via the lumen of the gut. Based on the results of experiments performed during the previous two years of grant support, it is planned to broaden these objectives to include the characterization of the fragments of administered antigen that appear in the systemic circulation after feeding and to determine some of their biologic functions. Further, it is planned to include under the concept of "injury", the broadening of anaphylactic sensitivity that may occur in animals as the result of enhanced uptake of bystander antigens from the intestine during the altered intestinal permeability which accompanies local intestinal anaphylaxis. Specifically, it is sought to: I. characterize the fragments of certain dietary antigens detected n the systemic circulation of adult rats and to test their role in the induction of systemic tolerance to ingested antigens. II. Determine whether the enhanced uptake of bystander antigens present in the intestine during local anaphylaxis contributes to the broadening of anaphylactic sensitivity to ingested antigens. III. Determine the site(s) and the consequence(s) of interaction of dietary antigen with IgG antibodies in rats prepared with or without IgE antibodies to the corresponding antigen. IV. Determine the histopathology of the local and remote tissue injury resulting from the ingestion of antigen in immunized rats.
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{
"pile_set_name": "NIH ExPorter"
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Increasing evidence suggests a critical role for epigenetic mechanisms in development, stem cell physiology and cancer. Polycomb Repressive Complex 2-mediated trimethylation of histone 3 on lysine residue 27 (H3K27me3) is catalyzed by the enzyme Enhancer of Zeste 2 (EZH2). Overexpression of EZH2 is correlated with clinical cancer progression in patients and forced expression of EZH2 can contribute to transformation of cultured cells. Inhibition of EZH2 by shRNA strategies has been correlated with decreased proliferative potential in several cancer models. Interestingly, haploinsufficiency for EZH2 has recently been described in human B-cell lymphomas. The functional consequences of EZH2 haploinsufficiency on cellular phenotype have not yet been characterized in experimental cancer models. We propose to study the role of Ezh2 in a genetically defined model of acute leukemia mediated by MLL-AF9 that is well characterized in our laboratory. Genetic loss of Ezh2 in homozygous and/or heterozygous settings will allow us to compare the effects of defined dosages of Ezh2 on tumor phenotype, including complete loss of activity. This technical advance will circumvent limitations associated with shRNA technology and will definitively clarify if Ezh2 is essential for AML. Our system will also allow for detailed characterization of chromatin and gene expression changes associated with loss of Ezh2, thus providing important mechanistic insights. Results obtained in the murine system will be compared with results obtained from the analysis of human cell lines and primary patient samples. The specific aims are: 1: Loss of Function and Forced Expression Studies to Define the Role of Ezh2 in Leukemic Self-Renewal 2: Molecular Characterization of Pathways Affected by Loss of Ezh2 in Murine and Human MLL-AF9 Leukemia Cells Dr. Tobias Neff, a pediatric hematology/oncology fellow at Children's Hospital Boston (CHB) has outlined a 5-year career plan that will build upon his background in clinical hematology/oncology and experimental hematology. Under the co-mentorship of Drs. Scott Armstrong and Stuart Orkin, recognized leaders in leukemia, hematopoietic stem cell biology, and transcriptional gene regulation, he seeks to utilize a definitive biologically relevant mouse model and genomic approaches to study the role of Enhancer of Zeste 2 (Ezh2) in acute myeloid leukemia. Dr. Neff will be mentored by an Advisory Committee of internationally recognized experts in the field. Finally, the plan is ideally carried out in the Division of Hematology/Oncology at Children's Hospital Boston, given its distinguished record for training physician-scientists in a rich and collaborative environment.
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{
"pile_set_name": "NIH ExPorter"
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The 5-year survival from lung cancer has kept pace with the improvement in 5-year survival from all cancers over the last 40 years. Yet, it remains disappointingly low at -15%. This improvement in 5-year survival is a result of heightened awareness, better technology for detection, better selection of patients for various therapeutic options, and the selective use of palliative interventions. However, lung cancer mortality remains extraordinarily high, and it is the benchmark by which future generations will judge our success in effectively combating this disease. The Moffitt Cancer Center SPORE in Lung Cancer will 1) elucidate mechanisms of action of crucial molecules in lung carcinogenesis and tumor progression and investigate their impact on therapeutic efficacy and 2) prospectively assess the clinical utility of the molecules for therapeutic and prevention interventions. Our ultimate goal is to change the standard of care for people at risk for and with lung cancer. The SPORE has five translational research projects, three cores, a developmental research program, and a career development program. The projects are: 1) E2Fs Impact on Therapeutic Efficacy (Cress, Chen &Bepler) 2) RRM1 in the Management of Lung Cancer (Bepler, Cress &Simon) 3) Antitumor Mechanisms of Src Inhibitors in Lung Cancer (Haura, Cheng &Pledger) 4) Lung Cancer Chemoprevention with Enzastaurin (Bepler &Alexandrow) 5) p53-Based Vaccine for Small Cell Lung Cancer (Gabrilovich &Antonia) They are support by: A) Tissue Procurement, Pathology, and Bioinformatics Core (Altiok &Fenstermacher) B) Clinical Trials &Biostatistics Core (Simon, Schell &Bepler) C) Administration &Patient Advocacy Core (Bepler, Cress &Quinn). The SPORE also has Developmental Research &Career Development Programs. This SPORE will be firmly integrated into the H. Lee Moffitt Cancer Center and Research Institute, a free-standing, NCI designated Comprehensive Cancer in Tampa, Florida.
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{
"pile_set_name": "NIH ExPorter"
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We have developed a method to identify non-histone proteins involved in organizing the nucleoprotein fiber of metaphase chromosomes isolated from HeLa cells. The approach is to remove histones from the metaphase chromosomes by competition with polyanions such as dextransulfate and heparin. The histone-depleted chromosomes can be isolated by sucrose gradient sedimentation. They contain a highly-folded DNA fiber, organized by repetitive attachment to a central matrix (chromosomal scaffold) consisting of non-histone proteins. The chromosomal scaffold can be isolated as a structurally independent entity following removal of most of the DNA and has the shape of a metaphase chromosome. The main focus of this proposal is to study the histone-depleted chromosome structurally and biochemically. We will identify the proteins involved, study the DNA-protein interactions and deduce the overall folding of the DNA fiber by electron microscopy.
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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. The capability for accurate mass measurements is an important attribute of Fourier transform mass spectrometry (FTMS). Unlike other instrumental methods in mass spectrometry, FTMS still offers significant opportunities to improve mass measurement accuracy (MMA), making it an area of research. Although FTMS can measure ion frequencies to nine significant figures, the conversion of frequency to m/z is difficult because frequency depends on the electric field and the space charge in the ion trap. We are working on methods to shim the electric field (electrical compensation) and to correct for space charge effects. We are also interested in the relation of mass calibration and the fundamentals of ion motion and space-charge effects, and we wish to to improve the basic calibration procedure particularly those that correct for effects of space charge. The advent of matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) have opened the door for FTMS to be used for analyzing at high performance biopolymers, including proteins, oligodeoxynucleotides (ODNs), oligosaccharides, and synthetic polymers. We will pursue accurate mass measurement in these areas and some practical ways to improve MMA.
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{
"pile_set_name": "NIH ExPorter"
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Even in the US, infectious diseases are the third most common cause of death, despite the widespread availability of antibiotics, vaccines, and advanced hygienic standards. The identification of new pathogens, the re-emergence of old pathogens, and the increasing incidence of antibiotic resistance reflect the globalization of humanity and microbes. As illustrated by the recent anthrax, SARS, and monkeypox outbreaks, new as well as old pathogens can be rapidly moved great distances and establish footholds in new niches. Strategies for the prevention, treatment, and control of infectious diseases will require fundamental bench research that takes advantages of rapid advances in genomics, proteomics, cell biology, and immunology to develop new diagnostics, therapeutics, and vaccines. The study of microbial pathogenesis is a rapidly advancing area of research, with unprecedented opportunity to influence health worldwide. Over the past 5 years, UCSF has created a strong training environment and established an outstanding program in Infectious Disease and Host Defense. There are 26 faculty already associated with the program and several new hires planned; space and resources to recruit 2-3 more faculty have been allocated; a course, journal club and research talks are now offered, and a yearly symposium established. This new T32 grant application describes the new program at UCSF in Microbial Pathogenesis and Host Defense that will train the next generation of scientists who can combat disease through advances in basic and translational research. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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DESCRIPTION: Ebola viruses (EBOV) and Marburg virus (MARV) are two genera of enveloped viruses that constitute the family of Filoviridae. Outbreaks of fulminant hemorrhagic fever in human and non-human primates in Central Africa caused by EBOV have reached mortality rates of up to 90% in the past and the mortality rate in the recent West Africa outbreak is 60%. No specific antiviral treatment or vaccine is approved for these deadly pathogens. There is thus an urgent need to develop effective antiviral therapies against infection by filoviruses. Targeting cell entry of enveloped viruses as an antiviral strategy has been proven effective against a wide range of viral diseases. However, the intracellular sequestration of filovirus fusion machinery makes it challenging to develop antivirals that block EBOV glycoprotein (GP)-mediated viral entry. We aim to overcome this challenge by adding a cell penetrating peptide sequence and conjugating a lipid moiety to fusion inhibitory peptides (C-peptides). We have shown that sustained plasma levels of our lipid-conjugated EBOV C-peptide are achieved after parenteral administration in mice. This C-peptide inhibits in vitro infection by EBOV with an IC50 of 0.2 ?M and efficiently protects mice from lethal EBOV infection. We propose to leverage these preliminary results to design, synthesize and evaluate novel EBOV C-peptide analogs that can be delivered intranasally or subcutaneously, and have enhanced efficacy against a broad range of filoviruses. Analogs will be evaluated for antiviral activity in infectivity assays, for cytotoxcity on human cells, and for GP2 subdomain-binding interactions. Promising analogs will be evaluated for in vitro antiviral activity against diverse filoviruses and for toxicity in mice. We ill also identify the molecular determinants of antiviral resistance. Selected analogs will be tested in challenge experiments in a mouse model. We anticipate that the knowledge gained from our proposed studies will significantly enhance our ability to address the threat of natural and intentional epidemics by developing potent antiviral drugs with feasible delivery routes for containing acute filovirus outbreaks. We propose the following specific aims: Aim 1. To use structure-guided mutagenesis and protein engineering to optimize the antiviral potency and bioavailability of EBOV C-peptide inhibitors. a) Design and synthesis of new C-peptide analogs; b) Characterization of antiviral activity against a range of filoviruses in cell culture; c) Characterization of determinants of viral resistance by in vitro virus evolution experiments. Aim 2. To evaluate the protection afforded by novel EBOV C-peptide inhibitors delivered intranasally or parenterally against lethal EBOV infection in mice. a) Analysis of in vivo biodistribution of improved EBOV C-peptide analogs; b) Evaluation of toxicity in mice; c) Assessment of in vivo potency and breadth of activity of fusion inhibitors in the mouse model of EBOV infection.
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{
"pile_set_name": "NIH ExPorter"
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The effect of immunostimulation will be tested as an adjunct to surgical resection in thoracic surgical patients with no evident disease after surgical removal of their tumors. As an extension of our prior studies with intrapleural BCG immunostimulation, patients with resectable cancers of the lung and esophagus will be treated with intrapleural BCG as regional nonspecific immunotherapy, followed by two vaccinations of autologous tumor cells. These cells will be reproductively treminated by x-irradiation, but will be viable, and will be administered in combination with BCG in an optimal ratio to elicit specific immunization. By this means we seek to protect from disseminated micrometastases. Patients will be randomly assigned to receive immune therapy after resection, or to enter the control group treated by surgery alone. They will be followed by clinical criteria and skin tests at regular intervals to determine time to recurrence or death. This clinical program will be in collaboration with the NCI Frederick Cancer Research Center. Mutual exchange of visits by the scientific staff of both institutions are planned to facilitate the continuing development and application of a relevant animal model of human cancer, and to catalyze the rapid translation of laboratory advances to the clinical setting.
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{
"pile_set_name": "NIH ExPorter"
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Animal replication-dependent histone mRNAs are the only eukaryotic mRNAs that lack a polyA tail ending instead in a conserved stemloop. In contrast mRNAs for histone variants, e.g. H3.3 and H2.v, are encoded by polyadenylated mRNAs. The genes for all five histone proteins are clustered in metazoan genomes, and factors required for histone gene expression are localized near the histone genes. We will determine the requirements for the coordinate expression of the replication-dependent histone mRNAs in vivo using Drosophila as a model system, and in particular the role of the Histone Locus Body in histone mRNA metabolism. We will use both biochemical and genetic approaches to determine 1. The structural requirements in the HLB components FLASH and NPAT required for efficient production of properly processed histone mRNA, 2. The sequences in the histone gene locus that specifies the formation of the HLB at the histone locus and 3. The composition of the histone cleavage factor, which contains polyadenylation factors Symplekin, CPSF73 and CPSF100, will be determined and the role of Symplekin in histone pre-mRNA processing in vitro and in vivo elucidated. PUBLIC HEALTH RELEVANCE: We will combine biochemical and genetic studies in the fruit fly to study the regulation of the synthesis of histone proteins, which are complexed with the DNA chromosome. Each time DNA is replicated, histones are synthesized in large amounts to package the new DNA into chromosomes. Factors controlling histone synthesis are critical for stable inheritance of genetic information and proper development of the organism; these factors are conserved between humans and flies and our genetic studies in flies will allow us to understand their function in humans.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary/Abstract Over the last two decades, many giant DNA viruses have been discovered, some of which are bigger than a small cell. How these giant viruses assemble their virion shell from thousands of simple protein building blocks so precisely is a mystery. However, the sheer size of these viruses poses a significant challenge to currently available techniques. This project will tackle this challenge by using a marine giant virus Cafeteria roenbergensis virus (CroV) as a model to decipher the assembly mechanism of giant viruses, and as an opportunity to develop technology to push the resolution limit of these gigantic structures to the atomic level. During the last five years, cryo-electron microscopy (cryo-EM) has become an increasingly powerful tool to study the structures of biological molecules at atomic resolution, earning its developers the 2017 Nobel Prize in Chemistry. We will collect higher quality images using state-of-the-art cryo-EM equipped with latest new hardware, such as energy filters, direct electron detectors and phase plates. Using these images together with new software algorithms, we will determine the structure of giant CroV to high resolution by image analyses and reconstruction. Structures of individual CroV proteins will also be solved to atomic resolution by cryo-EM using various methods and docked into the cryo-EM reconstructed maps. The resultant pseudo-atomic structure will allow characterization of the ultrastructural features and architecture of CroV, building the essential foundation to unravel the assembly of giant viruses. The structure information will be combined with classic biophysical, molecular dynamic simulation, mathematical modeling, and computational analyses to evaluate the novel assembly model of giant viruses. In the new assembly model, the protein shells of giant viruses are assembled continuously from the 5-fold vertices in an interesting spiral way instead of assembled from patches in a step-wise fashion previously assumed. Giant virus protein shell is assembled from protein building block similar to other viruses, including many human pathogens. Some giant viruses have been associated with human diseases such as pneumonia and cognitive functional change. Understanding these principles governing the assembly of giant viruses will improve the development of therapeutic agents to inhibit virus assembly, thus providing a new avenue for preventing and treating viral diseases in general. Elucidation of the molecular interactions that drive assembly of these giant viruses will also shed light on how to control protein-protein interactions effectively, facilitating the rational design of virus-like nanoparticles with a wide size range for biomedical and other nano-applications. Since some giant viruses are bigger than a small cell, techniques and methods developed in this project will push the limits of structural biology and provide new and useful tools to study even larger supramolecular assemblies and eventually the whole cell in the future.
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{
"pile_set_name": "NIH ExPorter"
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This application proposes a supplement to the NBER's ongoing program project on the Economics of Aging. In particular, it proposes a continuation of the subproject on "Improving Data Measurement in Surveys of the Elderly" for the final two years of the current PO1 award period. Accurate responses to surveys of elderly populations are essential for studying the health and economic circumstances of older persons and for tracking the impact of health and economic policy on economic status and behavior. To date, this project has developed and implemented experimental Internet survey techniques and has applied these techniques in evaluating and improving survey data. Project investigators have also studied in depth the limitations and errors and biases that may exist in survey data and how they relate to questionnaire format, sequencing and context. The proposed supplement project would extend this work, focusing on data about expectations and perceptions of future health, mortality and financial risks. The continuation of the subproject will involve the conduct and analysis of a new set of experimental surveys, which we have labeled the Retirement Perspectives Surveys (RPS). These surveys will include both mail-out and Internet components, using a sample of AARP members. The new surveys will explore in detail people's subjective perceptions of their future health, mortality, and financial risks, so that these perceptions can be related to how people behave--the decisions that they make--as they age. The RPS surveys will also draw on the special capabilities of the project's Internet Virtual Laboratory (IVLab) to vary format and sequencing and content of questions so that any biases resulting from these design features can be better understood. In addition, the Internet version of the RPS will be complemented with traditional mail surveys so that self-selection into the Internet can be evaluated, and so that the response variation across survey type can be explored. An auxiliary goal of the continuing project is to further develop the IVLab software for Internet experiments and to provide an easy-to-use implementation of experimental Internet surveys to the research community.
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{
"pile_set_name": "NIH ExPorter"
}
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Hepatitis C virus (HCV) often causes a prolonged and persistent infection, and an association between hepatocellular carcinoma (HCC) and HCV infection has been noted. Understanding the molecular basis of viral pathogenesis is a major challenge in the effort to gain insight into HCV associated disease progression. The pathogenesis of liver damage is likely to be related to both viral and immune mediated factors. HCV core protein displays numerous physicochemical and biological activities. The core protein exhibits in vitro RNA and ribosome binding activities, nucleocytoplasmic localization, a regulatory role on cellular and unrelated viral promoters, and interacts with heterogeneous nuclear ribonucleoprotein K and the cytoplasmic tail of the lymphotoxin-beta receptor. Furthermore, the core protein inhibits programmed cell death (apoptosis) under certain conditions, has a transforming potential on primary rodent cells, and induces hepatocellular carcinoma in transgenic mice. These interesting properties suggest the hypothesis that the core protein may play a critical role in cell growth deregulation, and thus contribute to virus mediated pathogenesis during persistent infection. This pilot research proposal is designed to (1) determine the role of HCV core gene expression in normal human hepatocyte growth and (2) map the core protein domain responsible for human hepatocyte growth regulation. The results from this pilot study will be extremely informative in the characterization of the functional role of HCV core protein in its natural host. The capacity to transform primary human hepatocytes will provide a model to examine in detail the core gene and associated cofactors as they relate to multistage hepatocarcinogenicevents in vitro. The long term goal of this study is to understand the molecular mechanism of HCV core protein mediated pathogenesis and to design effective strategies for intervention of HCV related disease in humans.
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{
"pile_set_name": "NIH ExPorter"
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this proposal will complete preclinical toxicity testing and safety assessment of i.t. Amitriptyline. We will test its effect on acute nociceptive pain alone, its interaction with iv alfentanil in acute nociceptive pain, and its efficacy alone against intradermal capsaicin-induced hyperalgesia and allodynia in humans. These effects will be correlated with CSF neurotransmitter and amitrip-tyline concentrations to provide presumptive evidence for mechanisms in humans.
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{
"pile_set_name": "NIH ExPorter"
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The objective of this program is to prepare post-doctoral trainees to establish independent research careers in the areas of gonadal function, early pregnancy and embryonic development through implantation. The training program will consist of three elements;independent laboratory research, group seminars and other functions held jointly with the Center for Reproductive Sciences, and formal course work in research ethics and animal welfare, as well as subjects that will enhance the trainee's research expertise. The independent research will occur in a laboratory selected by the trainee from a group of 12 scientists who are leaders in various aspects of reproductive and developmental biology. The members of this training group have well established NIH-funded programs and have trained more than 86 postdoctoral fellows. Areas of study include fertilization, pre-implantation development, trophoblast-uterine interactions, the immunobiology of implantation and early pregnancy, as well as regulation of gonadal function, gene expression during testis and embryonic development. The strength of this group is enhanced by collaboration among the members as evidenced by the record of joint publications, grants, and students. This program will take advantage of an existing Center for Reproductive Sciences comprising 21 principal investigators and 78 additional staff and trainees to provide an enhanced training environment. The Center for Reproductive Sciences sponsors an active seminar program and hosts a regional conference in reproductive biology, which provides additional opportunities for trainees to present their work. The recruitment, selection and guidance of trainees will be coordinated through an Internal Advisory Committee, with input from all participating faculty and in consultation with an External Advisor, who will meet with the group once each year. Applicants who have been awarded a Ph.D. degree in one of the biological sciences, or a D.V.M. or M.D. degree and who demonstrate a commitment to research in reproductive biology will be considered for entry into the program. An emphasis will be placed on recruiting under represented minorities into the program. The records of the six trainees selected during the first funding period are impressive, with two of the four trainees who have completed their training having assumed faculty positions, one tenure track and one research track. The overall success of the program will ultimately be measured by the contributions of these trainees after they have established independent research programs.
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{
"pile_set_name": "NIH ExPorter"
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PROJECT SUMMARY Experimental models and human studies have repeatedly linked persistent organic pollutants (POPs) such as perfluorinated alkylate substances (PFASs) and polychlorinated biphenyls (PCBs) to abnormal fat distribution, body mass index, and glucose and lipid metabolic abnormalities during early life. These are pervasive chemicals to which nearly all children are exposed during early life: PFASs are used in several consumer and industrial applications and PCBs accumulate in food chains. Evidence suggests that these abnormal cardiometabolic trajectories during early childhood may predict insulin resistance, metabolic syndrome, obesity, and even cardiovascular events in adulthood. However, no biomarkers are available to identify in the immediate postnatal period children who are at risk for abnormal cardiometabolic development, thus curbing opportunities for effective, targeted prevention. To address this gap, our long-term goal is to identify novel mechanistic biomarkers in breast milk that reflect environmental influences and predict the risk of abnormal cardiometabolic programming and childhood obesity. We will leverage groundbreaking evidence on the roles of breast milk extracellular vesicles (BMEVs) in metabolic programming. BMEVs are small vesicles that are released by luminal epithelial cells in breast milk. BMEVs have been proposed as conveyors of molecular signals from the mother to the child. In particular, BMEVs transport a cargo of microRNAs (miRNAs) that, once ingested by the child, integrate themselves in recipient cells in the child's body and can remotely affect the expression and translation of child's messenger RNAs. This process has been shown to be key to the child's metabolic programming in early life. To date, no studies have been conducted to identify the potential roles of BMEVs as part of the paths linking PFASs and PCBs exposure to its adverse effects on cardiometabolic trajectories and obesity during childhood. To achieve this goal, we will leverage the unique resources of the longitudinal Faroe Islands birth cohort, a prenatal cohort with biobanked breast milk (N=300, all from nursing mothers), extensive exposure data, and repeated postnatal cardiometabolic measures over 13 years of follow up. We hypothesize that BMEV number, BMEV size, and BMEV-encapsulated miRNAs are modified in response to prenatal (third-trimester of pregnancy) exposure to PFASs and PCBs (Aim 1) and that BMEV number, BMEV size, and BMEV- encapsulated miRNA predict cardiometabolic outcomes over 13 years of follow up (Aim 2). We will use advanced statistical modeling to integrate BMEV biomarkers in the paths linking exposure and abnormal cardiometabolic trajectories (Exploratory Aim 3). This study is a high-risk/high-reward and cost-effective project that will provide new noninvasive tools to identify and reduce the burden of abnormal cardiometabolic programming during childhood.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of the proposed project is to test a theoretical model of HIV/AIDS disclosure that simultaneously examines multiple factors hypothesized to predict and inhibit disclosure and tests the relationships between disclosure events and psychological and behavioral outcomes. To date, there exists no known model that simulates the hypothesized complexity of disclosure processes or relates disclosure events to post-disclosure psychological (e.g., negative affect) or behavioral (e.g., sexual risk) outcomes for people living with HIV/AIDS (PLWHA). The proposed model of HIV/AIDS disclosure will provide a parsimonious theoretical framework through which to understand how PLWHA make decisions about whom to disclose their positive serostatus and how they are affected by these decisions. The proposed study will employ a disclosure recipient-specific, longitudinal design in order to assess how these disclosure processes vary across disclosure recipients and time. Specific Aim 1 will examine the relative predictive utility of factors hypothesized to facilitate (i.e., negative coping, negative psychological effects of concealment, significance of relationship with disclosure recipient, and eco-concerns for well-being of others) and inhibit (i.e., anticipated negative reactions from disclosure recipient, ego-concerns for well-being of self, social norms against disclosure, and perceived positive serostatus of disclosure recipient) disclosure that are conceptually related with either the discloser or the disclosure recipient. Specific Aim 2 will examine how disclosure (or nondisclosure) decisions affect psychological and behavioral outcomes. It is hypothesized that the effect of disclosure on behavioral outcomes will be mediated by the effect of disclosure on psychological outcomes such that, to the extent that people experience positive psychological outcomes due to disclosure, they will also experience concomitant decreases in sexual risk and increases in adherence behaviors. This project has important conceptual and applied implications. The proposed model of HIV/AIDS disclosure will provide a framework for theorists and researchers to understand the complexity of disclosure processes and allow them to use this knowledge to inform interventions that assist PLWHA in identifying the optimal conditions and recipients for disclosure. Additionally, this framework has the potential to enhance the understanding of general disclosure processes across other types of concealed stigmatized identities (e.g., mental illness) and to emphasize both the potential psychological and behavioral effects of disclosure.
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{
"pile_set_name": "NIH ExPorter"
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During oogenesis and maturation of the sea urchin egg, large stores of mRNA are accumulated for subsequent use during early embryogenesis. Utilization of stored maternal mRNA occurs at a very low rate prior to fertilization. After fertilization, dramatic changes in mRNA availability and recruitment occur, allowing rapid increases in the synthetic rate of many proteins during early cleavage. Changes have been documented in both overall rates of maternal mRNA utilization and in the selection of particular mRNA species at different developmental stages. Our previous studies have allowed the development of cell-free translation systems from eggs and embryos that reproduce the overall differences observed in the intact system. This investigation proposes to utilize these cell-free systems, in conjunction with other approaches, to identify the cellular components involved in the utilization of maternal mRNA. Using the cell-free systems an inhibitor of eIF-4F activity has been discovered in the unfertilized egg, that decreases gradually in activity over the first 2-3 hr after fertilization. The identity and nature of the eIF-4F inhibitory activity will be characterized. The mechanism by which this activity inhibits the initiation sequence will also be investigated. The manner in which the inhibitor is inactivated after fertilization will be explored. To complement these studies, a systematic search for developmentally regulated mRNA-interactive proteins will be undertaken, using a protein blotting and in vitro binding technique, with capped 32p-labelled mRNA's synthesized in vitro.
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{
"pile_set_name": "NIH ExPorter"
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The trigeminal mesencephalic nucleus, main sensory nucleus, and rostral part of the spinal sensory nucleus of cats will be stereotaxically explored with extracellular microelectrodes to determine the location and firing patterns of single neurons which respond to passive movement of the condylar head of the mandible within the temporomandibular joint. Latency measurements will be made to determine the location of the cell bodies of first-order neurons by elctronically stimulating the TMJ mechanoreceptors. The response patterns of these neurons will be analyzed with a digital computer. The analysis will be compared statistically with similar analyses derived from single neurons in the ventral posteromedial nucleus of the thalamus. This nucleus is thought to contain third-order projection neurons for kinesthesia of the temporomandibular joint. The results will be used to illuminate the mechanisms whereby mechanoreceptors in the tempormandibular joint single relative position of the jaw during masticatory movements.
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{
"pile_set_name": "NIH ExPorter"
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The primary symptoms of hypoparathyroidism include muscle cramping, convulsions, intellectual disabilities, cataracts and abnormal heart rhythm. Symptoms are due to low serum calcium (hypocalcemia). Replacement of parathyroid hormone (PTH) has been explored to remedy the calcium deficiency, but maintaining an optimal calcium level has proven problematic because hypercalcemia can occur as a result of excess PTH. Multiple efforts are under way targeting either full-length PTH (PTH 1-84) or the active amino-terminal domain (PTH 1-34), but these molecules have undesirable pharmacokinetic properties for chronic daily management of calcium levels in patients with hypoparathyroidism. Eli Lilly scientists have identified a PTH receptor modulator (PTH-RM) that can normalize serum calcium. At fairly low doses, the PTH-RM was shown to normalize calcium levels in parathyroidectomized rats. The investigators are collaborating with TRND to develop this PTH-RM toward a Phase II proof-of-concept study for hypoparathyroidism by leveraging the existing data package. TRND scientists, in collaboration with researchers from Eli Lilly & Company and the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), have further developed the animal model of hypoparathyroidism to generate robust efficacy data. The team will execute the full preclinical development plan. TRND will support the preparation and filing of the Investigational New Drug application with the Food and Drug Administration, with NICHD providing support for subsequent clinical trials in patients.
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{
"pile_set_name": "NIH ExPorter"
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The long range goal of this work is to study the molecular basis of membrane-cytoskeleton associations and their role in the directed intracellular movements of membranes and organelles. Specifically, it is planned to investigate the idea that actin filament-membrane attachments are instrumental in the translocation of secretory vesicles to the plasma membrane during exocytosis. The approach taken here will be to demonstrate specific and regulated associations of actin with purified membrane components. The catecholamine-containing secretory vesicles (chromaffin granules) are an experimentally accessible system because they can be easily prepared in large quantities and high purity. Also, actin filaments can bind to a protein site on purified chromaffin granule membranes and these interactions are regulated by free calcium ion concentrations thought to mediate exocytosis in vivo (Fowler, V.M. and H.B. Pollared. 1982. Nature 205:336-339). However, using available methods of measuring actin binding to membranes, it was not possible to distinguish a small number of high affinity sites on the membranes from a large number of low affinity sites, nor to clearly distinguish effects of regulatory factors on actin binding from effects on actin polymerization or on filament self-association. Therefore, a primary goal of this proposal will be to quantitatively characterize actin binding to membranes, using define, radiolabelled actin species (e.g., monomer, convalently cross-linked dimer) at subcritical concentrations so that actin polymerization will not interfere with analysis of binding data. Subsequently, it is planned to identify and purify the actin-binding protein(s) from the chromaffin granule membrane, to characterize its interaction with monomeric and filamentous actin, and to determine its relationship to known actin-binding proteins, as well as its subcellular location in the cell. Regulatory features of actin-membrane association (e.g., [Ca++] free, phosphorylation) will be investigated at three levels: actin binding to membranes, interactions of the purified protein with actin, and during exocytosis from intact cells. It is also planned to determine ultrastructurally how actin filaments are attached to the chromaffin granule membrane in vitro, and in situ in the cell, and to determine whether immunologically cross-reactive proteins might also be present on membranes of other subcellular organelles.
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{
"pile_set_name": "NIH ExPorter"
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Talk given for the second MSA (Microscopy Society of America) Workshop on Multiphoton Excitation Microscopy (Atlanta, 12 July 1998). The topics described were aspects associated with multiphoton imaging down in the 700-800nm region: UV excitable probes, tissue autofluorescence and cell viability under the imaging conditions necessary to excite these types of fluorescence.
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{
"pile_set_name": "NIH ExPorter"
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(Adapted from the Applicant?s Abstract) The purpose of the core is to provide the resources and expertise for the efficient generation and delivery into embryos of recombinant expression viruses (adenovirus and retroviruses) containing genes of interest to the project investigators. (End of Abstract) This core provides the critical adeno-viral and retro-viral support for the projects in this proposal. Most of the projects propose to either use lineage tracing or expression of certain genes utilizing either adeno-viral or retro-viral transfection into mouse or chick embryo cultures. This core is central to the experiments outlined in the various projects. The principal investigator and co-investigators have experience in the adeno-viral and retro-viral techniques proposed. Their approach is straight-forward and appropriate. They will maintain cell lines necessary for these experiments and generate the high titer viral stocks necessary for the experiments. They propose to make the core laboratory available to the investigators for them to learn as well as provide this service. The plans to provide direct infection of whole embryos in culture (mouse and chicken) are reasonable, and the investigators have experience with such techniques. There are overall no glaring difficulties with the proposed approach. The types of experiments proposed are now rather routine and will be useful for the various experiments outlined in the individual projects. The ability to infect specific tissues in whole embryo cultures is rather innovative, although it has been used successfully by other investigators. Nevertheless, it is not a common skill and it is certainly a strength that this core will have that ability. The prinicipal investigator and co-investigators have good experience, as they have demonstrated, utilizing retro-viral and adno-viral techniques. They have done these sorts of experiments for other purposes successfully, and should have no difficulty in performing the tasks outlined. Dr. Kubalak has had experience with mouse embryo culture and this will certainly be an asset to the program project grant. The environment for the core is strong and they appear to already have all the necessary equipment and technology to provide the core services outlined. There are no problems with the environment. This is a very important core for this program project and appears to be designed appropriately without any major flaws. The core has many strengths and will be a critical one for most of the projects. The availability of Dr. Mikawa as a consultant adds furthur strength to the ability of this core to troubleshoot and be effective.
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{
"pile_set_name": "NIH ExPorter"
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Haemophilus influenzae is an important cause of human disease, producing both localized respiratory tract and systemic (bacteremic) infections. The initial step in the pathogenesis of disease due to this organism involves colonization of the upper respiratory tract. Despite the essential role of colonization, the determinants of this process remain poorly defined. We have developed an in vitro model for studying interactions between H. influenzae and human epithelial cells. In this model H. influenzae demonstrates efficient attachment and appreciable cell entry, properties that are likely to be important in colonization. The objective of this proposal is to define the bacterial and host cell factors involved in H. influenzae attachment and cell entry. Initially the genetic elements required for in vitro attachment and cell entry will be isolated. An attachment-deficient mutant and a recombinant strain endowed with the capacity for cell entry, both of which have already been isolated, will facilitate these efforts. Subsequently the attachment and cell entry proteins will be purified and their expression will be characterized. Following purification of these proteins, their affinity for epithelial cell surface molecules will be examined and these putative receptors will be isolated and characterized. Finally, to confirm a role for the attachment and cell entry proteins in natural colonization, H. influenzae mutants deficient in attachment and cell entry will be constructed and examined in an animal colonization model. From a practical perspective, results from these studies may form the basis for a novel approach to the universal prevention of H. influenzae disease. Perhaps more importantly, they may provide general insights into the nature of the host-microbial relationship.
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{
"pile_set_name": "NIH ExPorter"
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The objectives of these studies are to define which subcellular constituents of animal and human granulocytes possess biologically active substances affecting blood coagulation, to determine if subcellular fractions of macrophages have these same biological activities, and to examine the influence of endotoxin and certain drugs on the procoagulant activity of leukocytes. Blood leukocytes will be isolated by dextran sedimentation and differential centrifugation on ficoll-hypaque gradients. Lysosomes will be obtained from disrupted cells by differential centrifugation and separated into distinct populations on linear sucrose density gradients; these will be monitored by light and electronmicroscopy. Soluble extracts of lysosomes will be separated by column chromatography, electrofocusing and gel electrophoresis into purified fractions for characterization and biological testing.
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{
"pile_set_name": "NIH ExPorter"
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The goal of this research is to devise automated methods of analyzing the Auditory Brainstem Response (ABR) when it is recorded using three orthogonal channels. The resulting 3-Channel Lissajous Trajectory (3-CLT) shows both linear and planar portions, which will be detected using algorithms tested under realistic conditions of background noise. The 3-CLT methodology is likely to be more sensitive to pathological changes in the auditory nervous system than is the ABR, though the ABR has had extensive use in audiology and neurology as a measure of brainstem normality or abnormality. The automated methods will be designed for microcomputers which are likely to be the basis for evoked response recording systems in the future. At least six companies now produce such systems, primarily for clinical use, but the ABR systems cannot record or analyze the 3-CLT at present, and reliable algorithms are not yet known for the otherwise complex and time-consuming analysis that the 3-CLT requires.
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{
"pile_set_name": "NIH ExPorter"
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Iron is essential for cell growth and division, but it is also harmful when it accumulates in cells. Therefore, the intracellular level of iron must be precisely regulated. Iron-overload diseases are primarily caused by hereditary hemochromatosis (HH)-the most common genetic disorder in Europe and the United States. The well studied pathways for regulated iron uptake and storage are mediated by the transferrin/transferrin receptor complex and by the storage protein, ferritin, respectively. However, it appears that novel proteins can regulate iron metabolism by interaction with the transferrin/transferrin receptor complex and with other proteins involved in transferrin-independent iron-uptake pathways. Recently, the gene responsible for HH was cloned and characterized as a novel major histocompatibility complex (MHC) class I-like gene (HFE) that is distinguishable from the wild type by a single mutation that prevents its transport to the cell surface. The HFE protein is known to regulate iron uptake possibly by interaction with the transferrin receptor. HFE transcripts are not detected in resting lymphocytes, but are present at high levels in human tumor cells, especially those of lymphoid origin, implying that HFE may play a role in tumorigenesis. In order to define the role of HFE in iron-homeostasis and to determine if interferences with its normal expression play a role in diseases such as cancer, we propose (a) to characterize cellular proteins that interact with HFE, (b) to investigate the intracellular trafficking of HFE and its complexes, and (c) to determine how extracellular factors (e.g. iron, cytokines, viral infections) as well as intracellular factors (e.g. chaperones, heat shock proteins, viral proteins) regulate the trafficking and/or recycling of HFE complexes. Ultimately, by determining just how the HFE molecule functions to aid the growth of cancer cells, we hopefully will be able to develop drugs that interfere with the process and result in cancer cell death. Due to its specific binding to transferrin receptor, our recent studies suggest that it may be possible to exploit soluble HFE as a specific vector for the intracellular delivery of drugs.
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{
"pile_set_name": "NIH ExPorter"
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The contractor will perform laboratory assays to provide information on the relationship of virus and tumor associated antigens to the etiology of human cancer. Most of these studies which will be predominantly utilizing cell-mediated immunity assays, will be performed in collaboration with ongoing contracts within the Immunology-Epidemiology Segment of the Virus Cancer Program as directed by the Project Officer.
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{
"pile_set_name": "NIH ExPorter"
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My goal is to determine, using NMR, the structure of a vaccinia virus DNA sequence containing two extrahelical bases (G and C) on the same strand, separated by d(AA).d(TT). It appears at each terminus of the genome, with the extrahelical bases at corresponding locations but as complements. I am testing alternative structural models by restrained molecular dynamics, using the progrm MARDIGRAS, based upon both NOE- derived proton-proton distances and relaxation matrix-calculated NOESY cross peak volumes in both D2O and H2O. I will particularly emphasize assessment of the extent of pairing of the two AT base pairs between the extrahelical bases and the looping out or stacking of the extrahelical bases themselves. The analysis of my experiments is heavily dependent upon the resources of the Computer Graphics Laboratory, both for the analytical software, including molecular dynamics, and for molecular modeling of the calculated structures using MidasPlus. My research is biologically significant in many respects. First, the mismatched sequence is required for resolution in the replication of poxvirus DNA, and the structure is needed for a molecular explanation. Second, the structure forms during the extrusion of the imperfect palindrome of the poxvirus replicative intermediate (cloned into a closed circular plasmid vector) to form the hairpin structure of the mature virus genome. Experimentally, this mismatched structure requires a considerable free energy to extend the cruciform. Finally, terminal hairpin loop structures are characteristic of the DNA of several eucaryotic viruses and organelles, including the iridoviruses, parvoviruses, phycodnaviruses and poxviruses; and the rDNA from Tetrahymena, the mitochondrial DNA of Paramecium and the linear mitochondrial DNA of yeast. Many of these terminal sequences contain extrahelical bases as bulges, loops or mismatches. Examples, in addition to vaccinia virus DNA, are the DNAs from the minute virus of mice, the Autographa californica multinucleocapsid nuclear polyhedrosis virus, the human parvovirus B19, the Leporipoxvirus Shope fibroma virus, and the linear mitochondrial DNA of yeast.
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{
"pile_set_name": "NIH ExPorter"
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Significant progress was made during this reporting period. In collaboration with Dr. Blough, we continued to evaluate a large number of newly synthesized compounds using in vitro assays. The current focus is on DA/5-HT releasers based on the phenmetrazine structure, since these compounds have minimal activity at the 5-HT2B receptor. A key aspect of this project is to avoid making compounds that activate 5-HT2B receptors, since this property is associated with the development of cardiac valve disease in humans. We published In collaboration with Dr. Ananthan, who received extramural NIDA funding for further identification of dopamine transporter allosteric modulator ligands, we commenced our part of this collaboration. Five papers related to this project were published in the reporting period, two of which will be highlighted here because of their direct relevance to medication development efforts. Paper #1: In vivo effects of amphetamine analogs reveal evidence for serotonergic inhibition of mesolimbic dopamine transmission in the rat. J Pharmacol Exp Ther. 2011 Apr;337(1):218-25. Evidence suggests that elevations in extracellular serotonin (5-HT) in the brain can diminish stimulant effects of dopamine (DA). To assess this proposal, we evaluated the pharmacology of amphetamine analogs (m-fluoroamphetamine, p-fluoroamphetamine, m-methylamphetamine, p-methylamphetamine), which display similar in vitro potency as DA releasers (EC(50) = 24-52 nM) but differ in potency as 5-HT releasers (EC(50) = 53-1937 nM). In vivo microdialysis was used to assess the effects of drugs on extracellular DA and 5-HT in rat nucleus accumbens, while simultaneously measuring ambulation (i.e., forward locomotion) and stereotypy (i.e., repetitive movements). Rats received two intravenous injections of drug, 1 mg/kg at time 0 followed by 3 mg/kg 60 min later. All analogs produced dose-related increases in dialysate DA and 5-HT. All analogs increased ambulation and stereotypy, but drugs causing greater 5-HT release (e.g., p-methylamphetamine) were associated with significantly less forward locomotion. The magnitude of ambulation was positively correlated with extracellular DA (p <0.001) and less so with the ratio of DA release to 5-HT release (i.e., percentage DA increase divided by percentage 5-HT increase) (p <0.029). Collectively, our findings are consistent with the hypothesis that 5-HT release dampens stimulant effects of amphetamine-type drugs, but further studies are required to address the precise mechanisms underlying this phenomenon. Paper #2: Serotonin (5-HT) precursor loading with 5-hydroxy-l-tryptophan (5-HTP) reduces locomotor activation produced by (+)-amphetamine in the rat. Drug Alcohol Depend. 2011 Apr 1;114(2-3):147-52. Background, Evidence suggests that increases in synaptic serotonin (5-HT) can reduce the stimulant properties of amphetamine-type drugs. Here we tested the hypothesis that administration of the 5-HT precursor 5-hydroxy-l-tryptophan (5-HTP), along with the peripheral decarboxylase inhibitor benserazide, would decrease locomotor effects of (+)-amphetamine. METHODS: Drug treatments were administered to conscious male rats undergoing in vivo microdialysis in nucleus accumbens. During dialysis sampling, rats were housed in chambers equipped with photobeams to detect forward locomotion (i.e., ambulation) and repetitive movements (i.e., stereotypy). Extracellular concentrations of dopamine (DA) and 5-HT were measured by high-pressure liquid chromatography with electrochemical detection.RESULTS:5-HTP (10 &30 mg/kg, i.p.) plus benserazide (30 mg/kg, i.p.) caused dose-related increases in 5-HT but failed to alter other parameters. (+)-Amphetamine (0.3 &1.0 mg/kg, i.p.) produced dose-related increases in DA, ambulation and stereotypy. Combined administration of 5-HTP and (+)-amphetamine evoked large elevations in extracellular DA and 5-HT, but caused significantly less ambulation than (+)-amphetamine alone (50% reduction).CONCLUSIONS:Our results confirm that 5-HTP can decrease hyperactivity produced by (+)-amphetamine, even in the presence of elevations in dialysate DA. The data suggest that 5-HTP and (+)-amphetamine may be useful to broadly enhance monoamine function in the clinical setting, while reducing undesirable effects of (+)-amphetamine.
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{
"pile_set_name": "NIH ExPorter"
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The long-term objective of the proposed research is to elucidate the mechanisms by which neuronal activity shapes synaptic connections during development of the visual system in mammals. The goal of the present application is to examine the role of nitric oxide (NO) and its downstream signaling pathways in the formation of specific connections between retinal ganglion cell axons and cells of the lateral geniculate nucleus (LGN) in ferrets, during the sorting of retinogeniculate projections into on-center and off-center sublaminae. The hypothesis is that synchronously active retinal afferents activate N-methyl-d-aspartate (NMDA) receptors on LGN cells, allow the entry of calcium into the postsynaptic cell and stimulate the production of NO by neuronal nitric oxide synthase (NOS). NO is available simultaneously to both pre and postsynaptic cells, and is particularly suited to coordinate synaptic sites in both presynaptic terminals and postsynaptic dendrites. We propose that NO diffuses to the presynaptic terminal and acts as a retrograde messenger to activate soluble guanylyl cyclase (GC), elevating levels of the cyclic nucleotide cGMP and is associated cGMP-dependent protein kinase (PKG), thus stabilizing active synapses. In the postsynaptic cell, NO, acting through pathways that include cGMP and PKG signaling, and by virtue of the close coupling between nNOS and NMDA receptor-anchoring proteins, regulates transmission at retinogeniculate synapses. Specific aims are to examine: (1) The role in retinogeniculate arbor segregation of specific substrates that link NMDA receptor activation and the generation of NO; (2) The role of cGMP as a downstream effector of NMDA receptors and NO in retinal axon patterning; (3) The effect of NO on NMDA currents during retinogeniculate development, and on the ratio of NMDA to AMPA-mediated retinogeniculate responses; (4) The effect of NO on the developmental regulation of NMDA receptor subunits; (5) The role of NO and cGMP in retinogeniculate transmission and plasticity; (6) The role of the NO-cGMP pathway in shaping postsynaptic LGN cell morphology; (7) The dynamic regulation of pre- and postsynaptic structure by NO and cGMP. These experiments will utilize anatomical labeling of single presynaptic retinal axons and postsynaptic LGN cells, intracellular recording from LGN in slices, immunocytochemical labeling of intracellular signals and of glutamate receptor subunits, and confocal microscope imaging of pre-and postsynaptic elements in living slices. Such approaches are crucial to understanding the complex cellular mechanisms that underlie the development and refinement of connections during pattern formation.
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{
"pile_set_name": "NIH ExPorter"
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C-11 alpha-methyl-L-tryptophan (AMT) is being used to assess the turnover of serotonin, a major transmitter in the brain that is linked to depression and impulsive behavior. Studies are being conducted with the tracer in monkeys before its use in humans. The monkeys being studied have high or low levels of aggressive behavior, and it is hoped that they will show differences in AMT uptake. A chiral precursor to [C-11]AMT is commercially available. Several procedures have been reported utilizing this material for the synthesis of [C-11]AMT. Although the methylation reaction using [C11]methyl iodide are similar in these procedures, the hydrolysis steps leading to [C-11]AMT are different. In our attempts to synthesize [C-11]AMT, we did not obtain reproducible results by the method of hydrolysis using hydrogen iodide. More consistent results were obtained by the method of hydrolysis using trifluoroacetic acid/ION sodium hydroxide. We obtained [C-11]AMT with a radiopurity greater than 99 percent and a radiochemical yield of about 40 percent. No D-isomer could be detected. [C-11]AM4T (1.5 to 5 mCi) was injected in three monkeys to determine the brain uptake and absorbed radiation dose. Whole-body scans were obtained using the General Electric Advance. The average residence times for lung, liver, kidney, brain, and heart were 0.67, 1.44, 1.52, 0.22, and 0.21, respectively. The kidney is the critical organ at 0.022 mGy/mBy (0.083 rad/mCi). Time-activity curves in monkey brain were determined for cortical gray matter, subcortical gray matter, and white matter.
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{
"pile_set_name": "NIH ExPorter"
}
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This study addresses how parents'discipline strategies affect children's development. The first and second aims are to examine whether the association between harsh discipline and child adjustment is moderated by: 1) the normativeness of the discipline method as indicated by cultural acceptance of its use, and 2) the affective quality of the parent-child relationship. We hypothesize that under conditions of cultural normativeness and warmth within the parent-child relationship, there is a weaker association between harsh discipline and children's adjustment difficulties. The third aim is to examine whether the association between harsh discipline and children's adjustment difficulties is mediated by children's cognitive appraisals regarding the discipline. The way children interpret a discipline practice is hypothesized to explain, in part, the link between discipline and children's adjustment. As part of understanding children's interpretations of discipline practices, we will try to "unpack" culture into measurable contextual components and to understand specific characteristics of culture (e.g., beliefs, values) that affect parenting practices and children's interpretations of them. We will collect and analyze data in 8 countries (i.e., China, India, Italy, Kenya, Philippines, Sweden, Thailand, United States) and subgroups within these countries. Each country has been selected because of its potential to provide a unique contribution to understanding how parents'discipline behaviors affect children's adjustment. Structured interviews will be conducted with 100 children and their mothers and fathers in each cultural group. Longitudinal follow-up will occur 12 months and 24 months after the initial assessment. Analyses will be conducted through structural equation modeling and multilevel modeling in which measurement occasions are nested within families, which are nested within cultures. Findings regarding mechanisms through which parenting affects children's adjustment will have public health implications because of their potential to influence interventions designed to prevent adjustment problems, especially in maltreated and other high-risk groups. Understanding how parents'discipline practices relate to children's adjustment from multiple cultural perspectives has the potential to inform applications of developmental psychology in law, policy, and intervention.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this contract is to provide research and development support for the Unified Medical Language System. Specifically, the contractor will support (1) the definition of the functional components and algorithms needed: to relate the user's terms to information in the UMLS Knowledge Sources; to interact with the user to clarify the information needed; and to select and access the information sources relevant to the user's inquiry, and (2) evaluation of the utility of the UMLS Knowledge Sources and proposed functional components in a variety of environments.
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{
"pile_set_name": "NIH ExPorter"
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Cardiovascular disease (CVD) is the leading cause of death in both men arid women. Magnetic resonance imaging (MRI) is a highly promising imaging modality for the detection and characterization of CVD. 1 of the major MRI tests that has found widespread clinical application is the dynamic (cine) imaging of cardiovascular morphology and motion for assessment of cardiac function. A major requirement for successful cardiac MRI is to reduce potential image artifacts by compensating the motion of the heart during data acquisition. This has usually been accomplished by electrocardiogram (ECG) gating and breath-holding. ECG gating within the MR scanner is not always reliable or sufficiently sensitive to arrhythmias. Breath-holding is not feasible in all patients and limits image acquisition time therefore limiting temporal and spatial resolution as well as overall image quality. The objective of the project is to improve cardiac MRI by providing more robust cardiac cycle synchronization strategies and eliminating breath-holding. This will be accomplished using self-gating strategies which extract motion information from the imaging data itself. This may potentially allow for cardiac MR imaging in patients that is not feasible using conventional techniques. The specific aims of the projects are to validate the following hypotheses: Specific Aim 1: Self-gating can be used for robust retrospective synchronization of MR cine imaging data to the cardiac cycle with improved arrhythmia rejection capabilities over ECG gatirig. Specific Aim 2: Self-gating can be used for synchronization of image acquisition to the respiratory cycle allowing segmented cine MRI while free-breathing. Specific Aim 3: Self-gating can be used for free-breathing high resolution 3D whole heart cine MRI. Specific Aim 4: Self-gating can be used for prospective synchronization of MR image acquisition to the diastolic phase of the cardiac cycle.
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{
"pile_set_name": "NIH ExPorter"
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A study of radiation effects is proposed which will employ electron spin resonance (ESR) spectroscopy and electron-nuclear double resonance (ENDOR) spectroscopy to identify direct radiation damage in irradiated single crystals of biomolecules. A principal objective will be to synthesize and crystallize DNA oligomers of sufficient length for meaningful ESR-ENDOR studies of radiation damage in nucleic acids. Paramagnetic one-electron oxidation and reduction products are observed in this type of study. An objective of the study is to relate these primary species to the final products of direct radiation damage which form in subsequent reactions of the primary radical species. Another objective is the characterization of electrons trapped in defined positions in single crystals. V band spectroscopy has been developed to the point where not only nearby protons associated with the polar tapping groups but also distant protons can be detected by ENDOR. This development makes detailed characterization of electrons trapped in a polar environment possible.
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{
"pile_set_name": "NIH ExPorter"
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The goal of this project is to develop a c-jun N-terminal kinase 2/3 (JNK2/3) inhibitor that may be used in the treatment of neurodegeneration in Parkinson's disease (PD). Development of a drug that prevents dopaminergic neurodegeneration would be first in class for halting progression of the disease and a clinical complement to existing medication used in symptomatic treatment of PD. We have assembled a team of biochemists, cell biologists, medicinal chemists, structural biologists, pharmacologists, analytical chemists, and behavioral pharmacologists with extensive pharmaceutical experience to execute this drug development program. In Aim 1 (years 1-2) we will optimize JNK2/3 inhibitors to select a preclinical development candidate. By the end of year 2 we anticipate having one or more compounds that: 1) are potent and selective JNK2/3 inhibitors, 2) promote primary dopaminergic neuronal survival, and 3) have good pharmacokinetic properties and brain penetration. This aim will be accomplished by utilizing medicinal chemistry and structure-based drug design supported by biochemical and cell-based assays, and pharmacokinetics to develop structure-activity-relationships (SAR). During Aim 2 (year 3) we will demonstrate efficacy in MPTP-mouse models of PD for greater than or equal to three compounds, demonstrate lack of interaction with human CYP450s, and evaluate five to ten compounds in preliminary toxicity studies. The reason for evaluating five to ten of our most promising compounds is to optimize the chance for clinical success and mitigate the risk of developing a single molecule that may fail in development. In Aim 3 (years 4-5) we will conduct Investigation New Drug enabling genotoxicity, safety pharmacology, and toxicology studies aimed at helping select a clinical candidate that has the best metabolic profile and widest therapeutic index. Collectively these studies are intended at generating a lead clinical candidate (and back ups) that have sufficient data to meet Food and Drug Administration standards to support up through Phase II human clinical trials in Parkinson's disease.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of these studies is to develop a pig animal model of Spinal Muscular Atrophy (SMA) whereby the development and testing of therapeutics can be efficiently accomplished. SMA, the most common inherited motor neuron disease, occurs in ~1:8,000 live births and has a carrier frequency of ~1:40; however, no cure exists. Currently there are ongoing efforts to develop SMA therapeutics using the available mouse models; however, these mouse models have limitations when considering therapeutic applications. The rapid and progressive postnatal phenotype makes it difficult to evaluate compounds and antisense oligonucleotides (ASOs) and to deliver and achieve expression of virally delivered therapeutics before the phenotype is too severe to change course. These difficulties are routinely discussed within the field, and as a result efforts have been made to develop alternative animal models. In addition, the biological differences between mouse and human make translational approaches difficult and sometimes impossible. It is anticipated that since the pig has many biological and physiological similarities to the developing human that a pig SMA model will more closely mimic the human condition. The availability of a large animal model for the development and validation of therapeutics would be of real significance for SMA and other neurodegenerative diseases. A number of translational programs are accelerating in SMA research, including small molecules, gene therapy and ASOs. Small molecules have a relatively well-defined pathway for FDA approval; however, it is likely that novel biologics such as gene therapy, antisense RNAs and cellular therapies will be greatly enhanced once blood-brain barrier permeability, dosing, delivery, distribution, sustainability and the immune response can be examined in a larger-animal model of disease such as an SMA pig. With no large SMA model available, SMA investigators have been turning to wildtype monkeys and pigs to analyze delivery and distribution without the ability to demonstrate efficacy. And while wildtype large animals can be used for distribution studies, the cellular and tissue differences between SMA and wildtype individuals could significantly impact these outcome measures. The development of a pig SMA model would allow researchers the ability to perform efficacy and delivery-related studies in a single large model and efficiently traverse from bench to bedside. PUBLIC HEALTH RELEVANCE: SMA (Spinal Muscular Atrophy) is the most common inherited motor neuron disease and occurs in ~1:8,000 live births and has a carrier frequency of ~1:40; however, no cure exists. This proposal is aimed at developing a large animal model of SMA in order to better develop and understand the effectiveness of drug, stem-cell and gene therapies. Pigs are well-suited animal models for a number of critical reasons including the similarity between human and pig development, metabolism and organ systems; therefore, the development of a pig SMA model would provide unique benefits for the SMA community as the field pushes toward a cure.
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{
"pile_set_name": "NIH ExPorter"
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The mechanisms by which neuronal pathways are established are not well understood. One theory states that pathways are first laid out by pioneer neurons and that subsequent axons travel along them. This pattern has yet to be demonstrated in the vertebrate CNS. I will test this theory by examining development of descending tracts in the relatively simple caudal spinal cord of embryos and tadpoles of Xenopus laevis. First, I will use light and electronmicroscopy to determine the time of appearance and location of the earliest tract axons. I will then identify cells giving rise to them using retrograde transport of HRP from sites near the caudal tip of the spinal cord. Axons which enter the tracts later will be examined using light and electron microscopy and HRP backfilling from progressively more rostral levels. Once the earliest neurons have been identified, I will selectively delete them by backfilling with Lucifer yellow, a fluorescent dye which when exposed to blue light destroys cells containing it. My preliminary studies show that tracts develop by ingrowth of axons into preexisting spaces between adjacent neuroepithelial cells. When the caudal-most front of tract growth is compared with progressively more rostral levels it can be seen that spaces and, later, fascicles, appear in consistent sequence and position. HRP backfills of the most primitive tract level show that primary sensory and motor cells initiate the two earliest fascicles, one dorso- and one ventrolateral. Filling at more rostral levels shows that additional primary neurons are added before spinal interneurons and that axons from trunk and supraspinal levels appear only after many axons of local origin are already present. These pilot data suggest that in descending tracts local primary neurons may first guide each other in overlapping segmental arrangement and, later, guide spinal and supraspinal interneurones. Primary neurons may thus serve as "segmental pioneers." The proposed experiments will further define the components and sequence of tract development and test an important theory of axonal guidance. The paradigm developed for these studies can be applied in the future to comparison of normal and regenerative spinal cord development.
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{
"pile_set_name": "NIH ExPorter"
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DESCRIPTION: (Applicant's Abstract) The aim of this project is the development of theory-based strategies that foster construction of anti-drug communications tailored specifically to the particular psychology of the adolescent. Acknowledging the adolescent's growing need for individuation, control, and autonomy, this work makes use of insights derived from reactance theory and the elaboration likelihood model to structure the construction of messages that target children and adolescents at different developmental stages, and of different ethnic backgrounds. The goal of the research is to provide practitioners and scientists with information about the most efficacious means of persuading and informing adolescents about the dangers of drug initiation and use. The principles that guide the research will allow development of generalizations that may be usefully employed in a variety of drug-prevention contexts. Ultimately, our aim is to create a persuasive strategy that meshes with the peculiarities of our targeted sample. In this school-based project, we propose to present targets of different developmental stages (as inferred by grade in school: 4th, 7th, and 10th graders will be used) with anti-drug information via an involving, high tech, interactive, multi-media presentation. Marijuana and inhalants are the primary focus of the messages, whose development constitutes a major goal of the first year of this work. The target audience will be segmented by ethnic status. Messages are ascribed to, and presented by, either a peer (a young adolescent) or an adult source. Two message forms are employed: one emphasizes the controlling features of the source and message ("You must do this... you must behave in this way..."), the other the target's freedom of choice ("You decide..."). This 4-way design provides a sufficiently broad foundation to test a host of important mediating factors in adolescent acceptance of drug-prevention media.
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{
"pile_set_name": "NIH ExPorter"
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The current project examines the use of calcipitrol to improve diffusion of gemcitibine into the cancer. Studies examine the effect on vasculature and the potential increase in efficacy of gemcitibine to reduce cancer burden in mouse model. Cohorts of 10-15 mice each are nrolled ito preclinical trials by imaging with ultrasound to detect tumors of a specific size. Mice are treated with carrier, gemcitabine or calcipitrol, or the combination of both drigs. Tumor growth is monitored by ultrasound. The study is evaluated when contro tumors become life threatening.
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{
"pile_set_name": "NIH ExPorter"
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Neariy half of CAIR's extramural research is now in the international arena, chiefly in former Soviet countries. International HIV prevention studies have specialized research support needs, and CAIR's development of vigorous, successful line of internafional research, service, and training activifies[unreadable]all inifiated over the past seven years[unreadable]has been facilitated by resources of the International Core. Core resources have allowed CAIR to establish important relafionships with internafional HIV prevenfion investigators, NGOs, service providers, and health ministries. These collaborations have resulted in the award of 11 international research grants, publication of over 50 articles describing results of our international research, establishment of an HIV prevention research satellite infrastructure in Russia, and the provision of resources and services to institutions and AIDS NGOs in Eastern Europe. Core functions in the renewal period include resources for conducfing international pilot studies in areas of high public health priority, translafion and linguistic support resources, vehicles to enhance international HIV prevention informafion exchange, consultafion in AIDSrelated cultural issues in Eastern European countries, conduct of biannual ethics training seminars for investigators and institutions in post-Soviet countries, and international research logistics support.
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{
"pile_set_name": "NIH ExPorter"
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Insulin resistance and hyperinsulinemia are critical to the development of type 2 diabetes mellitus. The pathogenesis, however, of the multiple complications and conditions associated with this disease such as increased risk for and decreased recovery from stroke is not yet understood. Chronic inflammation is a prominent component of diabetes and inflammatory processes mediated by cytokines are critical to the pathogenesis of stroke. Studies from our laboratory have shown that hyperinsulinemia-induced serine phosphorylation of insulin receptor substrate (IRS) proteins blocks the ability of IRSs to serve as a Janus kinase substrates, which leads to impaired cytokine signaling. In that, anti inflammatory molecules such as insulin-like growth factor (IGF)-1 and the cytokines interleukin (IL)-4 and -10, have been shown to signal through IRS proteins, we hypothesize that the balance between pro- and anti-inflammation is likely perturbed in the developing and frankly diabetic subject. In order to demonstrate that diabetes exacerbates pro-inflammatory processes in the CMS, causing increased morbidity and mortality of stroke, we will determine if type 2-like db/db mice have an enhanced IL-1-beta-mediated response to hypoxia, a model of transient ischemic attack, and examine the ability of anti inflammatory cytokines to modulate IL-1 beta- mediated central inflammatory processes in diabetes.
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{
"pile_set_name": "NIH ExPorter"
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A small subpopulation within NINCDS Collaborative Perinatal Project children has been identified as being of mixed black and white parentage. Within the interracial group of matings there is no evidence that genetic or socio-economic differences are related to race of mother (or father). Thus socio-psychological influences, presumable operating through mother-child interactions, can be examined indirectly. Two papers have been published which indicate that, although early childhood differences are wholly negligible, children of white mothers eventually develop positive intellectual differences. A paper analyzing longitudinal differences in early growth rates will be published.
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{
"pile_set_name": "NIH ExPorter"
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This is an application to renew a K24 Mid-Career Investigator Award in aging-related patient-oriented research. The Awardee is a professor of psychiatry, neurology, and epidemiology with an established track record as a researcher, clinician, and educator. The Awardee's current research focus is on mild cognitive impairment (MCI), a variably defined condition generally regarded as a prodromal and/or risk state for Alzheimer's and other dementias. Her group's epidemiological work , like those of other population-based studies in North America and Europe, has demonstrated that current "diagnostic" criteria for MCI do not reflect the real-world distribution and outcomes in the typical community or primary care clinical setting, where the majority of older adults are served. In the community at large, MCI is a unstable and heterogeneous entity, particularly in individuals with significant comorbidity. For the concept to be clinically useful, it must have better predictive value for different outcomes. Broadly, the Research Plan aims to refine the current MCI criteria based on predictive validity for dementia, using data from a new, ongoing, representative cohort study that was begun during the current K24 award period. A specific additional proposed activity is to expand the focus on vascular risk factors which are hypothesized to improve prediction of MCI outcomes. Health history and neurological exam, blood pressure, total and HDL cholesterol, and APOE genotype are already being measured on all participants. The proposed new research will obtain measurements on additional peripheral vascular markers (glycosylated hemoglobin, Apo A1, ApoB, homocysteine, C-Reactive Protein, and cystatin) from subgroups of participants with normal cognition and with subtypes of MCI. The renewed K24 will also permit the development of an application for a brain structural MRI project to determine the extent to which global and regional atrophy as well as evidence of cerebrovascular disease improve the predictive value of the clinical data, in a sample of the representative community cohort. The Mentoring Plan remains targeted towards early career investigators in clinical and epidemiological research into cognitive aging and mild cognitive impairment. As appropriate to their backgrounds and goals, trainees will enhance their experience of patient-oriented research;participate in multidisciplinary consensus diagnosis;obtain supervised experience of analyzing, interpreting, and reporting data from the Awardee's ongoing and completed community studies;participate in group interactions around individual research projects;and take advantage of additional educational opportunities available at the institution. Trainees'productivity will be monitored with regard to development of research proposals and publications and maturation as independent investigators. The Awardee will also continue to organize trainee workshops in neglected areas such as project management, principles of neuropsvcholoav for non-psvcholoaists. and scientific writing skills.
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{
"pile_set_name": "NIH ExPorter"
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Our long term objective is to investigate the molecular mechanisms that control the proliferation and differentiation of human neural crest stem cells (hNCSCs) and use these cells for regenerative medicine. Embryonic NCSCs are pluripotent and NCSCs have been shown to persist in the adult in at least two differentiated tissues, the enteric nervous system of the gut and the skin and hair follicles. Hair follicles are a niche for NCSCs in the skin. Bone morphogenic proteins (BMPs) play a critical role in embryonic NCSC self-renewal and differentiation. NCSCs have been well characterized in chick and murine systems. Despite the importance of neural crest cell biology in development and disease, the isolation of NCSCs of human origin has been elusive. We hypothesize that human hair follicle NCSCs retain pluripotency and have similar functions as embryonic NCSCs. We have recently identified a stem cell population in human hair follicles. These cells are distinctively different from epithelial or melanocyte stem cells. They are clonogenic, capable of self-renewal and pluripotent. They express NCSC markers and BMP-4 as well as BMP receptors but not lineage specific markers. These cells are located in the bulge region of human hair follicles. We propose to characterize the bulge hNCSCs in vitro and in vivo. Aim 1: Characterize hNCSCs in human hair follicles. We will further characterize the pluripotent of these bulge hNCSCs, define their genetic signature and identify surface markers that can be used to directly isolate these cells from fresh tissues. We will study the role of BMP signal pathway in the proliferation and differentiation of bulge hNCSCs. Aim 2: Study the physiological function of hNCSCs in vivo. We will test the hypothesis that bulge hNCSCs have a similar biological function as embryonic NCSCs. We will study whether bulge hNCSCs follow microenvironmental neural crest cues to develop into appropriate cell types and their physiological functions in the skin and hair follicles in vivo. Aim 3: Determine the signals for hNCSCs to differentiate to melanocytes. We will study the mechanism underlying lineage determination for hNCSCs, particularly staged differentiation of hNCSCs to melanoblasts to mature melanocytes. We will study the interaction of Wnt3a, stem cell factor and endothelin-3 with Mitf during this process. We expect that bulge hNCSCs retain the pluripotency and function of embryonic NCSCs. They contribute to normal human skin and hair follicle homeostasis. The proposed studies will establish a novel model to study human NCSCs in the somatic tissues. This model can be used to study various signalling pathways that are essential for hNCSC self-renewal and lineage determination, the physiological functions of hNCSCs in adult tissue, and the potential use of these cells for regenerative therapy. Also, because many diseases, including some cancers, are known to involve neural crest cells, the models developed in these studies may be useful in testing the etiologies of these diseases.
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{
"pile_set_name": "NIH ExPorter"
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In the last two decades though the field of neuroprosthetics has gained tremendous momentum through the development of novel architectures for neural interfaces, however, till date there is no device which can be nomenclatured as the gold standard in the field. As a result the research and technology environment at each laboratory is unique and require a high-level of customization and integration. The objective of this application is to devise a one-in-all technology that encompasses the assets (such as ultra high aspect ratio, multi- channels, flexible substrate, interconnection etc.) of the silicon (such as the Utah and Michigan array) and polymer based arrays and mitigates their limitations. Most importantly it provides a highly customized and poor man's solution to research laboratories. In this application we present a platform technology, called Blackrock Micro Molding Arrays (BRMMA), based on micro molding technique, which enables devolvement of highly customizable and affordable micro devices without compromising the characteristics of the neural probe such as electrical and mechanical properties, reliability, and durability. BRMMA offers highly customized design rules which include surface and penetrating electrodes. It allow the end users for the first time to dictate their own electrode design (specified within the design rules of BRMMA) as per their hypothesis, and choose the number of active channels per shaft, location and material used for active sites, shape and size of the shaft etc. BRMMA developed by Blackrock Microsystems would produce relevant statistical number of devices to test in vitro and in vivo functionality for long term use BRMMA would be marketed to the neuroscience research community by Blackrock Microsystems.
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{
"pile_set_name": "NIH ExPorter"
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OBJECTIVES: a) The overall objective of this entire research project is to purify sufficient quantities of D-lactic dehydrogenase from two strains of Moraxella (ATCC19965, 19975) from one strain of Neisseria (ATCC8193) and anthranilate synthetase from Moraxella (ATCC19965) for the preparation of rabbit antibody specific against these enzymes. The antisera will then be used to establish the degree of antigenic similarity for these enzymes from these and other catalogued strains of Moraxella and Neisseria. The potential use of these antisera for routine determinative purposes will then be assessed by testing them against clinical isolates identified as various species of Moraxella, Neisseria, Acinetobacter, or Mima-Herellea. b) The goal for the second year of this project was to devise a purification procedure, using Moraxella osloensis (ATCC19965) as a source of D-lactic dehydrogenase (D-LDH), which could also be used for the purification of D-LDH from Moraxells non-liquefaciens (ATCC19975) and Neisseria (ATCC8193) and to scale up the procedure in order to obtain amounts (2-3 milligrams) sufficient for antiserum production in rabbits. A procedure has been devised for isolating cytoplasmic membranes from the organisms (ATCC19965) using lysozyme (750e.u.) in a Tris buffer (10 to the minus 2nd power M, pH 8) containing EDTA (10 to the minus 4th power M) and sucrose (20%). Washing the membranes with buffer removed cytoplasmic proteins as well as some envelope proteins. The membrane-bound D-LDH was recovered following solubilization of the membrane with 50 mM phosphate buffer (pH 7.5) containing Triton-X-100 (1%) and EDTA (10 micromolar). Enrichment of D-LDH is approximately 7 fold using this procedure compared to the specific activity of the enzyme observed in cell free preparations obtained using ultrasonic disruption techniques. A greater enrichment of initial D-LDH activity has been achieved by replacing EDTA with 10 micromolar MgCl2 in the membrane-solubilization buffer. Chromatography of the enzyme of DEAE cellulose and affinity chromatography have yielded preparations estimated to be approximately 70% pure as judged by polyacrylamide gel electrophoresis (PAGE). Both protein and activity stains have been used to assess enzyme purit (Text Truncated - Exceeds Capacity)
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{
"pile_set_name": "NIH ExPorter"
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: U6 small nuclear RNA (U6 snRNA) is a component of eukaryotic spliceosomes and is required for splicing of nuclear pre-messenger RNAs. It has been suggested that U6 RNA may be directly involved in catalysis during nuclear pre-mRNA splicing. U6 RNA, besides being the most conserved of all U snRNAs, is unique in many respects. First, it contains CH3-O-pppG cap instead of the trimethylguanosine (TMG) cap found in other U snRNAs. Second, while TMG cap-containing U snRNAs are transcribed by RNA polymerase II, U6 RNA is transcribed by pol III. Third, the transcription of mammalian U6 snRNA gene does not require any internal promoter and the transcription is dependent on 5' flanking sequences. Therefore, studies on U6 snRNA synthesis are important to understand the fundamental cellular events. Experiments will be carried out with the following specific objectives: (A). To purify the factors involved in the capping of U6 and 7SK small nuclear RNAs. (B). To study the mechanism of capping with purified components. Since U6 RNA has an unusual cap structure (gamma-monomethyl phosphate), it offers a model system towards elucidating the fundamental differences in the mechanism(s) of cap formation on U6 and 7SK snRNAs vs mnRNAs and other U snRNAs. (C). To obtain monoclonal and polyclonal antibodies specific to mepppG and mepppA cap structures. (D). To identify and characterize the minor methyl-capped snRNAs. (E). To identify and purify the factor(s) involved in the transcription of U6 snRNA gene.
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{
"pile_set_name": "NIH ExPorter"
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There is concern about the misuse and abuse of benzodiazepine anxiolytic/sedative drugs which are among the most widely used of all medically prescribed compounds. Caffeine is the single most widely used behaviorally active drug and there is increasing concern that a significant proportion of the population may be at health risk from excessive caffine consumption. Despite the fact that both benzodiazepines and caffine have been periodically identified as being drugs of abuse, the reinforcing and physical dependence producing effects of these compounds remain incompletely characterized. This project will investigate these aspects of benzodiazepines and caffine. In addition to providing specific information about behavioral and pharmacological mechanisms by which benzodiazepines and caffine come to capture and control behavior, this research should provide valuable insights into the general nature of the drug dependence process. A series of studies of baboons and rats is proposed. Two experiments will study environmental conditions which may modulate or enhance the intravenous and oral self-administration of benzodiazepines in baboons. A related experiment will use progressive-ratio schedules of drug self-administration to characterize the relative reinforcing effects of benzodiazepines and barbituates in baboons. Two experiments will characterize spontaneous and precipitated withdrawal in baboons exposed to low chronic doses of the benzodiazepine diazepam. Two other experiments will use baboons and rats to investigate the modulation of benzodiazepine physical dependence and/or tolerance by administration of a benzodiazepine receptor antigonist. The final set of experiments will focus on caffine in baboons. Two experiments will investigate optimal conditions for maintaining maximal intravenous and oral caffine self- administration and one experiment will characterize spontaneous withdrawal effects after high chronic doses of caffine.
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{
"pile_set_name": "NIH ExPorter"
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This project involves the study of the effects of antibiotics on bacterial infections in 100 women undergoing elective hysterectomy in the Yale-New Haven Hospital from 1973 to 1977. Antibiotics used were Penicillin and Cefazolin. Staff members of OBE are collaborating on the extensive data analysis, including multivariate analysis of variance for categorical data.
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{
"pile_set_name": "NIH ExPorter"
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Leucocytes from a total of 212 patients have been assayed against an antigen derived from pancreatic adenocarcinoma. Results, to date, have indicated that the leukocytes from 11 of 12 patients whose cancer are localized or confirmed to lymph nodes adjacent to the pancreas have identified the antigen, whereas leukocytes in 6 of 9 patients with more distant lymph node involvement have identified that antigen. Of patients with metastatic pancreatic carcinoma with either intra-abdominal spread or hematogenous spread to organs other than the liver, leukocytes from 10 of 12 patients have identified the antigen. In contrast, leukocytes from only 15 of 27 patients with metastatic pancreatic adenocarcinoma of the liver have identified these patients. There have been no false positives among the 17 patients with pancreatitis, 2 false positives among 16 patients with other diseases of the gastrointestinal tract and one false positive among 30 normal healthy controls. Among patients with carcinoma, there are 2 false positives among 71 patients with gastrointestinal tract cancer (both had cholangiocarcinoma) and 2 other false positives among 18 patients with carcinomas other than gastrointestinal origin. Preliminary studies evaluating serum "arming" of normal healthy leukocytes and the use of those "armed" leukocytes in LAI has shown concordant results in 76% of tests thus far. Further studies are underway to assess the value of "arming" and towards standardization of an antigen source.
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{
"pile_set_name": "NIH ExPorter"
}
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The major objective and principal scientific challenge of this proposal is the implementation of new methodology for the stereocontrolled synthesis of building blocks for a number of biologically important natural products, based on novel organomolybdenum chemistry. Some key targets chosen so as to highlight the considerable potential of this new approach to organic synthesis are a C30 diol obtained from messel shale kerogen, the side chain of vitamin E and the right hand segment of an important macrolide antibiotic, magnamycin B. When properly developed, the approaches described may be applied to the synthesis of unnatural analogs of these substances leading to compounds having beneficially modified biological activity. The method to be developed is based on the ability of the dicarbonylcyclopentadienylmolybdenum cation, when attached to a diene, to activate the diene to nucleophilic attack. Successive carbon nucleophile addition, reconversion of Pi-allylmolybdenum products to diene complexes followed by second nucleophile addition generates a stereodefined disubstituted - - allylmolybdenum complex which can be converted to organic products using novel methods developed in our laboratory. This unique approach to stereocontrol is to be used in six-, seven- and eight-membered rings to generate the proposed target molecules. Of particular significance is the influence which we can expect the dicarbonylcyclopentadienylmolybdenum moiety to have on the conformations of awkward ring sizes, thereby allowing controlled manipulations which are not possible without the aid of the metal.
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{
"pile_set_name": "NIH ExPorter"
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Recently, major advances have been made in elucidating both the genetics and biochemistry of chloroquine resistance as well as in elucidating the atomic level interactions between chloroquine and its principle target, uncrystallized heme. In the former, groundbreaking advances by the Wellems lab have expanded into a series of detailed studies conducted in our laboratories, as well as by others that have generated enormous data in a short period of time. In the later, major advances have very recently been made by members of this consortium in defining the physical chemistry of quinoline - heme interactions using both new solution NMR methods as well as cutting edge solid state NMR methods. In collectively analyzing these data new (previously unrecognized) concepts that assist the design of quinoline and acridine based antimalarial drugs become evident. Capitalizing on these, while remaining within antimalarial drug cost limitations, also requires significant advances in synthetic chemistry, including developing highly chemo- and regioselective cross-coupling reactions. Over the past 18 months, we have pioneered major advances in the synthesis of heme-targeted antimalarial (HTA) pharmacophores. Our discussions and collaborations in this regard have developed into highly synergistic drug discovery activities between the laboratories of the investigator, Dr. Roepe, and Dr. de Dios. We will combine the unique genetic, biochemical, physical chemical and synthetic chemistry expertise present among our groups to design, synthesize, and solve drug - target structures for new HTA drugs. Using the unique drug screening capabilities present in our consortium, we will analyze large libraries of these for antimalarial activity, both alone and in combinations. Our long term goal is the identification of novel, inexpensive, efficacious therapy for treating drug resistant malaria.
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{
"pile_set_name": "NIH ExPorter"
}
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Project Summary/Abstract Endometriosis, a chronic gynecological disorder defined as the presence of endometrial glands and stroma outside the endometrial cavity, is characterized by peritoneal inflammation, fibrosis, adhesions and ovarian cysts. Patients suffer from chronic pelvic pain, pain during intercourse, painful periods and infertility. This condition remains poorly understood with limited treatment, and continues to affect the productivity and lifestyle of millions of women around the world causing high levels of psychological and emotional distress. Our study aims to shed light on possibilities for an entirely new therapeutic approach for patients with endometriosis. Our previous studies have shown that exposure to stress and modulation of the hypothalamic- pituitary-adrenal (HPA) axis exacerbates the development and severity of endometriosis. The goal of this proposal is to extend our prior results by investigating the impact of exercise, a therapy that could be easily extrapolated to the patient population. There is already evidence that exercise can act as a stress buffer with beneficial effects in many chronic conditions, most likely via suppression of the immune system and HPA axis regulation. A recent systematic review and meta-analysis suggest that physical activity may reduce the risk of endometriosis. However, until now no specific experimental data exist regarding the impact of exercise as an intervention on the course of endometriosis. This study will test the hypothesis that physical exercise can ?realign/reset? dysregulation of the brain-gut axis caused by stress resulting in reduced endometriosis symptoms in an animal model. Under Specific Aim 1, we will expose our endometriosis animals to exercise before/during and after the induction of endometriosis under non-stress conditions, examining its impact on vesicle and intestinal pathology, cytokines, behavior, hippocampal expression of CRF, and the intestinal microflora. In Specific Aim 2 we will identify how exercise interventions can reduce or prevent the stress-induced exacerbation of endometriosis, examining the impact of timing on the endpoints outlined above. This study will contribute to the goals of the AREA program by strengthening the institutional environment and exposing students at various levels to multidisciplinary research areas, thus offering a unique training opportunity to encourage their continued involvement in biomedical sciences research. Our long-term goal is to implement behavioral and stress-reduction strategies as part of an integrated clinical management plan for endometriosis Outcomes from this study will also further enhance our understanding of the neuro-immune system in women?s health.
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{
"pile_set_name": "NIH ExPorter"
}
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The mission of the Developmental Therapeutics Program (DTP) is the development of novel synthetic and characterized natural products as potential anti-cancer agents. Towards this mission the contract will be devoted to the synthesis of small molecules needed by the various programs of the Division of Cancer Treatment and Diagnosis (DCTD), and other NIH divisions. This contract will provide essential synthesis support for the early stages of the drug development program. The small molecules assigned for the syntheses encompass a variety of structural classes;e.g., carbocyclics, heterocyclics, steroid analogs, and natural products. They originate from the Rapid Access To Intervention Development (RAID), Rapid Access to NCI Discovery Resources ( R*A*N*D) programs, Biological Evaluation Committee (BEC), Operating Committee (OC) recommendations, and other NCI, and NIH programs. About 90% of the synthesis work involved in this contract originated from the current RAID and R*A*N*D programs, and from extramural applicants. Under the scope of the R*A*N*D program, the synthesis of a limited series of analogs in order to improve the potencies and pharmacodynamic properties of the lead compound, where the core chemistry is either well established or less documented, were also undertaken. The remaining 10% of the work has originated upon the request of the OC and other NCI, NIH programs. The purity and identity of all resynthesized compounds and intermediates were established by state-of-the-art physico-chemical methods. The quantities synthesized would vary from 200 mg to 100 grams. The overall objective of the contract will be the [unreadable]Non-GMP[unreadable] chemical synthesis of compounds of good purity needed for the cancer program, such as synthetic compounds prompted by the RAID and R*A*N*D programs, and compounds arising from recommendation of the Biological Evaluation Committee. The existing contract is not utilized for the synthesis of combinatorial libraries or leads resulting from such libraries. The new contract will be utilized for the purpose of synthesis of active single compound leads and their analogs using parallel synthesis methodology, as sometimes required by the R*A*N*D program
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{
"pile_set_name": "NIH ExPorter"
}
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Approximately 60% of the U.S. population is overweight or obese, and obesity has been linked to numerous health problems. Physical activity has been recommended for overweight and obese adults to enhance weight- loss and/or weight-maintenance, and to reduce risk of chronic disease, including heart disease and cancers of the breast and colon. Despite the benefits of exercise, only 20% of overweight or obese adults meet the minimum national recommendations (150 minutes/week of moderate intensity exercise-e.g., brisk walking), and drop out in the first few months of exercise programs is as high as 50%. Given these data, there is a need to improve adherence to exercise programs, especially for overweight and obese adults. We propose to conduct a pilot study to test feasibility and proof-of-concept for two incentive programs to promote exercise among low-active overweight and obese adults. To address weaknesses in the literature, the programs will be (a) conducted in a community setting by the Greater Providence YMCAs and (b) financially sustainable, such that they do not require removal of the incentives after a specified period of time. In both incentive programs, participants will pay the standard monthly YMCA membership fee of $49/month. In the Rebate incentive program, participants will have the opportunity to earn $1/day in rebates on their membership fee for each day that they attend the YMCA (verified by objective swipe-card data), with a maximum of $5/week. In the Donation incentive program, participants will have the opportunity to earn $1/day (using the same incentive schedule) in donations to a registered local charity of the participant's choice. Thus, the proposed study will compare three experimental conditions: (a) Rebate incentives (n=25); (b) Donation incentives (n=25); and (c) Control (i.e., no incentives) (n=25). Our Primary Aims are to test (1) feasibility f the two incentive programs, (2) feasibility of the research methods to evaluate the preliminary efficacy of the two incentive programs, and (3) proof-of- concept for the two incentive programs (relative to control), through comparison of average number of incented sessions/week over one year. Secondary outcomes will be self-reported minutes per week of exercise over one year. The proposed research will provide a preliminary investigation into two community-based, financially sustainable incentive programs to promote exercise for adults who are at increased risk for cancer. Following demonstration of feasibility and proof-of-concept we plan to apply for an R01 to test the incentive programs in an RCT. Positive findings from such a trial would provide a fast-track for a community-based ready-to- implement exercise promotion intervention. Additionally, such findings would have significant implications for the use of financially sustainable incentive programs for exercise through other community organizations (e.g., privately-owned health clubs), healthcare organizations, or employers (e.g., employer fitness facilities), as well as providing a model for incentive programs for other health-related behaviors (e.g., smoking cessation, weight loss).
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{
"pile_set_name": "NIH ExPorter"
}
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Previous studies have demonstrated that intracellular Cs2+ overload plays an important role in myocardial ischemic injury, and that an important determinant of Ca2+ homeostasis is the control of intracellular Na+ (Na+). However, Na+ homeostasis during ischemia is still poorly understood. The proposed project will employ a unique combination of state-of-the-art NMR techniques to obtain a comprehensive assessment of how Na+I is controlled in the myocardium, during ischemia and hypoxia. Using the intact perfused rat heart model, interleaved 23Na and 31P NMR spectra will monitor changes in Na+I and cellular energy. In the same preparations, 87Rb NMR spectroscopy will be used to monitor Rb+ uptake and Na+/K+ ATPase activity. Using 7Li NMR spectroscopy, Li+ uptake will be measured to monitor voltage- gated Na+ channel activity. This methodology not only measures net Na+ accumulation, but is designed to also provide simultaneous information concerning how changes in unidirectional Na+ fluxes induce it. The following specific aims will be addresses: 1. To measure sarcolemmal Na+/K+ ATPase activity during control normoxic perfusion, hypoxia, and low-flow ischemia. The experiments will determine whether Na+ extrusion decreases or increases under the latter two conditions. Also, to examine Na+/K+ ATP activity in the context of the various metabolic modulators such as Na+, cellular energy status, pH, etc., in order to assess how functional alteration may occur during ischemia/hypoxia. 2. To determine the rate of unidirectional Na+ influx, during control normoxic perfusion, hypoxia, and low-flow ischemia, by using the Na+ and Na+ extrusion rate measurements. These measurements will determine if unidirectional Na+ influx is altered under the various conditions, and in particular, if it is downregulated during low-flow ischemia, as compared to control normoxia and hypoxia. Also, to compare these Na+ influx alterations with those obtained from measurement of Li+ uptake, which has specificity for Na+ channel activity. 3. To measure the rates of Na+ influx which occur via Na+/H+ exchange, during control normoxic perfusion, hypoxia, and low- flow ischemia, and to assess the importance of this mechanism in contributing to Na+ accumulation under various ischemic/hypoxic conditions. 4. To measure the rates of unidirectional Na+ influx which occur via Na+-HCO3- cotransport, during control normoxic perfusion, hypoxia, and low-flow ischemia, and to assess the importance of this mechanism in contributing to Na+ accumulation under various ischemic/hypoxic conditions. Also, to assess the possible interaction of this pH regulatory mechanism with Na+/H+ exchange (whether inhibition of one stimulates the other). 5. To measure the rate of Na+ influx which occur via non- inactivating Na+ channel current, during control, normoxia, hypoxia, and low-flow ischemia, and to assess the importance of this mechanism in contributing to Na+ accumulation under various ischemic/hypoxic conditions.
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{
"pile_set_name": "NIH ExPorter"
}
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Type 2 diabetes, obesity, and hypertension are highly prevalent in Western society and increasingly prevalent in the developing world. Abundant clinical and basic research suggests that insulin resistance (IR) underlies each of these disorders and emerges in response to the environmental stresses of Western diet and diminished physical activity. However, the precise linkage between these environmental changes and these diverse clinical disorders is only marginally understood. Others have reported that insulin delivery to muscle is the rate limiting step for its metabolic action in vivo. Our laboratory has been testing the hypothesis that vascular IR plays an important pathogenetic role in the development metabolic IR. Specifically, we have provided abundant data (supported by this grant) that skeletal muscle microvasculature is exquisitely sensitive to physiologic concentrations of insulin which act to expand the microvascular pool perfused. We have also reported that insulin is transported across the endothelial cell (EC) by a regulated process which is activated by insulin. In this manner, insulin can facilitate its own delivery and that of glucose and other nutrients to muscle. IR inhibits these vascular actions of insulin. In studies proposed here we will probe the linkage between environmental factors (elevated plasma free fatty acid concentrations, high fat diet, acute and chronic exercise) and vascular and metabolic insulin resistance. We will utilize techniques developed in our laboratory to assess the functional actions of insulin on the microvasculature, its biochemical signaling, and the role of oxidative stress and inflammation within the vasculature on insulin actions. This will be done addressing 3 specific aims : Aim 1-will test whether FFA promote an inflammatory response in the EC, the vascular smooth muscle cell or both in rat vasculature and thereby diminish muscle insulin delivery;Aim 2-will test whether enhancing nitric oxide availability or blocking either the AT1 or endothelin A receptor will diminish/prevent FFA-induced microvascular inflammation and dysfunction, and whether exercise training may have a similar effect;and Aim 3- will test whether different dietary lipids [saturated, omega-3, and omega-6 polyunsaturated fatty acids (PUFA)] will differentially affect muscle microvascular insulin responsiveness, delivery of 125I-insulin to skeletal muscle, muscle insulin sensitivity, and vascular inflammation. By addressing the origins of muscle microvascular dysfunction in IR should allow us to develop strategies to limit or reverse vascular IR and thereby improve metabolic IR. PUBLIC HEALTH RELEVANCE: Insulin resistance (IR) underlies multiple, prevalent, serious diseases. Muscle is a major insulin target tissue and insulin movement from the plasma to muscle interstitium is rate limiting for insulin's metabolic action and IR slows this. Using novel methods developed in our laboratory we will test whether vascular inflammation and increased production of reactive oxygen species induced by pro- inflammatory lipids play a critical role in producing skeletal muscle vascular and consequent metabolic IR and if this can be reversed.
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{
"pile_set_name": "NIH ExPorter"
}
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Research suggests that clinical research subjects often fail to appreciate significant elements of what their informed consent because of the "therapeutic misconception." The therapeutic misconception involves patient/subjects failing to grasp the difference between participating in a clinical research project and receiving ordinary clinical care. We do not know how: 1) prevalent it is, 2) with which personal and situational variables the therapeutic misunderstanding is associated, 3) how seriously this misconception affects other aspects of a decision to participate in research, 4) what its effects are on patients' experience of research participation and 5) how it affects the rate of recruitment to, and dropout from clinical research. This proposal aims to remedy these lacks of knowledge by interviewing in depth 200 subjects from 10 different clinical research projects and then reinterviewing them after they have completed their participation in the project. This will facilitate new approaches to gaining consent and reduce potential sources of public disaffection with clinical research.
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{
"pile_set_name": "NIH ExPorter"
}
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The long-term objective of this project is to obtain detailed biochemical information and understanding of the metabolism of acetate in the strictly anaerobic microorganism Methanosarcina barkeri. Specific goals are concerned with determination of structural and catalytic properties of a high molecular mass multienzyme complex that, as previous results of this project have demonstrated, catalyzes a critical, early event in acetate decomposition--namely, cleavage of the carbon-carbon bond of acetate (as acetyl-CoA). This project has shown that tetrahydrosarcinapterin (H4SPT) is required as a specific acceptor of the methyl group of acetyl-CoA, and that the carbonyl group is oxidized to carbon dioxide. Previous results have shown also that the reaction catalyzed by the enzyme complex is as follows: Acetyl-CoA + H4SPT + H2O -> CoA-SH + CH3-H4SPT + CO2 + 2[H] Currently, insight on the mechanism of this reaction has been gained by UV-visible and EPR spectroscopic analyses of reactions of the enzyme complex with CO and CO plus CH3-H4SPT. Catalysis of a related reaction in the direction of acetyl-CoA synthesis was shown to proceed as follows: CO + CH3-H4SPT + CoA-SH -> H4SPT + acetyl-CoA Partial reactions with CO alone caused stepwise reduction of enzyme Fe/S and corrinoid centers. Addition of CH3-H4SPT caused rapid methylation of the reduced corrinoid component. After previous reduction of the enzyme complex, further interaction with CO generated an EPR detectable adduct. Based upon observed quenching effects of CH3-H4SPT on the EPR radical signal, evidence was obtained supporting the hypothesis that the EPR signal may emanate from a one-carbon enzyme-bound precursor of the carbonyl group of acetyl-CoA. In the absence of CoASH, it is postulated that formation of an acetyl-enzyme intermediate is responsible for quenching of the EPR signal by CH3-H4SPT.
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{
"pile_set_name": "NIH ExPorter"
}
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Immunotoxins combine the activity of a powerful toxin with an antibody capable of delivering the conjugate to a specific target cell. These compounds have proven effective in the in vitro elimination of a number of cell types, including leukemia cells and the bone marrow cells implicated in graft vs. host disease. Immunotoxins may be an effective way of controlling a number of diseases attributable to a population of antigenically distinct cells. The in vivo use of immunotoxins is currently limited by an unacceptably high level of non-specific toxicity. This probably results from the ability of the toxic component to enter the cytosol without antibody help. The goal of this research is to develop immunotoxin components that retain their intracellular toxicity, but are incapable of transmembrane relocation unless bound to antibody. Five ribosome-inactivation proteins capable of functioning as immunotoxin components will be enzymatically or chemically cleaved into smaller peptides. It is hoped that by this process functional domains of the toxins can be found that retain the ability to catalytically inactivate ribosomes, but are incapable of self-directed entry into the cytosol. The activity of these peptides will be tested in vitro in whole cell and cell-free assays of protein synthesis, as well as in vivo where the LD50 of active peptides will be compared to that of the intact toxin. The amino acid sequence of active peptides will also be determined.
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{
"pile_set_name": "NIH ExPorter"
}
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The Child Care and Development Block Grant (CCDBG) Act of 2014 incorporated several statutory changes intended to promote child well-being. One of those changes, a longer redetermination period, was intended to lead to longer program participation spells, more stable usage of high quality childcare arrangements, and more stable parental employment spells. Identifying the actual effects of the new redetermination policy is a priority for the Illinois Department of Human Services (IDHS) and its partner agencies. With funding from this planning grant, IDHS will build on its existing partnership with Chapin Hall at the University of Chicago to develop a comprehensive research and evaluation agenda to answer whether and how extending the CCDF redetermination period from six months (three months if self-employed) to twelve months will affect broad outcomes in three domains: 1) parental employment; 2) continuity of care, and; 3) access to childcare quality. Funding from this planning grant will be used to design a comprehensive, well defined, and achievable research plan to answer those questions. The plan will draw on the Illinois Early Learning Council's research agenda and will incorporate feedback from Illinois program and policy experts in the ECE field as well as technical research experts. This process will ensure that grant activities are informed by a diversity of critical stakeholders both within DHS and in partner organizations so that deliverables and products are methodologically rigorous, optimally policy relevant and can provide actionable policy recommendations that reflect the realities of Illinois' complex early learning and development system. IDHS will leverage resources and deliverables from this planning grant to enhance its own internal capacity to manage and conduct CCDF research and evaluation. Four main outcomes are expected under this grant: 1) A literature review; 2) An inventory of data sources; 3) Creation of a research advisory committee, and 4) Development and validation of an evaluation research proposal. These components of the planning strategy will provide IDHS with a clear, high quality, and actionable research plan to evaluate the impacts of the new redetermination policy on Illinois children and families participating in the CCAP program.
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{
"pile_set_name": "NIH ExPorter"
}
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This proposal is to implement the techniques of sensitivity analysis as a means to 1) quantatitvely assess the relationship between the potential and the observables calculated using molecular mechanics and dynamics; and 2)to quantitatively assess the relationship between the function of a biomolecule and its structure. These techniques will be used in the quantitative evaluation of the of reliability modelling.Phase I showed the significance of these concepts and Phase II will develop their capabilities for biomolecular modeling.
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{
"pile_set_name": "NIH ExPorter"
}
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The goal is to develop improved diagnostic imaging agents for human melanoma based on a radiolabeled monoclonal antibody. P97 is an oncofetal antigen present on human melanoma tumor cells. Studies in nude mice have shown there is active concentration of anti-p97 antibody and Fab fragments directed against p97 antigen of human melanoma. The uptake is antigen specific, as documented by comparisons with radiolabeled control immunoglobulin and immune fragments. Ten patients have been studied with whole anti-p97 antibody, and uptake has been documented in metastatic lesions. About 88% of documented metastatic lesions have been imaged, and antigen-specific uptake in tumor has been documented by comparison to radiolabeled control immunoglobulin. Maximum target-to-nontarget ratios observed from human tumors, biopsied at 72 hours after intravenous administration of the anti-p97 antibody, are 7 to 1. Thirty-three patients with advanced malignant melanoma were studied on 47 occasions with I-131-labeled Fab (anti-p97). In 20 of these, a simultaneous administration of an I-125 labeled nonspecific Fab was performed. Antigen-specific localization was observed with average ratios of specific:nonspecific uptake of 3.7 (48 hours) and 3.4 (72 hours); uptake was correlated with tumor p97 concentrations. Twenty patients had positive scans. Antibody injections were well tolerated with the exception of four reactions: one episode of transient chills and fever; flushing and hypotension; and two skin rashes. All these reactions responded promptly to symptomatic therapy. Human antimouse antibodies were documented in the two patients who developed skin rashes.
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{
"pile_set_name": "NIH ExPorter"
}
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Integrins are transmembrane heterodimeric glycoproteins composed of 1 and 2 subunits that mediate the interactions between cells and extracellular matrix (ECM). In humans there are 18 1 and 8 2 subunits, which combine in a restricted manner to form dimers, each of which exhibit different ligand binding properties. Integrins can be classified by these ligand binding properties into collagen, laminin and RGD binding receptors. The principal laminin binding integrins are 1321, 1621 and 1624. Integrin-mediated cell interactions with basement membranes are critical for normal development of the kidney collecting system. Key components of basement membranes are laminins which are trimers consisting of 1, 2 and 3 subunits. Although much work has been done to show the requirement of laminins in the development and function of the glomerulus of the kidney;their role in renal collecting system development in vivo is poorly defined. In this context, until recently only the 15 chain containing laminins were known to play a role in ureteric bud (UB) development in vivo. In the last funding cycle we showed that the 13, 31 and 32 chains of laminins are also required for UB development. There is controversy as to the role of the laminin binding integrins in the developing kidney collecting system. Although all 3 principal laminin binding integrins are expressed, their roles are not well defined. Only integrin 1321 has been shown to play a role in UB development in vivo, however we have shown that the integrins 1621 and 1624 play a role in UB development in vitro. During the last funding cycle we demonstrated that deleting the 21 integrin subunit in the UB in vivo resulted in significantly worse developmental abnormalities than when the 13 subunit was deleted, suggesting that other 121 integrins are important for this developmental process. In this proposal we aim to delineate the specific roles of the laminin receptors 1321, 1621 and 1624 in UB development in vivo and determine the mechanisms whereby these integrins regulate this process. We will delete the 13, 16, and 24 integrin subunits either alone or in combination in the developing UB by crossing integrin-floxed mice with hoxb7cre mice to test the hypothesis that the three laminin-binding integrins, namely 1321, 1621 and 1624, are required and act synergistically for the UB to develop and function normally in health and disease. This hypothesis will be tested in the following 3 aims. 1) Determine the role of the specific laminin receptors in UB development and function. 2) Determine the roles of the 13, 16and 24 integrin subunits in maintaining the structural integrity of the renal collecting system. Aim 3) Determine the mechanism whereby the specific laminin receptors regulate renal epithelial cell branching morphogenesis. PUBLIC HEALTH RELEVANCE: We anticipate that this study will generate novel insights into the role of laminin receptors in the development of the collecting system of the kidney. This knowledge is fundamental to our understanding of how the renal collecting system functions and why there is an increased incidence of renal abnormalities in patients with bullous skin diseases.
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{
"pile_set_name": "NIH ExPorter"
}
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The project represents a continuation of the investigation into the variables related to the development of the optical and refractive characteristics of the human and sub-human primate eyes. Since the work to date indicates that nearwork plays a considerable role in the development of myopic refractive characteristics, the present project will involve several approaches to the evaluation of its role. One is the longitudinal study of optical and refractive characteristic changes in Warm Springs, Oregon Indian children beginning at age five. The results obtained have been analyzed statistically and regression equations have been partially developed. As more information is gathered these equations will be modified in order to permit prediction of those individuals who will be come myopic and the point in time at which they will become myopic as well as the amount of myopia that will be achieved. The second approach involves the study of optical characteristics of monkeys subjected to different conditions of accommodation and/or convergence. When the intraocular pressure is monitored by an implanted pressure transducer of the radiosonde type, preliminary evidence suggests that there is an increase in vitreous chamber pressure which is linearly related to accommodation. We hope to be able to factor out the relationship and the contribution of both accommodation and convergence to this possible relationship.
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{
"pile_set_name": "NIH ExPorter"
}
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The majority of breast cancers are diagnosed as breast cancer of no specific type (or ductal carcinoma), a heterogeneous category of tumors that includes curable neoplasms as well as highly aggressive cancers. This classification is inadequate for clinical management of breast cancer patients as well as for designing laboratory and clinical research protocols. Although there are clearly a number of different tumor phenotypes within this diagnostic category, traditional morphology and even current molecular markers of prognosis are incapable of clearly discriminating different subtypes of ductal cancer from one another. In this project, we propose to distinguish biologically different groups of ductal breast cancer from one another using gene expression profiles. This is based on the concept that gene expression profiles represent an objective and quantitative measure of a neoplasm's phenotype. We have in preliminary experiments identified a number of genes that are differentially expressed among different ductal cancers, consistent with our hypothesis that will use a custom breast cancer array that we developed to represent the genes most capable of differentiating breast cancers, and then use this array to analyze gene expression profiles in microdissected breast cancer samples. Groups of tumors that share expression profiles identified through cluster analysis of data will then be examined for pathological and clinical similarities to develop more specific hypothesis of ductal cancer classification. Finally, we will design case-control studies to test and refine these classification hypotheses. Through this stepwise development and testing of classification hypotheses, we expect to identify biologically distinctive categories of breast cancer. A sound molecular classification of this nature will greatly benefit clinical management of breast cancer patients and will provide a useful reference for the design of breast cancer research.
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{
"pile_set_name": "NIH ExPorter"
}
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The Women's Health Initiative is a randomized controlled clinical trial conducted in 40 clinical centers across the United States addressing some of the most common causes of death, disability, and impaired quality of life in postmenopausal women ages 50-79; cardiovascular disease; breast and colon cancer; and osteoporosis. The three components of this disease prevention trial are: (1) hormone replacement therapy with a primary outcomes of coronary heart disease and stroke and secondary outcomes of hip and other fractures; (2) low fat dietary intervention with primary outcomes of breast cancer and colorectal cancer and a secondary outcome of coronary heart disease; and (3) calcium and vitamin D supplementation with a primary outcome of hip fractures and secondary outcomes of colorectal cancer and combined fractures. A concurrent observational study will provide estimates of health effects of known risk factors and new predictors for disease. Each clinical center will collect data through questionnaires and biological specimens over 8-12 years. Additionally, this center is one of three performing bone mineral density scans to correlate changes with risk of osteoporosis and fractures.
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{
"pile_set_name": "NIH ExPorter"
}
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