text
stringlengths 1
32.7k
| meta
dict |
|---|---|
Fucosylation plays an important role in many cellular processes. Fucosylated oligosaccharides in the cell are involved in many biochemical recognition processes, microbial infections, toxin entry, and cancer cell metastasis. These properties make fucosylated molecules valuable for pharmaceutical and drug discovery needs but current production methods are very expensive and impractical. Most notably is the expense and difficulty in producing the activated sugar, GDP-fucose. Our goal is to increase accessibility of GDP-fucose and fucosylated molecules such as oligosaccharides so that the research community can better understand the role of these compounds in human health, develop novel antimicrobial, anti-inflammatory and anti-cancer agents, and develop strains suitable for large-scale production of various oligosaccharides and fucosylated molecules. Current methods described to date for the production of GDP-fucose using either chemoenzymatic synthesis or modified E. coli and S. cerevisiae strains all yield only small milligram quantities of material or are overly complicated and can't be scaled. Here we propose to develop an entirely new yeast-based method for production of GDP-fucose. There are two main advantages to this this yeast-based system. First, it uses an inducible promoter, in the presence of glucose, to overexpress two enzymes capable of converting a naturally abundant source of GDP-mannose to GDP-fucose. Second, it utilizes a nucleotide-sugar transporter for the extracellular release of GDP- Fucose. The system also allows the possibility of using additional enzymes for in vivo synthesis of target molecules. In Phase I we demonstrated the feasibility of using this approach by developing a yeast strain that can produce GDP-mannose at high yields, overexpressing the enzymes necessary to convert GDP-mannose to GDP-fucose and demonstrating the ability to produce GDP-fucose at high yields. We have also demonstrated the ability to transport GDP-fucose out of the cell and have determined the initial conditions for fermentation. In Phase II we will further engineer and optimize the production of GDP-fucose and demonstrate its utility by testing the production of several important fucosylated molecules such as human milk oligosaccharides and fucosylated proteins from starting materials that are readily available to us. Finally, Phase III commercialization will involve selling GDP-fucose, licensing the system for use in a variety of applications, and using the system to produce custom fucosylated oligosaccharides, small molecules, and proteins.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Preclinical and clinical studies have shown that Photodynamic Therapy (PDT) augments the host immune response by unknown mechanisms. Previous studies, which have largely been descriptive, have shown that 1) the tumor response to PDT is augmented by an intact immune system; 2) PDT promotes the formation of immune memory cells; 3) PDT enhanced tumor immunity is mediated by T cells. We have made the novel discovery that the direct effects of Photofrin (Pf)-PDT on tumor cells is sufficient to stimulate the host anti-tumor response and that the ability to stimulate an antitumor response can be correlated to activation of antigen-presenting cells (APCs). We have also shown that alterations in tumor immunogenicity by PDT are dependent upon the inherent tumor immunogenicity and the PDT protocol employed. These data have led to the hypothesis that PDT treatment results in the expression of immune mediators that are able to activate APCs, which stimulate tumor specific T cells and trigger the initiation of a cell-mediated antitumor immune response. We further hypothesize that the ability to activate APCs is dependent upon the photosensitizer, the PDT protocol employed and the immunogenicity of the tumor. The overall goal of this proposal is to understand how PDT enhances tumor cell immunogenicity and the mechanisms that lead to the augmentation of the host immune response. In Specific Aim I we will examine the role of HSP and inflammatory cytokines in the ability of PDT to enhance tumor immunogenicity. In Specific Aim II we will examine the role of direct and secondary effects of PDT on the activation of the host immune response in vivo as a function of PDT protocol and will examine the ability of the enhanced immune response to combat distant disease. In Specific Aim III we will use transgenic mouse models and tetramer and ELISPOT analyses to determine the kinetics and characteristics of tumor specific T cells that have been activated in response to PDT. Finally, in Specific Aim IV we will expand upon our original findings that Photofrin-PDT-generated tumor cell lysates are effective anti-cancer vaccines by determining whether PDT vaccines are able to augment the host immune response against established tumors and thus promote tumor control. The studies outlined in this proposal are primarily mechanistic and will point the way toward the design of PDT protocols with optimal immune enhancing capabilities, which could lead to enhancement of long-term control of tumors within and outside the treatment field.
|
{
"pile_set_name": "NIH ExPorter"
}
|
a. Specific Aims Mitochondrial oxidative phosphorylation is a fundamental process in biological energy, transformation, and disruption of this process leads to serious human health problems, including mitochondrial myopathies, degenerative diseases, and aging. However, it has not previously been possible to measure the actual rate of electron transfer between key redox centers in this process. Millett and Durham introduced a new method to initiate electron transfer by exciting ruthenium complex with a nanosecond laser flash. This method is being used to study the pathway and kinetics of electron transfer within the cytochrome bc1 complex and cytochrome oxidase. However, the mechanism of energy coupling between cytochrome oxidase and ATP synthase is not well understood, particularly regarding the pathways of proton translocation. We propose a multi-disciplinary, collaborative approach to this problem which combines rapid kinetics techniques, bacterial model systems, and structure determination. The specific aims will be: 1. Carry out a detailed study of electron transfer in cytochrome oxidase from Bacillus firmus that combines rapid kinetics, site-directed mutagenesis, and structure determination. Major goals will be determine the pathway and kinetics of electron transfer from cyt c through CuA and heme a to the heme a3 CuB binuclear center, as well as coupled proton uptake and release. 2. Carry out a detailed study of proton translocation and ATP synthesis in the alkaliphile B. firmus OF4 to assess the significance of alkaliphile- specific sequence motifs in the membrane-bound subunits of the Ro- ATPase. 3.Use NMR spectroscopy to study the structure of the membrane-bound subunits of the alkaliphile Fo-ATPase, primarily focusing on the isolated c subunit. Potential structural manifestations of the unusual primary amino acid sequence of the alkaliphile Fo-ATPase will be investigated.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This phase I single-center, open-label, randomized study will evaluate the effects of phenobarbital on the defluorination of sevoflurane and recovery from sevoflurane in healthy male volunteers. We will also evaluate & compare immediate post-anesthetic & residual recovery effects on psychomotor performance in subjects who have or have not been subjected to hepatic enzyme induction with phenobarbital.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The millisecond time resolution of event-related potentials (ERPs) gives them a unique advantage in studying brain function, but ERP research is seriously limited by the lack of statistical methods to address the complexity and variability of ERP data. In this project, we will develop a new statistical approach to decomposition of ERP waveforms, analysis of the sources of variability, and estimation of the effects of experimental conditions and disease states. We will evaluate the new methods using simulated ERPs and many animal and human ERP data sets, including ERPs acquired from schizophrenic and stroke patients who also were studied using structural magnetic resonance imaging (MRI). The new statistical methods will combine time series modeling using the wavelet transform with nonlinear mixed effects models. Wavelet analysis decomposes ERPs by time and frequency. We have already validated our wavelet models in applications to simulated data, cat auditory evoked potentials, and human P300 potentials. Wavelet analysis separated superimposed components, yielding realistic condition effects and topographies, even in difficult cases in which principal components analysis failed. Our nonlinear mixed effects models will provide a parsimonious representation of the variability among individuals (human subjects or experimental animals) and single trials (responses to single stimulus presentations). They will yield valid significance tests and confidence intervals, extending familiar linear statistical procedures to complicated nonlinear time series. The specific aims of this project are to develop, evaluate, and apply the following statistical methods. 1. The Single Channel Wavelet Model will separate superimposed components in single channel average ERPs, and yield significance tests for condition effects on the amplitude and latency of each component. 2. The Topographic Wavelet Model will extend the single channel wavelet model to multichannel data, and provide estimated of the topography of each component. Regularization of the topography will allow analysis of ERPs from dense electrode arrays. 3. The Trial-Specific Wavelet Model will extend the single channel wavelet model to include both inter-individual and inter-trial variability, allowing estimation of the relationships among ERP components and between ERP components and trial-specific variables such as reaction time and subjective intensity.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Molecular Genetics of Retinal Degenerations Clinical Protocol support contract designed to attain extramural support for developing, designing, interpreting, and evaluating clinical trials, epidemiologic and natural history studies. In addition, it will provide for outcomes research involving eye diseases and visual disorders and some preclinical studies. The focus shall be on the design of studies and the collection, analysis, and interpretation of data emanating from these studies, as well as support, and monitoring patient safety and follow-up. Contractor shall also provide analytical and data management support, as described in the work statement, for specified clinical research data bases, cost-effectiveness and economic analyses, quality of life assessment and outcomes research. This will include, but not be limited to, the following areas: analysis of Medicare and other health care databases;evaluation of existing NEI databases such as, centralized NEI Intramural Research database, the Eye Disease Case Control Study, Early Treatment Diabetic Retinopathy Study, Framingham Eye Study, and intramural AIDS and uveitis databases. Objective of Clinical Protocol: This multinational study will investigate the inheritance of genetic retinal degeneration in families of different nationalities and ethnic backgrounds in order to identify the genes that, when altered, cause retinal degeneration. The retina is a light-sensitive membrane lining the back part of the eye. It relays vision signals to the brain, which the brain interprets into sight. When the retina degenerates, vision is altered and possibly lost. The findings of this study should help improve diagnosis and methods of treatment for these disorders. Participating institutions include: the National Institutes for Health in Bethesda, Maryland;the University of Miami in Florida;the Casey Eye Institute in Portland, Oregon;the Byrd Health Sciences Center in Morgantown, West Virginia;the University of Texas Southwestern Medical School in Dallas, Texas;the University of Tennessee Health Sciences Center in Memphis;the Prasad Eye Institute in Hyderabad, India;National Center of Excellence in Molecular Biology in Lahore, Pakistan;and the Jules Gonin Hospital in Lausanne, Switzerland.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Liver transplantation has failed to reach its full potential for saving lives due to the inadequate organ supply. Every year thousands of potentially salvageable patients die awaiting an allograft, hundreds of others receive a `marginal? donor organ, often in desperate circumstances, and a many times this number of potential recipients are never offered listing for lifesaving organs due to the need to ration this precious but limited resource. Liver xenotransplantation offers a potential solution to the organ shortage but clinical application has been stymied by four principle hurdles: 1) risk of zoonotic infection of humans, 2) the vigorous immune response mounted to xenogeneic tissues, and 3) physiologic incompatibilities due to species divergence arising in a rarity of protein: protein interactions, and if these obstacles can be consistently overcome, the 4) need to identify a clinically applicable IS regimen. A new tool with unprecedented potential to address these barriers to widespread application of xenotransplantation is found in technology such as CRISPR-Cas9 that dramatically increases gene editing specificity and efficiency. A powerful example is found in recent work by our industry partner, eGenesis, who used CRISPR-Cas9 to rid the pig genome of 62 copies of functional porcine endogenous retroviruses (PERVs), essentially eliminating the risk of PERV transmission to an organ recipient. In the current proposal, we explore the ability of advanced gene editing to address immunologic and physiologic barriers that cause immediate graft dysfunction of liver xenografts when transplanted into a translational baboon model. With the general goal preventing initial xenograft dysfunction (IXD) to gain long term survival of pig liver xenografts in baboon recipients, studies in Aim I will focus on two recently identified potential impediments to xenograft survival: 1) ischemia reperfusion injury, and 2) platelet consumption. Recent findings in cardiac xenotransplantation studies have exposed the critical contribution of IR injury to early heart xenograft demise. Our wealth of experience with ex vivo liver perfusion and also xeno liver perfusion with human blood and the recently reported technique for ischemia free liver transplants should yield a definitive answer to this question. Aim II will focus on understanding the impact of gene edits that: 1) eliminate expression of the three major antibody targets of preformed human anti-pig antibodies, 2) gain expression of human proteins designed to address complement and coagulation dysregulation occurring with porcine liver xenotransplants in NHP, and 3) mitigate innate and cell-mediated immunity and inflammation. To accomplish this, we will take advantage of liver transplant, ex vivo liver perfusion with human blood and in vitro assays well established in our lab and a panel of CRISPR modified pigs with varied expression of genetic edits.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Abstract: The majority of American women transitioning through the menopause will experience hot flashes. However, safe and effective treatments for hot flashes are currently lacking. Investigation of new treatments for hot flashes has been limited by the measurement of hot flashes. Current gold standard physiologic measures of hot flashes have key limitations, including electrode design, monitor size, and data storage capabilities that result in poor user interface, discomfort, interference in activities and sleep, and short monitoring durations. The first Phase I specific aim is to develop an improved ambulatory sternal skin conductance monitoring device that is highly wearable and unobtrusive, has increased data storage, and has a simplified user interface. This device would utilize novel electrodes, designed with the capability to be easily detached and reattached by the user. The device would include a wearable, sensor module that processes the sternal skin conductance signal. Data will be stored on a flash card and signal processing will be done to detect criterion skin conductance changes. The second phase I specific aim is to validate the measurement device in a laboratory and ambulatory setting relative to the existing gold standard physiological measurement device for hot flashes. This validation will include establishing criterion skin conductance changes and the laboratory sensitivity and specificity of device. The development of a valid, unobtrusive hot flash measurement device suitable for long-term monitoring can serve as a tool for the accurate measurement of hot flashes. This device can thereby aid in the subsequent development of effective treatments for hot flashes to improve the quality of life of the majority of women.
|
{
"pile_set_name": "NIH ExPorter"
}
|
PROJECT SUMMARY Childhood obesity is a national epidemic that disproportionately burdens low income and ethnic minority communities. By preschool, nearly one-third of low income children are already overweight or obese, setting the stage for a myriad of adverse health conditions in adolescence and adulthood. Experts agree that effective, scalable, and sustainable obesity prevention efforts are urgently needed and must shift their focus to early childhood. Preschools offer a very promising environment for implementation and wide dissemination of interventions targeting large numbers of young children. However, limited rigorous implementation research targeting low income and ethnic minority preschool age children has left a major evidence gap, severely constraining the tools available to educators and policymakers to address the obesity epidemic. Individually- focused and single-component interventions have generally failed to produce long-term changes in obesity- related outcomes. Evidence increasingly supports creating entire environments (e.g., preschools) where the healthy choice is the default option. Therefore, we propose to conduct a pragmatic cluster randomized trial, based on our Healthy-By- Default framework, to evaluate the effectiveness and implementation of a multi-level, multi-component intervention that integrates healthy eating and physical activity into preschool routines. Our intervention will involve 1) establishment of organization-level policies to support healthy nutrition and physical activity, 2) integration of nutrition and physical activity into the preschool curriculum, 3) modeling of healthy behaviors by preschool teachers and staff, and 4) engagement of parents as partners in this effort. The study is a collaboration between UCLA and the Child Care Resource Center (CCRC), a large non-profit organization dedicated to ensuring that all children receive high quality, comprehensive preschool/child care experiences, particularly in low resource, diverse communities. We will randomly assign 60 preschools served by CCRC, in underserved areas of Los Angeles County, to the intervention or a usual practice, wait-list control condition. Our aims are to: 1) implement a multilevel obesity prevention intervention for preschool settings that builds on existing evidence-based approaches, stakeholder input, and our pilot research; 2) evaluate the effectiveness of the intervention on child BMI z-scores (primary outcome), and parent-reported child nutrition and physical activity (secondary outcomes); and 3) systematically assess intervention reach, adoption, implementation, adaptation and maintenance, guided by the RE-AIM Framework, which we have tailored for our specific study focus and setting. By simultaneously evaluating both effectiveness and implementation of our intervention, we will efficiently generate new knowledge vital to informing widespread dissemination. Given the staggering long-term societal costs of childhood obesity, rigorous studies to identify effective and scalable obesity prevention interventions are needed now.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. 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. Visible light-absorbing polynuclear charge-transfer sites anchored on the pore surface of high surface area mesoporous silica afford splitting of carbon dioxide to CO. Polynuclear water oxidation catalysts can be covalently linked to metal centers embedded in the silica pore surface for oxygen evolution under visible light. Evidence for covalent anchoring on the silica surface and connection of the metal centers through an O bridge was obtained by FT-infrared, FT-Raman and optical spectroscopy. In order to reach our long term goal of coupling the photocatalytic components of the half reactions to build integrated units in these nanoporous scaffolds that reduced CO2 by H2O under visible light, a more detailed structural understanding of the redox sites is essential. Therefore, XAS studies are proposed with the goal of elucidating the coordination environment of the metal centers of the binuclear charge-transfer chromophores, and the precise structure of the linkage between the donor metal center and the catalytic core of the water oxidation catalyst.
|
{
"pile_set_name": "NIH ExPorter"
}
|
There are no approved medications for preventing relapse to cocaine abuse. Previous clinical evaluations of drug candidates have been disappointing. Stress has often been implicated in relapse to drug abuse. Converging evidence from genetic, neuroanatomical and pharmacological studies suggest that the kappa opioid receptor (KOR) system interacts with the hypothalamic-pituitary-adrenocortical axis and modulates the effects of stress on behavior. Importantly, KOR agonists have been reported to exacerbate, and KOR antagonists have been reported to attenuate the behavioral effects of environmental stressors. We have synthesized a novel, KOR antagonist, (3R)-7-Hydroxy-N-{(1S)-1-{[(3R,4fR)-4-(3-hydroxyphenyl)-3,4-dimethyl- 1-piperidinyl]methyl}-2-methylpropyl}-1,2,3,4-tetrahydro-3-isoquinolinecarboxamide (JDTic) which has shown high potency and selectivity for the KOR and potent and extremely long-lasting KOR antagonist activity. Recently, we found that JDTic was able to block footshock-induced stress reinstatement of cocaine-seeking in rats, an effect hypothesized predictive of medications which would be useful in treating cocaine relapse. These observations, and others, have led us to propose JDTic as a candidate medication for treating relapse to cocaine abuse. The purpose of this project is to evaluate four JDTic analogs identified in Project 1. In this project, the four analogs will be evaluated at least three doses each in footshock-induced reinstatement procedures using Long-Evans hooded rats with histories of self-administering 0.5 mg/kg cocaine during daily, 2-h experimental sessions. Candidates demonstrating an ability to significantly attenuate the effects of footshock-induced stress will then be evaluated in cocaine-prime reinstatement procedures to assess their specificity and to delineate their range of effectiveness. Candidates attenuating stress-induced reinstatement of cocaine seeking which minimally affect cocaine-prime reinstatement or attenuate it will be promoted onto toxicity studies for further evaluation as a backup to JDTic. JDTic is an exciting compound which offers a novel mechanism for potentially treating relapse to cocaine abuse, an unfilled therapeutic niche. This project will identify successors to JDTic in the event future studies during its development necessitate a replacement for it.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This career training proposal is to train Michael D. Twa, OD, MS as an independent clinician-scientist. A five year training program is proposed, consisting of formal coursework in vision science, specific training in computer science and image processing, and mentoring in the application of these skills to clinical outcomes research in glaucoma. In September 2003, NIH announced a new "Roadmap" to accelerate advances in biomedical research for the 21st century. Three areas listed in this Roadmap are relevant to this research proposal: (1) Interdisciplinary research training. (2) Clinical research informatics. (3) Development of enabling technologies for improved assessment of clinical outcomes. The Roadmap emphasizes coordinated strategies to develop both technological and human resources to take full advantage of multidisciplinary and translational research opportunities. This proposal addresses the stated training objectives at an individual level. Glaucoma is a leading cause of blindness. Visual field assessment and optic nerve head imaging (confocal scanning laser tomography) are commonly used to diagnose the disease and monitor its progression, yet there is considerable controversy about how to interpret and make best use of this information. Currently, raw data from these observations are reduced to statistical indices that are meant to summarize clinically meaningful features and provide a basis for classifying test results as normal or not. Unfortunately, these indices may sacrifice other relevant features in the data for interpretability. We will use mathematical modeling methods (polynomial modeling, spline fitting and wavelet analysis) to quantify patterns in visual field data and topographic images of the optic nerve head. We will use features derived from these modeling methods to apply novel pattern recognition techniques from computer and information sciences-decision trees and non-linear regression analysis-and then compare these techniques to current methods to identify glaucoma. By improving current methods of analysis we can provide a more quantitative basis for clinical decisions, and offer greater consistency and objectivity on data interpretation. The long-term objective of this proposal is to translate advances in computer and information sciences to the analysis of clinical outcomes research in glaucoma and other eye diseases.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The overall aim of the proposed program is to develop, fabricate and clinically test an easy-to-use affordable diagnostic instrument that can non-invasively detect subsurface inflammations and estimate the temperatures of such inflammations. The design of this instrument will be based on a novel fast-reading, self-calibrating, dual-frequency digital radiometer developed under an SBIR grant from NIH for use in two-dimensional eonformal hyperthermia array antennas. By taking advantage of recent advances in micro-miniaturization of microwave devices, the front-end of the radiometer will be integrated with the receiving antenna. This will simplify the radiometer circuitry, increase its sensitivity, and reduce calibration and acquisition times. The examining health professional will be able to move the small, integrated antenna assembly (Thermoscope) to any point on the body of a patient with the same ease as moving a stethoscope and quickly and accurately determine skin temperatures and two subsurface tissue temperatures averaged over two different tissue depth. The specific aims of the proposed Phase I program are to design, fabricate and test an experimental model of a Thermoseope equipped self-calibrating diagnostic dual-frequency digital microwave radiometer and write software for the radiometer that will enhance its diagnostic uefulness.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Fibrosis leads to organ dysfunction and is characterized by an excessive production of extracellular matrix (ECM) components, namely type I and III collagens. Although activated hepatic stellate cell (HSC) remains the principal cell type responsible for liver fibrosis, other liver cell type of fibroblast lineage derived from portal and centrolobular veins also have fibrogenic potential. The overall hypothesis is that liver fibrosis can be treated by systemic administration of a1(I) collagen gene promoter specific triplex forming oligonucleotides (TFOs) conjugated with mannose 6-phosphate-bovine serum albumin (M6P-BSA) via a disulfide bond. In preliminary studies, antiparallel phosphorothioate polypurine TFOs specific for CI region formed triplexes, inhibited transcription of a1(I) collagen promoter and improved rat liver fibrosis. TFOs rapidly distributed throughout the body after systemic administration, with the highest accumulation in the liver. TFO accumulation in the liver was decreased when injected into liver fibrotic rats. Kupffer, sinusoidal endothelial and hepatic stellate cells accounted for approximately 70% of the liver uptake, and the remaining 30% in the hepatocytes. Bioconjugation with M6P-BSA significantly enhanced the cellular uptake of the TFOs by HSC-T6 cells in vitro, leading to enhanced inhibition of type a1(I) collagen transcription. TFO delivery to the liver and to the HSCs was significantly increased when M6P-BSA-TFO was injected intravenously into fibrotic rats. Our specific aims are to determine whether i) conjugation of targeting ligands to TFOs affect triplex formation; ii) TFOs inhibit fibrosis by inhibiting transcription of a1(I) collagen and/or blocking inflammation and activation of liver fibrogenic cells; and iii) M6P-BSA-TFO can be delivered efficiently to liver fibrogenic cells and prevent fibrosis. The significance of this research is that the proposed targeted TFO delivery to liver fibrogenic cells will inhibit disproportionate accumulation of a1(I) collagen, which is essential for the treatment of liver fibrosis. The data will also be beneficial to successful treatment of other organ fibrosis. [unreadable] [unreadable] [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
Two of five abnormal protein bands in sera of febrile individuals appear to share antigenic determinants with the Amyloid A protein extracted from tissues of patients with chronic inflammatory disorders. This relationship will be further explored by paired electrophoretic and immunologic studies. The possibility that two further bands are antigenically related to immunoglobulin light chains will be tested by immune-absorption from purified fractions on anti-kappa and anti-lambda columns. Antigenic comparison between the most cathodal abnormal protein in serum and urine and a similar protein in CSF will be made using antiserum raised against the purified urinary protein. Tests for pyrogenicity of purified protein fractions from serum and urine will be made with the assistance of Dr. Phyllis Bodel of Yale University.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The major focus of this project is to evaluate and treat patients with malignant brain tumors. Treatment protocols are designed with the specific input of laboratory-based researchers. This clinical-laboratory interaction allows the rapid introduction of novel treatment strategies with the intended long-range goal of improved survival and quality of life for patients with primary malignant tumors of the central nervous system. Modalities of therapy will include surgery, gamma knife radiosurgery, interstitial brachytherapy, radiation therapy, radiosensitizers, chemotherapy, and gene therapy alone or in combination. The approach is multidisciplinary and includes the expertise of neurosurgeons, neuro- oncologists, neurologists, radiotherapists, medical oncologists, neuropathologists, clinical research nurses, and laboratory-based scientists. Protocols are jointly developed with clinical and laboratory researchers and the data captured and analyzed with the support of biostatiticians and data managers. For this renewal period, we will emphasize the use of radiation in the treatment of these patients. Protocols in progress or under development include randomized trials with interstitial brachytherapy and hyperthermia, the use of experimental radiosensitizers such as the halogenated pyrimidines and the polyamine inhibitors, treatment using altered fractionation schemes such as hyperfractionated radiotherapy and accelerated hyperfractionation, and radiosurgery using the gamma-knife. Adjuvant chemotherapy and phase II drug studies, including gene therapy, for recurrent tumors will also remain as an integral component of our total approach to the patient. Correlative data support for laboratory studies that are a part of this grant will come from clinical trials. Tumor and normal tissue stored in the tissue bank will serve as the source of material for other projects in this grant that will evaluate the radiosensitivity and molecular basis for radiation response. The clinical laboratory correlations that develop will serve as the substrate for newer, more individualized specific treatment recommendations for patients that will hopefully control disease with minimal or no toxicity.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This is a continuing study in comparative oncology, designed to identify and characterize neoplasms in lower animals including cold-blooded vertebrates and the invertebrates. By studying neoplasms in animals phylogenetically more remotely removed from man, it is hoped to gain a more complete understanding of the basic biology of neoplasia at all phylogenetic levels. It is expected that such studies will reveal properties of neoplasia not previously suspected to exist because of the excessively narrow scope of studies limited to the higher animals and man. Important differences in the immunologic, biochemical, and regenerative characteristics of the lower animals should serve to emphasize the relevance of these characteristics to the development of and the susceptibility to neoplasia. In addition, by study of neoplasms in lower animals under feral conditions, these neoplasms may be found to serve as indicators of environmental tumorigenic agents. Neoplasms are collected, either in the field or through submittal from outside sources and are studied morphologically and biologically. Light and electron microscopy, tissue culture, transplantation, and tumor induction methods are applied.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Research programs in this Center are maintained to implement our mission in Primate Behavior, Reproduction, and Neuroscience. The research related to each of these major components of the mission is distributed over three divisions comprising more than one laboratory or professional core staff appointment within the Center. Thus, work on Primate Behavior includes studies on social organization and communication, Reproductive Behavior, and behavioral adaptations to natural environments. Research on reproduction includes studies of folliculogenesis, implantation, corpus luteum of pregnancy, and endocrine events at parturition, pituitary function during different reproductive stages of male and female, and steroid biosynthesis and metabolism in target tissues. The Neurosciences are represented by research on neurophysiological correlates of pituitary activity and of feedback actions of steroids on neural tissues, studies of the ultrastructural localization of releasing factors, and studies of steroidal influences on brain enzymes and catecholamines.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Complex biological systems within a single cell are governed by complex coupled nonlinear dynamical processes. Such systems respon to perturbations in nonintuitive ways and can be best understood by synthesizing a coherent description of the signaling system as a set of coupled nonlinear ordinary differential equations that can be solved numerically. Such a system can be used to synergistically design experiments to confirm and extend our theoretical understanding of the biology. This core module consists of a dedicated server that can accessed by COBRE participants for use in computer modeling to help them better predict the results of there experiments.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Adolescence is a developmental stage in humans that is characterized by dramatic changes in an individual's biology and their behavior. It is also a period during which individuals may begin using psychostimulant drugs, whether for therapeutic or recreational purposes. Repeated exposure to these drugs is associated with deficits in memory, decision making, impulse control, and reward processing, and these adverse consequences on cognition may persist through extended periods of drug abstinence. Thus, it is critically important to understand the neurobiological processes that mediate drug-induced changes in behavior and to determine how adolescents, compared to adults, are particularly vulnerable. Our long-term goal in these studies is to understand the neuroadaptations induced by amphetamine in corticolimbic regions of the adolescent brain and determine how these changes can be prevented or reversed. In the proposed studies, we will use behavioral, pharmacological, and electrophysiological techniques in animal models of adolescence and adulthood to address two aims. In Aim 1, we will determine if changes in dopamine and NMDA receptor function in the mPFC are responsible for the enduring deficits in cognitive behavior induced by amphetamine exposure during adolescence. In Aim 2, we will determine the basis of the long-lasting functional changes in mPFC neurons that are observed in adolescent- compared to adult-exposed individuals. Our working hypotheses are that, 1) adolescent-exposed rats, when tested as adults, will be more sensitive to drug-induced deficits in cognitive function and to selective manipulations of dopamine and NMDA receptors, compared to those exposed as adults; 2) the effects of repeated amphetamine treatment on the intrinsic firing properties, NMDA-dependent long term potentiation, and dopamine receptor-mediated responses of mPFC neurons are enhanced in adolescent- compared to adult-exposed individuals; and 3) the effects of this exposure on the in vivo responses of mPFC neurons to amphetamine and dopamine or NMDA receptor selective drugs will be greater in adolescent- compared to adult-exposed individuals. These hypotheses are consistent with our preliminary studies, which show that that exposure to amphetamine during adolescence impairs behavior on an mPFC-sensitive working memory task and alters the intrinsic firing properties of layer V pyramidal cells recorded in vitro. Through the research proposed in this application, we seek to fill the large gaps in our knowledge about what makes the brain and behavior of adolescence so uniquely different from adults and increases their vulnerability to the adverse consequences of repeated drug exposure. By understanding the unique plasticity of the adolescent brain, we will likely identify targets for preventative or therapeutic strategies aimed at ameliorating the adverse consequences of repeated amphetamine exposure during adolescence. In addition, we anticipate our results will move the field towards a clearer understanding of the unique effects of psychostimulants during this critical period of neural and behavioral development.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The major objective of the proposed research is to conduct in-depth studies on the mode of action of several compounds representing three groups of insecticide synergists (substituted methylenedioxybenzenes, 1,2,3-benzothiadiazoles and imidazoles) known to act by inhibition of microsomal mixed-function oxidation. In vitro studies will be carried out with microsomes from control and induced phenobarbital, 3-methylcholanthrene) rat livers and midgut tissues from control and induced (pentamethylbenzene) armyworm (Spodoptera eridania) larvae. Emphasis will be placed on studying the formation of irreversible, inhibitory product adducts with cytochrome P-450 in NADPH-reduced microsomes and on the possibility that the synergists may interact with specific types or pools of cytochrome P-450. The formation of product adducts will be studied by difference spectroscopy and/or the binding of appropriately labeled synergists. The properties of any rsidual "non-reactive" cytochrome P-450 will be studied by enzymatic assay epoxidation, hydroxylation and N-demethylation and a variety of spectral techniques. In vivo studies with rats (or mice) and armyworm larvae will be conducted to determine the formation and reversibility of synergist-cytochrome P-450 complexes in the intact animal and to evaluate the effects of these complexes on microsomal oxidation. Equilibrium dialysis will be used to study the binding of labeled 1-phenylimidazole to microsomes and to evaluate this ligand as a potential tool for qualitative and quantitative measurements of different types of cytochrome P-450 in intact microsomes.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Rho family GTPases Rac, Rho and Cdc42 are central coordinators of the signal transduction pathways that regulate the actin cytoskeleton, cell migration, gene expression and cell proliferation. Our past work demonstrated that integrins, which are the major receptors for extracellular matrix proteins, regulate Rho family GTPases in two ways. First, integrins controls GTP loading (i.e., activation). There is a separate level of regulation in which integrins control membrane targeting of the activated GTPases. Both steps, activation and membrane targeting, are essential for GTPases to activate downstream effectors. In the current grant application, we propose to pursue two projects to elucidate these two mechanisms. First, we will study a new nucleotide exchange factor for Cdc42 that we have recently cloned. It's protein binding partners, mode of regulation and roles in signal transduction will be investigated. Second, we will investigate the mechanism by which integrins control the membrane targeting of active Rac. The project will be based on preliminary data suggesting that effects of integrins on lipid rafts and/or caveolae mediate this effect. Thus, the regulation of lipid rafts/caveolae by integrins and their role in Rac signal transduction will be investigated. These studies will contribute to our fundamental understanding of how integrins regulate Rho family GTPases. These regulatory events are central to cell migration, growth and function in normal physiology and human diseases.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This proposal aims to contribute to an understanding of the organization of elementary genetic units in higher organisms and the regulation of the expression of such units in development. The rosy locus of chromosome 3 in Drosphila melanogaster carries the structural information for a peptide which, as a homodimer, functions as xanthine dehydrogenase in this organism. Mapping experiments have served to map the borders of the structural element, and variants which map outside and just to the left of the structural element appear to be associated with regulation (i.e., they are alterations in a cis acting control element. The direction of experiments in the current proposal are (1) mutagenesis experiments designed to make several classes of control variants, (2) analysis of such variants, and (3) examination of polygenic clustering about the rosy locus. BIBLIOGRAPHIC REFERENCES: Chovnick,A., M. McCarron, W.M. Gelbart and J. Pandey. 1975. Electrophoretic variants as a tool in the analysis of gene organization in higher organisms. In Ioszymes IV. Genetic and Evolution. Ed. C. Markert. Academic Press, New York. pp. 609-622. Duck, P. and A. Chovnick, 1975. Resolution of an equivocal genetic element in Drosphila melanogaster: organization of the maroon-like locus. Genetics 79:459-466.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The main theme of the UNMC Nanomedicine Development Center is the protein misfolding phenomenon. Protein misfolding leads to spontaneous formation of protein nanoparticles of different morphologies, nanofibrils in particular. Misfolding and aggregation of proteins is a common thread behind a number of important human health problems. A fundamental understanding of molecular processes leading to misfolding and self-aggregation of proteins involved in various neurodegenerative diseases will provide critical information to help identify appropriate therapeutic routes to control these processes. Despite crucial importance of the protein misfolding and its self-assembly into aggregates, very little is currently known about the molecular mechanism underlying these processes. Factors that lead to protein aggregation are poorly understood not to mention the formation of the protein nanoparticles with different morphologies, the nanopores in the first place. It is unclear why aggregation even of the same protein sometimes leads to fibrillar, pore-like and amorphous aggregates and how aggregation may be prevented. Understanding the molecular mechanism of aggregation could have tremendous practical importance. It will facilitate the rational design of approaches aimed at inhibiting and controlling protein aggregation, modeling and manipulating the aggregation process within cells. The ability to control these processes increases the efficiency of drugs delivery, and eventually will lead to prevention and cure of protein deposition diseases. To achieve these goals we will develop novel nanotechnology based approaches for observation and measuring the interaction between individual molecules, manipulating with interacting molecules in vitro and will develop practical solutions for controlling protein aggregation process within cells. Our ultimate goal is the development of nanomedicine approaches to repair defective machines or to replace them with artificial nanosystems. We assembled a multidisciplinary team of 12 investigators with broad expertise in nanobiology, imaging, spectroscopy, single molecule biophysics, proteomics, drug delivery and medicine.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Core E has several investigators with extensive expertise in drug delivery, synthetic/bioconjugation chemistry, and nanotechnology. The central function of Core E is to provide a tool box comprised of various types of delivery systems to accommodate different radiation protectors/mitigators that either have been or will be developed from different research projects in our CMCR. Specific services will be provided to 1) improve the bioavailability of testing compounds that are non-orally active and poorly water soluble by formulating them in nanoparticulates for testing of in vivo activity and toxicity; 2) selectively deliver radiation mitigators to the cytoplasm or mitochondria of critical cell types in vital organs through the use of lipidic systems or lipid-coated biodegradable carbon nanotubes; and 3) to achieve controlled/slow release of the active ingredients through either the emulsion particles with an oil core that is made of fat with high melting temperature or through triggered release from the nanotubes
|
{
"pile_set_name": "NIH ExPorter"
}
|
The ClpAP protease from E. coli is composed of a proteolytic component (ClpP) and a regulatory ATPase component (ClpA). Ultracentrifugation has been conducted at pH 7.5 (0.1-0.3 M KCl +/- 10% glycerol) and 4 or 20 degreesC to determine the macromolecular size and shape of the ClpAP components. Without nucleotide, ClpA (subunit MW of 84,150) exists in a monomer-dimer equilibrium with log K = 5 (apparent s/20,w = 8.7 S). With the non-hydrolyzable analogue adenosine-5'-O-(3-thiotriphosphate) (ATPgammaS) present, ClpA associates to a hexamer with MW = 505,000 +/- 3000 and s/20,w = 17.2 S. The inactive pro-ClpP(SA) (13.2 S) and mature ClpP (12.2 S) have identical subunits of 23,164 and 21,558 MW, respectively, and appear as two rings of seven subunits in electron microscopic image reconstructions (M. Kessel et al., J. Mol. Biol. 250, 587-594, 1995). Ultracentrifugal studies confirm that both the pro-Clp(SA) mutant and ClpP are 14-mers with molecular weights of 324,000 and 302,000 +/- 5%, respectively. Nucleotide-bound, hexameric ClpA is required for association with ClpP to form the active ClpAP protease; log K = 8.5 /(M ClpA hexamer or M ClpP tetradecamer) under assay conditions. Sedimentation velocity experiments in the presence of ATPgammaS, Mg(II), and varying molar ratios of hexameric ClpA to tetradecamer ClpP show two complexes: s/20,w = 21.0 +/- 0.4 S and 27.3 +/- 0.7 S. These correspond to 1:1 and 2:1 complexes of hexameric ClpA and oligomeric ClpP, with MW approximately 807,000 (frictional ratio = 1.6) and MW approximately 1,310,000 (frictional ratio = 1.7), respectively. Both complexes are active in casein or propeptide degradation and both are seen in electron micrographs. Recently, it has been established that ClpP undergoes a temperature- dependent, reversible dissociation of the two rings (containing seven subunits/ring) in 0.1 M sodium sulfate at pH 7. At 5 _C, the protein sedimented as a monodisperse boundary corresponding to the size of half molecules of ClpP (s/20,w = 7.9 S). Warming from 5 to 20 _C, produced complete reassociation of ClpP to its original size (s/20,w = 12.2 S). Cooling from 20 to 5 degreesC and centrifuging at 52,000 rpm within 1 h, produced two boundaries (approximately 8 and 12 S/20,w), which suggests that dissociation is slower than reassociation kinetics. Dr. Zolkiewski has initiated studies on the oligomeric structure of E. coli ClpB, a protein homologous to ClpA. ClpB, like ClpA, has protein activated ATPase activity but unlike ClpA, ClpB does not interact with ClpP. The sizes of ClpB complexes at different protein concentrations in the absence and presence of nucleotides (ATP, ATPgammaS, and ADP) are being determined by sedimentation velocity, sedimentation equilibrium, HPLC, gel filtration, and chemical cross linking with glutaraldehyde.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Regulation of intracellular activities by a variety of extracellular signals often involves cell-surface receptors which interact with guanine nucleotide-binding regulatory proteins (G-proteins). It is generally known that G-proteins regulate other proteins, yet little is known about the regulation of the expression of G-proteins themselves. The purpose of this project is to investigate how the levels of G-protein expression may be regulated in select preleukemic (K562, U937, and HL-60) cell lines, and whether altered levels of specific G-protein subunits correlate with physiologic end-points such as cell growth, proliferation, or differentiation. This correlation could provide insight on defining the molecular mechanisms underlying changes in cellular concentrations of G-protein subunits. Western immunoblotting will be used to identify both the types and relative amounts of G-proteins expressed in K562, U937, and HL-6O hematopoietic cells. Northern blotting, western immunoblotting, nuclear run-on assays, and other molecular techniques will be performed to establish which G-proteins and their transcripts are expressed as different growth conditions are imposed on the cells.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Glioblastoma is a highly invasive tumor with a recurrence rate of 98%, which is in most cases rapidly fatal. The failure of current clinical and experimental therapies to eradicate disseminated glioma cells results in tumor recurrence and a median survival of 3?6 months. Furthermore, currently available imaging technologies are not sufficiently sensitive for detection of small tumor satellites, which are responsible for recurrence. Recent research advances have established mesenchymal stem cells (MSCs), neural stem cells (NSCs), and neural progenitor cells (NPCs) as promising therapeutic delivery vehicles for gliomas. These stem cells track down cancer cells in the tumor mass as well as migratory cancer cells into the brain parenchyma. Because of this unique ability, the use of MSCs and NSCs/NPCs as therapeutic and imaging modalities has been explored. Several recent studies demonstrated many advantages of NSCs over MSCs. Specifically, NSCs have a unique ability to suppress tumor growth in vivo, and exhibit greater and more specific tropism to gliomas. The disadvantage of NSCs is the lack of an adequate NSC source. Current alternative approaches to produce NSCs by induced pluripotent stem cell (iPSC) technologies are fraught with incalculable risks. Recently, we have been able to generate neural stem/progenitor cells from human bone marrow and adipose derived MSCs using a purely chemical approach (or small molecule approach) that has several advantages over currently known technologies. Our recent studies demonstrated that these chemically induced neural progenitors (CiNPCs) exhibited higher migratory capacity to malignant glioma D54-GM cells compared to hMSCs which was further increased by pre-exposure of these cells to D54-MG conditioned medium (GCM). Thus, CiNPCs produced by our recently developed safe, fast, reproducible, and cost-effective reprogramming approach can be efficiently used as a drug delivery vehicle for the treatment of brain tumors and/or as carriers for magnetic resonance imaging (MRI) contrast agents for visualization of gliomas. The goal of this SBIR phase I proposal is to investigate whether GCM pre-exposed and magnetically labeled CiNPCs could be utilized for visualization of intracranial D54-MG glioma cells by MRI. Phase II studies will investigate the potential use of these cells for MRI visualization of different types of gliomas. Commercial and clinically compatible research products emerging from Phase I/II work includes technology for large-scale clinical grade production of magnetically loaded CiNPCs that can be used for visualization of different gliomas by MRI. To achieve these goals the following Specific Aims are proposed for Phase I studies. Specific Aim 1 will test the hypothesis that GCM pre- exposed CiNPCs loaded with ferumoxide-protamine sulfate complex (FE-Pro) will retain their viability and tropism to D54-MG cells in vitro. Specific Aim 2 will test the hypothesis that GCM pre-exposed and FE-Pro- loaded CiNPCs will exhibit tropism to D54-MG glioma xenografts in vivo, which can be monitored by MRI.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Gender disparity has been recently reported for the severity of inflammatory bowel disease (IBD), with female patients with Crohn's disease (CD), one of the idiopathic IBDs, likely to experience more severe disease manifestations compared to males, and with symptom activity associated with hormonal surges during puberty, pregnancy and the post-partum period. Although the precise etiology is currently unknown, it is well accepted that IBD results from a dysregulated immune response to environmental factors (e.g., gut microflora) in genetically-predisposed individuals. However, the mechanism(s) underlying sex differences in IBD is an understudied area of investigation and little is known regarding how the orchestration of genetics, host immunity, as well as the gut microenvironment leads to gender disparity, specifically in Crohn's patients. Based on our extensive preliminary data, the central hypothesis of the present proposal is that gut microbial ligands signaling through TLR9 promote phenotypic and functional diversity in mucosal Treg populations that contribute to sex differences in experimental CD, such as that observed in SAMP mice, a spontaneous model of CD-like ileitis, and in patients with CD. This hypothesis will be tested by performing two specific aims: 1) Determine the impact of the gut microbiota on sex differences in the phenotypic and functional properties of the mucosal Treg population/subpopulations, and 2) Determine the mechanism(s) as to how TLR9 activation specifically results in sex-associated gut inflammation. The proposed studies are designed to elucidate potential mechanism(s) underlying sex differences in spontaneous, experimental Crohn's-like ileitis, and identify potential, early diseas pathways that can lead to the development of novel, gender-targeted strategies to improve treatment of patients suffering from this devastating disease. PUBLIC HEALTH RELEVANCE: Crohn's disease (CD), one of the idiopathic inflammatory bowel diseases (IBD)s, is characterized by a chronic, relapsing inflammation of the GI tract that is generally thought to result from uncontrolled immune responses to environmental factors in genetically predisposed individuals. Recent evidence shows that clear gender disparity in CD exists, with females displaying more severe disease manifestations than males that is associated with hormonal surges during puberty, pregnancy and the post-partum period; however, sex differences in mucosal immunity, in general, and IBD, in particular, is an understudied area of investigation. The present proposal will investigate one potential mechanism that gut microbial ligands promote phenotypic and functional diversity in mucosal Treg populations that contribute to sex differences in the pathogenesis of chronic intestinal inflammation, such as that observed in IBD.
|
{
"pile_set_name": "NIH ExPorter"
}
|
It is the purpose of the proposed research to develop and test specific hypotheses regarding the central control of desynchronized sleep (D) at the level of single neurons by the method of chronic extracellular microelectrode recording. Results in hand indicate that the giant neurons of the cat pontine tegmentum (FTG), a region whose intrinsic anatomy and experimental destruction had suggested a role in D sleep, satisfy better than other cell groups three physiological criteria of a central control system: (1) greater selectivity of discharge during D; (2) earlier anticipatory tonic rate change in transition to D; and (3) earlier anticipatory phasic rate change in association with the eye movements of D. Long-term recordings show FTG neuronal activity to be (1) consistently selective across repeated D cycles; (2) stereotyped, for each cell, within each cycle; and, (3) periodic when a series of cycles is considered. We have recently discovered that other cells, in the region of the locus coeruleus, fire in a tonically reciprocal fashion with respect to the FTG. All of these findings together suggest that FTG neurons function as output elements in an oscillatory brain stem system during D sleep. We plan to extend these findings and to test their implications in several new ways. First, we propose to define more precisely the axonal domain of the FTG by anatomical and physiological techniques. Second, we propose to make simultaneous recordings from two or more individual neurons within and across the cell fields that we presume to be interacting. Third, we propose to combine lesion and stimulation techniques with extra-cellular unit recording to clarify the nature of the interaction between elements in the control system.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This proposal concerns the mechanisms by which thin films of pulmonary surfactant form solid structures in the lungs. Surfactant films stabilize the small air spaces of the lungs by lowering the surface tension of the thin liquid layer that lines the alveoli. When compressed by the decreasing surface area during exhalation, the surfactant film reduces surface tension to extraordinarily low levels. In vitro, compressed monolayers containing the complete set of surfactant constituents fail to reach such low surface tensions because above an equilibrium density, constituents collapse from the interface. The ability of the two dimensional films to flow into the third dimension indicates fluid behavior. In the lungs, the persistence of the surfactant films at the interface when compressed to densities well above equilibrium indicates a solid structure. The classical model of surfactant function contends that only a film in the highly ordered tilted-condensed (TC) phase can be compressed to the low surface tensions observed in the lungs. Of the components in lung surfactant, only its most prevalent constituent, dipalmitoyl phosphatidylcholine (DPPC), can form the TC phase at physiological temperatures. Therefore the classical model explains the transformation of the surfactant film from fluid to solid structures in terms of a change in composition, with elimination of constituents other than DPPC, and a consequent transition between equilibrium phases. Our recently published and preliminary unpublished data, however, disagree with the predictions of the classical model. Our results suggest instead that the fluid surfactant films transform to solid structures by a process analogous to the supercooling of three dimensional liquids to form glass. The experiments proposed here will test first if fluid films transformed to solid structures by supercompression in vitro replicate the behavior of pulmonary surfactant in physiological settings, and then the extent to which the supercompressed fluid monolayers fit predictions of the analogy with supercooled liquids.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The present research project has two specific aims: (1) to gather information using survey methods concerning the development of self-awareness and empathic tendencies during middle and late adolescence, and (2) to determine how these personality characteristics add to the current understanding of personal and social adjustment in adolescence. As such, this proposal not only represents the first longitudinal study of self-awareness and empathic tendencies, but also is the first attempt of its kind to test theoretical links between these parameters and measures of adjustment having to do with parent-child and peer group relations. It is proposed that dispositional self-awareness and empathy indirectly affect adolescent adjustment through their effect on interpersonal behavior known to be related to adjustment. The proposed program of research would investigate this general model over a four year period of time employing as measures of adjustment the adolescents' self-reports of loneliness, sociometric indexes of friendship networks within the school, and subjective versus objective perceptions of self. Preliminary results of the first year's measurements of adolescents in the proposed target high school are discussed, with specific attention given to the effects that self-awareness and empathy have on loneliness.
|
{
"pile_set_name": "NIH ExPorter"
}
|
There are three classes of recoding: ribosomal frameshifting, stop-codon readthrough, and bypass of internal mRNA region. Recoding instructions may include an RNA structure within the mRNA. These structures can kinetically alter the mRNA passage through the ribosome, can interface with the ribosome directly, and can bind proteins or other co-factors and bring them into place to direct the recoding event. Within the universal code, the UGA codon signals the termination of protein synthesis. This event can be circumvented in three ways, including co-translational insertion of selenocysteine at this triplet (a special case of readthrough). For selenocysteine insertion at stop codons in eukaryotes, there are four requirements: (1) two enzymes essential for the synthesis of the selenocysteine amino acid; (2) a special tRNA; (3) an RNA stem-loop structure, termed the selnocysteine insertion element (SECIS), is required in the 3'-UTR of the mRNAs and; (4) a protein which may act as the homologue to the prokaryotic SelB protein (SBP) is thought to bind the SECIS and program the ribosome for selenocysteine insertion. An NMR structural investigation of this structure will help to understand how an element in the 3'-UTR of a specific mRNA "informs" ribosomes translating that same mRNA that all UGA codons, with the exception of the terminal codon, should specify selenocysteine insertion. Two models of the SECIS secondary structure are available. Both models identify two stems separated by an internal loop, in addition to an apical loop. Phylogeny identifies three consecutive adenosines in loop II, and a run or 4 base-pairs in the "core" region that are essential for the function of SECIS. The base pairing register of stem II differs in the two models by one nucleotide including two tandem G-As which are proposed to be formed using N6-A to the N3-G, and a N7-A to N2-G H-bonds, allowing room for the restricted geometry of the U-U basepairs that flank the tandem. These intriguing helical arrangements are ideal sites for specific recognition of the SelB homologue (SBP) protein for eukaryotic selnocysteine insertion. A43 nucleotide RNA corresponding to stemII/loopII has been produced for NMR studies using runoff T7 transcription in conjunction with double ribozyme cleavage to produce homogeneous RNAs. Simino resonances in the 1D 1H spectra of this RNA lie within the chemical shift range expected for G-A base pairs. This attribute strongly suggests model B to be more consistent with the stemII/loopII region of this SECIS RNA. A smaller RNA has been purified, which corresponds to stemII with a UUCG tetraloop replacing loopII, to simplify the assignment of stem resonances. This RNA has indirectly confirmed the base pairing register to be consistent with model B, by forcing this stem arrangement using the tetra-loop junction, and observing a nearly identical imino spectrum. Homonuclear 2D 1H data have suggested the same base-pairing arrangement of model B, but have been limited by degeneracies in the 1H spectra. Isotopic enrichment of these RNAs will be essential for more complete assignments required for the determination of a high resolution structure. These include complete random isotopic enrichment of RNAs in both 15N and 13C, and selective enrichment of specific nucleotides.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This is the second competing renewal application for Yale University's Vascular Research Postdoctoral T32. Of fellows who have completed training to date, 86% have obtained academic faculty positions, and 86% have entered independent scientific careers, strong indicators of program success. The continued goal is to provide laboratory and translational research training for highly qualified physician (M.D. and M.D. /Ph.D.) and Ph.D. postdoctoral fellows in vascular biology, in preparation for careers as independent investigators in blood vessel biology- and medicine-related disciplines. Selection of trainees will be based on a commitment to vascular biology and strong prior research experience or potential of same. Applications will be encouraged from clinical and basic science departments, with a nationwide competition for 7 yearly slots. Minority applicants will be specifically solicited. The training will be mentor-based, also including advisory committees and didactic courses. The minimum duration of training will be 2 years, with the majority of fellows remaining in the program for 3 years. The Cardiovascular Medicine Division and Yale's Interdisciplinary Program in Vascular Biology and Therapeutics will be the foundations for the program. A key asset to this training program is Yale's interdisciplinary strength in vascular biology. Departmental affiliations for participating faculty include Cardiovascular Medicine, Pulmonary and Critical Care Medicine, Immunobiology, Pharmacology, Molecular Cellular and Developmental Biology, Pathology, Genetics, Bioengineering, Physiology, Epidemiology and Public Health, and Cardiothoracic Surgery. This is a testimony to the wide spectrum of strong vascular biology laboratories at Yale and the program's institutional nature. The faculty was chosen based on impressive histories of mentorship, ongoing vascular research productivity, strong extramural support and commitment to serve as mentors within the program. Examples of research opportunities include: (1) molecular determinants and consequences of leukocyte-endothelial cell interactions; (2) molecular imaging of angiogenesis and vascular remodeling utilizing nuclear and MR imaging in animal models; (3) mapping and identification of genes that contribute to the development of vascular disease (arterio-venous malformation, coronary artery disease) in humans; (4) engineering of vascular biomaterials, and molecular determinants of healing responses post-implantation; and (5) generation of angiogenic gene regulators for use in clinical trials. This represents a wide range of disease-related vascular research, with key translational components. Trainee progress will be monitored by each mentor, the trainee's advisory committee and the Program Director. It is the expectation that we will train future national and international leaders in vascular research. Cardiovascular disease is the leading cause of death in the U.S., and a major cause of mortality world-wide. This program will provide important opportunities for individuals from multiple disciplines to eventually lead high impact efforts at reducing the incidence of, and improving outcomes in, cardiovascular disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Cerebral amyloid angiopathy (CAA) is an age-associated condition in which amyloid is deposited in the medial layer of primarily small- and medium-sized arteries and arterioles of the cerebral cortex and leptomeninges and is present in most patients with Alzheimer's disease (AD) and certain related disorders including hereditary cerebral hemorrhage with amyloidosis Dutch-type (HCHWA-D). The overall hypothesis of this proposal is that age and other risk factor-related changes in cerebrovascular smooth muscle cells renders them more susceptible to the pathogenic effects of Abeta resulting in CAA-related pathology. We plan to utilize novel and unique cell culture and transgenic model systems to investigate the role of age and other risk factors in the development of the smooth muscle cellular pathology of CAA. The studies outlined in this proposal will shed light on the roles of specific molecules, and age-related changes in these molecules, that could contribute to CAA pathology. The specific aims are as follows: First, we will determine if cultured cerebrovascular smooth muscle (CSM) cells from younger and aged comparisons with cultured CSM cells from young and aged non-human primates that develop pronounced or mild CAA (squirrel monkey or rhesus monkey, respectively). Second, we will determine the pathogenic effects of Abeta in cultured CSM cells obtained from control non-transgenic and wild-type or HCHWA-D mutant human AbetaPP transgenic mice. This will ascertain if over-expression of human Abeta PP affects the pathologic leads to earlier and/or more extensive cerebrovascular Abeta deposition and accompanying CAA-related pathology in our human Abeta PP transgenic mice. Lastly, we will determine if over-expression of RAGE influences the pathogenic effects of Abeta in cultured CSM cells obtained from RAGE transgenic mice and RAGE/human Abeta PP pathologic responsiveness of these cells to Abeta. These combined investigations will provide information about how age and changes in key molecules in CSM cells are involved in the development of CAA-related pathology.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Queens Cancer Center (QCC) Minority-Based Community Clinical Oncology Program (MBCCOP) is the first National Institute of Health (NIH) funded grant, awarded to the public hospital system of New York City known as the Health and Hospital Corporation (HHC). QCC is part of the Queens Health Network (QHN) which is comprised of the only two public hospitals in Queens, Queens Hospital Center (QHC) and Elmhurst Hospital Center (EHC). Since the population of QHN is 92% minorities, we are uniquely qualified for this minority program. Since its inception in 2002, the QCC has been dedicated to bringing the best possible cancer care to the people in Queens who were previously underserved, including minority patients, recent immigrants and patients without insurance or means. The population served by the Queens Health Network has an unfortunate history of late stage cancers of all kind and. poorer survival. The mission of QCC is to reduce the rate of late stage disease and to improve the care for cancer. The strategies to accomplish this includes education of the community, introduction of cancer control trials, outreach for early detection programs and state of the art cancer care including access to cancer treatment trials. The patient population of QHN is overwhelmingly from minority groups who usually do not have access to clinical trials. The QHN is the only health facility in Queens, a borough more than 2 million people that offers access to multi-institutional treatment trials. Since the MBCCOP was awarded (September of 2010) the infrastructure was put in place to combine the resources of both centers (QHC and EHC) to assure access of patients to appropriate clinical trials. The process has been slow and is still evolving. It required hiring research staff and developing a quality training program in areas where previously no research was in place. We now have the staff and infrastructure in place to carry out the mission of MBCCOP going forward to increase access to clinical trials for our patients.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The purpose of our studies is to develop new methods of gene transfer into hematopoietic stem cells using the genetic disease leukocyte adhesion deficiency or LAD as a model. For these studies we are using the canine form of LAD first to identify the number of hematopoietic stem cells required to reverse the disease phenotype and second, to test new retroviral vectors, conditioning regimens, and immunosuppressive agents required to enable sufficient numbers of gene modified hematopoietic stem cells to engraft and reverse the disease phenotype. The canine form of this disease is an optimal model for these studies since: a) the defect involves a membrane receptor on the surface of leukocytes, flow cytometry allows fascile detection and analysis of the number of gene corrected cells, b) low levels of gene corrected cells are likely to result in reversal of the disease phenotype, and c) studies in the canine model have been predictive of success in humans in the field of hematopoietic stem cell biology in that the background genetic heterogeneity between non-inbred dogs is similar to the genetic diversity of affected human populations, and d) the presence of a human counterpart to CLAD, namely LAD, allows the results from the animal model to be directly extrapolated to humans. We have now generated affected CLAD animals. CLAD animals with a matched donor are receiving non-myeloablative, allogeneic transplants to establish the number of CD18 positive cells required to reverse the disease phenotype. Those animals without donors will receive infusions of autologous, gene-corrected cells. These studies should provide the basis for future clinical approaches to LAD.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. 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 skeletal dysplasias are a heterogeneous group of disorders which result in disproportionate short stature and/or skeletal deformities. This study involves a multidisciplinary investigation of the clinical, genetic, morphologic, biochemical and molecular characteristics of the skeletal dysplasias. The International Skeletal Dysplasia Registry (ISDR) is involved in defining new skeletal (bone and cartilage) disorders and has been a unique national and international resource in the genetics community. The study objective is to collect information about skeletal disorders in order to better understand these conditions and help doctors and patients with their diagnosis and treatment. The study also identifies new types of skeletal disorders, their characteristics, their causes and determines how they are inherited. By collecting information and material from individuals with skeletal disorders we are able to elucidate a better understanding of the natural history of these disorders, as well as identify the genes that are defective for many of these diseases. There are approximately 200 different disorders in this group and, at present, many of the genes for this group of disorders are yet to be identified and studied. In the ISDR we conduct research in several ways. Each case undergoes a review of clinical information, as well as an evaluation of the x-rays. In addition, for a subset of the cases, a blood or tissue (bone, skin, cartilage etc.) sample for detailed analysis is collected and stored. In some cases, these materials are shared with other researchers studying the skeletal dysplasias. We potentially enroll approximately 100 subjects a year at Cedars-Sinai, and an additional 400-500 subjects from throughout the United States. Subjects with skeletal disorders and their families are invited to participate. There are no exclusion criteria - subjects are not excluded based on gender, age, race or ethnicity. The longstanding involvement and national reputation of the ISDR in all aspects of care and research on skeletal disorders has allowed us to collect a great deal of information regarding these conditions. We feel that our efforts have contributed a great deal to the current understanding of the skeletal dysplasias and expect that the efforts under this study will continue to significantly contribute to the understanding of skeletal disorders for the foreseeable future. In the future the GCRC will continue to assist in specimen collection for these molecular studies.
|
{
"pile_set_name": "NIH ExPorter"
}
|
During my career I have worked on several projects creating software (FASTLINK, CASPAR, rh_tsp_map,[unreadable] PedHunter, etc.) and a database (Anabaptist Genealogy database) for genetic studies.[unreadable] My association with this software and a past track record of effective collaboration[unreadable] with wet lab scientists leads to more and more such collaborations.[unreadable] Two highlights from the past year are:[unreadable] -- publication in late 2007 in the New England Journal of Medicine that heterozygous[unreadable] mutations in STAT3 cause hyper-IgE syndrome[unreadable] -- the completion of a dense genome-wide radiation hybrid map of the horse;[unreadable] the map is already available in NCBI's MapViewer and will be formally[unreadable] published soon in Volume 122, issue 1 of Cytogenetic and Genome Research.[unreadable] [unreadable] The immunology gene hunting work was done in collaboration with Steven Holland[unreadable] (NIAID/NIH), Bodo Grimbacher (University College London), and Jennifer[unreadable] PucK (UC San Francisco).[unreadable] [unreadable] The horse map work was done in collaboration with Richa Agarwala (NCBI/NIH),[unreadable] Edward Rice (former student at NCBI/NIH), Terje Raudsepp (Texas A&M) and[unreadable] Bhanu Chowdhary (Texas A&M).
|
{
"pile_set_name": "NIH ExPorter"
}
|
Helicobacter pylori, a bacterium that infects the stomach, is related to a form of low grade, B cell, non-Hodgkin's lymphoma described as mucosa- associated lymphoid tissue (MALT) lymphoma. Recent studies indicate that antibiotic treatment of H. pylori infection can cause the regression of MALT lymphoma in some patients. These clinical observations are supported by preliminary laboratory findings that tumor B cell proliferation in MALT lymphoma may be dependent on growth stimuli, such as cytokine growth factors, produced by H. pylori-induced immune accessory (inflammatory) cells present in the MALT lesion. Withdrawal of the proliferative stimulus by eradication of H. pylori is postulated to cause tumor cell death and tumor regression. These findings raise a number of important questions regarding: i) the nature of the malignant lymphoid cells in the MALT lymphoma lesion; ii) the role of the inflammatory response to H. pylori, in driving or sustaining the neoplastic (MALT lymphoma) cells; and iii) the relationship of H. pylori to the immune accessory cells and the possible role of H pylori in inducing the production of growth and differentiation factors required by MALT lymphoma. The proposed laboratory experiments are designed to address these questions by defining: 1) the nature and biology of the MALT lymphoma cells; 2) the response of MALT lymphoma cells to specific B cell growth stimulating cytokines (e.g.IL14, other BCGFs and Interleukins) and co-stimulatory factors for B cells (e.g. CD40 ligand, gp39); 3) the role of H pylori in generating and sustaining the lesions in which MALT lymphomas arise; and 4) the potential therapeutic effects of depriving the lymphoma cells of the cytokine-rich milieu generated by the HP-induced inflammatory response. A clinical trial of antibiotic treatment of patients with MALT lymphoma, conducted in parallel to the in-vitro experiments, will provide the appropriate tissue specimens as well as the opportunity for clinical and laboratory correlations. These studies may contribute to understanding the pathophysiology of MALT lymphoma in relation to H. pylori, as well as provide insights into therapeutic strategies for these patients.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This proposal is for support to develop a specialized cancer center at Purdue University whose primary objectives are to correlated and integrate existing programs in cancer research at Purdue University, to stimulate new research in selected areas, to contribute to advanced training in cancer research, and to develop interactions among Purdue researchers, local physicians, hospitals and clinics and cancer researchers at other universities and research laboratories within the State of Indiana. Commitment is to the development of a multidisciplinary program of experimental cancer research with focus on the biological and biochemical events involved in cell transformation as the basis for new approaches to the prevention, diagnosis and chemical therapy of cancer.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This project uses different systems to develop objective and subjective methods to monitor and document opacities in the human lens. We are actively recruiting patients with and without cataracts for reproducibility studies on the objective system-the Scheimpflug cameras (Zeiss and Topcon), retroillumination camera (Neitz), specular microscope (Keeler) and laser light-scattering spectroscope (Kowa). Tests of other systems will use sound (ultrasonography) and nuclear magnetic resonance (magnetic resonance imaging). Our study of subjective systems or methods such as the effects of cataracts on visual perception, contrast sensitivity, and glare may be useful in identifying additional parameters for monitoring cataract presence, progression, or regression.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The New York Academy of Science is sponsoring a conference entitled: "The Parahippocampal Region: Basic Science and Clinical Implications" in Washington, D.C. in September, 1999. The principal investigator and chair is Helen Scharfman, Ph.D., Department Head, Neurology Research Center, Helen Hayes Hospital and Assistant Professor, Departments of Pharmacology and Neurology, Columbia University. The cochairs are Menno Witter PhD., Professor, Department of Anatomy, Vrije Universiteit, The Netherlands and Robert Schwarcz, Ph.D., Head, Neuroscience Program, Maryland Psychiatric Research Center, and Professor, Departments of Psychiatry and Pharmacology, University of Maryland. Several structures in the temporal lobe play a critical role in brain functions, such as learning and memory, emotions, and complex behavioral processes. These include the hippocampus, the most frequently studied structure in the temporal lobe, and the juxtaposed parahippocampal region, which is comprised of the entorhinal cortex, perirhinal, and parahippocampal cortex. During the last decade, it has become increasingly clear that pathological changes in the parahippocampal region occur during the early stages of several catastrophic neurological and psychiatric diseases, such as Alzheimer's disease, schizophrenia, and epilepsy. Thus, it is quite likely that parahippocampal neuropathology contributes to dysfunction in these, and perhaps other brain disorders. Notably, pathology in the parahippocampal region may be a critical element of several of these diseases, regardless of the state of the hippocampus proper. This meeting is intended to integrate past studies with the latest information about the structure and function of the parahippocampal region. A recurrent theme will be the use of this information to advance the diagnosis and treatment of debilitating brain disorders. Anatomical, biochemical, physiological, behavioral, and pathological studies in both laboratory animals and humans will be described by the leaders in the field, and will provide a comprehensive, up-to-date review of this important part of the brain. The presentations will be targeted to a wide range of basic and clinical researchers and to clinical practitioners. Special emphasis will be placed on attracting women and minorities, and on including individuals at a wide range of educational levels. Such a diverse audience will provide optimal opportunities for cross-fertilization of ideas, and thus ensure a dynamic and stimulating atmosphere, in which exciting new concepts and hypotheses can be generated.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Crystallization, followed by subsequent structure determination, is a major step in understanding the structure-function relationship of macromolecules. Understanding macromolecule structure has become a key part in the development of new pharmaceuticals, and is a major area of NIH research. Crystallization however is also the rate limiting step, despite technological efforts to automate the set-up and crystallization data acquisition processes. The major objectives of the proposed effort are two fold. First is the implementation of a low cost epifluorescence and transmission microscopy system for the automated documentation of macromolecule crystallization plate outcomes. Second is the development of software for the rapid scoring of the images of the crystallization screen outcomes obtained by the microscopy system. The goals of the scoring process in this work are the rapid identification of likely crystals based upon the image pixel intensity due to the fluorescence from trace fluorescently labeled macromolecules (<0.5% of the molecules labeled with fluorescent probe), and the graduated scoring of precipitation outcomes that reflects the experimentally determined propensity of those outcomes to be optimized to crystallization conditions. Experimentally, trace fluorescently labeled protein will be subjected to crystallization screens and the outcomes periodically imaged. Intensity-based image analysis will be carried out using the software developed for this effort. Precipitated conditions which show high scores based on the image analysis will be subjected to optimization screening, and we propose there will be a correlation between the scores obtained and subsequent crystallization. Previous research has shown that fluorescence can be a powerful aid in finding and identifying crystals in screening plates (Judge et al., 2005;Forsythe et al., 2006;Groves et al., 2007;Pusey et al., 2008). Crystallization gives the most densely packed state for a protein, and therefore trace fluorescently labeled protein will have the greatest fluorescence intensity relative to clear or precipitated outcomes. The probe, being covalently attached, serves as a reporter to the protein's response to the solution conditions. Some precipitates showed 'bright spots'of fluorescence, and these conditions could subsequently be optimized to crystallization conditions (Pusey et al., 2008). It is further proposed that intensity-based scoring of precipitation outcomes may be used to better discriminate between non-productive and potentially productive precipitation results. Preliminary tests indicate that fluorescence intensity-based crystallization screen scoring should be very fast, with processing times likely to be 5 seconds per image. The capabilities to be developed are necessary for the subsequent introduction and sale of crystallization screening kits that have a balanced incomplete factorial (IF) approach to searching 'precipitation space'during the screening process. This relies upon accurate scoring of the outcomes, which is time consuming and expensive in labor if done by hand. Larger laboratories or research groups can afford the efforts and equipment needed to develop automated crystallization plate image documentation and scoring systems. Smaller groups cannot. Affordable IF implementation is seen as advantageous to improved crystallization and to our business long range development efforts. PUBLIC HEALTH RELEVANCE: Successful crystallization and X-ray data analysis provides important three-dimensional information on the macromolecules structure-function relationship. Many proteins that are potential drug targets or key components in diseases are only available in trace quantities, or are difficult to obtain. This proposal is to expand the data returned during protein crystallization process and the information that can be derived from it, by putting a powerful but affordable results documentation and analysis tool into the hands of crystallization laboratories.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The specific aims of this study will be to determine if individuals at risk for developing Type II diabetes mellitus (DM) have insulin resistance of glucose transport and phosphorylation within skeletal muscle using positron emission tomography (PET). Secondly, to determine if a) obese offspring of parents with type 2 diabetes mellitus (DM) have impairments in glucose transport and/or phosphorylation and b) if weight loss can improve glucose transport and phosphorylation. Three groups of subjects will be studied: 1) obese subjects with Type II DM, 2) obese subjects with a family history of Type II DM and normal glucose tolerance, and 3) obese subjects with no family history of Type II DM and normal glucose tolerance. This study will test the hypothesis that weight loss improves both glucose transport and phosphorylation, and that the effect of weight loss on these parameters is similar across the three groups.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Dental caries is one of the most common chronic infectious diseases in the world. The occlusal (biting surface) and interproximal (between teeth) areas are where there need for quantitative imaging of tooth decay and depth of cracks in the USA. In addition a majority of the caries load in USA is carried by different groups, such as Alaska Native people who have many times the national rate of tooth decay. Health care disparities in this population are exacerbated by rural remoteness. The average number of dental fillings per Alaskan Native person is double that of urban dwellers. To provide safe, high-quality dental care in these remote areas, an x- ray replacement is desired which provides high-resolution optical images of cracks and decay deep within teeth that does not require ionizing x-ray radiation. The current problem is that the optimal wavelengths of infrared light (1310 and 1460 nm) that can be used to see decay in teeth, do not have small high-resolution cameras for pediatric clinical use. Thus, an alternative method of forming infrared images will be developed and tested that produces the highest contrast of the lesion, which is scanning laser light either i transmission, reflection, and in a right-angle configuration (for example, scan the occlusal surface and detect above the gum line). The smallest laser-based imaging device has been developed at the University of Washington (UW), being the diameter of a round toothpick (1.2-mm) while producing high-quality video images at multiple wavelengths. The scanning fiber endoscope (SFE) technology has been tested by Dr. Joel Berg in children at The Center for Pediatric Dentistry with shared administration from UW Pediatric Dentistry and Children's Hospital, Seattle, WA. Although this device used visible light for testing the health of the enamel the change to infrared is straightforward, allowing much deeper imaging of cracks and carious lesions using optimal laser wavelengths for high-contrast transmittance and reflectance imaging. The small size of the scanner allows the two imaging modes to run concurrently if needed to find the extent of a crack, maximum depth of caries, and even total volume of caries under surveillance with non-surgical medicinal therapies. The hand piece will be smaller than any bitewing radiographic sensor used in children now, while providing new features of imaging during restorative work, possibly under tele-medicine conditions. The SFE technology is fundamentally low in cost because it uses laser and detectors that are fully developed in the telecommunications industry. The low- power scanned laser light imaging can be made portable and carries no significant risks. This project will fabricate a prototype device, determine its optimal imaging modes (wavelength &configuration), develop new 3D imaging algorithms, test with extracted teeth and verify with a pilot study with children 6 to 12 years of age.
|
{
"pile_set_name": "NIH ExPorter"
}
|
DESCRIPTION [unreadable] This is an application for renewal of NIOSH support for the graduate industrial hygiene (IH) training program at San Diego State University (SDSU) in San Diego, California during the academic years 2005 - 2010. The program offers a Master of Science (MS) degree with an Industrial Hygiene concentration. It is administratively housed within the Division of Occupational and Environmental Health (DOEH) of the Graduate School of Public Health (GSPH) at SDSU. [unreadable] [unreadable] The overall objective of the masters program in Industrial Hygiene at SDSU is to train practitioners. The program serves the Southern California region, including the border areas and practical experience in the curriculum is strongly emphasized. The GSPH has a multicultural student body. Since many of the IH students are already employed in IH and other related fields, evening courses are offered to accommodate students who work. [unreadable] [unreadable] Applicants to the Industrial Hygiene program must have a B.S. degree in basic sciences and/or adequate backgrounds in biological, chemical, and physical sciences. Additional requirements for admission are a GPA > 3.0 and a GRE > 1000 in verbal and (> 550) quantitative aspects. Highly qualified graduates in chemistry, physics, and engineering may also be accepted in the program after completing courses in the biological sciences including human anatomy and physiology. [unreadable] [unreadable] The MS degree is a two-year program that requires 42 semester credit hours of which 26 are in required core courses as follows: [unreadable] [unreadable] Course # Course Title Semester Hrs [unreadable] PH 601 Epidemiology 3 [unreadable] PH 602 Biostatistics 3 [unreadable] PH 732 Principles of Industrial Hygiene 4 [unreadable] PH 638A Principles of Toxicology 3 [unreadable] PH 737A Chemical Agents in the Workplace 3 [unreadable] PH 735 Industrial Ventilation 3 [unreadable] PH 636 Hazardous Waste Management 3 [unreadable] PH 700 Principles of Ergonomics 2 [unreadable] PH 737B Physical and Biological Agents in the Workplace 2 [unreadable] [unreadable] The IH core courses and the ventilation course have laboratory components and/or field trips. In addition to the core, there is a minimum of seven credits in prescribed elective courses. [unreadable] [unreadable] In addition to the core and elective courses, there is a minimum requirement of 3 credits (60 contact hours per credit) of field practice (PH 650 Field Practice 3 credits) in a designated internship facility. [unreadable] [unreadable] The program director is Behzad S. Samimi who has guided the program since its inception. He is a tenured professor, has certification in the practice of industrial hygiene, and is active in AIHA. He also teaches several of the core courses, and advises and supervises the majority of the IH students. [unreadable] [unreadable] The program has a 9 member advisory board (the Occupational Health Training Advisory Board or OHTAB), of which Dr. Samimi is a member and which he consults on student recruitment, research and field training. [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
Our lab is interested in the process of chromosome segregation and how defects in this process can affect the development of a multicellular organism. Over the past few years we have focused on the meiotic divisions that produce haploid gametes. We have been studying a class of temperature-sensitive (ts) embryonic lethal mutants from C. elegans that arrest in metaphase of meiosis I. In wildtype animals, oocytes in prophase of meiosis I are fertilized by sperm. Following fertilization, the oocyte chromosomes undergo two meiotic divisions, discarding the extra chromosomes in the polar bodies. These first meiotic divisions are important as any errors in chromosome segregation at this stage can lead to embryos with an abnormal number of chromosomes, which would likely lead to lethality. In our mutants, the oocyte chromosomes arrest in metaphase of meiosis I and never separate their chromosome homologs and never extrude polar bodies. Our meiotic mutants define five genes; they encode subunits of the Anaphase Promoting Complex or Cyclosome (APC/C). This complex serves as an E3 ubiquitin ligase that targets proteins for destruction (by the 26S proteasome) during the metaphase to anaphase transition of the cell cycle. We have named these mutants mat for their defects in the metaphase to anaphase transition during meiosis I. [unreadable] [unreadable] To identify extragenic regulators or substrates of these APC/C subunits, we have carried out a genetic suppression screen using a mat-3 mutant. The majority of our 27 suppressor mutations are dominant. These suppressors have been mapped using single nucleotide polymorphism (SNP) technology and define at least 9 complementation groups. One allele is a second site mutation within the mat-3 gene itself. A large number of alleles represent mutations in three spindle checkpoint components. These are the C. elegans orthologs of MAD1, MAD2, and MAD3. The spindle checkpoint prevents the metaphase to anaphase transition when chromosomes are not properly attached to the mitotic spindle. Our results suggest that this checkpoint also operates during meiosis. We identified one allele in the mdf-1 (the C. elegans Mad1 ortholog), two alleles in the mdf-3 gene (the Mad3 ortholog), and 12 alleles in the mdf-2 gene (the Mad2 ortholog). We believe that our mat mutants are not triggering the checkpoint, but rather that the checkpoint normally operates during meiosis as a negative regulator of the APC/C. Perhaps the checkpoint functions to regulate the proper timing of the meiotic divisions. We also identified three dominant suppressors that were mutations in a positive regulator of the APC/C. This gene is called fzy-1 and is the Cdc20/Fzy ortholog. These three mutations cluster in a small region of the protein thought to be important for its interaction with MDF-2. These mutations presumably disrupt the interaction with MDF-2 and thus prevent MDF-2 inhibition of the APC/C. We are currently mapping the remaining suppressors and anticipate finding novel molecules that shed light on how the APC/C is regulated during meiosis. To address whether our suppressor mutations have phenotypes on their own, we have crossed them away from the original mat-3 mutation. For some of our mutants, there is a significant reduction in brood size. However, we have not observed any obvious embryonic lethal phenotypes for these genes, suggesting that our screen uncovered viable alleles of essential spindle checkpoint genes.[unreadable] [unreadable] We are currently characterizing the meiotic defects associated with the strict paternal-effect lethal mutant spe-11. SPE-11 is a sperm-specific factor that is contributed to the oocyte upon fertilization. Homozygous spe-11 mutant males produce dead embryos when mated with wild-type hermaphrodites or females. These embryos fail to extrude their polar bodies, secrete an eggshell, and initiate proper mitosis and cytokinesis. As a result, the embryos die as 1-cell embryos with eggshell and cytokinesis defects. We are further characterizing the defects associated with the mutant spe-11 alleles in order to address what function spe-11 plays in influencing the meiotic divisions of the oocyte. We intend to screen for other genes that phenocopy the spe-11 phenotype using classical genetic screens and RNAi screens in order to identify other factors that operate in the SPE-11 pathway.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The benefits of current pharmacological approaches to treating heart failure (HF) are near their limits and new adjunctive therapies are needed. Recent results from this PPG suggest that low carbohydrate/high fat diets may prevent the development and progression of HF. The fatty acid composition of the diet may have a major impact on development and progression of HF. Intake of polyunsaturated fatty acids (PUFA) has a more favorable effect on cardiac gene expression, mitochondrial function, and cardiomyocyte apoptosis than a high saturated fat diet. We also observed that high sugar intake may accelerate the development and progression of HF. This Project investigates the mechanistic effects of dietary macronutrient composition on the progression of HF. We hypothesize that 1) high intake of PUFA (18:2n-6 and 18:3n-3) or the EPA+DHA) prevents and reverses deterioration in mitochondrial function, apoptosis, cardiomyocyte hypertrophy, fibrosis, chamber remodeling and contractile dysfunction in HF; and 2) a high sugar diet activates insulin-stimulated growth and ROS production and accelerates the progression of HF. Studies will be performed in established models of HF. Underiying mechanisms are addressed with targeted alterations in specific pathways using nutritional, pharmacological, and genetic approaches. The specific aims are: 1) Assess the ability of high fat diets comprised of saturated fatty acids or PUFA to stop or reverse the progression of HF compared to a standard high carbohydrate diet. 2) Determine the effects of dietary supplementation of a standard high carbohydrate diet with UJ-3PUFA (EPA+DHA) on the progression of established HF. 3) Elucidate the role of insulin signaling in the adverse effects of a high sugar diet in HF. 4) Determine the role of ROS generation in mediating the adverse effects of high sugar diet in HF.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Cellular proliferation and differentiation are responses elicited by interaction of extracellular molecules with the cell surfaces of eukaryotic cells. In Dictyostelium extracellular cAMP acts similarly to paracrine hormones in mammalian cells by interacting with a specific cell surface receptor to stimulate the synthesis of intracellular second messages. Our results indicate that there are at least two independent mechanisms involved in the developmental regulation of gene expression by cAMP in Dictyostelium. Conditions which allow intracellular synthesis of cAMP promote the normal regulation of a gene known to be repressed in conjunction with cAMP signalling. In contrast, expression of genes which exhibit maximal activity after aggregate formation depends upon accumulation of extracellular cAMP signalling. We have additionally shown that extracellular cAMP can be at least partially bypassed in permeabilized cells exposed to inositol tris phosphate (IP3), a mobilizer of intracellular calcium ion; we suggest that IP3 acts as a second messenger in Dictyostelium to mediate certain developmental processes. Finally, we have provisionally isolated a gene for the cAMP cell surface receptor and have also cloned a gene which has homology with the Alpha subunit of the GTP-binding, N-regulatory protein and with the ras family of proto-oncogenes. Studies now focus on their expression and function in order to understand the molecular mechanisms of signal transduction.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Endometrial cancer (EC) is the fourth most frequent cancer in women, ranking first in incidence and second in mortality among female genital tract tumors. Although several genetic lesions associated with EEC have been identified, numerous grey areas still exist, especially regarding the mechanism of action of these mutations and their relative contribution to the progression from hyperplasia to metastatic cancer. A relationship between excess estrogen and type I EC (EEC) has been recognized for decades; however little is known about the molecular basis of this association. The broad, long- term objective of this project is to determine the role of the PI3K/PTEN/AKT pathway in endometrial hyperplastic and neoplastic transformation, and to elucidate the mechanisms relevant to these functions. As with most other neoplastic diseases, the elucidation of the pathogenetic basis of EC initiation and progression at the molecular level are greatly facilitated by laboratory models. Emerging reagents and strategies allow us to use the mouse in more and more sophisticated ways to define the molecular, cellular and physiological events that lead to cancer initiation and progression. We have developed a mouse strain that closely recapitulates several features of the progression from endometrial hyperplasia to neoplasia. We propose to utilize a combination of direct in vivo genetic approaches and ex vivo and in vitro experiments to carry out the following specific aims: Aim 1, To test the hypothesis that AKT activation in EEC causes a physiologically relevant alteration of ER1-mediated signaling. Aim 2, To identify and characterize, in mutant mice and cells, novel specific pathogenetic mechanisms activated by Pten loss during EEC initiation. The proper diagnosis and management of endometrial cancer are still major challenges in clinical oncology, as we still do not understand clearly the mechanisms that are altered during the progression from hyperplasia to cancer. This proposal is designed to dissect several specific biochemical and genetic pathways controlled by PTEN in its tumor suppressive function in the endometrial epithelium, and to define novel mechanisms involved in neoplastic transformation using engineered model systems that closely mimic the molecular events taking place in human lesions.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Longitudinal bone growth occurs at the growth plate, which consists of three principal layers: the resting zone, the proliferative zone, and the hypertrophic zone. Studies in our laboratory indicate that stem-like cells in the resting zone differentiate into rapidly dividing chondrocytes of the proliferative zone. The proliferative chondocytes then terminally differentiate into the nondividing chondrocytes of the hypertrophic zone. [unreadable] [unreadable] To explore the molecular switches responsible for this two-step differentiation program, we developed a microdissection method to isolate RNA from the resting, proliferative, and hypertrophic zones of growing rats. Microarray analysis followed by real-time PCR analysis identified genes whose expression changed dramatically during the differentiation program, including multiple genes functionally related to bone morphogenetic proteins (BMPs). BMP-2 and BMP-6 were found to be upregulated in hypertrophic zone compared with resting zone and proliferative zone. In contrast, BMP signaling inhibitors, including BMP-3, gremlin, and growth differentiation factor-10, were expressed early in the differentiation pathway, in the resting and proliferative zones. Our findings suggest a BMP signaling gradient across the growth plate, which is established by differential expression of multiple BMPs and BMP inhibitors in specific zones. We have previously shown evidence that BMPs can stimulate both proliferation and hypertrophic differentiation of growth plate chondrocytes. Therefore, taken together, our findings suggest that low levels of BMP signaling in the resting zone may help maintain these cells in a quiescent state. In the lower resting zone, greater BMP signaling may help induce differentiation to proliferative chondrocytes. Farther down the growth plate, even greater BMP signaling may help induce hypertrophic differentiation. Thus, BMP signaling gradients may be a key mechanism responsible for spatial regulation of chondrocyte proliferation and differentiation in growth plate cartilage. [unreadable] [unreadable] Fibroblast growth factor (FGF) signaling is also essential for endochondral bone formation. Mutations in FGF receptors cause skeletal dysplasias including achondroplasia, the most common human skeletal dysplasia. To explore the role of FGF signaling in the postnatal growth plate, we quantitatively analyzed expression of FGFs and FGF receptors (FGFRs). Rat proximal tibial growth plates and surrounding tissues were microdissected, and specific mRNAs were quantitated by real-time RT-PCR. To assess the FGF system without bias, we first screened for expression of all known FGFs and major FGFR isoforms. Perichondrium expressed FGFs 1, 2, 6, 7, 9, and 18 and, at lower levels, FGFs 21 and 22. Growth plate expressed FGFs 2, 7, 18, and 22. Perichondrial expression was generally greater than growth plate expression, supporting the concept that perichondrial FGFs regulate growth plate chondrogenesis. Nevertheless, FGFs synthesized by growth plate chondrocytes may be physiologically important because of their proximity to target receptors. In growth plate, we found expression of FGFRs 1, 2, and 3, primarily, but not exclusively, the c isoforms. FGFRs 1 and 3, thought to negatively regulate chondrogenesis, were expressed at greater levels and at later stages of chondrocyte differentiation, with FGFR1 upregulated in the hypertrophic zone and FGFR3 upregulated in both proliferative and hypertrophic zones. In contrast, FGFRs 2 and 4, putative positive regulators, were expressed at earlier stages of differentiation, with FGFR2 upregulated in the resting zone and FGFR4 in the resting and proliferative zones. Thus, this analysis identified ligands and receptors not previously known to be expressed in growth plate and revealed a complex pattern of spatial regulation of FGFs and FGFRs in the different zones of the growth plate.[unreadable] [unreadable] Previous studies of the insulin-like growth factor (IGF) system gene expression in growth plate using immunohistochemistry and in situ hybridization have yielded conflicting results. We therefore studied the spatial patterns of mRNA expression of the IGF system in the rat proximal tibial growth plate quantitatively. IGF-I mRNA expression was minimal in growth plate compared with perichondrium, metaphyseal bone, muscle, and liver. In contrast, IGF-II mRNA was expressed at higher levels than in bone and liver. IGF-II expression was higher in the proliferative and resting zones compared with the hypertrophic zone. GH receptor and type 1 and 2 IGF receptors were expressed throughout the growth plate. Expression of IGF-binding proteins (IGFBPs) -1 through -6 mRNA was low throughout the growth plate compared with perichondrium and bone. These data suggest that regulation primarily depends on IGF-II produced by chondrocytes, and IGF-I produced by surrounding structures.[unreadable] [unreadable] With age, growth plate chondrocyte proliferation slows down, causing longitudinal bone growth to slow and eventually stop. This functional change in the growth plate is accompanied by structural changes; with age, the number of resting, proliferative, and hypertrophic chondrocytes decreases as does the size of the individual hypertrophic cells. The chondrocyte columns also become more widely spaced. We have termed this developmental program, growth plate senescence. Growth plate senescence appears to be caused by a mechanism intrinsic to the growth plate. To explore the molecular mechanisms responsible for growth plate senescence, we analyzed how gene expression patterns change in the growth plate during postnatal life, as the rate of longitudinal bone growth decreases. [unreadable] [unreadable] In particular we analyzed the insulin-like growth factor (IGF) system because IGFs are capable of potently regulating growth plate chondrocyte proliferation and differentiation. With increasing age (3-, 6-, 9-, and 12-week rats), IGF-I mRNA levels increased in the proliferative zone but remained at least tenfold lower than levels in perichondrium and bone. IGF-II mRNA decreased dramatically, 780-fold, in proliferative zone whereas, type 2 IGF receptor and IGF binding proteins (IGFBPs)-1, -2, - 3, and -4 increased significantly with age in growth plate and/or surrounding perichondrium and bone. These findings suggest that growth plate senescence, including the decrease in growth velocity that occurs with age, may be caused, in part, by decreasing expression of IGF-II and increasing expression of type 2 IGF receptor and multiple IGFBPs. [unreadable] [unreadable] We also analyzed temporal changes in fibroblast growth factor (FGF) expression in the growth plate. We identified several changes in FGF and FGFR expression that may contribute to growth plate senescence. In the growth plate, FGFRs 2 and 4, both implicated as positive regulators of growth, undergo a decline in expression with age. In perichondrium, we observed increases in FGFs 1, 7, 18, and 22 mRNA with age. Increasing levels of these ligands, interacting with constant levels of FGFR3 in growth plate might contribute to growth plate senescence.[unreadable] [unreadable] These studies have begun to elucidate the regulation of gene expression that is responsible for the complex spatial organization of the growth plate and for the temporal changes of growth plate senescence. Combined with previous functional studies performed in our lab and by others, the findings indicate a highly complex system involving BMPs, FGFs, and IGFs, their receptors and other interacting proteins.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Lymphocytic choriomeningitis virus and Tacaribe virus (another arenavirus) have been found to produce a destructive retinopathy following intracerebral inoculation in the neonatal rat. These lesions are immunologically mediated as indicated by the sparing effects of antilymphocyte serum. These models, delineated by prior work in this laboratory, provide a superb opportunity to study in detail a prototype of virus-induced retinal immunopathology. It is now proposed to exploit these models to pursue several long term goals: to establish other animals of virus-induced ocular disease; to clarify the pathogenetic mechanisms which contribute to the development of viral retinopathy; and to dissect the intimate immunological and pathological mechanisms which mediate these diseases using current techniques of cellular immunology.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The purpose of this project is to determine the role of glucagon in health and its contribution to the endogenous hyperglycemia of diabetes mellitus. The role of glucagon in health would be determined in normal humans and by dogs by somatostatin-induced suppression of glucagon during conditions of glucose need, e.g. stress, starvation and exercise. The importance of glucagon in the diabetic state would be tested by sudden insulin withdrawal in totally depancreatized humans, by studying the effect of exogenous glucagon administered to adult type and juvenile diabetics, and by studying glucagon's relationship to glycemia in the insulin-independent Houssay dog. The mechanism of the insulin-independent evanescence of glucagon's action on the liver would be studied to determine whether this "fade", demonstrated thus far only during constant rate infusions of exogenous glucagon, represents "down-regulation" of glucagon receptors and, if so, if it occurs with the phasic hyperglucagonemia of endogenous glucagon release in diabetes. The nature of the A-cell dysfunction in human diabetes would be studied to determine whether or not insulin can correct the defects or whether there is a lesion unresponsive to normal levels of plasma insulin that requires an alternative means of glucagon suppression. The role of "big plasma glucagon" (BPG), a 180,000 dalton IRG with apparent glucagon-like biologic activity and a familial distribution, would be characterized and its tissue of origin searched for. The role of the D-cells in health and in diabetes would be studied in isolated perfused dog pancreas preparations and an attempt to characterize their function in the normal and diabetic pancreas. Finally, to determine if agents which reduce the abnormal secretory patterns of glucagon that characterize human diabetes provide a useful adjunct to conventional therapeutic methods, trials of glucagon-suppressing analogues of somatostatin would be conducted.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Abstract This R13 proposal seeks support for a symposium and workshop at the Forsyth Institute October 11-12, 2018 on ?The Uncultivable Bacteria?. The overall objective of the Symposium is to increase understanding of the reasons 30 percent of the bacteria in the human oral cavity, and the majority of bacteria in the environment, remain ?uncultivable? using standard microbiological methods. The problem of ?uncultivability? is a major impediment to both host associated and environmental microbial research. The Candidate Phyla Radiation (CPR) is a recently recognized subdivision of bacteria representing 15% of bacterial diversity on earth. The CPR bacteria occur in essentially all environments, including the human oral cavity. To date, no members of the CPR have been cultured except for 10 isolates representing 5 species of human oral Saccharibacteria cultured by Forsyth investigators. The information to be presented at the symposium represents a paradigm shift in approach to culturing CPR and other as-yet-uncultivated groups of bacteria. This symposium and workshop will train a new cadre of investigators to tackle cultivation rather than attempt to rely solely on sequencing and in silico analysis. The conference will create a community of investigators who have previously resided in scientific silos that did not meet or communicate with each other: dental research, environmental microbiology, microbial ecology, and the life cycles and biology of obligate parasites. The Symposium will include poster presentations by attendees and talks by: Jill Banfield, Stephen Giovannoni, Kim Lewis, William Wade, Jeffrey Mclean, Floyd Dewhirst, Jessica Mark-Welch, Pallavi Murugkar, Xuesong He and Angela Douglas. The second day of the conference will be a workshop with live demonstration of the isolation and cultivation of human oral Saccharibacteria in coculture with appropriate host bacterial species. Detailed protocols will be distributed so that all attendees should be able to return to their laboratories and culture Saccharibacteria from their own mouths. Dissemination of approaches and protocols may enable widespread cultivation of taxa in the many phyla comprising the CPR. For environmental, industrial an d medical microbiology, this would be a transformative advance. For example, culturing CPR bacteria would greatly enhance our understanding of global carbon cycles and the development of biofuels by being able to utilize isolates of previously uncultured members of microbial dark matter. Knowledge presented in the symposium will enhance attendee?s approach to cultivating as-yet-uncultured members of the human oral cavity. Raising the percentage of cultivated organisms would allow a higher percentage of g enomes to be obtained for oral bacteria. The genome information provides the amino acid sequence of all genes. This increased information would greatly facilitate investigators mapping proteome and transcriptome data to species. This would significantly strengthen microbiome studies in dental research.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The long-term objective of this research is to understand the factors which influence and regulate the permeability of cardiac gap junctions. We will test the hypothesis that the regulatory pathways which operate to maintain junctional permeability under normal conditions fail during ischemia, leading to arrhythmias and irreversible injury. Primary cultures of neonatal rat myocytes and isolated gap junctions will be used to examine the regulation by calcium and protons of intercellular coupling and channel conductance, respectively. The impact of ischemia on intercellular coupling will also be examined. Specifically, in the intact cells the effects of several interventions designed to increase intracellular (Ca2+), (H+), or both on junctional permeability, and cellular calcium and proton levels will be addressed using pharmacological interventions to inhibit the organelles' functions. Alteration of intracellular coupling will be measured using dual cell voltage clamping techniques, or by measurement of dye transfer rates. Using patch clamp techniques, the effects of these ions and of lipid composition on the conductance of isolated channels will be determined, and the results compared to what is observed in the intact cells. The effects of ischemia on intracellular ion levels and cellular contractile activity will be correlated with alterations of junctional conductance and ion fluxes. Finally the role of lipid peroxidation in the generation of the ischemic effects will be examined. These studies will test several hypotheses. 1) That cardiac gap junctions are sensitive to both Ca2+ and H+, and that these ions can act in a synergistic fashion to block permeability. 2) That intracellular organelles serve to protect the junctions from elevated cytoplasmic levels of Ca2+ and H+. 3) That failure of gap junctions contributes to the onset of post-ischemic arrhythmias. 4) That post-ischemic failure of gap junctions occurs as a result of high intracellular levels of Ca2+ and/or protons or to damage to cellular membranes (sarcolemma, mitochondria, sarcoplasmic reticulum). Realization of our objectives will lead to a better understanding of the factors which influence gap junction function and of the role of gap junctions in the occurrence of arrhythmias.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Genetic changes related to carcinogenesis are being studied using hybrids of human lung carcinoma cells with normal human bronchial epithelial cells. Initial studies suggest that a limited population doubling potential (mortality) is the dominant genetic trait in hybrids cells. Other hybrid cell lines have been isolated and are being characterized for doubling potential, karyotype and tumorigenicity in athymic nude mice. The effects of individual chromosomes are being assessed by fusion with mini-cells containing single marked chromosomes.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Hypoxia inducible factor 1-alpha (HIF-1alpha) is a component of the transcription factor HIF-1. Cell culture studies and mouse gene knockouts demonstrate HIF-1alpha transactivation of genes controlling angiogenesis, cellular metabolism, invasion and apoptosis. In addition to hypoxia. HIF-1alpha protein expression is regulated by growth factor signaling and survival pathways, and viral oncogenes. HIF-1alpha expression is induced in cancers and high-grade premalignant lesions. However. HIF-1alpha function in each stage of epithelial carcinogenesis or metastasis is unknown. We created transgenic mice expressing either wild type or a constitutively active mutant HIF-1alpha lacking the "oxygen-dependent degradation domain' about in basal squamous epithelium (K14-HIF-1alpha and K14-HIF-1alpha delta ODD transgenic mice). We have engineered Cre-loxP mediated HIF-1a deletion in basal squamous epithelium without a discernable phenotype. Now we test whether HIF-1alpha plays a fundamental role in multistage epithelial carcinogenesis with these Specific Aims 1. Determine the stage-specific effect of gain or loss of HIF-1alpha function on Ha-ras initiated epidermal carcinogenesis. 1.1. Test whether gain of HIF- 1alpha function promotes two-stage epidermal chemical carcinogenesis induced by dimethylbenzanthracene and tetraphorbolmyristate acetate in Kl4-HIF-1alpha and Kl4-HIF-laz\ODD transgenic mice. 1.2. Test necessity of HIF-1alpha in two-stage chemical carcinogenesis in mice with Cre-loxP mediated epidermal HIF-1alpha deletion. 2.0. Determine biology of gain of HIF-1alpha function on multistage carcinogenesis in skin and cervix induced by the HPV16 early transforming region. 2.1. Test alteration of each stage of HPV16-induced epidermal carcinogenesis by gain of HIF-1alpha function in KI4-HPV16:HIF-1alpha or HIF-1alpha delta ODD double transgenic mice. 2.2. Test cooperation between HIF-1alpha gain of function and estrogen in HPV16 induced cervical carcinogenesis in mice transgenic for the HPV16 and HIF-1alpha or HIF-1alpha delta ODD. 3.0. Test biology of HPV16 E6-HIF-1alpha coexpression, and determine p53 dependent or independent functions of E6 in conjunction with HIF-1alpha. 3.1. Create Kl4-E6HIF-1alpha or K14-E6:HlF-1alpha delta ODD double transgenic mice and determine alterations in the biology of skin and cervical carcinogenesis. 3.2. Create Kl4-E6 HIF-1alpha or K14-E6:HIF-1alpha delta ODD double transgenic mice and determine alterations in the biology of skin and cervical carcinogenesis. Our possession of all relevant animal models, positions us to determine the precise role of HIF-1alpha in carcinogenesis mediated by either activated oncogenes or inactivation of tumor suppressor genes.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Focal cerebral ischemia (ischemic stroke) is the most common cause of disability and the fourth leading cause of death in the United State. Despite of the efforts on developing the pharmacological and surgical treatments of the disease, tissue plasminogen activator (tPA) is the only effective therapy at present. A better understanding of the pathological process and the discovery of new targets and therapies will significantly advance the field. There have been little studies on lncRNAs in cerebral ischemia. Through an lncRNA array analysis in a rat model of focal cerebral ischemia/reperfusion (IR), we have identified CAMK2D-associated transcript 1 (C2dat1) and 2 (C2dat2) (C2dat1-2) as two novel IR-induced lncRNAs that specifically regulated the expression of CaMKII? in rat and mouse models of focal cerebral IR. In our pilot study, C2dat1-2 mRNAs were upregulated in a time-dependent manner in mouse cortical penumbra after focal ischemic brain injury, which was accompanied by increased expression of CaMKII? at transcript and protein levels. The expression patterns of C2dat1-2 and CAMK2D were confirmed in mouse Neuro-2a cells in response to in vitro ischemia (oxygen-glucose deprivation/re-oxygenation, OGD/R). Knockdown of C2dat1 resulted in a significant blockade of CaMKII? expression, and potentiated OGD/R-induced cell death. Mechanistically, reduced CaMKII? expression upon silencing C2dat1 inhibited OGD/R-induced activation of the NF-?B signaling pathway. Further analysis showed that the downregulation of IKK? and further inhibition of I?B? degradation accounted for the inhibition of the NF-?B signaling activity by depleting C2dat1-2. Thus, C2dat1 appears to promote neuronal survival through regulating the NF-?B signaling pathway. Therefore, lncRNAs may be potential targets for therapeutic intervention of ischemia brain injury. Based on these preliminary findings, we hypothesize that C2dat1-2 are novel IR-induced lncRNAs that regulate the expression of CaMKII? to promote neuronal survival through the activation of the NF-?B signaling pathway. The primary goal of this application is to determine the function and signaling mechanisms of C2dat1-2, as well as the associated CaMKII? in IR- induced neuronal injury. The long-term goal is to gain more insights into the molecular bases of IR-associated biological processes and to identify novel therapeutic targets that confer neuroprotection during IR. Two specific aims are proposed: Specific Aim 1. Define the role of C2dat1-2 as novel ischemia-induced lncRNAs that promote neuronal survival by modulating CaMKII? expression in mouse primary neuronal cultures. Specific Aim 2. Determine if knockdown of C2dat1-2 potentiate IR-induced cell death in mouse model of focal cerebral ischemia and if the effects are mediated through down-regulation of CaMKII?.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objectives of this research are: (1) to find an adequate method for the production of antibody against several carcinogenic mycotoxins, including aflatoxins, patulin, penicillic acid, and a group of cocarcinogenic mycotoxins, i.e., ochratoxins; (2) to develop a radioimmunoassay (RIA) or an immunochemical assay for mycotoxins; (3) to investigate the interaction of mycotoxins and their analogs with homologous antibody and; (4) to study the possibility of neutralization of the mycotoxic effect through immune systems. Different mycotoxins and their analogs will be coupled to proteins using a water soluble carbodiimide as the coupling agent and by mixed anhydrous method. The best preparation (high mycotoxin to protein ratio) will be used for immunization in rabbits or goats. Production of antibody will be followed by a binding assay method using radioactive ligands. In order to improve the sensitivity of RIA, efforts will be made to improve the antibody titers, develop the solid state RIA, and to synthesize high specific activity radioactive ligands. Different physiochemical methods will be used to characterize the interaction of mycotoxins (and their analogs) with homologous antibody, and to determine the binding constant. The IgG against specific mycotoxin will be tested for the possibility of neutralizing certain inhibitory effect of the mycotoxins in specific biochemical systems. In addition, the possibility of neutralization of mycotoxins in vivo will be tested by immunizing small animals, e.g. guinea pigs, or rats, with mycotoxin-protein carrier, and subsequently challenging these animals with mycotoxins 4-5 weeks after immunization. The LD50 of mycotoxin of the immunized animals will be compared with that of controls.
|
{
"pile_set_name": "NIH ExPorter"
}
|
[unreadable] It is currently appreciated that both insulin resistance and beta-cell dysfunction are early and essential events in the development of type 2 diabetes. Our current knowledge of factors that influence beta-cell function is lacking, despite research in this field having been conducted for several decades. To this end, we have recently shown that the heterotrimeric G-protein alpha-subunit, G{alpha}z, modulates an endogenous signaling pathway that is inhibitory to glucose-stimulated insulin secretion in an insulinoma cell line [Kimple et al. (2005) J Biol Chem 280:31708]. These results led to the hypothesis that loss of G{alpha}z activity would result in increased insulin secretion and improved beta-cell function in vivo, possibly protecting against the development of type 2 diabetes. In support of this hypothesis, we have performed preliminary experiments in which G{alpha}z-null mice, when compared to wild-type littermate controls, display increased plasma insulin concentrations and correspondingly decreased blood glucose levels during glucose tolerance tests. Furthermore, the increased plasma insulin levels observed in G{alpha}z-null mice are likely a direct result of enhanced insulin secretion, as pancreatic islets isolated from G{alpha}z-null mice exhibit significantly higher glucose-stimulated insulin secretion than those from wild-type mice. To further address our hypothesis, and our understanding of the role of G{alpha}z signaling in insulin secretion and islet cell function, we propose the following Specific Aims: (1) to delineate the signaling pathways upstream and downstream of G{alpha}z that are important for its inhibition of insulin secretion, (2) to determine at which step in the stimulated secretion process G{alpha}z is acting, and (3) to determine whether loss of G{alpha}z is protective against the development of diabetes, both age-induced and high-fat diet-induced. The results of these studies are expected to yield important new insights into the regulation of insulin secretion and beta- cell function at a molecular level, and may point to G{alpha}z as a potential new target for therapeutics aimed at ameliorating beta-cell dysfunction in Type 2 diabetes. Relevance: This proposal aims to delineate the specific pathways by which a protein involved in the regulation of insulin secretion functions. How much insulin is secreted into the blood is one determinant of blood glucose levels; therefore, this project has direct relevance to diabetes. [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
Project Summary The oral cavity represents a unique healing environment that is relatively unexplored. Regeneration within the oral mucosa occurs rapidly, with wounds closing at four times the rate of the skin, and the inflammatory reaction to wounding is relatively suppressed. Chronic wounds within the oral cavity compromise the ability of patients to obtain nutrition and are prone to debilitating infections, thus constituting an important oral health problem. Understanding how the oral mucosa is able to achieve such regeneration, and therefore how wound healing is disrupted in disease environments, would be a significant achievement for the field of craniofacial research. The specific cellular mechanisms used by the oral mucosa to facilitate its distinct regenerative phenotype remain elusive and are addressed in this proposal. Studies comparing dermal and oral wounds have shown that oral wound healing displays a short inflammatory phase followed by a rapid transition into wound closure via epithelial cell proliferation, migration and differentiation. Chronic inflammation in the oral cavity is the cause of many pathologies, such as periodontitis, oral mucositis and osteonecrosis of the jaw, but how inflammation leads to epithelial atrophy or lingering open wounds in the oral mucosa is unknown. Given that the oral epithelial progenitor cell (OEPC) population mediates re-epithelialization, this proposal will test the hypothesis that inflammation acts as an important coordinator of wound closure and re-epithelialization via direct regulation of the OEPC population. Wound healing will be assessed in animals under three inflammatory states: normal, low and high. The effects of attenuated and exacerbated inflammation on regeneration will be established by scoring for time to wound closure, epithelial layer integrity and proliferation and apoptosis within the entire tissue (Aim 1). Next, the cellular and molecular mechanisms of inflammatory regulation on the OEPC population will be determined using single cell RNA sequencing under the three conditions, and direct regulation of the epithelium by the immune system tested using an in vitro co-culture system (Aim 2). Data from these aims will determine how inflammation regulates wound closure and elucidate key mechanisms through which inflammation influences re-epithelialization, which will yield valuable insights into the mechanisms of wound closure within the oral mucosa. These research goals will be conducted in conjunction with a comprehensive training plan designed to develop the applicant?s career as a dentist-scientist. The training includes structured mentorship from two highly qualified clinician-scientist sponsors, and scientific and technical training through individual and group meetings, seminars, journal clubs, classes and departmental events. Research and training will take place at the University of California, San Francisco, which offers both an outstanding research environment and an excellent dental school for clinical training.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Neurons in the adult mammalian central nervous system (CNS) do not regenerate after injury. One major factor reported to contribute to reduced regenerative potential is the inhibitory effect of chondroitin sulfate (CS) and keratan sulfate (KS)-containing proteoglycans (CS/KSPGs) on neuronal outgrowth and neural plasticity. Of note, the effects of CS and KS in aggrecan are neither additive nor synergistic, which suggests that CS and KS may act together in a single or closely related set of pathways. Disruption of either glycosaminoglycan (GAG)-target interaction should increase neuronal regeneration and rehabilitation. We propose to develop new tools to understand in detail the structure-activity relationship (SAR) of CS and KS oligosaccharide responsible for modulation of the neuronal outgrowth inhibition in vitro. For this, we will exploit in our research sulfated glycan oligosaccharides of unique and regular structures. The extensive library of structurally defined GAG and GAG-mimetic oligosaccharides (especially those derived from marine sources) will allow us to disrupt the CS/KS-dependent down-regulation effect in neural outgrowth. At the end of this project, besides understanding the structural requirements, particularly sulfation patterns and chain lengths, of sulfated glycans in the process of axon inhibition, we will be also able to present a library of sugars that can be beneficial to modulate the inhibitory process of neural growth when exogenously applied in the system. This project not only adds new information to the community regarding the actual contribution of CS and KS of aggrecan in neuronal outgrowth inhibition but will also offer new molecular tools to be used for controlling this process.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The identification of mutations causing Mendelian diseases has revolutionized the understanding of diseases of every organ system. While over 3,000 such diseases have been solved at the molecular level, with 21,000 genes in the human genome and about 15% embryonic lethal loci, it is clear that many remain to be discovered. This includes both described and presently undescribed human traits that contribute to both health and disease. With the spectacular 6-log drop in the cost of DNA sequencing over the last 12 years, it has become apparent that selectively sequencing all of the genes in the genome, which comprise only ~1 % of the human genome represents a very cost-effective means for discovering the basis of new Mendelian diseases. We have pioneered the development of the exome sequencing method as well as the tools for analysis, and have shown that both are scalable, with current cost under $1,500 per exome and expected to be under $1,000 in the near future. We have demonstrated the utility of this approach with the identification of a range of disease genes that were previously intractable due to difficulties in gene mapping owing to high locus heterogeneity, de novo mutations, or small one-of-a-kind families. These considerations motivate new efforts to efficiently solve substantially all Mendelian traits using these technologies. To this end we have established the Yale Center for Mendelian Disorders which will ascertain and acquire samples from patients and families with known or suspected Mendelian diseases, sequence exomes to high coverage sufficient to call 95% of all variants with high specificity and use new analytic approaches we have devised to identify new Mendelian trait genes. We will make all sequences available to the research community as allowed and will establish a Web interface to enable physicians and investigators to submit research samples and retrieve annotated results. These studies will rapidly expand our understanding of the genes and pathways underlying human disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
N-methyl D-aspartate receptors are a subtype of glutamate receptors that mediate excitatory signal transmission in the mammalian central nervous system. Their primary function involves converting the chemical signal into an electrical signal, i.e. glutamate binding to an extracellular domain in the receptor triggers the formation of cation permeable transmembrane channels in the receptor. Given the importance of these receptors in mediating a number of physiological processes and the need to modulate their function in disease states, the primary questions are how does the agonist activate the protein and how can this mechanism be modulated. The NMDA receptors are modulator in structure consisting of an amino terminal domain, agonist binding domain, channel segments and the C-terminal domains. Here we propose to study the communication between the domains and their role in dictating activation and allosteric modulation. Specifically we will investigate the role o the interactions between GluN1 agonist binding domain with the GluN2 subunit in controlling agonist binding domain dynamics and extent of activation (specific aim 1) using a combination of luminescence resonance energy transfer, smFRET, and electrophysiology. We will also investigate the pathway for allosteric modulation by establishing the conformational changes starting at the amino terminal domain through the agonist binding domain and study the changes in dynamics in the extracellular domains during the allosteric modulation (specific aim 2). The spectroscopic investigations will be complemented by electrophysiological measurements investigating the changes in function induced by alterations at the interface between the domains. These functional and structural investigations will provide a comprehensive understanding of the conformational pathway as well as role of dynamics in activation and allosteric modulation in NMDA receptor function.
|
{
"pile_set_name": "NIH ExPorter"
}
|
A training-induced increase in muscle capillarity appears to contribute to the reduced risk of cardiovascular disease and type two diabetes associated with aerobic fitness. Vascular endothelial growth factor (VEGF), a potent initiator of angiogenesis, is upregulated in cardiac and skeletal muscle by exercise. Estrogen also enhances VEGF expression, but the interactive effect of estrogen and exercise on VEGF in muscles has not been studied. The proposed study tests the theory that estrogen status affects exercise-induced upregulation of VEGF in cardiac and skeletal muscle. Four specific hypotheses will be addressed: 1. The levels of VEGF mRNA and protein are lower in the cardiac and skeletal muscles of estrogen-depleted, sedentary female rats than sedentary females rats with normal estrogen levels; 2. Exercise-induced upregulation of VEGF mRNA and protein are attenuated in the cardiac and skeletal muscles of estrogen-depleted female rats; 3. The levels of VEGF receptor (Fit-1 and FIk-1) proteins are attenuated in the cardiac and skeletal muscles of sedentary and exercised estrogen-depleted female rats; 4. Exogenous estrogen replacement restores exercise-induced VEGF mRNA and protein upregulation to control levels in the cardiac and skeletal muscles of estrogen-depleted female rats. These hypotheses will be tested using six groups of 6 mo. female Fischer 344 x Brown Norway rats (n=7 per group): sedentary control; sedentary ovariectomized (OVX), exercise control, exercise OVX, sedentary OVX with estrogen replacement, and exercise OVX with estrogen replacement. Rats in the exercise groups will be run once for 45 min at 20 m/min, 10 degree incline, an intensity previously shown to induce a 4 to 6-fold increase in VEGF mRNA in 6 mo. female Fischer 344 rats. VEGF mRNA levels in the heart and gastrocnemius will be analyzed by real-time PCR. VEGF and VEGF receptor protein levels will be assessed by Western blot. Analyses will be done using two-way ANOVA (exercise status, estrogen status). The results of this study will contribute to the assessment of the risk/benefit ratio for hormone replacement therapy, particularly in the context of those at risk for cardiovascular disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Abstract Elimination of excess dendritic spines on cortical pyramidal neurons during adolescence is critical for excitatory/inhibitory (E/I) balance in adult cortical circuits, and its impairment can lead to altered spine density in autism spectrum disorders (ASD) and schizophrenia. This proposal seeks to illuminate a novel mechanism for dendritic spine remodeling in the developing mammalian frontal cortex, mediated by immunoglobulin (Ig)- class cell adhesion molecules of the L1 family and class 3 Semaphorins. Aims focus on NrCAM (Neuron-Glial Related Cell Adhesion Molecule) and Close Homolog of L1 (CHL1), which are associated with ASD, schizophrenia, and intellectual disability. The central hypothesis to be investigated is that NrCAM and CHL1 form holoreceptor complexes with Neuropilin1/2 and PlexinAs for secreted class 3 Semaphorins, which signal through Rho GTPases to prune dendritic spines of cortical pyramidal neurons during adolescence. Aim 1. Developmental Regulation of NrCAM and Role of CHL1 in Dendritic Spine Remodeling A new conditional mutant mouse (Nex1Cre-ERT2: NrCAMf/f) that inducibly deletes NrCAM from cortical pyramidal neurons will be studied to define the developmental timing of NrCAM function in dendritic spine morphogenesis, spine dynamics, and cortical excitability. A novel function for CHL1 in spine remodeling will be identified by analysis of spine/synapse morphogenesis and cortical excitability in CHL1 null mutant mice. Aim 2. Structural and Functional Interactions of the Sema3F Holoreceptor A structure-function approach using mutagenesis will be undertaken to probe an innovative molecular model of the Sema3F holoreceptor complex, in which the NrCAM extracellular domain interacts with Npn2 to induce Sema3F-induced receptor clustering and spine remodeling. NrCAM cytoplasmic domain interactions with Synapse-associated protein 102 (SAP102) and cytoskeletal adaptors (Ankyrin-B, -G, and Doublecortin-like kinase 1) will be analyzed for promoting receptor clustering and signaling at the nascent postsynaptic density in cortical neuronal cultures and in vivo. Aim 3. Molecular Mechanism of Sema3F Signaling through Small GTPases A novel dual signaling pathway will be investigated in which Sema3F-induced signaling through RhoA (Rho Kinase-Myosin II) generates contractile force that exerts tension on actin filaments assembled through Rac1 signaling (Tiam1-Rac1-PAK-LIMK1-Cofilin1) to regulate spine elimination/protrusion. The role of intrinsic activity in Sema3F-mediated spine pruning will be evaluated by activity blockade in cortical neuron cultures. This project is expected to have sustained overall impact as it will delineate a novel molecular mechanism for regulating excitatory synapse development in the frontal neocortex, advance mechanistic understanding into pathology associated with neurodevelopmental disorders, and may reveal new therapeutic targets for intervention in adolescence, a window of opportunity for influencing cortical networks.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Administrative and Clinical Core will be responsible for overseeing and coordinating all activities of the Program Project. Dr. Luzuriaga, the Principal Investigator of the Program Project will serve as the PI of the Administrative Core. She will be assisted by a Scientific Advisory Committee, consisting of the Project Pi's (Drs. Selin and Rothman) and two UMMS advisors (Drs. Kenneth Rock and Mario Stevenson). The Administrative and Clinical Core will provide the following services of benefit to all Program Project Investigators and Core Leaders: 1) Scientific leadership and administration of Program business operations (including grant administration, budgets, purchase or service contracts); 2) Oversight of the Acute Mono Clinic at the University of Massachusetts, Amherst Student Health Services, specimen processing, and access to clinical specimens; 3) Organization and administration of monthly meetings to facilitate interactions among investigators and allow for discussion of scientific data and progress; 4) Organization of a seminar series; 5) Access to EBV serology and performance of molecular HLA Class I typing on blood from study subjects.; 6) Communication with our NIH Program Officer and coordination of progress reports for submission to the NIH.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Autosomal dominant polycystic kidney disease (ADPKD)is a major cause of disabling morbidity and is the fourth leading cause of end-stage renal failure in the world, affecting more than 500,000 U.S. citizens and millions more worldwide. Researchers at the University of Alabama, Emory University, University of Kansas, Mayo Clinic and Washington University St. Louis joined together in 2000 to create the Consortiumfor Radiologic Studies of Polycystic Kidney Disease (CRISPI). The primary objectives of this investigation were to: (1) to develop and test the accuracy and reproducibility of imaging techniques to monitor changes in renal cyst size and parenchyma! involvement, (2) to establish and maintain a database of uniformly and accurately collected information, and (3) to maintain and make available such data to facilitate the planning and implementation of clinically appropriate interventions in the near future. The goals of CRISPII are to extend the observations of CRISPI in order to: 1) draw unequivocal linkage between the rate of kidney/cyst enlargement and qualitative and quantitative end-points;2) to provide a marker of disease progression (kidney volume) sensitive and accurate enough to be used as a primary outcome marker in clinical trials aiming to forestall disease progression;3) to develop and test other biomarkersof disease progression in ADPKD. The specific aims are: Aim 1: Extend the preliminary observations of CRISPIto ascertain the extent to which quantitative (kidney volume and hepatic and kidney cyst volume) or qualitative (cyst distribution and character) structural parameters predict renal insufficiency. Aim 2: Extend the preliminary observations of CRISP! to ascertain the extentto which age and sex-adjusted measurements of renal blood flow byMR technology predict the rate of renal growth;and, renal blood flow and kidney volume predict the rate of renal function decline in ADPKD.Aim 3: Exhaustively analyze the living database and stored biologic samples derived from CRISPI and the CRISPII extension to develop and test new metrics to quantify and monitor disease progression. Given that hypertension appears to be a predictor of greater rates of renal and cyst growth during CRISP I, in CRISP II we will obtain ambulatory blood pressure data on normotensiveCRISP II participants and continue to collect circulatory markers of the renin-angiotensin-aldosterone system (plasma renin activity, plasma aldosterone concentration and urinary aldosterone excretions) in all CRISPII participants.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The purpose of this study is to analyze the reasons for the decline in fertility in postwar Japan. The empirical evidence gathered through questionnaires administered in six prefectures in Japan, along with macro-economic data and demographic data published by various branches of the Japanese government, will be analyzed with the aid of economic and demographic theories. The objective of this study is to seek interrelationships between fertility measured in various ways and selected economic and social variables in post-1945 Japan.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The CAPRISA Clinical Trials Unit for AIDS/Tuberculosis Prevention and Treatment (CAPRISA CTU), strategically located in KwaZulu-Natal, South Africa, at the epicenter of one of the world's most severe HIV and tuberculosis (TB) epidemics, has recently made significant scientific contributions on tenofovir gel and pre-exposure prophylaxis, HIV-TB treatment, prevention of breastfeeding transmission, and immunity with broadly neutralizing antibodies. The scientific team includes investigators who have served at the highest levels in Network leadership. During the current funding cycle, 2,854 participants have been enrolled in 25 protocols to date, with retention rates usually exceeding 90% and data quality scores exceeding 95%. The CAPRISA CTU comprises 4 components; firstly, experienced leadership, senior investigators and CTU coordinator; secondly, an administrative and governance component; thirdly, 10 research support cores; and fourthly, 4 Clinical Research Sites (CRSs). The eThekwini CRS for vaccine, microbicide and adult HIV treatment studies, is located in central Durban adjoining a local clinic that annually treats 40,000 patients with sexually transmitted infections (HIV prevalence of 59.3% (CI: 56.5-62.0) and HIV incidence rate of 6.4 per 100 person-years (CI: 2.6-13.2)) as well as 3,500 new TB cases, two thirds of whom are co-infected with HIV. At the rural Vulindlela CRS for integrated prevention, vaccine and microbicide studies, the HIV prevalence among young women is 35.7% (CI: 32.7-38.8) and the HIV incidence rate is 6.5 per 100 person-years (CI: 4.4-9.2). In this community, HIV incidence is 10.2 per 100 person-years (CI: 4.1-20.9) in women under 20 years. The Springfield CRS for adult HIV/TB treatment studies is located at a regional referral hospital that treated 2,359 MDR-TB and 206 XDR-TB patients (71% co-infected with HIV) in 2010. The Umlazi CRS for maternal/pediatric HIV studies is at a 1,200 bed hospital that provides pre-natal services to about 12,000 women annually (HIV prevalence: 39% (CI: 36.7-41.5)). The vertical transmission rate is 2.4% (CI 1.1-4.5) at birth but increases to 6.6% (CI 5.3-8.2) at 6-8 weeks post-partum due to breastfeeding. The CAPRISA CTU headquarters is located at the Nelson R Mandela School of Medicine and houses the Administration as well as the laboratory, pharmacy, data management and IT, community engagement, evaluation and quality assurance, financial management, bioethics, communication, regulatory compliance, and training Cores. The CTU's organizational structures (Leadership Group, Executive Committee and Community Advisory Board) and communication tools (regular meetings, video conferences, monthly newsletters and website) enable effective communication, management and governance in the unit. Overall, the CAPRISA CTU has diverse well-characterized high-risk populations, well established clinical facilities, accredited laboratories, pharmacies, and data management systems, strong community linkages, and extensive experience in conducting clinical trials, together with a track record of scientific innovation available to support the 5 Networks in developing new approaches to HIV and TB prevention and treatment.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Microscopy Core will support each of the 4 projects with state-of-the-art electron microscopy, confocal microscopy and fluorescence microscopy equipment, services, and support. These will include: 1. Electron Microscopy and High Resolution Immunolocalization of Proteins. The Core will perform electron microscopic sample preparation and imaging as a service to the projects. Conventional embedded section microscopy will be available, and we are one of the few centers capable of resinless section EM. We have a stereo imaging capability for both techniques. Facilities for digitization of micrographs and for digital image processing will be provided. Training will be available for investigators wanting to do their own sample preparation and imaging. 2. Localization of Proteins and Nucleic Acids by Confocal Microscopy. The Microscopy Core will provide a dedicated Leica TCS SP1 microscope and will train users in its use. Several Leica SP2 confocals are available on a shared basis. Confocal microscopy as a service will be available to occasional users. Image processing software and training in its use will be provided. 3. Live Cell Imaging. The Core will assist users with the imaging of fluorescent proteins in living cells by both fluorescence and confocal microscopy. This will include time lapse studies, FRAP to measure mobility and binding, and FRET to detect protein-protein interactions. 4. Localization of Proteins and Nucleic Acids by Computer Deconvolution Microscopy. Each project will have access to the Digital Imaging Center. This center performs deconvolution of microscopy image stacks and provides sophisticated digital image processing and analysis. 5. Microscopy Consultation. The Core will provide assistance with experimental design, microscopy protocols and instruments, analysis of data, and the preparation of micrographs and figures for publication and presentation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Environmental public health tracking (EPHT), a type of public health or epidemiologic surveillance, is the ongoing monitoring for and identification of local disease or injury excesses attributable to environmental hazards and the dissemination of these data for development of interventions to prevent and/or reduce the frequency and severity of these occurrences. This proposal seeks to undertake a regional, multi-institutional EPHT project to develop collaborations among neighboring EPHT Partners in the northeast US (e.g., Maine, New Hampshire, Massachusetts, Connecticut, New York and New York City) for surveillance and epidemiologic research, to demonstrate new applications and developments in methods for data linkage, pattern detection and assessment of exposure-disease associations, and to train students and other researchers in the use and interpretation of these approaches for the study of the possible environmental causes of disease. Specifically, in addition to conducting a review of current surveillance methodology, we plan to investigate and validate the addition of new data fields relevant to environmental problems to current reporting systems, such as birth certificates. We will review the current applications of ecologic analysis methods to EPHT, noting limitations and opportunities for improvement, develop training materials illustrating key points with concrete, non-mathematical logic and examples, and provide practical guidance for implementation. We also will explore new methods that exploit the spatial coherence of local environmental exposures, such as geographically weighted regression (GWR) and multivariate methods, conducting specific analyses to demonstrate their strengths and weaknesses for EPHT. We will work with EPHT partners to identify and develop a regional EPHT epidemiologic study focusing on a problem of local importance, such as the reproductive effects of exposure to arsenic in drinking water or air pollution, or assessment of the spatial pattern of childhood cancers and related environmental exposures. Initially, our focus will be on facilitating collaboration and data sharing while accommodating concerns about confidentiality. To provide guidance, we will convene an advisory committee of representatives from partner health agencies, partner environmental agencies, industry, NGOs and the public. We see these efforts as an important step from individual EPHT partner programs towards national EPHT network and collaboration. [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
Marijuana use negatively impacts the life course of many of the adolescents who use it; it has been associated with increased risk for multiple psychosocial, mental health, and physical health problems. Racial minorities may be more likely to experience adverse outcomes subsequent to marijuana use, including problems such as: substance use disorders, employment problems, and involvement with the criminal justice system. Neighborhood-level factors represent important but understudied potential determinants of marijuana use and subsequent adverse outcomes, highlighting a critical gap in knowledge that limits our ability to identify effective primary prevention strategies targeted at the neighborhood level. The neighborhood context is uniquely relevant for low-income urban youth, as they are persistently exposed to drug activity, physical disorder, and violence in their neighborhoods, all of which may influence marijuana use and adverse outcomes related to use. With this background, this K01 seeks to: (1) fully characterize patterns of marijuana use among adolescents, cross-sectionally and over time; (2) examine the association between perceptions of neighborhood problems and patterns of marijuana use, cross-sectionally and over time, (3) examine the prevalence of adverse psychosocial outcomes in early adulthood by patterns of marijuana use in adolescence, (4) characterize the spatial association between chronic and severe neighborhood violence, such as homicides and shootings, and patterns of marijuana use in adolescence, and (5) investigate the association between objectively-measured neighborhood-level factors and patterns of marijuana use in adolescence. I will use latent variable analyses to characterize patterns of marijuana use, and the Neighborhood Inventory for Environmental Typologies (NIFETY) tool to assess neighborhood-level factors (i.e., physical disorder, drug activity). Data will come from Dr. Nicholas Lalongo's Baltimore Prevention Program dataset, a longitudinal study of youth in Baltimore, MD, a low-income, urban setting. Supplemental data include violent crime records of Baltimore Police Department, and objective data on neighborhood-level drug activity and physical disorder collected as part of Dr. C. Debra M. Furr-Holden's NIFETY project. The findings will identify modifiable neighborhood-level factors associated with substance use, and will be strategically disseminated so as to inform primary prevention efforts. This research project will be embedded in a comprehensive training program designed to further my strengths and to develop new research skills, under the direction of a team of experts. Research and training activities will assist me in achieving my career goal as a leading, independent investigator in social epidemiology and prevention science, focused on investigating the neighborhood-level determinants, use patterns, and consequences of adolescent marijuana use among low- income, urban youth. This goal reflects my core belief that neighborhood factors - including community violence, drug activity, and physical disorder - negatively impact urban youth and make them more vulnerable to initiate marijuana use, and to experience the adverse effects of use. This K01 will afford me the time to acquire new statistical skills, further develop content expertise in substance use, conduct research, write manuscripts, participate in professional meetings, prepare grant applications based on initial findings, and participate in targeted academic and professional development activities. My training goals for this career development period are to: (1) expand my knowledge of adolescent substance use, particularly among low- income, urban youth; and (2) develop skills in the following four methodological areas: analysis of correlated data, latent variable analysis, spatial analysis and mapping, and environmental assessment. I will achieve these goals through formal didactic experiences (i.e., workshops, directed readings, and coursework), and by conducting research under the mentorship of a team of experts in relevant fields of study. In the final stages of this K01 award period, I plan to submit an R01 to NIDA to further investigate the association between neighborhood-level factors and adolescent marijuana use, ideally within the setting of Boston, MA. In addition to building toward an R01 application, I will also use the protected time afforded by this career development award to develop additional grant ideas in this field of research.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The risk of End Stage Renal Disease (ESRD) due to diabetes has tripled in recent decades. This has occurred despite widespread implementation of treatment with antihypertensive drugs and ACE inhibitors. This epidemic of ESRD is due to a real increase in the proportion of diabetic patients developing renal function loss rather than a consequence of improved survival of these patients. To contain this epidemic, research efforts are urgently needed to identify the determinants and mechanisms of renal function loss in diabetes so that new preventive programs can be developed. Particularly lacking is knowledge about the initiation and promotion of the early renal function decline. Recently, we found that renal function begins to decline in a large proportion of patients with type 1 diabetes once microalbuminuria (MA) develops. This early renal function decline was unrelated to further increases in the level of urinary albumin excretion but was associated with elevated levels of urinary chemokines. Preliminary proteomic analysis of urine from these patients revealed the presence of specific proteins in the urine of individuals with MA and early renal function decline that were absent in the urine of individuals with MA and stable renal function. These unknown urinary proteins represent candidates for exposures that injure the proximal tubules of patients with MA and are responsible for the elevated urinary chemokines. We aim to identify the proteins most associated with early renal function decline. Furthermore, we propose to use methods of proteomic analysis to characterize the urinary chemokines that distinguish patients with MA who are at risk of early renal function decline from those with stable renal function. These questions will be examined in both type 1 and type 2 diabetes. The Specific Aims of this proposal are: 1) To determine the frequency of significant early renal function decline in two cohorts of individuals with MA and type 1 diabetes (n=300), and type 2 diabetes (n=500). 2) To identify urinary protein(s) that cause early renal function decline in both cohorts by comparing urinary protein profiles between cases with early renal function decline and controls with stable renal function using proteomics analysis based on mass spectrometry. 3) To identify urinary and plasma cytokine/chemokine profiles that predict early renal function decline in the two cohorts using a targeted proteomics approach and Luminex technology. 4) To develop an etiologic model of early renal function decline in individuals with type 1 and type 2 diabetes and MA incorporating all the findings from these studies. The proposed research on the mechanisms and determinants of early renal function decline is novel and will provide data for the development of effective methods of prevention of renal function loss in diabetes. [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
While the presence of the immune system in virtually all organ systems is widely recognized, little attention has been paid to the influences of immunity on the inner ear. Research in human subjects demonstrates that hearing loss is successfully treated with corticosteroids in some patients, presumably due to their anti-inflammatory and immunosuppressive effects. Unfortunately, we possess limited knowledge of the cellular mechanisms that influence interactions between inflammatory processes and normal function of the inner ear. Because steroid therapy provides such hope for successful treatment of hearing loss, the specific mechanisms of how inflammation affects hearing warrant further study. The long-term goal of our research is to identify the role of inflammation in cochlear maintenance and injury. In prior work, we have demonstrated that hair cell injury evokes a strong, rapid recruitment of professional phagocytes in the form of monocytes and macrophages into the mouse cochlea. The present studies will use novel pharmacological methods and knockout mouse strains to examine the role of macrophages in cochlear pathology and recovery from injury. In the first set of experiments, we use liposomally-encapsulated clodronate to deplete cochlear macrophages in vivo, in order to determine whether recruited macrophages are protective or detrimental in cochlear injury. Next, we will examine the role of the chemokine receptor CX3CR1, expressed in cochlear monocytes and macrophages, in ototoxic injury. Our studies have shown that genetic deletion of CX3CR1 leads to both increased macrophage entry into the cochlea and enhancement of ototoxic injury. Proposed experiments will determine whether suppressing macrophages in CX3CR1-knockout mice can reverse this effect. Additional studies will examine the involvement of the cardinal scavenger receptor, CD36, in the phagocytosis of injured hair cells. We have shown that CD36 is expressed by both cochlear macrophages and cochlear supporting cells, suggesting that both cell types may be involved in phagocytosis of hair cell debris. We will use CD36 knockout mice to determine whether CD36 is essential for the recognition and removal of apoptotic hair cells from the organ of Corti. We will also use radiation bone marrow chimeras to isolate the effects of CD36 expression on leukocytes versus cochlear supporting cells. A final experiment will examine the dynamic role of macrophages in ototoxicity by time lapse confocal imaging of the live mouse cochlea in culture. Using this technique, we can follow the movement and activity of macrophages to determine how they interact with hair cells in ototoxic injury. These experiments will broadly study the role of leukocytes in the inner ear after hair cell injury and will allow us to begin understanding how immunosuppression affects normal processes as well as pathological processes in the inner ear.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goal of this pilot project is to construct and evaluate a new culture technique for mammalian cells called the "aging-machine". The new aging-machine is similar to the original cellular baby-machine in that it involves the immobilization of a cell population on a surface such that newborn cells are shed from the surface as a consequence of division of the attached cells. The difference with the aging-machine is that the cells are attached to the surface by a means, differential pressure, such that the attachment is permanent and one of the two sister cells formed at division is always released. As a consequence, newborn cells are released indefinitely, enabling automatic analysis of the cell cycle, and numerous other cell growth properties, as a function of the age of the attached cells. The pilot research support will be used to assemble the instruments and to examine the growth characteristics of the immobilized and released cells, including growth rates and quality of synchronous growth, the duration of growth in the instruments, and the maximum cell yields. The tests will be performed initially with mouse L1210 cells, but applications to other cell types will also be evaluated. The Utility of the instruments for investigating cell cycle parameters and partitioning of components between cells at division in relation to replicative aging will be established. One of the major advantages of the aging-machine is that it permits cell cycle studies on essentially undisturbed, or very minimally disturbed, populations. If the instrument operates as easily and as well as anticipated, then it could be used for routine laboratory maintenance of cell populations in a continuous culture system which also automatically generates newborn cells throughout the lifespan of the immobilized population.
|
{
"pile_set_name": "NIH ExPorter"
}
|
I propose to study the polarity of axonal regeneration in a primitive vertebrate, the sea lamprey, Petromyzon marinus. My goal is to understand how neurons maintain their normal polarity following axotomy; i.e., how they regenerate their axons at the axon stump, by-passing other sites within the cell. The lamprey is ideal for addressing this question, as it permits direct access to and visualization of mature identified neurons in a vertebrate CNS in situ. Furthermore, the site of axotomy of certain identified neurons (ABCs) determines whether axonal regeneration occurs from the axon stump (following axotomy distant from the soma) or from ectopic sites in the dendrites (following axotomy close to the soma). I will therefore compare the effects of close .and distant axotomy on factors that preliminary experiments suggest may be important in maintaining normal polarity (i.e., the distribution and phosphorylation of neurofilaments, the stability of microtubules and the influx of Ca++ at the site of axotomy). The importance of each of these factors in maintaining normal polarity following axotomy will be assessed by (1) comparing post lesion changes in these factors that correlate with the occurrence and distribution of sprouts, and (2) perturbing neurofilament transport and phosphorylation, microtubule stability and the intracellular free Ca++ level with specifically acting drugs, and then examining the response of treated ABCs to axotomy. The results of this work should illuminate the cellular mechanisms that maintain axonal and dendritic identity following injury. They should also be relevant to the important health related problems of CNS axonal regeneration and the loss of aspects of dendritic and axonal identity that occur in Alzheimer's disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Obsessive-compulsive behaviors (OCB) are common in children and adolescents. In addition to being the core features of obsessive-compulsive disorder (OCD), OCB are often associated in youth with tic, grooming, generalized anxiety, and autistic spectrum disorders. This competitive renewal application combines the unique clinical assessment, magnetic resonance imaging, and genetics expertise of three performance sites: Wayne State University (WSU), University of Michigan (UM), and the Hospital for Sick Children, affiliated with the University of Toronto (UT). The overall goal of this project - which extends prior and existing NIMH-funded research including Drs. Rosenberg and Diwadkar's neurodiagnostic imaging-genetic studies (K24MH02037; R01MH59299), Dr. Hanna's family, molecular genetic, and action monitoring studies (R01MH53876; R01MH59299; K20MH01065; R01MH101493) and Dr. Arnold and colleagues' extensive genetic studies in pediatric OCD (R01MH59299; R01MH101493) - is to exploit the emerging field of imaging genetics to 1) determine the relationship between alterations in functional connectivity of fronto-striatal-thalamic circuitry (FSTC) as measured by task and resting state functional magnetic resonance imaging (fMRI) and childhood OCB; 2) identify common, rare, and novel genetic variants associated with alterations in connectivity of FSTC as measured by fMRI; 3) clarify whether fMRI measured alterations in FSTC are potential intermediate phenotypes of OCB by determining whether they mediate the effects of genetic variants on OCB; and 4) combine structural MRI data from this study and our previous imaging genetics study to identify genetic variants associated with anterior cingulate volume and other FSTC structures in 1000 youth. The Child Behavior Checklist Obsessive-Compulsive Scale (CBCL-OCS) shows substantial heritability in pediatric twin studies. Heritability of structural and functional abnormalities in STC has also been demonstrated in OCD patients and their unaffected relatives. By using a research design consistent with the Research Domain Criteria (RDoC), targeted high field (3 Tesla) fMRI at WSU will be combined with comprehensive genomic assessment in 200 child psychiatric outpatients with CBCL-OCS scores = 5, 200 child psychiatric outpatients with CBL-OCS scores < 5, and 200 matched healthy controls aged 8-18 years. We will examine for common and rare genomic variants associated with FSTC dysregulation and conduct whole genome sequencing (WGS) in 60 subjects with FSTC dysregulation in the highest 10% of the distribution and compare to 60 subjects with FSTC dysregulation in the lowest 10% of the distribution to identify rare and novel variants of possible clinical significance. This unique study enacts the call for translational approaches to mental illness outlined in PAR- 14-165 by examining multiple genomic variants in FSTC in a spectrum of common, but understudied disorders in youth. Our work will provide a better understanding of the impact of genetic variants on FSTC dysregulation in the pathogenesis of OCB and lead to new diagnostic, treatment, and prevention strategies.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Developmental Research Program of the University of Pittsburgh Cancer Institute Lung Cancer SPORE will be carried out for the purpose of identifying and facilitating innovative new pilot projects in lung cancer research. The overall goal of the Developmental Research Program is to provide seed funding to investigators for novel research in lung cancer to further the basic, clinical and translational research priorities of the Lung Cancer SPORE. The specific goals of this Program are : 1) to provide seed funding opportunities for initial investigation of promising novel research in lung cancer: 2) to stimulate basic, clinical, and translational lung cancer research in areas of high priority to the Lung Cancer SPORE;3) to facilitate development of pilot projects into full project status in the Lung Cancer SPORE or to be competitive for independent investigator-initiated proposals to other funding agencies;and 4) to increase the visibility of Lung Cancer SPORE activities and increase participation among the institution's clinicians and researchers. Our design for the Developmental Research Program in the renewal is similar to the process used during the first grant period. Drs. William L. Bigbee and Joel S. Greenberger will provide Program leadership for basic and clinical research, respectively. The Developmental Research Program will utilize institutional web-based resources and printed announcements to notify the research community at the UPCI, the University of Pittsburgh Medical Center (UPMC), and the affiliated Carnegie Mellon University of the availability of research funds to support pilot projects in lung cancer. In response to the new SPORE guideline encouraging inter-institutional collaboration, we will also make the program available to our sister Cancer Centers in Philadelphia, who are part of the statewide Pennsylvania Cancer Control Consortium (PAC-3) initiative. A standing Developmental Research Program Committee, together with ad hoc reviewers with specific expertise as needed drawn from the UPCI, UPMC, the Lung Cancer SPORE External Advisory Board, or other SPOREs will provide rigorous and consistent peer review of the solicited project proposals. The proposed Developmental Research Program budget will include $70,000 per year in funds from the NCI through this proposal, together with an additional $50,000 per year in matching funds from the UPCI. Proposals will be solicited and reviewed on an annual basis and grants of $30,000-$50,000 per year for 1-2 years will be awarded to the most meritorious proposals. These activities of the Developmental Research Program will stimulate innovative research toward meeting the translations goals of the Lung Cancer SPORE: reduction in the incidence, morbidity, and mortality from lung cancer.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Sex hormones, such as estrogens, are believed to play a major role in gender-based differential immune competence and autoimmunity. One mechanism by which estrogens may influence the immune system is by regulating cytokine levels. We have recently reported that estrogen-treated wild type C57BL/6 mice have increased IFNgamma, mRNA and protein levels. This grant proposal is aimed at mechanistically studying how estrogen alters the production of lFNgamma and the molecular consequences of increased IFNgamma. Estrogen-induced IFNgamma is significant, since IFNgamma is a "master" cytokine with physiological effects on nearly all cells of the immune system: it is involved in resistance against intracellular infections, and in pathological effects of many autoimmune and inflammatory diseases. The hypothesis of this proposal is that increased IFNgamma in estrogen-treated mice is due to the promotion of increased numbers of specific IFNgamma secreting cells, an enhanced response to IFNgamma-promoting cytokines and/or co-stimulatory signals. A consequence of this increased IFNgamma will be altered cellular and molecular functions of IFNgamma target cells. This may be evident as increased expression of IFNgamma responsive genes and molecules, altered patterns of apoptosis and changes in susceptibility to autoimmunity. AIM (1) of this proposal will examine the molecular basis for estrogen-induced IFNgamma. AIM (2) will determine whether responsiveness of IFNgamma-target cells to IFNgamma is altered in estrogen-treated mice, with regard to STAT1 activation, IFNgamma-responsive genes, and expression of IRF-1, IRF-2, Cox-2, and MHC molecules. AIM (3) will investigate whether increased survival of lymphocytes from estrogen-treated mice is due to IFNgamma inducible nitric oxide, by using estrogen-treated wild type, IFNgamma knockout, and iNOS knockout mice. AIM (4) will address whether estrogen treated non-autoimmune mice are prone to develop selected types of induced-autoimmunity, and whether this is due to IFNgamma. This proposal is novel since it will provide a mechanistic-based understanding of how estrogen promotes IFNgamma and its consequences at molecular, cellular, and organismal levels. The proposal will benefit the future understanding of human health, especially with regard to gender-based immune diseases.
|
{
"pile_set_name": "NIH ExPorter"
}
|
DESCRIPTION (Applicant's Description): Most premature morbidity and mortality from cancer is preventable, through primary prevention involving interventions to reduce cancer risks or secondary prevention involving early detection and treatment of localized disease. Despite the clear need for cancer prevention research training, there has been very little support for such programs. This proposal requests continued support for a cancer control and epidemiology research training program at the University of Southern California (USC), where program and collaborating clinical faculty are nationally and internationally recognized for their research in cancer epidemiology/ etiology, primary prevention, early detection, and treatment compliance. Their successful record of extramural funding and collaborative investigations coupled with the resources of the USC-Norris Cancer Center provide an exceptionally rich training environment which includes public and private hospitals and medical centers which are training branches of the USC School of Medicine; a SEER-funded Cancer Surveillance Program; international, national and regional data bases; computer and statistical resources; and association with community organizations, school districts, work sites, and health departments. Funds are requested for four predoctoral and four postdoctoral trainees. Predoctoral positions are allocated to behavioral cancer research coordinated within the Ph.D. program in Health Behavior Research (HBR). NCI support establishing external predoctoral training funds for the first time will facilitate the growth of the training effort and heighten research contributions and development of new technologies for reducing cancer incidence. NCI predoctoral trainees are selected from among the most promising HBR graduate students and will receive three to four years of training culminating in original dissertation studies in cancer prevention and control. Predoctoral training includes structured course work (e.g., research methods, biometry/ statistics, epidemiological methods, biological basis of disease, theories of prevention, compliance/adaptation, foundations of health behavior), weekly research seminars, directed research, journal clubs, and writing groups. Postdoctoral positions are allocated to behavioral and epidemiology research training emphasizing collaborative studies. Since 1985, 27 NCI postdoctoral fellows from the behavioral, public health, and biomedical sciences have received training. The postdoctoral program requires course work or the equivalent in the biological basis of cancer, cancer epidemiology, and statistical methods. Emphasis is placed on involving fellows in on-going research projects and assisting them in their own research. Other training activities include journal clubs, writing groups, and weekly research seminars. All trainees attend a mini- course on research ethics. Minority recruitment plans have been developed.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This is a competitive renewal application where we propose to study cortical network differences between neuropathic and inflammatory chronic pain conditions, and determine the re-organizational properties of these networks as a function of time and of therapy. In the last funding cycle we developed noninvasive brain imaging techniques tailored specifically to study clinical pain conditions. With these tools we studied two populations of chronic neuropathic pain conditions, and showed that the brain network underlying chronic pain is distinct from that of acute pain, that it may be reorganizing in time and also undergoing a neurodegenerative process. In this cycle 3 groups of chronic pain patients are studied, compared and contrasted to matched normal volunteers: chronic inflammatory or neuropathic back pain (CLBP), chronic post herpetic neuropathy (PHN), and chronic osteoarthritic knee joint pain (OAP). [unreadable] [unreadable] In Aim 1 we determine the cortical network underlying ongoing, spontaneously fluctuating, pain perception in chronic neuropathic and inflammatory conditions, by comparing fMRI results between CLBP, PHN, & OAP. In Aim 2 we determine the cortical network underlying stimulus-evoked chronic neuropathic and inflammatory conditions, by comparing fMRI results for tactile allodynia in PHN to joint stimulation-evoked pain in OAP. In Aim 3 we attempt to find a causal link between brain gray and white matter density decreases and gray and white matter chemical decreases, mainly of N-acetylaspartate (NAA), as a result of chronic pain, differentiating between neuropathic and inflammatory conditions. These entail MRI-based morphometric studies, MRI spectroscopic measures of brain gray matter and white matter chemistry, and water diffusion based studies of white matter tracks. These studies are done in a cross-sectional and longitudinal design. In Aim 4 we study the reversibility of cognitive deficits, chemical concentration decreases and gray matter density decreases in OAP and PHN patients before and after pain therapy. [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
Duchenne Muscular Dystrophy (DMD) is caused by mutations in the gene coding for dystrophin, which functions to maintain muscle fiber structure and function, preventing it from being damaged by muscle contraction. Presently, there is no definitive treatment for DMD patients. Current therapies focus on prolonging survival and improving quality of life. Definitive treatment will require that functional dystrophin protein is restored in all affected muscle groups. Possible approaches include cell therapy, gene therapy or a combination of the two. We hope that transplantation of a particular type of muscle repair cell will help us to develop new therapeutic approaches to this disease. Muscle stem cells, termed satellite cells, isolated from healthy donors or patients should be able to provide dystrophin and repair muscle damage in DMD patients. For efficient therapy of DMD, satellite cells which maintain the self-renewing ability are necessary. Therefore, in this proposal, (1) we will focus on the study of how cultured satellite cells maintain their self-renewal capacity. In addition, systemic injection of satellite cells is an essential protocol that will provide healthy ells into all affected muscle tissues in DMD patients. Thus, (2) we will attempt to improve systemic delivery methods for satellite cells using a novel homing receptor expression. Furthermore, we need to use cells that are not rejected by a patient's immune system. (3) We are able to generate an unlimited number of satellite cells from induced pluripotent stem cells (iPSCs) derived from the patient's myoblasts. In combination with gene transduction, this concerted approach will help us to make satellite cells that can be transplanted into patients for a definitie cure of DMD. PUBLIC HEALTH RELEVANCE: Duchenne Muscular Dystrophy (DMD) is caused by mutations in a gene called dystrophin, which acts to maintain muscle fiber structure and function, preventing it from being damaged by muscle contraction. Presently, there is no definitive treatment for DMD patients and current therapies focus on prolonging survival and improving quality of life. In combination with gene transduction of a functional dystrophin protein this concerted approach will enable us to make satellite cells that can be transplanted into patients for a definitive cure of DMD.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objectives of the research described in this application are to investigate the fate of the adrenal medulla chromaffin granule after it has released its secretory products by exocytosis. Biochemical and pharmacological procedures will be used in the perfused isolated cow adrenal gland to study the ability of the empty chromaffin granule to function in the a) synthesis, b) storage and c) release of newly synthesized catecholamines and chromogranin-proteins. Incorporation of radioactive precursors will be used to follow the synthesis of chromaffin granule components and to estimate the turnover of chromaffin granule components. Morphological studies in in vivo rabbit adrenal medullar and in perfused isolated cow adrenal glands will be carried out in an attempt to provide a morphologic interpretation of the biochemical findings obtained in this study. It is felt that a combination of biochemical and morphological approaches offers the best opportunity for gaining an understanding of the full life cycle of the chromaffin granule.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Class II MHC (Ia) gene products play critical roles in a variety of T and B lymphocyte responses. A combination of immunological, molecular genetic, and biochemical appraoches is being used to study the relationship between Ia structure and function. The defect in expression in most cases of "allelically mismatched" Alpha and Beta chains lies in the failure of assembled heterodimers to efficiently exit the endoplasmic reticulum. Many of these poorly transported dimers can be "rescued" at least in part by co-expression of adequate amounts of the non-MHC encoded invariant chain, a process that is affected by the intracellular localization signal(s) on the cytoplasmic tail of the invariant chain. This same region also controls the efficiency of class II peptide acquisition. In vitro experiments with purified class II demonstrate that conformation is dictated by whether or not appropriate peptides are bound by the NH2-terminal domains of the AlphaBeta dimer, and that the most stable structure of class II includes such bound peptides. The use of direct peptide binding techniques, together with transfectants expressing mutant class II molecules, has permitted the definition of a local peptide binding subregion in the murine I-E molecule, and the role of specific allelically polymorphic residues in controlling the quantitative and qualitative binding of peptide to this molecule. Finally, mutational analysis has begun to define the site(s) of interaction of Ia with CD4, a molecule that functions as a co-receptor along with the T cell receptor in recognition of class II molecules during T cell activation. Together with the studies on class II folding, transport, and control of peptide binding by the polymorphic domain, these experiments will provide new insight into the molecular mechanisms involved in antigen recognition by and activation of T lymphocytes.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Despite the availability of antibiotics to combat Tuberculosis (TB), it is one of the leading causes of death due to infectious disease. Mtb is a successful pathogen because it survives within immune cells and effectively establishes and maintains a latent tuberculosis (TB) infection. Treatment of latent TB is lengthy in part because quiescent or slowly replicating Mtb acquires a drug-tolerant phenotype. Therefore, understanding the mechanisms underlying the establishment or maintenance of dormancy can inform new strategies for TB therapeutics. MmpL (Mycobacterial membrane protein large) transporters are dedicated to the transport of cell wall lipids. MmpL proteins are therefore crucia players in mycobacterial physiology and pathogenesis. MmpL3 is essential to mycobacterial viability; and MmpL4, MmpL5, MmpL7, MmpL8, MmpL10 and MmpL11 contribute to Mtb virulence. Our goal is to elucidate the function of MmpL11. Our data show that MmpL11 transported monomeromycolyl diacylglycerol and a mycolate ester wax to the M. smegmatis cell wall. These lipids belong to classes of lipid species that are sometimes referred to as storage lipids and are associated with dormant bacteria in vitro and accumulate in granulomas of TB patients. Therefore, it appears that MmpL11 plays a role in a clinically relevant, but poorly understood, aspect of Mtb pathogenesis. Our working model is that MmpL11 is a conserved transporter of mycolic acid-containing lipids. We hypothesize that the biosynthetic pathway for the MmpL11 substrate can be revealed by defining the MmpL11 interactome. This project will define proteins that interact with MmpL11TB demonstrate the biosynthetic pathway leading to production of the MmpL11 substrate. Our preliminary data indicate that MmpL11 interacts with LpqN and TesB, and mutants lacking these proteins share phenotypes with mmpL11 mutants. Both MmpL11 and the biosynthetic pathway leading to the MmpL11TB substrate are potential targets for therapeutic intervention.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This research will develop an optical instrument to aid in the noninvasive evaluation of skin affected by injury, surgical procedures and/or disease. The instrument will use characteristics of light absorption by hemoglobin species to delineate skin regions compromised by ischemia, hypoxia or necrosis. This research is based on the working hypothesis that reflectance spectra of pigments in skin provide information that can be used to locate and evaluate compromised regions of skin. During Phase I a skin imaging system will be constructed to obtain reflectance images at visible and near-infrared wavelengths from skin. Wavelengths will be chosen to monitor changes in hemoglobin absorption and tissue light scatter. Phase l studies will demonstrate feasibility of the working hypothesis using the rat pedicle skin flap as a model and provide design parameters for instruments to be constructed during Phase ll for clinical trials. During Phase lithe imaging system will undergo clinical testing to determine the system's ability to assist in the evaluation of vascular disorders, burns, wound healing and reconstructive surgery. The instrument will provide clinicians with a noncontact diagnostic method that is truly noninvasive and requires no injections or skin contact. It will provide rapid, easily interpreted image displays and be technically easy to use.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Antigenic variation underlies a large number of the most critical problems now facing vaccine development. Influenza virus is an ideal subject for tackling this issue: it is the most variable pathogen against which we can effectively vaccinate, but its principal surface antigen varies so rapidly that new vaccines must be introduced almost yearly. We propose to use innovative analyses of B-cell repertoires and new possibilities in structural studies to understand the structural and immunological mechanisms underlying cross reactivity and immunodominance, as a foundation for designing immunogens that will elicit a more broadly neutralizing response than those currently in use. We will test the following three hypotheses, to which the three Aims of each Project correspond. (1) The B-cell repertoires elicited by inactivated vaccines and by infection differ in degree of polyclonal activation and breadth of neutralization; the repertoires elicited by immunization with adjuvanted and non-adjuvanted TIV differ because adjuvant increases the breadth of recognized epitopes. (2) Broadly reactive Abs, including those recognizing the heterosubtypic stem epitope, are less frequent after true primary than after secondary TIV immunization due to broadening of the response by multiple HA stimulations. (3) Efficient induction of Abs against a desired epitope requires: (a) proliferation of a favorable germline Ab and (b) an affinity maturation pathway to a desired final specificity; there are preferred germline precursors and maturation pathways for Abs targeting particular epitopes. The group of investigators that joins in this proposal brings together strengths in immunology, virology, structural biology, and vaccine development. The principal investigators have worked closely and effectively together in past collaborations.
|
{
"pile_set_name": "NIH ExPorter"
}
|
In 1994 we discovered that yeast can have prions, infectious proteins analogous to the transmissible spongiform encephalopathies of mammals. We showed that the non-mendelian genetic element, URE3, is a prion of the Ure2 protein, and that PSI+ is a prion of Sup35p (1). We found the first biochemical evidence confirming our discovery (2) and defined the prion domain of Ure2p (2). These prions are amyloids of the respective protein (reviewed in 3). Unexpectedly, shuffling the prion domain amino acid sequence of Ure2p or Sup35p did not alter the ability of these domains to support prion formation, suggesting that the amyloid structure is parallel in-register (4). We have shown by solid-state NMR (in collaboration with Rob Tycko of NIDDK) that the amyloids of Ure2p, Sup35p and Rnq1p are indeed in-register parallel beta sheets (5-7). It has not escaped our notice that this in-register parallel beta sheet structure can explain how a given protein sequence can encode any of several biologically distinct prion variants based on biochemically distinct self-propagating amyloid structures (8). We have examined the URE3 prions based on Ure2 proteins from non-cerevisiae species of Saccharomyces, and have demonstrated a similar species barrier to that seen among mammals of different species in their transmission of spongiform encephalopathies (9). We showed that the variant properties, as defined by species barrier, are maintained even during passage through a different species. We also noted that the Ure2p of Saccharomyces castellii cannot become a prion (9). We find that the Candida albicans Ure2p can form a URE3 prion in S. cerevisiae, but that of Candida glabrata cannot, even though the prion domain of glabrata is closer in sequence to that of cerevisiae than is that of abicans (10). Thus the conservation of sequence in the Ure2 prion domains is not for prion-forming ability, but must reflect the function of this domain in protecting the full length protein from degradation in vivo (11). We find that the C. albicans Ure2 protein or its prion domain each readily form amyloid which is highly infectious for yeast, and, like the other yeast prions, has a parallel in-register beta sheet architecture (12). While the disaggregating chaperone Hsp104 plays a dominant role in prion propagation, it was not known to have a role in prion generation. We found that overexpression of Hsp104 increases the frequency of generation of the C. albicans URE3 prion by 70 fold or more (13). This effect appears to be mediated in part by effects on the PIN+ prion, and in part by preventing formation of amorphous aggregates. We have shown that PSI+ does not occur in wild strains, as it certainly would were it advantageous (14). To further examine the biology of PSI+, we designed a method to screen for a lethal (Suicidal) PSI+ which efficiently incorporated all of the essential Sup35 protein into amyloid, should such a variant exist. We indeed found that lethal variants of PSI+ and those which produce extremely slow growth comprise more than half of total isolates (15). We also found that common variants of the URE3 prions cause extremely slow growth, although deletion of the URE2 gene in these strains did not slow growth (15). This toxic URE3 therefore cannot be due to a simple deficiency of Ure2p, but must be a due to a pathogenic amyloid. Current efforts are directed to understanding the nature of this toxicity. These results confirm the pathologic nature of the yeast prions PSI+ and URE3. Understanding their mechanisms of pathogenesis may be useful in understanding human amyloidoses. 1. Wickner RB (1994) URE3 as an altered URE2 protein: evidence for a prion analog in S. cerevisiae. Science 264: 566 - 569. 2. Masison DC &Wickner RB (1995) Prion-inducing domain of yeast Ure2p and protease resistance of Ure2p in prion-containing cells. Science 270: 93 - 95. 3. Wickner RB, Edskes HK, Ross ED, Pierce MM, Baxa U, Brachmann A &Shewmaker F (2004) Prion Genetics: New Rules for a New Kind of Gene. Ann. Rev. Genetics 38: 681-707. 4. Ross ED, Minton AP &Wickner RB (2005) Prion domains: sequences, structures and interactions. Nat. Cell Biol. 7: 1039-1044. 5. Shewmaker F, Wickner RB &Tycko R (2006) Amyloid of the prion domain of Sup35p has an in-register parallel b-sheet structure. Proc. Natl. Acad. Sci. USA 103: 19754 - 19759. 6. Baxa U, Wickner RB, Steven AC, Anderson D, Marekov L, Yau W-M &Tycko R (2007) Characterization of b-sheet structure in Ure2p1-89 yeast prion fibrils by solid state nuclear magnetic resonance. Biochemistry 46: 13149 - 13162. 7. Wickner RB, Dyda F &Tycko R (2008) Amyloid of Rnq1p, the basis of the PIN+ prion, has a parallel in-register b-sheet structure. Proc Natl Acad Sci U S A 105: 2403 - 2408. 8. Wickner RB, Shewmaker F, Kryndushkin D &Edskes HK (2008) Protein inheritance (prions) based on parallel in-register b-sheet amyloid structures. Bioessays 30: 955 - 964. 9. Edskes HK, McCann LM, Hebert AM &Wickner RB (2009) Prion variants and species barriers among Saccharomyces Ure2 proteins. Genetics 181: 1159 - 1167. 10. Edskes HK, Engel A, McCann LM, Brachmann A, Tsai H-F, Wickner RB. Prion-forming abilityof Ure2 of yeasts is not evolutionarily conserved. Genetics 2011;188:81 - 90. 11. Shewmaker F, Mull L, Nakayashiki T, Masison DC, Wickner RB. Ure2p function is enhanced by its prion domain in Saccharomyces cerevisiae. Genetics 2007;176:1557 - 65. 12. Engel A, Shewmaker F, Edskes HK, Dyda F, Wickner RB. Amyloid of the Candida albicans Ure2p prion domain is infectious and has a parallel in-register b-sheet structure. Biochemistry 2011;50:5971 - 8. 13. Kryndushkin DS, Engel A, Edskes HK, Wickner RB. Molecular chaperone Hsp104 can promote yeast prion generation. Genetics 2011;188:339 - 48. 14. Nakayashiki T, Kurtzman CP, Edskes HK, Wickner RB. Yeast prions URE3 and PSI+ are diseases. Proc Natl Acad Sci U S A 2005;102:10575-80. 15. McGlinchey R, Kryndushkin D, Wickner RB. Suicidal PSI+ is a lethal yeast prion. Proc Natl Acad Sci USA 2011;108:5337 - 41.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The purposes of the project are to investigate the biological roles of members of the chemokine family of cytokines, to use chemokine receptors to understand the relationships between the trafficking patterns and broader biological functions of subsets of effector/memory T cells, and to understand the contributions of the chemokine system to infectious and inflammatory/autoimmune disease and cancer. Chemokines and their receptors are critical for leukocyte trafficking, and our experiments are directed to understanding how manipulating the chemokine system could be used to treat diseases in which leukocytes play a critical role. In addition, some chemokine receptors are expressed by cancer cells, and these receptors can potentially be exploited for diagnosis and as targets for therapy. In FY 2017, we have continued to investigate mouse models of skin inflammation that have features of psoriasis. One model involves injection of a cytokine, IL-23, which appears to have a role not only in psoriasis, but also in other immune-mediated diseases, such as Crohns disease. We and others had described that the chemokine receptor CCR6 is expressed by IL-23-dependent T cells that produce the cytokines IL-17 and IL-22. IL-22 and IL-17 are important in producing disease in the mouse psoriasis model, and are thought to be important in causing tissue injury in some autoimmune diseases. We have shown previously that mice lacking CCR6 are resistant to the IL-23-induced disease, but the mechanism has been unclear. We further characterized the origins and types of monocyte/macrophage and dendritic cell subsets that infiltrate the skin and contribute to the psoriasis-like inflammation. We have identified a subset of inflammatory dendritic cells that enter the outer layers of the skin (epidermis), determined that these and other dendritic cells important for the inflammatory response are derived from infiltrating monocytes, and studied how these cells are recruited. We have found that CCR6 supports the recruitment of the monocytes that give rise to monocyte-derived dendritic cells that are important for IL-23-induced dermatitis. We also determined that the initial responding cells are resident gamma/delta T cells that produce IL-17. Most recently, we have focused on the role of the skin microbiota in lymphocyte homeostasis in the dermis and epidermis and the effect of the microbiota on inflammation in psoriasis-like dermatitis induced by the TLR7/8 agonist, imiquimod. We have been characterizing the roles of chemokines and their receptors in microbiota-dependent positioning of lymphocytes in the skin. In the last year we have continued studies of the process whereby effector/memory T cells migrate from the blood, across the layer of endothelial cells that line the inside off the blood vessel, into a site of tissue infection or inflammation. We have characterized subsets of CD8alpha+ T cells that are particularly efficient at migrating across the endothelium, and we have identified some of the molecular features of the surfaces of these cells, including the combination of chemokine receptors, that make them so efficient. We have continued work on a transcription factor that regulates a number of genes that encode proteins important for the ability of these cells to migrate across endothelium into inflammatory sites. Most recently, we have begun to extend these studies to subsets of human CD4+ (helper) T cells, initially characterizing the relationships among patterns of expression of chemokine receptors, selectin ligands, and transcription factors of interest. We have found that some of the chemokine receptors that are critical for transendothelial migration are shared between subsets of CD8+ and CD4+ T cells. In the last year we have completed a series of studies on the expression of the chemokine receptor CXCR4 on a range of cancers. We found expression of CXCR4 on significant percentages of major types of kidney, lung, and pancreatic adenocarcinomas, and, notably, on metastases of clear cell renal cell carcinoma and squamous cell carcinoma of the lung. We found particularly high expression of CXCR4 on adrenocortical cancer (ACC) metastases. A first-in-human study using 64Cu-plerixafor for PET in an ACC patient prior to resection of metastases showed heterogeneity among metastatic nodules and good correlations among PET SUVs, CXCR4 staining, and CXCR4 mRNA. Additionally, we were able to show that CXCR4 expression correlated with the rates of growth of the pulmonary lesions in this patient.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This application seeks to understand mechanistic elements of the myosin motor of striated muscle and to determine how specific human mutations disrupt myosin function and lead to skeletal muscle disease or cardiomyopathy. We will employ a multidisciplinary approach that takes advantage of the powerful genetic tools available for Drosophila melanogaster along with a broad range of expertise that allows us to study myosin in an integrative manner, from its crystal structure and kinetic function through its effects upon muscle ultrastructure, fiber mechanical properties and locomotion. For Aim 1, we will build the first models of human distal arthrogryposis syndromes, which cause skeletal muscle contractures of varying severity. Using transgenic Drosophila, we will assess effects upon myosin structure, ATPase activity, in vitro motility, indirect flight and jump muscle ultrastructure, fiber mechanics and organism locomotion. We will test the hypothesis that distal arthrogryposis mutations cause abnormal interactions with nucleotides yielding enhanced ADP binding, reduced sliding velocity, slowed relaxation dynamics and hypercontraction and that the severity of the defects correlates with the severity of the human syndromes. For Aim 2, we will examine the mechanistic basis of myosin dysfunction caused by two hypertrophic cardiomyopathy mutations. We will construct organisms expressing point mutations that change the charge of the disease-causing residues and then test predicted suppressor mutations for functional rescue in vitro and in vivo. This will reveal interactions altered by the initial mutatins and provide direct insight into protein-protein interactions that are critical for myosin function. We will test the hypothesis that mutations in residues that cause human hypertrophic cardiomyopathy alter contacts between the strands of the central ?-sheet of myosin or interactions between the N-terminal domain of myosin and the lever arm, resulting in increases in ATPase activity, actin sliding velocity, fiber power levels and myofibrillar disarray. For Aim 3 we will examine the ability of the Drosophila heart to hypertrophy as a result of expressing myosin mutations known to cause human hypertrophic cardiomyopathy. This will be the first attempt to determine how the simple Drosophila heart tube, which is known to exhibit dilated, constricted or hypertrophic phenotypes, reacts to human contractile protein mutations that cause hypertrophy. We will monitor effects upon RNA transcription, calcium handling, ultrastructure and cardiac physiology and determine whether manipulating signaling pathways by gene knockdown or pharmacological intervention ameliorates observed defects. We will test the hypothesis that myosin mutations that cause human hypertrophic cardiomyopathy result in significant wall thickening, abnormal myofibrillar arrays, arrhythmias, disrupted calcium signaling, and transcript profiles that mimic the human condition, and that these pathologies can be suppressed by pharmacological treatment or genetic intervention. Overall our project will take advantage of an innovative experimental system to test significant questions regarding basic myosin function and disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The ADAMHA/MIRDP program at the University of Puerto Rico, Rio Piedras aims to advance the growth of high quality and rigorous research on alcohol, drug abuse, mental health and AIDS through: 1) Involving faculty members in conducting pilot work leading to a competitive research grant on alcohol, drug abuse, mental health, and AIDS (ADM) problems, specially evaluation, and treatment outcome research; 2) Supporting research development activities for faculty involved in ADM research; 3) Enhancing and strengthening the institutional infrastructure for conducting ADM research; and 4) Increasing the participation of graduate and undergraduate students in ADM research. The rationale for the program is based on the need to develop a parallel structure of support to carry out independent ADM research by minority faculty at a minority institution. The proposed program for faculty research development UPR-RP consists of two components: 1) developmental phases of faculty research activities; and 2) faculty development activities. A unique feature of the program is the organization of current institutional resources in a clear, coherent, and integrated plan of career development in support for ADM research. Faculty members are involved in carrying out their research from the moment the program begins and participate in a developmental individualized plan that supports the faculty member in a step by step approach toward the objective of producing a competitive research proposal. The participation of undergraduate and graduate students in the development of faculty research is another aspect of the program aimed at increasing the training opportunities offered to students in ADM research. Eleven faculty members from four graduate programs (Psychology, Sociology, Social Work) at the College of Social Science and the College of Education (Department of Graduate Studies) will participate in the proposed MIRDP.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Colorectal cancer (CRC) has served as a unique platform for durable insights into and advances in molecular pathogenesis, chemoprevention, diagnosis, and therapy, not only in this venue but expanded into other disease states. The elucidation of these comprehensive aspects of CRC has been facilitated through the identification of key genes and molecular pathways. It is well accepted that the majority of sporadic colorectal cancers feature the chromosomal instability pathway, involving alterations in key tumor suppressor genes (APC and p53, but also SMAD4) and oncogenes (especially Ras, but also EGFR, c-myc, B-Raf) and their downstream effectors. A subset of sporadic colon cancers undergo microsatellite instability (MSI) pathway, typically right-sided with B-Raf mutations but without Ras mutations. Recently, it has been advocated that certain colorectal cancers feature hypermethylation. We have genetically and physically mapped a region of loss of heterozygosity (LOH) on chromosome 22q13.31 as a basis to understand the role of new genes in colorectal carcinogenesis (and other cancers). We now have discovered a new telomeric region of LOH, which harbors two microRNAs, namely Let-7a3 and Let-7b, and these microRNAs are downregulated in up to 40% of colorectal cancers. MicroRNAs have become increasingly recognized for their pivotal, diverse roles in development, differentiation, proliferation, and cancer, which is achieved the degradation or interference with translation with messenger RNAs. We hypothesize that the Let-7a3 and Let-7b microRNAs are instrumental in colorectal cancer progression, and this may be achieved by targeting the Ras oncogene, and also, the c-myc oncogene. This novel hypothesis will be pursued by the following interrelated Specific Aims: (1). To identify the roles of the Let-7a3 and Let-7b microRNAs in colorectal cancer. A. To determine the relationship between these microRNAs and clinical parameters and molecular parameters; B. To determine the function(s) of Let7a and Let7b in colorectal carcinogenesis through genetic /overexpression/knockdown studies of these microRNAs with determination of effects upon cellular proliferation, apoptosis, migration and invasion, which are critical features of the colon cancer initiation and progression. (2). To determine if the Ras and c-myc oncogenes represent targets of Let-7a3 and Let- 7b. This will be achieved through examination of Ras transcriptional and protein activities in the setting of genetic approaches, namely Let-7a3 and Let-7b overexpression/knockdown. (3) To determine the functional consequences of restoration and inhibition of Let-7a3 and Let-7b in colon cancer cells in vivo. This will be assessed by innovative three-dimensional organotypic cultures, which mimic the colon cancer microenvironment, and in athymic, irradiated nude mice through bioluminescence of tumor growth. In aggregate, our studies have the means to unravel new pathways of regulation of the Ras and c-myc oncogenes by the Let-7a3 and Let-7b microRNAs in colon cancer, especially where sporadic colorectal cancers (chromosomal instability pathway) do not harbor Ras mutations. PUBLIC HEALTH RELEVANCE: Colorectal cancer (CRC) serves as a paradigm for the investigation of basic mechanisms and translation into novel chemopreventive, diagnostic and therapeutic strategies. The underlying pathogenesis of CRC may involve the Let-7a3 and Let-7b microRNAs, which in general are critical regulators of normal cellular processes to malignant transformation. Our studies hope to reveal new paradigms in CRC pathogenesis, and offer opportunities for exploiting this knowledge in novel diagnostics and therapeutics. [unreadable] [unreadable] [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
We have reported that cultured mast (RBL-2H3) cells generate growth-promoting cytokines constitutively (e.g. TGFbeta) and inflammatory cytokines (e.g., TNFalpha and IL6) when stimulated. Production of inflammatory cytokines is mediated by mobilization of Ca2+, activation of protein kinase C (PKC), and a third synergising signal, tentatively identified as the p42 MAP kinase (ERK2) pathway (see last years report). Secretion of these cytokines is dependent on Golgi, mobilization of Ca2+, and activation of PKC and is readily blocked by selective disruption of any one of these three processes by pharmacologic agents. We have now positively identified the ERK2 pathway as a necessary signal for production of TNFalpha as follows. Of all stimulants tested, antigen is by far the most potent stimulant of this pathway and TNFalpha production. Both are inhibited equally well by dexamethasone, the kinase inhibitor, quercetin, and the MEK inhibitor, PD098059, at concentrations that have minimal effects on other stimulatory signals in RBL-2H3 cells. The small amounts of TNFalpha that are formed are secreted, consistent with previous indications that the MAP kinase pathway regulates production but not secretion of TNFalpha. Production of TNFalpha, however, is not affected by wortmannin, which inhibits phosphoinositide 3- kinase and, as a consequence, various other kinases including the JNK family of MAP kinases. In contrast to TNFalpha, the constitutive production of TGFbeta is unaffected by inhibitors of the MAP kinases and PKC, although a small additional increase in production TGFbeta can be elicited by antigen and this increase is suppressed by such inhibitors to indicate regulation of this inducible component of TGFbeta production. Thus, both constitutive and inducible pathways exist for production of cytokines and the inducible pathways can be selectively suppressed by pharmacologic agents. The studies with the above inhibitors reveal parallel effects on the phosphorylation of a cytosolic phospholipase A2 (cPLA2) by ERK2 and release of archidonic acid. These and previous studies permit the following conclusions. The ERK pathway, which is activated through PKC, calcium, and a Shc/Ras pathway that provides sustained signals for activation in antigen-stimulated cells, is required for phosphorylation and activation of cPLA. In antigen-stimulated cells, PKC causes transient stimulation of cPLA and arachidonic acid release, via the ERK pathway. Inhibitors of PKC thereby delay, but do not block, release of arachidonic acid whereas inhibitors of the ERK pathway suppress this release. Our studies provide for the first time plausible explanations for the actions of several experimental and therapeutic agents on release of inflammatory mediators from mast cells.
|
{
"pile_set_name": "NIH ExPorter"
}
|
In the first period, work in this BRP demonstrated that MP4 (PEG-modified hemoglobin) overcomes the most significant hurdle to the development of modified hemoglobin-based blood substitutes, namely vasoconstriction. MP4 promotes tissue oxygenation through a combination of O2 transport and maintenance of functional capillary density in spite of its counterintuitive properties, including increased O2 affinity, viscosity and oncotic pressure. MP4 is superior to blood in its ability to resuscitate animals from severe, uncontrolled hemorrhage, and it has been shown to be safe in human clinical trials. In this present application we will test the hypothesis that MP4 is an effective carrier of the heme ligands O2, carbon monoxide (CO)and nitric oxide (NO), and will carry out the related physiological studies in order to understand its effectiveness as a blood substitute and identify new clinical applications for its use. We will test the hypothesis that PEG-Hb formulations are vasodilators which interact with the circulation in ways not related to NO scavenging by Hb. We propose that MP4's properties are in part due to an increased nitrite reductase activity and NO transport. We will develop a procedure for using MP4 to deliver CO and exploit that CO-MP4 is exceptionally stable, even at elevated temperatures, making it valuable in field use for trauma. Project 1 has a GMP facility that provides MP4 of consistent quality and properties. It will develop and produce new PEG-Hb compounds with goals to optimize concentration without increasing colloid osmotic pressure and augment O2 delivery capacity so as to increase the applicability of MP4 to a wider range of clinical uses. Properties will be screened via biochemical analysis, mathematical modeling, and systemic experiments in rats including the effect on myocardial infarction. Project 2 will examine PEG-Hbs' effects on the glycocalyx integrity, how PEG-Hbs' presence influences reactive O2 species (ROS), and will investigate the combined effect of PEG-Hb and enhanced plasma viscosity. MP4 will be used as a delivery vehicle for CO to provide cellular protection during ischemia & hemorrhage. Microcirculation studies will use the awake hamster window chamber model, with direct measurements of O2 and NO levels, flow and diameter in blood vessels and functional capillary density. This research combines physiological analysis of transfusion, fundamentals of engineering transport processes and mechanotransduction with the expertise of two laboratories with more than 12 years of collaboration. Effective blood substitutes will significantly increase the safety and efficacy of blood transfusions in civilian and military settings, streamline and simplify transfusion medicine. Applications for a new type of blood substitute will be developed, one that provides a fundamentally different treatment of ischemia, based on enhancement of microvascular flow, cardiac and renal functions, repair of the endothelium and delivery of heme ligands (O2, NO and CO).
|
{
"pile_set_name": "NIH ExPorter"
}
|
The ion, oxalate, and its salt, calcium oxalate, are of special interest to scientists who study bacteria, fungi, plants, animals, and humans. Scientists from other scientific disciplines including physical chemistry, biochemistry, crystallography, membrane physiology, molecular biology and genetic engineering have had interest in oxalate and calcium oxalate. Through the ideal medium of the Gordon Research Conference, these many varied disciplines can meet with a common theme to share ideas and knowledge that potentially will be beneficial to all. The first two Gordon Conferences on Calcium Oxalate were judged extremely successful by the participants and the Gordon Conference organizers. With this background and enthusiasm the third Gordon Research Conference on Calcium Oxalate will be held June 10-14, 1991 at Plymouth State College, Plymouth, New Hampshire. Scientists from throughout the world will meet to share their knowledge including the measurement of oxalate in biological fluids and tissue; the metabolic origin and roles of oxalate in biology; oxalate transport by biological membranes; and the many processes and factors involved in the crystal formation of calcium oxalate. Scientists from the varied disciplines will be called on to present papers and discuss in open forums in the informal atmosphere of a Gordon Research Conference in these important areas of mutual interest. In addition to the oral presentations and discussions, poster sessions will be featured allowing a large number of participants, especially young investigators, to share their work with the other attendees.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This application is to provide the candidate with scholarly training, mentorship, and protected time with the goal of becoming an independent clinical researcher. Dr. Salehi will study the functions of gastrointestinal (Gl) hormones and neural signaling in glucose metabolism by focusing on the physiologic changes following Roux-en-Y gastric bypass (RYGB). RYGB has been reported to reverse type 2 diabetes (T2DM) immediately after surgery before any significant weight loss. In addition, a growing number of patients have been recognized with life-threatening hyperinsulinemic hypoglycemia several years following their surgery. These findings suggest that the RYGB procedure alters the regulation of (-cell function to promote insulin secretion. While the mechanisms by which RYGB improves glucose metabolism or alters islet cell function in patients after RYGB are not understood, recent studies suggest that increased secretion of Gl hormones, primarily glucagon-like peptide 1 (GLP-1), may contribute to enhanced insulin secretion in general, and to a greater extent in patients with hypoglycemia. However, there is also reason to suspect that changes in neural signaling may influence islet cell function in concert with Gl hormones after RYGB, since these patients have an earlier insulin peak and a larger glucagon secretion in response to food ingestion. The proposed research is designed to address the role of RYGB on insulin secretion by determining 1) the contribution of stimulatory factors (neural and Gl hormone) on islet cell function;and 2) the islet cell responsiveness to the physiologic stimulatory factors, in RYGB patients with and without hypoglycemia and non-operated controls. The first goal will be achieved by comparing the effects of cholinergic or GLP-1 blockade on the release of postprandial islet hormones. The second goal will be accomplished by measuring beta-cell responsiveness to exogenous GLP-1, sham feeing, or oral glucose ingestion. To achieve her career objectives, the candidate proposes a five-year training program to learn skills in laboratory research methods, statistical analysis, and islet cell physiologic studies focusing on the role of Gl hormones and neural regulatory factors. Her primary sponsor is highly accomplished investigator in glucose metabolism and Gl hormones and the secondary sponsor in neural signaling and islet cell physiology. Understanding the physiologic mechanisms whereby RYGB influences islet cell function, both improving glucose metabolism and causing hypoglycemic syndrome can lead to the development of new therapeutic options for T2DM as well as surgical refinements or medical therapies for those patients who develop hypoglycemia. PUBLIC HEALTH RELEVANCE: Better understanding of metabolic consequences of RYGB is critical given the rising use of gastric bypass surgeries due to dramatic increase of obesity in the population. The results of this project will provide insights to apply to treatment of diabetes as well as modification of bariatric techniques to reduce the late-complications of hypoglycemia in the groups of patients at risk.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objectives of this revised renewal proposal (RO1 CA 102431A2) are to determine whether protein kinase C epsilon (PKC5) signals ultraviolet radiation (UVR)-induced development of squamous cell carcinoma (SCC) via increased expression of the pro-inflammatory cytokines tumor necrosis factor alpha (TNF1) and granulocyte colony-stimulating factor (G-CSF) which may directly promote the proliferation of hair follicle stem cells, the targets of SCC. Chronic exposure to UVR is the most common etiological factor linked to the development of SCC, a nonmelanoma form of skin cancer that can metastasize. PKC5 is among the six isoforms (1, 4, 5, 7, 5, 6) expressed in both human and mouse skin. To determine the in vivo functional role of PKC5 in mouse skin carcinogenesis, we generated PKC5 transgenic mouse (FVB/N) lines 224 and 215 that overexpress approximately 8- and 18-fold respectively PKC5 protein over endogenous levels in basal epidermal cells and cells of the hair follicle. We reported that epidermal PKC5 level dictates the susceptibility of transgenic mice to the development of SCC elicited either by the repeated exposure to UVR or by the DMBA - TPA tumor promotion protocol. Our studies to elucidate mechanisms of PKC5-mediated development of SCC, either by DMBA-TPA or UVR, indicated a common converging point- elevated release of TNF1 and G-CSF. To conclusively determine whether TNF1 is essential for the development of SCC in PKC5 transgenic mice, we generated TNF1-knockout//PKC5 transgenic mice by crossbreeding TNF1 knockout mice with PKC5 transgenic mice. Deletion of both TNF1 alleles in PKC5 transgenic mice only partially inhibited (50%, p = 0.0136) the development of SCC by the DMBA-TPA protocol. Deletion of the TNF1 gene in PKC5 transgenic mice also partially suppressed UVR-induced skin damage and inflammation. These results indicate that, in addition to TNF1, other UVR-induced cytokines such as G-CSF may play essential roles in the development of SCC. We hypothesize that: 1) TNF1 cooperates with G-CSF in PKC5 signal transduction pathways to the development of SCC by UVR, 2) PKC5-induced TNF1 and G-CSF mediate induction of SCC through direct effects on stem cells in the mouse hair follicle and 3) PKC5 interacts with MAPK cascade to regulate UVR- induced TNF1 and G-CSF gene transcription. We propose three specific aims to test these hypotheses: Specific aim #1: To determine, using gene knockout approach, whether both TNF1 and G-CSF contribute to the development of SCC in PKC5 transgenic mice by UVR. Specific Aim# 2: To define, in intact mouse skin in vivo, whether the roles of TNF1 and G-CSF in UVR-induced development of SCC involve the activation and proliferation of putative hair follicle stem cell. Hair follicle stem cell proliferation will be determined by quantitations of label retaining cells (LRCs) using BrdU and FACS analysis using cell surface markers (CD34, 16-integrin). Specific Aim #3: To define the mechanism, using gene-silencing techniques, by which PKC5 signals UVR-induced TNF1 and G-CSF gene expression.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The mesolimbic dopamine system, which consists of dopaminergic neurons in the ventral tegmental area (VTA) and their projections to the nucleus accumbens (NAc) and other forebrain regions, is an important neural substrate for the reinforcing and locomotor activating properties of several types of drugs of abuse, including opiates, cocaine and other stimulants, and ethanol. We have demonstrated a series of common, chronic biochemical actions of these drugs in the VTA and NAc, which could underlie part of the long-term changes in mesolimbic dopamine function that characterize drug addiction. These changes include an upregulation of tyrosine hydroxylase (the rate limiting enzyme in dopamine biosynthesis) in the VTA and an upregulation in the cAMP pathway in the NAc. Moreover, chronic administration of these drugs of abuse reduces levels of neurofilament proteins and increases levels of glial filament proteins specifically in the VTA, changes suggestive of neural insult or injury. These findings raise the possibility that neurotrophic factors, which exert a neuroprotective effect in many systems, may counter these and other actions of drugs of abuse. We have recently obtained direct support for this possibility. Infusion of certain neurotrophic factors directly into the VTA can prevent and reverse the ability of morphine and of cocaine to produce many of their typical biochemical actions in the mesolimbic dopamine system. One major objective of this Project is to further characterize such novel pharmacological actions of neurotrophic factors in this neural pathway. A second major objective of this Project is to investigate the related possibility that some of the previously identified biochemical changes that drugs of abuse produce in the mesolimbic dopamine system occur via drug regulation of neurotrophic factor signaling pathways in these brain regions. Support for this possibility comes from preliminary investigations wherein chronfc administration of morphine or cocaine has been shown to regulate levels of specific proteins involved in the neurotrophic factor signaling cascades, including ERK (also referred to as MAP kinase), phospholipase C-gamma-l, and JAK (a cytoplasmic protein tyrosine kinase). The proposed studies will further investigate drug regulation of these proteins, as well as their possible role in mediating morphine and cocaine regulation of some of the other biochemical adaptations identified in the VTA and NAc. For example, recent experiments in which levels of ERK1 were selectively reduced in the VTA by an antisense oligonucleotide strategy support the hypothesis that morphine regulation of ERK1 may mediate its upregulation of tyrosine hydroxylase in this brain region. Together, the proposed studies have the potential to reveal important new information concerning the molecular mechanisms of action of drugs of abuse on the mesolimbic dopamine system, which could contribute ultimately to the development of novel pharmacotherapies for addictive disorders.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.