text
stringlengths 1
32.7k
| meta
dict |
|---|---|
The object of this research is to determine how information carried on the pulmonary CO sub 2(t) and PO sub 2(t) signals effect period and depth of breathing. And to perfect ways of studying biological control systems with minimal disturbance of the normal balance of the feedback control loops. A unique feature of this research is a unidirectionally ventilated bird in which air is forced into the trachea, passes through the lung and air sacs and then out through surgically placed exit tubes to the outside. Air flow is kept high and constant and the CO sub 2 and O sub 2 concentrations independently forced and respiratory period and tidal volume monitored with a whole body plethysmograph. Since the birds' respiratory movements no longer influence significantly the concentrations of pulmonary CO sub 2 and O sub 2 it is an open-loop system. This system may be made into an externally closed-loop one by using an analog computer to set the CO sub 2 (or O sub 2) level. Observing the input (CO sub 2, O sub 2)-output (respiratory period, tidal volume) relationship provides constraints for modeling this biological control system.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The ability of the motor system to respond appropriately in the presence of disturbances is a fundamental property known as impedance. Specifically, impedance relates a dynamic positional displacement to the corresponding torque about the joint. Many interesting studies investigating impedance have been published that elucidate numerous properties of motor control. However, there have been no studies that explore the dynamic impedance of lower-limb joints during dynamic tasks. In this study we propose to determine the dynamic impedance of the ankle during walking. We will use a custom built robotic platform to perturb the ankle about its center of rotation and measure the ground reaction force. Using mechanics, we can determine the torque about the ankle during the perturbation, which establishes the input (angular position displacement) and the output (torque) to be used in a system identification technique. We will employ a quasi-static, linear identification technique to determine the impedance of the ankle, subsequently analyzing the impedance in terms of mechanical components.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The general goal of Project 1 is to develop flexible and practical methods for synthesis of deoxyoligonucleotides to which carcinogens have been attached in a structurally defined manner. The project will play a pivotal role in the Program Project, i.e., it will provide DNA oligomers containing adducts of important carcinogens for use in biophysical and biochemical studies. Specific goals to be addressed involve preparation of adducts of polycyclic aromatic hydrocarbons (PAH) at exocyclic amino sites on guanine and adenine in oligodeoxynucleotides. Adducts of the PAH diol epoxides derived from benzo[a]pyrene, benzo[c]phenanthrene, benz[a]anthracene, and 7,12-dimethylbenz[a]anthracene will be prepared. Two novel, indirect strategies will be employed for the synthesis of these adducted oligomers. One will involve differential protection of potentially nucleophilic sites on the oligomer followed by (1) selective deprotection of the target site and then (2) reaction with the diol epoxide as a solution to the regiospecificity problem that plagues direct adduction strategies. The other will involve halogen-substituted nucleosides which will be incorporated into oligomers; adducts will be formed in a completely regiospecific and sterospecific manner via displacement of the halogen by aminotriols derived from the PAH diol epoxides.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goal of the Administrative Core Is to maintain and enhance an organization that coordinates and Integrates Center activities, attracts new Investigators of outstanding ability to environmental health sciences (EHS), contributes to career development, facilitates interdisciplinary activity, creates and promotes new cutting-edge research opportunities, and provides a two-way avenue of community interaction.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This initiative seeks to expeditiously test and analyze novel interventions (i.e., compounds) and their molecular and/or clinical targets for treating clinical dimensions of psychopathology (e.g., social engagement, rigidity or inflexibility) comprising the core features of ASD. Of particular interest are features of ASD as described in the current DSM-IV-TR diagnostic entities, but not typically identified as the primary target of current clinical therapeutics. As described above, there is interest in the study of mechanisms that cut across traditional disorder categories and where relevant mechanisms and clinical targets are assessed directly rather than being inferred through assignment of a particular diagnosis. The NIMH has great interest in studies and clinical trials assessing clinical mechanism and demonstrating target engagement. The outcome of this initiative is expected to lead to enhanced understanding of specific target engagement by such novel interventions, leading to development of innovative treatment approaches for clinical dimensions of psychopathology associated with ASD. In this context, novel interventions (i.e., compounds) may refer either to new chemical entities (NCEs) or to compounds that are being considered for re-purposing from other indications. Testing of compounds that have been FDA-approved for other indications (re-purposing) is of interest if recent basic research discoveries suggest that the compound(s) have the potential to affect a biological mechanism contributing to mental disorders and that has previously been untested in clinical studies. Compounds acting on molecular targets that replicate those of currently marketed psychiatric pharmaceuticals are not of interest for this contract.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The overall objective of this proposal is to develop a new type of peptide molecular conjugates that can effectively and selectively deliver transgenes into target cells. These conjugates will be produced by a combinatorial approach through rational molecular design of independent modules, characterized systematically through modeling and experimental approaches. This proposal is based on the hypothesis that these peptide molecular conjugates will have the potential for considerable flexibility, versatility and selectivity for gene delivery. The ultimate goal of this preliminary research is to develop a peptide molecular conjugate system to selectively deliver genes into target cells in vivo. It has been widely recognized that DNA delivery is the bottleneck impeding gene therapy from reliable clinical implementation. Without an efficient and selective gene delivery technology, the promise of gene therapy can never be fully realized. This proposal aims to develop an enabling technology to systematically address this issue, combining biochemical and biomedical engineering with molecular and cell biology. Because gene therapy is a broad-based medical technology that can be applied to a variety of cell and tissue systems, different applications will require a wide range of special delivery vehicles with high efficiency and selectivity. A new type of DNA-binding self-assembling peptide system (DSAPS) as a molecular conjugate gene delivery vehicle will be developed. These molecules will have three distinctive segments: 1) a receptor recognition ligand for a specific cell surface receptor; 2) a membrane fusion segment, either from a viral fusion segment or a variety of synthetic fusion peptides; and 3) a DNA binding segment with nonspecific DNA binding motif found in histones. One of the most important features of this type of delivery vehicle is that it has no DNA packaging limit. Efforts will focus on developing some model vehicles with several independent modules to deliver genes. The combination of protein engineering and bioengineering approaches represents a new paradigm in constructing delivery vehicles.
|
{
"pile_set_name": "NIH ExPorter"
}
|
DESCRIPTION: (Applicant's Abstract) This project explores how drug craving is influenced by pharmacologic and associative factors, and develops a human laboratory model of associative drug craving. Drug craving is thought to be important in maintaining regular drug use in the current drug user and in leading to relapse in the former user. A complete understanding of drug craving and successful treatments that reduce drug craving are likely to involve both pharmacologic and associative factors. Pharmacologic factors that influence drug craving might be agonist or antagonist administration. Associative factors that influence drug craving might be stimuli that are related to drug delivery. This laboratory-based project uses cigarette smokers and cigarette craving to understand better how subjective craving and drug self-administration are influenced by agonist (nicotine) and antagonist (mecamylamine) administration, and by drug-related stimuli (e.g., smoke, cigarette, etc.). The project consists of four studies of cigarette smokers. The first study systematically examines the influence of nicotine and mecamylamine on subjective cigarette craving and on cigarette self-administration. The two medications will be presented alone and in combination, as recent data suggest that the combination may reduce craving more than either medication alone. The second study uses a Pavlovian extinction procedure to determine the associative nature of craving reductions elicited by denicotinized cigarettes. The study is based upon data from the animal laboratory demonstrating that extinction of the responses elicited by drug-related stimuli can require an extended period of unreinforced exposure. The third study examines the combined impact of pharmacologic and associative factors on cigarette craving. This study uses the most effective nicotine/mecamylamine combination from the first study to examine pharmacologic craving reduction while smokers smoke denicotinized or nicotinized cigarettes. The fourth study explores a human laboratory model of associative nicotine craving by pairing nicotine administration with environmental stimuli. In sum, this project examines how pharmacologic and associative factors influence subjective craving and drug self-administration using nicotine as a model drug, and cigarette smokers as model drug users. This rigorously designed project uses controlled, safe, and convenient laboratory methods, multidimensional measures, and powerful within-subjects designs. Data from this project will yield specific knowledge regarding cigarette craving and, more generally, will guide theoretical understanding of drug craving and influence medications development for drug abuse treatment.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This is a continuing project directed toward obtaining a better understanding of lung function. The primary objective is to investigate the biosynthesis and metabolism of prostaglandins by the lung and to relate changes in steady-state concentrations of the various prostaglandins to biological effects. In addition, the effects of various environmental agents (SO2, O3, NO2, O2, etc.) on the ability of the lung to synthesize and degrade prostaglandins will be investigated. An attempt will be made to correlate the biological effect of pollutants and modulation of prostaglandin levels.
|
{
"pile_set_name": "NIH ExPorter"
}
|
We established that cells from Ewing tumors form neurites in response to Wnt-3a and have begun to define the mechanisms that account for this effect. Frizzled3 (Fzd3) was identified as the primary Wnt receptor that mediates the process, which also requires Dishevelled-2 (Dvl-2), Dishevelled-3 (Dvl-3), and amino-terminal c-Jun kinase (JNK). We showed that Dickkopf-1 also promotes neurite outgrowth in these cells, apparently by facilitating Fzd3/JNK activation by endogenous Wnts. Neurite outgrowth induced by Wnt-3a was associated with Dvl-2/3 phosphorylation; both neurite formation and Dvl phosphorylation were blocked by the casein kinase 1 delta/epsilon (CK1d/e) inhibitor, IC261. Knockdown of CK1d with small interfering RNA suppressed Wnt-3a-dependent neuritogenesis, whereas knockdown of CK1e stimulated neurite formation in the absence of exogenous Wnt-3a. CK1d but not CK1e was detected at the centrosome, an organelle associated with neurite formation. Deletion analysis mapped the centrosomal localization signal (CLS) of CK1d to its carboxyl-terminal domain. A fusion protein containing the CLS and EGFP displaced full-length CK1d from the centrosome and inhibited Wnt-3a-dependent neurite outgrowth. In contrast to wild-type CK1e, a chimera comprised of the kinase domain of CK1e and the CLS of CK1d localized to the centrosome and rescued Wnt-3a-dependent neurite outgrowth suppressed by CK1d knockdown. These results provide strong evidence that the centrosomal localization of CK1d is required for Wnt-3a-dependent neuritogenesis. Knockdown of the atypical PKCiota also blocked Wnt-3a-dependent neurite outgrowth. Wnt-3a stimulated the phosphorylation of PKCi. Dvl2 co-immunoprecipitated with PKCi, and this interaction did not occur when CK1 phosphorylation sites in Dvl2 were replaced with alanine residues. These results suggested that Dvl2 phosphorylation by CK1 was required for Dvl2-PKCi binding, which in turn might be necessary for neurite outgrowth. This work is significant not only because it provides insights about mechanisms involved in the formation of neurites. Many of the factors that participate in neurite outgrowth contribute to cell polarity in other contexts such as the formation of cellular extensions critical for cell migration. Moreover, we have evidence that CK1d also participates in the formation of primary cilia. Defective primary cilia are responsible for several disorders including neural tube defects, polycystic kidney disease and situs inversus. Aberrant Wnt signaling also can elicit these abnormalities. Thus, our studies of CK1d and Dvl may provide new insight about the ways in which Wnt signaling controls embryonic development and its dysregulation contributes to pathogenesis.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The overall goal of the research described in this grant application is to better understand a very bizarre and novel process of transcript maturation that involves the often very extensive insertion (and occasional deletion) of U residues from primary transcripts to achieve the mature RNA sequences. This "RNA editing" has thus far been shown to be utilized by mitochondrial RNAs of trypanosomes but it may well be used by other organisms as well and is thought to provide clues to the early evolution of RNA metabolism This U insertion/deletion is "guided" by short complementary RNAs (gRNAs) which evidently also serve to donate their U residues to the mRNA through formation of a chimeric intermediate with the mRNA portion downstream of the site of editing. But how these gRNAs do the guiding and what is the mechanism of formation of the chimeric intermediate and of the entire reaction are some of the crucial, unresolved questions about RNA editing that we plan to address. Using a trypanosome mitochondrial extract recently shown to form such gRNA/mRNA chimeras in vitro, we will investigate the mechanism/catalytic basis of this chimera formation. To help determine whether the in vitro chimera formation occurs by transesterification or by an endonuclease/ligase route, we will identify the precise site of the junction and the stereochemical considerations imposed on the junction phosphate of the chimer~ We will further analyze chimera formation by studying known co- fractionating activities and by further purification of the activity. We will also attempt to recreate an entire editing reaction in vitro (preliminary very encouraging results have now shown specific cleavage of the gRNA/mRNA chimera and reformation of an intact partly edited mRNA!) and then will study this reaction. Analysis in permeabiIized cells should provide much needed information about the kinetics of the editing process, including the production of the surprisingly abundant aberrant edited molecules. In complementary studies of the steady state mitochondrial RNA population, we will clone and analyze various intermediates in the editing process, asking whether these molecules correspond to predictions from transesterification or nuclease/ligase models of editing, whether editing occurs in strict 3'-> 5' order or shows some flexibility, and whether activities suggested from the in vitro analysis are utilized for editing in vivo. We will use in situ hybridization analysis to localize where within the trypanosome mitochondria the unedited, the partially edited, and possibly the fully edited RNAs are located. This information could allow determination of whether editing occurs in the kinetoplast mitochondrial body or in the long mitochondrial tubule that extends from it along the length of the trypanosome. Through these studies we hope to gain a better understanding about this very ancient and novel form of RNA maturation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goal of this Phase 1 clinical study is to evaluate the impact of patients' knowledge and expectancy that healing efforts on their behalf are being made from a distance on objective and subjective measures of wound healing, psychosocial functioning, and physiological symptoms following breast reconstruction surgery. People undergoing surgery are among those who make frequent use of distant healing modalities. Patients undergoing elective reconstructive surgery will be randomized into three groups (n=26 in each group), which will be compared with each other and with a normative comparison sample. The first two groups will follow a standard double blind randomized trial model assessing the effects of Distant Healing (DH) on wound healing as measured by collagen deposition. This design will allow for traditional assessment of the effects of DH on wound healing by comparing a blinded treatment group with a blinded no-treatment placebo group. The third group will be an open label treatment arm in which patients receive DH and are told directly that they are receiving it. Subjects will be assessed at baseline and 14 days post-surgical follow-up. Baseline attitudes toward the efficacy of DH will be assessed, as will belief about assignment of treatment condition (in blind groups) and confidence that DH is actually occurring on their behalf. If direct knowledge of healing efforts is beneficial, the open-label treatment group should fare better than both blind groups and the normative comparison sample. If DH has an effect, the blind treatment group should show more healing than the blind no-treatment placebo group and the normative comparison sample. If expectancy and DH have an additive booster effect, differences between the open-label vs. blind treatment group should exceed those of the blind treatment vs. blind no-treatment group. Finally, if attitudes toward DH and confidence in being healed are associated with greater healing, these measures should correlate with healing in all groups.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The principal objective of this research is to develop an injectable, bioerodible drug delivery device that will release the narcotic antagonist naltrexone by an erosion-controlled process by kinetics that approach zero order. Primarily because of size constraints, the designed lifetime will be 30 days. This system will be developed using an ointment-like bioerodible poly(ortho ester), recently developed in our laboratories. The hydrolysis of this polymer is very well understood, and the ultimate degradation products are the toxicologically benign 1,2,6-hexanetriol and a carboxylic acid. The ointment-like consistency of the polymer allows incorporation of high concentrations of naltrexone by a simple mixing procedures at room temperature without the use of solvents, thus minimizing potentially adverse naltrexone-polymer interactions and maximizing the amount of drug that can be incorporated into the polymer. The actual delivery system will consist of a polymer drug mixture, microencapsulated within a bioerodible, macroporous membrane. This encapsulation will confine the ointment-like material to a fixed geometry, allow administration by means of a conventional hypodermic syringe, and prevent removal by the patient. We have selected an interfacial polycondensation procedure to surround the polymer with a macroporous polyamide membrane produced by the condensation of a diamine and a diacid chloride. The ointment-like material is first mixed with dry ice and then ground to produce a fine powder. This is then added to ethanol, maintained at -40 degrees C, the diamine added, and this mixture dispersed in n- hexane, containing the diacid chloride, also maintained at -40 degrees C. Because polyamides are essentially nondegradable and contain a toxic diamine, a special diamine prepared from L-alpha-alanine and a diol will be prepared. This diamine is degradable by virtue of the ester groups and is based on a nontoxic amine. The resulting 90 micromoles capsules can be injected through an 18-gauge needle. Toxicology and pharmacokinetic data will be obtained to show biocompatibility and functionality of the system. The specific studies proposed are mutagenicity testing, an intravenous biodeposition study with polymer hydrolysate in rats, a tissue biocompatibility in rabbits, and a pharmacokinetic study in rabbits.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The long term objective of the proposed research is to understand the role of the plasma membrane (PM) proteins and the cytoskeleton in mediating the formation of apical and basolateral domains of the plasma membrane in cells that exhibit a transcellular polarity. Examples of such cells are epithelial cells which are bounded on one side by a fluid phase and on the other by an extracellular matrix. In vitro adhesion of HeLa cells (a transformed epithelium easily grown in suspension) to an extracellular matrix protein, gelatin, will be used as a model to study the formation of the apical and basolateral PM. In this system the initial stages in the formation of these PM domains are cell attachment and spreading. HeLa cells attached to gelatin coated culture dishes are induced to spread rapidly and form an apical PM exposed to the culture medium and a basal PM adjacent to the culture dish. The apical and basal domains can be isolated and redistribution of proteins between the domains during spreading can be quantitated. We have identified five gelatin receptors on the HeLa cell surface. The kinetics of cell attachment and spreading indicate that spreading is a cooperative process leading to the hypothesis that the mechanism for spreading is the segregation of the cell surface extracellular matrix receptors into the basal PM with subsequent clustering of the receptors into oligomers inducing them to bind to the cytoskeleton. The hypothesis will be tested by concentrating on several specific aims: 1, determine which of the known gelatin receptors mediate attachment and/or spreading; 2, determine which receptors redistribute and become segregated among the apical, basal and internal PM domains during cell adhesion; 3, determine if gelatin receptor clustering is correlated with cell spreading; 4, determine if receptor binding to the cytoskeleton is regulated by receptor clustering and/or covalent modification; 5, determine which cytoskeletal proteins bind to the gelatin receptors and; 6, determine if the receptors are transmembrane proteins present in oligomeric complexes and if they have common polypeptide fragments. Results from the specific aims will undoubtedly increase our understanding of the molecular mechanism of cell adhesion to an extracellular matrix and the formation of transcellular polarity.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Cytotoxic T cells have a critical role in eliminating intracellular pathogens and tumors. They arise when naive CD8 T cells are exposed to mature dendritic cells (DCs) that present an activating antigen (Ag). Upon priming by DCs, naive T cells differentiate into mostly short-lived cytotoxic effector cells and relatively few long-lived memory cells. Several memory cell subsets with distinct tissue tropism exist, of which the central memory cells are particularly important for sustained immune protection. We have developed in vitro methods to generate homogenous antigen-specific effector and central memory cell populations using T cells from newly constructed transgenic mice in which each subset expresses distinct genetically encoded fluorescent tags. These cells will be studied in homing experiments and by intravital microscopy to investigate the traffic signals that guide them to normal and inflamed tissues. Aim 1 is based on the hypothesis that upon antigen stimulation CD8 T cells acquire the capacity to migrate to certain non-lymphoid tissues, especially sites of inflammation. We will characterize these default changes in CD8 T cell trafficking that are evoked by TCR stimulation and cytokines without tissue-specific signals. We will dissect the molecular mechanisms by which in vitro generated effector and central memory cells migrate to normal and inflamed tissues (subaim 1.1) and examine the role of the lipid chemo attractant leukotriene B4 and its receptor BLT1 (subaim 1.2). Subaim 1.3 will determine where and when activated T cells acquire effector functions in vivo. In addition to the antigenic stimulus, the route of immunization also influences memory cell migration. We have discovered that lymphoid organ-specific DCs instruct T cells how to migrate to those tissues that are most likely to contain their cognate Ag. Aim 2 will explore how a particular tissue context modulates T cell differentiation induced by DCs and investigate the mechanisms of DC-mediated tissue-specific imprinting. We will ask how DCs from Peyer's patches induce effector cell migration to the small intestine (subaim 2.1) and whether DCs from cutaneous lymph nodes target effector cells to the skin (subaim 2.2). In subaim 2.3 we will dissect the mechanisms that allow skin- and gut-tropic effector cells to migrate into uninflamed tissues and determine how inflammation affects tissue-selectivity.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This protocol is designed to compare different methods for managing a patient in whom a central venous catheter has been inserted and to determine the optimal clinical life span of such catheters. The primary goal is to assess whether the incidence of infection differs when catheters are replaced only when clinically necessary compared to replacement on a regular (i.e., weekly) schedule. A second goal is to examine whether sequential replacement of catheters over guide wires results in a different incidence of infection than sequential replacement at new sites. The study population consists of Clinical Center patients who require central venous catheters for clinical indications; the study randomizes the frequency and manner of catheter replacement. The outcome variables are catheter insertion site infections and catheter related bacteremia or fungemia. The design of the study also will allow re-evaluation of the use of semiquantitative catheter cultures as a means of identifying catheters sufficiently colonized to be a source of septicemia. More than 100 patients have been randomized into this study within the first year of its implementation. It is anticipated that another Washington-area hospital will implement the same study protocol during 1994.
|
{
"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. With our current Startup allocation we have completed the initial port of Desmond 2.2 to the Cray XT3 and Altix 4700 (Bigben and Pople) and made these available to users at the PSC. We now request a renewal of this Startup allocation to continue our efforts to develop Desmond on these platforms. The initial implementation of Desmond on the XT3 uses native MPI. We would now like to look at the possibility of creating an even more efficient implementation of Desmond on the low-level Portals network interface, as had been done with Infiniband networks. In addition, as users at PSC start using Desmond we anticipate that there will be some as yet undiscovered performance issues that will have to be addressed. Also, Desmond is under active development and will likely include improved or additional capabilities that we will want to provide to users of PSC resources.
|
{
"pile_set_name": "NIH ExPorter"
}
|
In response to RNA #RR-00-003, entitled Centers of Biomedical Research Excellence (COBRE), the University of Puerto Rico (UPR) submits this multi-disciplinary application entitled Center for Molecular Developmental and Behavioral Neuroscience. This proposal is part of a UPR systemwide initiative to move the institution into the Research I category according to the Carnegie Classification. The University is nurturing and promoting the formation of these systemwide efforts under its initiative to create Multi-campus and Multi-disciplinary Research Institutes. The COBRE proposal could serve as the foundation for the development of one of these institutes. The prime objectives of this proposal are to: 1) create a competitive research center in neuroscience that will optimize the capacity of the UPR system to compete nationally, 2) develop young scientists into competitive RO1 researchers under the leadership of a Center Director and the scientific guidance of an External Advisory Committee; and 3) expand the capacity of a neuroscience research in Puerto Rico. The goals of the proposed collaborative research projects are to advance the scientific understanding of the molecular mechanisms underlying neuronal injury, emotional memory, cocaine- seeking behavior, and the expression of maternal behavior. These goals and objectives will be achieved by: 1) hiring an established researcher in the field of neuroscience to provide leadership and guidance to the Center; 2) hiring three additional competitive researchers to develop a cadre of neuroscience competency in PR; and 3) providing a supportive and nurturing environment (with the full backing of the administration) for the developing young faculty. The Advisory Committee, chaired by Dr. Edward A. Kravitz, George Packer Berry Professor of Neurobiology at the Harvard School of Medicine, will provide scientific guidance for the development of a strong thrust in neuroscience at UPR and the development of RO1 researchers. The University will provide the resources ($2.5 M in direct match funds plus four professional lines) for recruiting and hiring the Permanent Center Director and three new scientists and provide them the necessary startup funds to help them establish their laboratories.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Research Translation Core (RTC) of the UW Superfund Basic Research Program is well positioned to assist SBRP investigators in disseminating important research findings and new technology to a wide spectrum of stakeholders. The Specific Aims of the Core address the required components described in the SBRP request for applications: Government Agency Partnerships, Technology Transfer, and Communicating to Broad Audiences. The core will continue to: 1) develop communication strategies to exchange information regarding government agency priorities and emerging SBRP research findings. An Agency Seminar Series has been set-up to support experts who can help agency staff address local hazardous waste and contamination issues, 2) assist investigators in moving UW SBRP research findings into application through partnerships with the UW Tech Transfer office. The Core co-director and staff meet with SBRP investigators regularly to identify potential patents and leverage open-source approaches to advance research findings, 3) provide critical information to individual stakeholders by identifying key communication venues and materials with government agencies and community groups. Procedures and materials will be developed for communities concerned about impacts from environmental contaminants, local and state agencies concerned about their community outreach, and for disease advocacy groups and health professionals interested in environmental health education. Annual workshops and print/web materials will used to increase dissemination of current research findings between SBRP investigators, agency staff and community members, 4) partner with Community Groups. The UW SBRP has a long history of working with Tribal Nations, community groups, health professionals, and individual stakeholders to address environmental health concerns. The scope of activities will broaden to include new collaborations, and 5) work with the Administrative Core to refine a Research Translation and Outreach Tracking System to provide documentation and reporting of Core activities on an ongoing basis. The UW SBRP focuses most intensively on developing early biomarkers of adverse effects to human health and the environment from neurotoxic chemicals. Developing early indicators of health effects from exposures to neurotoxic compounds is a theme that resonates with the scientific community, governmental agencies who are involved with hazardous waste sites and the public as a whole.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Two neural circuits that we propose to study control appetitive Pavlovian learning and sleep. Abnormalities in these neural circuits are linked to drug addiction and sleep disorders, two major human health problems with severe negative consequences to human productivity. We focus on a class of neurons that originate from one of the five known progenitor domains in the ventral neural tube. These neurons are called the V2a neurons and are generated throughout the hindbrain and spinal cord. Following genetic ablation of V2a neurons, mice show significant deficit in appetitive Pavlovian learning and sleep. Which V2a neurons are recruited to neural circuits that regulate appetitive Pavlovian learning and sleep? Do V2a neurons project to and connect with known elements of the neural circuitry underlying appetitive Pavlovian learning and sleep? Finding answers to these questions is significant at three levels. (1) It would lead to important new insights for understanding the evolution and adaptability of the nervous system and how microcircuits are assembled. (2) It would for the first time demonstrate that inputs from pontine nuclei are needed for appetitive Pavlovian learning and that V2a neurons and their connections to the mesolimbic system might play a critical role in drug addiction. (3) It will provide neural basis of sleep induced muscle atonia and sleep disorders caused by its malfunction. To answer the aforementioned questions we have devised a genetic strategy to target pontine V2a neurons that originate from rhombomere 1 (r1-V2a neurons). We propose to use a mouse genetics approach to test three hypotheses. (1) The r1-V2a lineage generates neurons that populate functionally diverse pontine tegmental nuclei. (2) The r1-V2a neurons regulate appetitive Pavlovian learning via direct projections to the RMTg. (3) The r1-V2a neurons regulate sleep architecture via direct projections to interneurons in the ventral spinal cord. Successful completion of these studies will demonstrate that embryonic V2a neurons originating from r1 are recruited to distinct neural circuits that regulate appetitive Pavlovian learning and sleep.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The overall objective of this research is to test the hypothesis that monoclonal antibodies to radiation-induced tumor neoantigens cause therapeutically beneficial immune responses in cancer models. We will study the efficacy of a mouse monoclonal antibody to a radiation-inducible neoantigen and analyze immune effector cell activation to control cancer. This strategy complements the use of existing therapeutic antibodies by inducing additional cell surface tumor antigens for simultaneous targeting by multiple antibodies. Presently, therapeutic antibodies for cancer are limited to antigens that are either specific to cancer or are over-expressed in cancers. Not only is the number of such antigens limited, but they also tend to be over-expressed in a small percentage of patients (e.g. 30% breast cancers express Her2/neu). In contrast, radiation-inducible antigens are expressed in nearly all cancers tested thus far and expand the number of therapeutic targets for antibody development. The principles of radiation-inducible neoantigens are that cancer cells over-express certain intracellular proteins including TIP-1, the focus of this proposal. Cancer cells respond to ionizing radiation by transporting these proteins to the cell surface. In order to investigate the potential of therapeutics targeted to radiation- induced tumor antigens, we developed a monoclonal antibody to the radiation-induced antigen, TIP-1. We propose studies to test the hypothesis that the antibody to this antigen induces therapeutic anti-tumor immune responses. Tax interacting protein-1 (TIP-1) is a membrane-associated protein that is over-expressed in poor prognosis cancer including non-small cell lung cancer. We recently found that TIP-1 undergoes radiation- induced translocation to the cell surface. Cancer specific antibody binding is achieved in vivo for several days by anti-TIP-1 antibody administration after irradiation, and optical imaging and immunohistochemical staining indicated that this antibody (1A6H14) achieves specific binding to irradiated cancer in mouse models. As noted above, the proposed approach is dependent on the induction of anti-tumor immune responses (antibody- dependent cellular cytotoxicity, ADCC; antibody-dependent cellular phagocytosis, ADCP; and perhaps an adaptive T cell response) based on opsonization of tumor cells by the anti-TIP-1 mAb and recognition of the Fc by Fc receptors on natural killer (NK) cells, dendritic cells (DC) and other cell types. Importantly, it has been shown that several approved anti-cancer therapeutic mAb, including trastuzumab, cetuximab and rituximab, have portions of their mechanisms of action through ADCC and ADCP. PUBLIC HEALTH RELEVANCE: Because of the poor prognosis of non-small cell lung cancer even with current therapies, there is a need for new treatments. The proposed studies will investigate a novel approach to cancer treatment: the use of monoclonal antibodies to radiation-induced tumor antigens.
|
{
"pile_set_name": "NIH ExPorter"
}
|
UTSA's proposed RCMI Center for Interdisciplinary Health Research (CIHR) will expand research infrastructure and expertise to establish a program of excellence in basic and translational health research that will advance both the strategic goals of UTSA and RCIVII/NCRR. The RCMI CIHR is greatly needed for UTSA to develop present areas of scientific strength into competitive health research programs to advance our strategic mission. The proposed program will foster and guide the development of UTSA faculty whose research has great relevance to human health and will support translational research projects. The Administrative Core will provide leadership and support for activities to advance the center's mission, develop research cores, and provide administrative and academic support for investigators. It will coordinate the lAC/EAC, establish intra- and inter-institutional collaborations, and execute a Faculty Development Plan and Evaluation Program. A Biophotonics Core will acquire cutting-edge imaging technologies to study biological processes at the molecular level in live cells with exceptional detail. A Computational Systems Biology Core will build high-performance computing infrastructure for simulation of biological systems, live cell imaging, and protein biomarker research. A Protein Biomarkers Core will build infrastructure for protein biomarkers discovery for disease diagnosis and targeted therapy, including biomarkers that are specific to minority populations. A Nanotechnology and Human Health Core will build infrastructure to synthesize nanomaterials for diagnostics, drug/gene delivery, tissue engineering, and for use with electron microscopy to produce 3-D structural information at the finest level. Five pilot project Pis will also be developed. Dr Lorenzo Brancaleon: Photo-induced unfolding of cancer-specific membrane receptors;Dr Thomas Forsthuber: Biomarker discovery in glucocorticoid resistance in experimental autoimmune encephalomyelitis;Dr Fidel Santamaria:Structural basis of biochemical information processing in neurons;Dr Tao Wei: Molecular mechanisms of A. baumannii biofilm formation;and Dr. Jianqiu Michelle Zhang: Advanced Data Processing for Capillary LC/MS data.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goal of Academic Nephrology Training Program at the University of California San Francisco (UCSF) is to prepare trainees for a successful academic research career through expert and comprehensive mentoring in laboratory or clinical research pertinent to kidney disease. The rationale for the program is the pressing nationwide need for well-trained adult and pediatric nephrology faculty, and for skilled investigators in the pathophysiology and epidemiology of kidney disease, its systemic consequences, and its treatment. The program proposed in this application has the faculty expertise, infrastructure, and research opportunities to provide outstanding training. The major emphasis of this application is the training of MD postdoctoral fellows who plan to pursue careers in academic adult and pediatric nephrology, but will also support the training of some non-clinical postdoctoral fellows for careers in academic nephrology, or in basic science related to nephrology. The twenty-eight training faculty are mostly from Nephrology, but include UCSF faculty from different basic and clinical departments to provide greater scope and depth of training opportunities. The faculty has been selected to accommodate the increasingly complex and technologic nature of basic research training, and to provide mentoring in the rigorous biostatistical analysis, complexities of study design, and ethical considerations required for clinical research. The training program emphasizes thematic and interdisciplinary research pertinent to kidney disease. Areas of investigation include renal epithelial cell biology and transport, kidney development, basic immunology and immunologic renal disease, epidemiology of complications of chronic kidney disease and dialysis, and analysis of novel transplant immunosuppressive regimens. Trainees will have full access to the graduate school basic science and biostatistics and epidemiology curriculum, the Advanced Training in Clinical Research curriculum supported in part by the UCSF K30 with the option of full MPH training. Trainees are required to participate in a weekly formal research conference in which trainees, mentors, and outside speakers present their research, and progress is monitored by Fellowship Research Committees, selected jointly by the trainee and primary mentor. The program actively recruits women and minority applicants, and uses feedback from trainees, mentors, and external reviewers to enhance and improve the training experience.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Our goal is to develop a better understanding of central nervous system characteristics of persons at risk for alcoholism. Despite a large clinical literature on alcoholism, far less is known about the preclinical characteristics of persons at greatest risk for the disorder. The Oklahoma Family Health Patterns Project studies healthy young adults with and without a family history of alcoholism. Persons with a positive family history are four times as likely to develop alcohol use disorders as those with no such history, and this risk doubles in persons who also have antisocial and disinhibitory characteristics. The two tendencies are coinherited. Persons with both a positive family history and personal evidence of behavioral disinhibition are accordingly considered to be at High Risk of future alcoholism, and those lacking these factors are considered at Low Risk. Our major hypothesis is that High Risk persons have altered brain mechanisms that serve to produce normal emotional responses to the environment and have deficient regulation of overt behavior. While evidence points to altered communication between the limbic system and prefrontal cortex, confirmatory neuroimaging is lacking. This revised application pursues our recent finding that persons with a family history of alcoholism have differences in regional brain glucose metabolism in the resting state as compared to their counterparts with no such history. Preliminary findings are that FH+ have greater FDG uptake than FH- in structures involved in obtaining visual information (Rt. Middle &Sup. temp. gyrus) and obtaining rewards or assessing reward value to make relevant decisions (Left Cingulate and Caudate). This set of structures and functions is compatible with the greater activation seen in the hypothalamus, which may play a related role in preparation for obtaining such rewards. In contrast FH- have greater FDG uptake than FH+ in functions involving prefrontal regulatory controls: The right inferior frontal gyrus is involved in regulation of emotional and autonomic expression. This region is closely associated with functions involving conscious autonomic regulation in conjunction with the orbital frontal cortex having inputs to the hypothalamus and brainstem. The right middle frontal gyrus is involved in executive functions via extensive connections to the dorsolateral prefrontal cortex. This study will expand the resting database to allow a network analysis of resting metabolic activity of a network of brain areas concerned with resting metabolic differences in High-Risk persons. The present study will carry out a seed- based correlational analysis of default-mode and anti-default-mode function in our two risk groups. The planned studies are expected to yield new information concerning altered functioning in brain regions involved with cognition, decision-making, and behavioral regulation in young adults at high risk for future alcoholism. PUBLIC HEALTH RELEVANCE: Alcohol abuse and dependence are behavioral disorders that are a burden to society. Despite a large literature on persons with alcohol dependence, much less is known about the preclinical characteristics of persons at greatest risk for the disorder, especially those with inherited risk factors. The present study examines resting brain metabolism in persons with a positive family history of alcoholism who also have disinhibited and antisocial tendencies that place them at High Risk of future alcoholism. Studies of High Risk individuals are valuable in characterizing behavioral characteristics of those who are vulnerable to alcoholism free of central nervous system effects secondary to heavy drinking. The proposed positron emission tomography study will provide new and useful information on the underlying brain functional characteristics of these High Risk individuals.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The estrogen receptor is an important component in development, differentiation and growth. It is thought to play a role in the development of some breast cancers. Elucidation of the mechanism of estrogen regulation of transcription is important in understanding the cause of these cancers and in the design of effective treatments. An initial step in estrogen regulation of transcription is the binding of the receptor to a specific DNA sequence called an estrogen responsive element (ERE). A consensus ERE has been derived which is a perfect palindromic sequence. This consensus ERE has been shown to confer specific estrogen responsiveness. The interaction of the hormone-activated receptor with the ERE influences the activity of RNA polymerase II by an unknown mechanism. The objectives of this project are to determine the mechanism by which hormone activated-estrogen receptor recognizes EREs which contain deviations from the consensus ERE and the mechanism by which the receptor activates RNA polymerase II. There is evidence which suggests that components, that is, flanking sequences and proteins in addition to the estrogen receptor are required for imperfect EREs to be functional in vivo. The hypothesis will be tested that these components allow imperfect EREs to function in vivo by increasing the binding affinity of the estrogen receptor for the ERE. The imperfect ERE and flanking sequences from the upstream region of the rat luteinizing hormone B gene will be analyzed in either transient transfection experiments into HeLa cells or gel mobility shift assays. The binding of the receptor to the ERE does not appear to be sufficient for signal transduction. There is evidence which suggests that steroid receptors appear to compete for common limiting transcription factors which are not components of the basal transcription initiation machinery. The hypothesis will be tested that proteins which specifically interact with the hormone activated-estrogen receptor aid the receptor in influencing RNA polymerase 11 activity. These estrogen receptor-binding proteins will be identified by either gel mobility shift assays or immunoprecipitation with an isolated epitope tagged N-terminal domain of the estrogen receptor or by use of an estrogen receptor affinity column. The function of these proteins will be analyzed in either "transcriptional interference" experiments or steroid receptor-dependent in vitro transcription.
|
{
"pile_set_name": "NIH ExPorter"
}
|
To date, the use of marketed continuous glucose monitoring systems (CGMs) are limited to glucose trend analysis by patients with insulin-dependent type 1 and type 2 diabetes. Accuracy limitations, particularly in the hypoglycemic range, and low signal-to-noise ratio have precluded expansion of this promising technology into screening applications to identify early signs of diabetes, prior to clinical onset of disease, so preventive measures may be taken. The proposed research is focused on developing an in vitro model of glycemic oscillations in yeast using a combined yeast-biosensor hybrid. The information derived from this model will be used to determine if manifestations of glycemic oscillations can be measured and characterized within the interstitial fluid (ISF) of humans. So far, the phenomenon of in vivo interstitial fluid glycemic oscillations has not been reported. Preliminary in vivo interstitial fluid glucose data, measured in the dermis of swine, revealed an oscillatory phenomenon that may be indicative of intracellular glycolysis. Included in the Specific Aims is the development of an in vitro yeast-biosensor hybrid system that models the in vivo ISF environment. This unique hybrid, continuous glucose monitoring system will be used to develop a model of glycemic oscillations in yeast. Data from the in vitro model will be used to develop pattern recognition algorithms to characterize in vivo ISF oscillations specific to type 1 and type 2 diabetes. Successful completion of the proposed research will provide advances in continuous glucose monitoring technology that have the potential to resolve inaccuracy issues encountered by patients using CGMs. It is our belief the proposed research will break-down technical barriers that preclude the use of this technology as a stand-alone system with expanded utility for screening and preventative applications that may bring the artificial pancreas on step closer to reality. PUBLIC HEALTH RELEVANCE: The potential for continuous glucose monitoring systems (CGMs) to detect glycemic oscillations in interstitial fluid will advance the use of this technology into the realm of preventative medicine by providing an inexpensive screening tool, capable of identifying people with abnormal glucose metabolism long before overt clinical symptoms are apparent. The outcomes of the proposed research will allow physicians to intervene early in the progression of diabetes symptoms with low cost diet and lifestyle changes as opposed to high cost therapeutic intervention. Early stage intervention will have a significant impact on reducing the over $200 billion annual cost to the US healthcare system to treat the chronic complications of diabetes.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Specific Aim of the project is to improve the survival of cultured endocrine allografts by (1) Modifying culture conditions to increase the selective destruction of passenger leucocytes without damage to parenchymal cells, (2) Determining the mechanism of graft rejection, and (3) Determining the mechanism of the active tolerance that develops in long term recipients of cultured allografts. The first aim will be accomplished by first reducing the minimum oxygen pressure required for abrogation of rejection through methods that improve oxygen diffusion, and then by increasing the maximum oxygen pressure tolerated with antioxidants. The second aim will be approached by experiments that will (1) determine if MHC-incompatible grafts are selectively spared or rejected, (2) determine if a correlation exists between the suppression of cytotoxic (CTL) production in vivo and the protection against graft rejection, and (3) demonstrate whether the DNA fragments characteristic of CTL killing that appear in rejecting grafts come from host or graft cells. The third aim will be studied by the approaches now under development that permit a quantitative measurement of tolerance both in vitro and in vivo. In vitro studies are based on the measurement of CTL responses with and without the addition of IL-2 and the addition to cell cultures of immunoglobulin from tolerant animals. In vivo studies are based on an analysis of the suppressed CTL response of tolerant animals to the I.P. injection of allogeneic tumor cells.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objective of the proposed research are as follows: (1) Test the hypothesis that 5-hydroxytryptamine (5-HT) is involved as a nerurotransmitter in the non-adrenergic inhibitory innervation of the gut. (2) Determine the properties of 5-HT mechanisms in the myenteric plexus such as transport, storage and synthesis of the amine, and how these are affected by drugs. (3) Compare 5-HT mechanisms in the myenteric plexus with those of 5-HT containing, neural-crest-derived endocrine cells such as enterochromaffin cells of the gut and parafollicular cells of the thyroid. (4) Determine the fuction of 5-HT in the thyroid and its relationship to the polypeptide calcitonin, which appears to share storage vesicles with 5-HT. (5) Establish the role of pontine 5-HT neurons in the regulation of electrical activity, characteristic of REM, in oculomotor and visual systems (PGO waves). (6) Apply what is learned from studying the myenteric plexus to the PGO system to help establish the validity of using the myenteric plexus as a model for the less accessible 5-HT neurons of the central nervous system.
|
{
"pile_set_name": "NIH ExPorter"
}
|
In single celled eukaryotes, the pathways that monitor nutrient availability are central to regulating the meiotic program and spore development. However, how metabolic inputs influence meiotic progression and gametogenesis remains poorly understood in metazoans. Our current studies focus on understanding how metabolism influences meiotic progression and oocyte development. Target of rapamycin complex 1 (TORC1) is a master regulator of metabolism in eukaryotes that integrates information from multiple upstream signaling pathways. The GATOR1 complex inhibits TORC1 in response to amino acid limitation and is required for meiotic entry in yeast. To define the role of the GATOR1 complex in Drosophila, we generated null alleles of all three GATOR1 components, nprl2, nprl3 and iml1 using CRISPR/CAS9. Analysis of these mutants revealed that, as is observed in yeast, the GATOR1 complex down-regulates TORC1 activity to promote the transition from the mitotic to the meiotic cycle. Moreover, the delayed entry into meiosis can be suppressed by feeding the females the TORC1 inhibitor Rapamycin. Finally, we determined that mutations in Tor kinase, result in premature meiotic entry. Thus in Drosophila females, high TORC1 activity inhibits the mitotic/meiotic transition and gamete development while low TORC1 activity promotes these events. Currently, we are using genetic strategies to determine what downstream targets of TORC1 mediate the meiotic delay. To complement this line of investigation, we are examining if the GATOR1 complex has critical functions in the early stages of meiosis. Intriguingly, we determined that in nprl2, nprl3 and iml1 mutants the formation of meiotic double-stranded breaks is deregulated resulting in a dramatic increase in the number of meiotic double-stranded breaks and in the activation of the transcription factor p53. In mammals, all three members of the GATOR1 complex function as tumor suppressor genes. However, the precise role of the GATOR1 complex in the regulation of metabolism in metazoans remained poorly defined. We have shown that that the central importance of Nprl2, Nprl3 and Iml1 in the response to amino acid starvation has been conserved from single cell to multicellular animals. We find that in Drosophila, Nprl2, Nprl3 and Iml1 physically interact and are targeted to lysosomes and autolysosomes. Using oogenesis as a model system, we demonstrated that the GATOR1 complex inhibits TORC1 signaling in response to amino acid starvation. Moreover, the inhibition TORC1 by the GATOR1 complex is critical to the preservation of female fertility during times of protein scarcity. In young egg chambers the failure to down-regulate TORC1 in response to amino acid limitation triggers apoptosis. Thus, our data suggest the presence of a metabolic checkpoint that initiates a cell death program when TORC1 activity remains inappropriately high during periods of amino acid and/or nutrient scarcity in oogenesis. Going forward, we will continue to use Drosophila as a model system to study how the GATOR complex helps integrate developmental and metabolic inputs.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This Institutional National Research Service Award is designed to develop skilled investigatorswith research-oriented careers directed at solving basic and clinical problems in lung disease. The structure of the program is based on the premise that training requires 1) a multidisciplinary approach, 2) a close relationship between the student and mentor, and 3) a training environment with breadth and depth in both clinical and basic sciences. Training in this program spans a variety of disciplines, including cell and molecular biology, immunology, microbiology, toxicology, biochemistry, pulmonary and critical care medicine, neonatology, and imaging sciences. Major research themes include lung immunology and inflammation, the molecular basis of lung injury, health effects of gaseous and particulate air pollutants, vascular function and biology, and imaging techniques in animal models of pulmonary inflammation and fibrosis. Four pre-doctoral and four post-doctoral trainees will be supported each year of the program. Pre-doctoral students who have demonstrated an interest in lung research will be accepted for training through the Graduate Education in Biomedical Sciences (GEBS) clusters, after they have passed their preliminary examinations. Pre-doctoral trainees will be supported for three years. Post-doctoralMD candidates from training programs in adult Pulmonary and Critical Care Medicine, Neonatology,and Infectious Disease will be considered based on potential success in a pulmonary research-oriented career. Support will be for two years, and will commence after the bulk of their clinical training is completed. Post-doctoral PhD trainees involved in lung-related research are also candidates for training, and are identified and recruited by faculty members of this training program. A total of four post-doctoral trainees will be supported. A direct research experiencewith research mentors forms the primary mechanism for training, supplemented by didactic courses, seminars, conferences, journal clubs, and instruction in research ethics, and human and animal experimentation. Trainees will develop research protocols under the close supervision of their mentors and a of a committee from the interdisciplinary faculty with expertise in the appropriate field of research. This training program will help to meet the increasing need for scientific investigators in lung biology and disease. Research by these young investigators wjll improve our understanding of how the respiratory system responds to injury and environmental challenges, and will improve public health by developing new and improved avenues for the prevention and treatment of lung disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The research provides converging experimental studies designed to investigate the role of motivational, perceptual, and learning processes in the development of adaptive exploratoy behaviors of premature infants during the first year of life. Operant learning procedures are employed to assess the responsiveness of premature infants to differet classes sensory reinforcement and to assess the development of learning and perceptul capacities. The reliability and validity of performance measures on experimental tasks as early measures of individual subject differences, as well as early measures of perceptual and learning deficits as a function of birth status, are examined. Studies on the effects of compensatory stimulation programs are designed to investigate specific changes in motivational and learning processes that may be the result of early exposure to routine programs of hospital care and restricted patterns of stimulation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The long-term goal of the proposed research is to clarify the molecular mechanisms underlying the detection and discrimination of chemicals through contact chemosensation in the fruit fly, Drosophila melanogaster. Contact chemosensation allows flies to distinguish sweet from bitter molecules, as well as nonvolatile pheromones. Insect gustatory organs express a diversity of candidate molecular detectors. These include gustatory receptors (GRs), TRP channels, ionotropic receptors (IRs) and odorant binding proteins (OBPs), the latter of which promote the detection of chemicals by receptor proteins. However, the functions of most of these candidate gustatory receptors and binding proteins are unknown, or are understood poorly. We propose experiments to dissect the mechanisms underlying contact chemosensation in flies using a multidisciplinary approach that includes electrophysiology, behavior, genetics, and cell biological approaches. During the last few years, the concept that GRs are required broadly for sensing sugars and bitter-tasting compounds has been confirmed. However, the biochemical functions of GRs are unclear. Aim 1 is to test the hypothesis that GRs are tastant-activated cation channels. Aim 2 addresses one of the longstanding questions in taste sensation- the nature of sour receptors. These receptors are thought to be cation channels, and many candidates have been suggested. We propose experiments to dissect whether an IR is required for the responses to most sour tastants, which cells require the IR, and investigate the contributions of two other IRs that are expressed primarily in gustatory receptor neurons. Aim 3 is devoted to characterizing the roles for odorant binding proteins (OBPs) in contact chemosensation. OBPs are known primarily for promoting the detection of certain olfactory stimuli. We found that some OBPs are highly enriched in taste organs up to 800- fold over other tissues. Experiments are proposed to test the hypothesis that gustatory OBPs promote behaviors mediated by nonvolatile pheromones. Finally, the goal of our last aim is to dissect the molecular, cellular and biological basis for selective taste plasticity. We present preliminary data indicating that flies exposed to camphor reduce their repulsion specifically to camphor and not other bitter tastants, and this taste plasticity results from a reduction in the level of a TRP channel. Aim 4 is to distinguish between different possible models underlying this taste plasticity, and to address the biological basis for adaptation to some, but not all, unappealing tastants. An additional long-term goal of this research is to apply the findings to the control of insect pests that spread disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
In this project we propose to develop statistical methods for the analysis of microarray and RNA-sequencing data for expression QTL mapping. Our project is designed to address a number of important methodological issues, with particular relevance to the forthcoming GTEx study. We propose to extend a Bayesian hierarchical model for cis-eQTL mapping to enable simultaneous mapping in multiple tissues, and to improve the use of external biological information. We also propose to develop methods to allow more sensitive detection of trans-acting eQTLs that are correlated with networks or modules of co-regulated genes. Finally, we aim to develop methods for estimating transcript abundances from RNA sequencing data, for use in QTL mapping. PUBLIC HEALTH RELEVANCE: The purpose of this project is to develop new tools for analyzing and interpreting eQTL (expression quantitative trait loci) studies. We will develop analytical tools for both microarray-based and RNA-sequence-based measurements of gene expression.
|
{
"pile_set_name": "NIH ExPorter"
}
|
A specific marker protein for olfactory receptor cells has been isolated from mouse and rat tissues. This marker protein has been used to prepare an antibody. By using the peroxidase-antiperoxidase immunohistological technique we have demonstrated the presence of the marker protein in the perikaryon region of mature olfactory receptor cells and in the entire length of axon to its termination in the olfactory bulb. In developing rats the protein is first demonstrable on the l8th embryonic day only in those olfactory receptor cells the perikarya of which are located most superficially in the epithelium; the entire length of some axons also contains marker protein. In mice this is true as early as the 14th embryonic day. In the latter species, this time corresponds to the stage of development when synapses between receptor cell and second order neurons are first demonstrable in the olfactory bulb. This suggests a relationship between marker protein synthesis and synaptogenesis in the primary olfactory pathway. Presently studies are under way to determine; 1) At what developmental age synapse formation between receptor cells and second order neurons occurs in rats and whether this age corresponds with the time when the marker protein is first demonstrable. 2) Localization by electron microscopy of olfactory marker protein in receptor cells. 3) Organ culture of combined explants of olfactory mucosa and olfactory bulb at various developmental stages. This will provide a model system that can be experimentally manipulated.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Cocaine remains a major public health concern due to the continued high level of emergency room admissions involving cocaine use. Cardiovascular complications represent a large proportion of these admissions. Recent research has focused on determining whether proposed treatment agents will alter the cardiovascular effects of cocaine in conscious squirrel monkeys. Although not planned, it is likely that some individuals will use a treatment agent in combination with cocaine. Squirrel monkeys were seated in standard restraint chairs and placed in sound attenuating chambers 5 days per week. Twice per week the monkeys were administered cocaine, a treatment agent, or the two in combination about 30 min into the session. Cardiovascular parameters were monitored for an additional 60 min. In conscious squirrel monkeys cocaine produces clear increases in BP and HR. Drugs that interact with dopamine have been tested for cardiovascular effects both alone and in combination with cocaine. GBR 12909, a selective dopamine uptake inhibitor, produced small increases in blood pressure and heart rate, but when given in combination with cocaine, no potentiation of the cocaine effect was observed. Similarly, the dopamine D1 agonist SKF 82958 produces increases in blood pressure and heart rate, but did not potentiate cocaine?s effects at the doses tested. The partial D1 agonist SKF 77434 did not increase blood pressure and heart rate when given alone, and appeared to attenuate the effects of cocaine. The dopamine D2 agonist quinpirole increased heart rate and decreased blood pressure. When given in combination with cocaine, sub-additive effects were observed. In contrast, the dopamine autoreceptor/D3 antagonist UH 232 produced increases in blood pressure and heart, and when given in combination with cocaine additive effects are observed. No significant effects on ECG were observed for any of the drugs. Therefore, of the drugs tested thus far, only UH 232 would appear to represent a significant risk when used in combination with cocaine. The contribution of the metabolites of cocaine to its cardiovascular effects is of interest as complications to cocaine use have been reported hours after administration. Further, cocaine is often used repeatedly for many hours, so metabolite concentrations can be elevated. This was studied in squirrel monkeys by the systemic administration of various cocaine metabolites. Neither benzoylecgonine nor ecgonine methyl ester produced significant cardiovascular effects. The minor metabolite norcocaine produced increases in blood pressure, but only at doses higher than cocaine and the effects where relatively brief. In contrast, cocaethylene, a metabolite produced when alcohol is co- administered with cocaine, produced cardiovascular effects with potency similar to cocaine. These results suggest that the metabolites of cocaine do not contribute to its overall cardiovascular effect, with the possible exception of cocaethylene.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The normal-hearing ear processes complex sounds, such as speech, by stimulating different groups of neurons distributed along the length of the basilar membrane with different frequency components. Cochlear implants (CIs) attempt to mimic this spatial separation of frequency by allocating certain spectral components within the acoustic signal to different intracochlear electrodes. A limitation of this processing is that any two electrodes may stimulate overlapping groups of neurons. We can use forward masking of electrically evoked compound action potentials (ECAPs) to measure the degree of overlap in individual CI recipients, including those who can't participate in behavioral or speech perception testing. Unfortunately, these peripheral measures of spatial resolution have not been found to correlate with speech perception, which limits our ability to use these measures to evaluate and optimize the benefit a recipient receives from a CI. We will evaluate two hypothesizes relative to these limitations: (1) The integrity of the central auditory system is not reflected in ECAP forwar masking but is involved in speech perception, and (2) Spatial resolution, as measured by a limited set of electrode pairs, may not appropriately characterize how a complex signal, such as speech, stimulates multiple intracochlear electrodes. To address these issues, in addition to extensive peripheral measures of spatial resolution (ECAP forward masking), we propose using two electrophysiological measures of processing at the auditory cortex: a measure of spatial resolution (discrimination between two electrodes) and a measure of spectral resolution (discrimination between two spectrally complex acoustic signals). Three cumulative experiments have been proposed. Experiment 1 examines the relationship between peripheral/central measures of spatial resolution (i.e. electrode selectivity) within individual CI recipients. For Experiments 2 and 3, three novel CI programs will be used to effectively vary the spatial resolution within individual CI recipients. Experiment 2 examines the relationship between the processing of simple and complex stimulation. Experiment 3 examines the relationship between the three electrophysiological measures and vowel perception. Simple and multiple regression analyses will be performed using mixed models as needed to account for repeated measures. The results should more accurately define the relationship among these three electrophysiological measures and their ability to predict the way individual CI recipients perceive vowels.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The epidermal growth factor receptor (EGFR) tyrosine kinase pathway has been demonstrated to be a key molecular target for cancer therapeutics including TKIs such as Iressa (gefinitib) and Tarceva (erlotinib). These compounds are small molecule ATP mimetics, selectively binding to the intracellular kinase of EGFR, that have recently received FDA approval for treatment of non-small cell lung carcinoma (NSCLC). An ever growing body of data indicates that the presence of mutations in EGFR play an important role in response to TKI therapy and drug resistance. There are a subset of patients (~10%) who have a remarkable response to Iressa and Tarceva that have now been correlated to several specific activating EGFR mutations arising within the tumor. A second mutation has been identified in relapsed patients who were initially drug responsive and more recently this mutation has also been found in a family with a genetic predisposition for developing lung cancer. Recent studies from our lab and others suggest an emerging new paradigm for modulation of downstream signaling. These studies show that autophosphorylation of RTKs occurs in a specific sequential order, creating a temporal and dynamic phosphorylation pattern of different intermediate sets of phosphotyrosines. Accordingly, these discrete sets of phosphotyrosines can orchestrate the dynamic recruitment of specific downstream signaling partners in a temporal fashion that may well play an important role in the proper regulation of protein signaling. The underlying molecular mechanisms of the oncogenic activating and resistance mutations in EGFR and differential sensitivity to Iressa and Tarceva are not understood. Our preliminary data suggest that the underlying causes may be linked to alterations in nucleotide affinity for the receptor, kinase catalytic activity, and/or alterations in phosphorylation patterns that result in differential protein signaling. The studies outlined in this application will use a combination of in vitro biochemical studies and complementary cellular studies to identify specific molecular alterations that may distinguish wild type (WT) and oncogenic forms of EGFR harboring Iressa/Tarceva sensitive activating mutations as well as mutations associated with drug resistance. The overall objectives are to define the molecular mechanism of protein signaling in the EGFR pathway under normal and aberrant oncogenic processes and provide an understanding for the modulating effects of Iressa/Tarceva. This information will ultimately aid in the design of more potent and selective compounds for precisely targeted cancer therapy. Growth factors bind to their corresponding receptors located on the surface of cells and give signals for cell growth. In a cancer cell, the signals for cell growth have aberrant signaling and growth and proliferation have become out of control. The newest, most promising cancer therapies such as Iressa and Tarceva, targets these uncontrolled signaling pathways in cancer cells. Some patients with non- small cell lung carcinoma having particular mutations in their growth factor receptors respond very well to these drugs, however the molecular events are not understood. The studies described in this application are designed help unravel these molecular details to allow us to differentiate cancer cells from normal cells and assess the usefulness of current and new cancer therapies.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This proposal seeks to clarify the role of TGF-alpha, EGF and TGF-beta3, and their receptors, in lung morphogenesis and in lung repair following oxygen injury. These factors mediate autocrine/paracrine signalling between the respiratory epithelium and pulmonary mesenchyme. The specific aims are based on preliminary data demonstrating that 1) expression of TGF-alpha in the respiratory epithelium of transgenic mice causes marked alveolar hypoplasia and pulmonary fibrosis; 2) that the pulmonary lesions seen in TGF-alpha expressing transgenic mice are ameliorated by the expression of the dominant/negative EGF receptor in Type Il epithelial cells; and 3) that TGF-beta1 blocks prenatal alveolarization, in association with inhibition of epithelial and mesenchymal cell differentiation. The specific aims will discern the effects of TGF-alpha, TGF-beta and receptors in embryonic and postnatal lung morphogenesis and fibrosis seeking to identify the cellular mechanisms involved in the epithelial mesenchymal cell signalling pathways involved in the pathogenesis of chronic lung disease in neonates and in adults with fibrotic lung disease. Relationships between TGF-alpha and TGF-beta mediating cell signalling, lung cell proliferation and differentiation will be assessed in transgenic mice expressing TGF-alpha and TGF-beta, the latter under conditional control of a tetracycline regulatable lung-specific transgene. Transgenic EGF-R deficient, wa-2/wa-2, and TGF-beta1 gene targeted mice, as well as transgenic mice bearing EGF-R dominant/negative and TGF-beta receptor 2 dominant/negative receptors will be used to inhibit receptor signalling in specific subsets of respiratory cells. These animal models will be used to determine the role of TGF-alpha, EGF-R, and TGF-beta in acute and chronic pulmonary oxygen injury. These studies will determine whether EGF and TGF-beta receptor pathways are involved in lung remodelling and pulmonary fibrosis in the neonatal and postnatal lung. Morphological, biochemical, immunocytochemical and physiologic studies will be used to clarify the cellular and biochemical alterations mediated by these receptor systems in developing and injured lung. Potential role of EGF and TGF-beta receptor signalling in the pathogenesis of acute and chronic lung injury (RDS and BPD) and in adults with fibrotic lung diseases (ARDS, idiopathic pulmonary fibrosis and sarcoidosis) will be established.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The effect of steroid therapy in septic shock is evaluated in baboons (Papio spp.). Of special interest is the effect of endotoxemia on the myocardium. Instantaneous pressure measurements in the left ventricle of an awake intact baboon are digitized. Current indices of myocardial contractility are derived in an attempt to determine what role steroids may play in the treatment of Sepsis.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Multiple myeloma (MM) is an incurable malignancy that afflicts 15,000 new Americans each year. Patients typically die 3.5 years after diagnosis, experiencing bone destruction leading to spine deformities and pain. Despite its transformation, the malignant clone is often dependent on growth factors in its environment, not for growth - MM is a slowly proliferating disease - but for survival. The objective of this study is to critically test the hypotheses that neurotrophin signaling contributes to these survival signals, and that inhibition of neurotrophin signaling will control MM progression. Trk receptor tyrosine kinases and their neurotrophin ligands are expressed by MM cells, creating a signaling loop that promotes the survival of MM cell lines and primary MM cells in vitro. Blockade of Trk signaling, using a soluble Trk-Fc decoy receptor, inhibits MM growth in a xenograft model. Neurotrophins are also expressed by bone marrow stroma, by endothelial cells, by osteoblasts, and thus contribute to the support of MM within its favored environment. These observations suggest a neurotrophin-Trk axis in MM tumor progression, and led to preclinical evaluation of cep701 as anti-MM therapy. Cep701 is a derivative of the indolcarbazole, K252a, with an IC50 of 3 nM for Trk and for Jak2. It specifically kills both primary MM cells and MM cell lines in culture, and inhibits growth of MM cell lines implanted into the subcutaneous tissue of NOD-SCID mice. The ability to target both Trk and Jak2 likely underlies its potent anti-MM activity. This study will evaluate the role of Trk signaling in MM disease progression. Specifically, we will: 1) Determine the importance of neurotrophin: Trk signaling to MM tumor survival, by assessing cell viability after disrupting Trk activation alone and in combination with Jak2 inhibition. 2) Delineate the signaling cascades that are critical to the pro-survival effects of neurotrophin:Trk activation in MM. 3) Identify the prevalence of Trk and neurotrophin expression by MM, and correlate this expression with disease characteristics including stage, prior therapies, immunoglobulin isotype, and cytogenetics, 4) Establish whether dual Trk/Jak2 targeting using cep701 will control MM disease progression in the SCID-hu model of MM. [unreadable] [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
Impaired memory is an important component of diseases such as Alzheimer?s disease, temporal lobe epilepsy, depression, and schizophrenia that collectively affect over twenty million Americans. Our long-range goal is to contribute to a better understanding of the neural mechanisms that underlie memory processes, in order to bring us closer to developing new therapies for these disabled patients. The objective of this proposal is to characterize neural signals that support a prototypical form of memory, recognition memory. Recognition memory is the ability to perceive a recently encountered item as familiar. This ability is impaired following lesions of medial temporal lobe structures, including the hippocampus and the underlying perirhinal and entorhinal cortices. Neural signals that may support recognition memory have been described in the perirhinal and entorhinal cortices. However, despite extensive research, there is currently little evidence for the existence of recognition memory signals in the non-rodent hippocampus. This apparent inconsistency between the findings from lesion and physiology studies fuels a current controversy regarding the contribution of the hippocampus to recognition memory and prevents a full understanding of the organization of memory. Based on preliminary data, we hypothesize that single neurons, the local field potential (LFP), and synchronized ensembles of neurons in the hippocampus display modulations in activity that may be used for recognition memory. The experiments proposed here will directly test this hypothesis, using multi-electrode recordings of spiking activity and LFPs during a recognition memory task. We will examine modulations in single-unit firing rates, amplitude and power in the LFP, and spike-field neuronal synchronization with respect to performance on the Visual Preferential Looking Task, which is known to be highly sensitive to lesions of the hippocampus. The proposed experiments have the following potential outcomes: to resolve the apparent inconsistency between lesion and neurophysiological studies regarding the role of the hippocampus in recognition memory;to determine the functional significance of oscillatory activity, including theta-band oscillations, in the hippocampus;and to identify neuronal synchronization as a potential mechanism underlying memory formation. PUBLIC HEALTH RELEVANCE Impaired memory is an important component of diseases such as Alzheimer's disease, temporal lobe epilepsy, depression, and schizophrenia that collectively affect over twenty million Americans. Our long-range goal is to contribute to a better understanding of the neural mechanisms that underlie memory processes, in order to bring us closer to developing new therapies for these disabled patients. The objective of this proposal is to identify neural mechanisms in the hippocampus and subjacent cortex that may underlie memory formation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goals of this project are to define conserved epitopes in the V1/V2 domain of HIV-1 gp120 that mediate virus neutralization and enhancement, and to use this information to develop an HIV vaccine based on modified V1/V2 miniproteins. The structures of the alternate V1/V2 conformers will be determined by MALDI analysis of proteolytic fragments of the purified isomers. A number of human sera have been identified that contain antibodies that react with conserved V1/V2 epitopes, including some that recognize alternate conformational forms of V1/V2. Also available are sera from V1/V2 immunized macaques that possess either neutralizing or enhancing activities for specific M-tropic HIV-1 isolates. These antibodies will be fractionated by immunoaffinity chromatography and characterized for specificity for V1/V2 conformers, crossreactivity against distantly related sequences and for neutralizing (or enhancing) activities against various viruses. Novel monoclonal antibodies (Mabs) directed against functional targets in the V1/V2 domains will be isolated from hybridomas prepared from mice immunized with various antigens, including V1/V2 miniproteins, recombinant gp120/140 molecules, and mouse cells expressing native HIV-1 Env proteins. These studies will also utilize transgenic XenoMouse strains developed at Abgenix that produce human immunoglobulins, in order to isolate human Mabs; because of the human nature of these antibodies, any Mabs with potent neutralizing activities that are isolated in these experiments could have direct utility as passive immunotherapeutic agents in humans. Hybridomas will be generated by standard technologies, and screened against the relevant immunogens. These Mabs will be used to map epitopes that mediate viral neutralization and enhancement. Finally, a panel of mutant V1/V2 miniproteins, based on both the CaseA2 and SF162 sequences, will be prepared; mutations to be studied will include N-linked glysosylation sites, deletions and alanine substitutions. The immunoreactivities of the mutant proteins will be measured with available Mabs and fractionated human and macaque sera, and the distribution of neutralization and enhancement epitopes on the mutant proteins determined by absorption of human and macaque antibodies that possess such activities. Selected mutant proteins which preferentially express neutralization epitopes will be used to immunize rats, and the neutralizing and/or enhancing activities of the resulting sera quantitated. Modified V1/V2 miniproteins that induce effective neutralizing responses against clinically relevant HIV isolates would provide the basis of a V1/V2-based vaccine against HIV-1.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Vascular cognitive impairment (VCI) is a heterogeneous disorder that is a major cause of dementia. Selection of patients with small vessel disease, subcortical ischemic vascular dementia (SIVD), provides a more homogenous population that would facilitate clinical trials. Pathological studies in SIVD show thickening of the basal lamina around blood vessels with fibrinoid necrosis and hyalinosis secondary to hypertension, diabetes mellitus, and congenital vascular diseases. Ischemia/hypoxia leads to the production of matrix-degrading metalloproteinases (MMPs) as part of both an inflammatory response and tissue remodeling after injury. We have shown that brain tissues from patients with VCI immunostain for MMPs in reactive astrocytes and macrophage/microglia cells, MMPs disrupt the matrix around blood vessels and open the blood-brain barrier (BBB). In addition, MMPs break down myelin, which could contribute to the vascular demyelination seen in SIVD. Recently, we reported that increased levels of MMPs are found in the CSF of patients with VCI, but not in patients with Alzheimer's disease (AD). We hypothesize that MMPs are secreted by ischemic/hypoxic brain cells and contribute to the pathobiology of VCI. The specific aims are: 1) to develop a SIVD rating scale based on rnultimodal testing, including clinical evaluation, magnetic resonance imaging (MRI) and multivoxel proton spectroscopy (1H-MRS), neuropsychological testing, and CSF studies; 2) to identify the source of MMPs in the CSF by indexing the MMPs in the CSF and blood to albumin; 3} to determine the involvement of the BBB qualitatively with Gadolinium-DTPA enhancement and quantitatively with the Patlak Graphical Method and to correlate the opening with MMP levels; and 4) to correlate the levels of MMPs in a blinded fashion with the results of the SIVD rating scale that has been validated by long-term follow-up of the patients. These studies will determine the role of protease-mediated nflammation in the progressive form of SIVD, and could lead to novel treatments to suppress proteases. [unreadable] [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objectives of MBRS program at Selma University are, 1) to train and motivate undergraduate students who are capable of advancing for further studies in biomedical sciences, b) to enhance research capability in biomedical sciences at Selma University, c) to provide faculty with opportunities and facilities to develop biomedical research potential, and d) to attract minority students and faculty into biomedical research. Selma University proposes three research projects and one project on the enrichment activities in biomedical science. The proposed research activities will train 7 students each year in research areas related to metal toxicity, membrane biology, calmodulin effect, lipid peroxidation, enzyme mechanism, microbiology- immunology pathogenicity, immunogenicity, drug sensitivity to microorganisms, pesticide evaluation, medical entomology and statistical methods. The enrichment activities involves seminars, lecturers and workshops on hands-on-research techniques by invited guest scientists, a course in methods in biomedical science, summer research experience and an opportunity to attend scientific meetings. An exciting experience for students will be bio- computation-application of computer in biomedical research. The program will provide 4 facility opportunity to research along with their students. It is expected that proposed program will provide a wide variety of research related experiences which should not only motivate but also train the students to accept greater challenges in field of biomedical research. The expected success of our MBRS program will be a 'step-forward' in achieving the National MBRS mission to preparing competitive biomedical research scientists who are also ethnic minorities.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Patients with chronic granulomatous disease (CGD) frequently develop pulmonary infections. In many patients with CGD an accurate diagnosis is difficult to establish due to the scarcity of organisms in pathologic specimens. This study retrospectively assesses all procedural as well as diagnostic data in patients with CGD and pulmonary disease. The study assesses the utility of sputum, bronchoscopy (lavage and transbronchial biopsy), computer tomography-guided transthoracic needle aspiration, and open lung biopsy in establishing either a definitive or presumptive pulmonary diagnosis. The goal is to establish a diagnostic algorithm for pulmonary procedures in patients with CGD based on relative invasiveness and diagnostic yield. Data are being collected from chart reviews for analysis from 65 patients over a 15-year period.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Molecular Medicine Training Program of the Case Western Reserve University School of Medicine is specifically designed to train pre-doctoral students to take scientific advances from bench to bedside. It takes advantage of four factors to create an environment particularly suited to this purpose. First, this Program is located in the Lerner Research Institute where physicians and scientists from both the Cleveland Clinic and the School of Medicine have historically worked closely together to focus cutting edge research upon the causes and treatments of human illness. Second, a novel core curriculum has been designed throughout the first year to provide a solid foundation in basic scientific principles presented in the context of human organ systems and disease. Research presentations and rotations, seminars journal clubs, and a student retreat reinforce the translational application of the basic scientific principles throughout training. Third, the Training Program contains a broad range of trainers heading outstanding translational research laboratories in which our trainees perform rotations and thesis work. The range and strength of these laboratories, their collaborative interactions, and the commitment of the trainers to support of the Training Program is evidence of the collegial scientific environment within the Training Program. Finally, to ensure that each of the trainees comes to understand the clinical aspects of their area of investigation, each selects a Clinical Mentor who helps organize an individual clinical experience. Through this experience trainees are introduced to patient diagnosis and treatment on a first hand basis. To further emphasize the connection to clinical practice, students in the Training Program take a joint course with medical students of the Lerner College of Medicine. Experience has shown that these medical students who are in training to be physician scientists, and our trainees who are finding clinical application to basic scientific observations, have much to learn from one another. This proposal seeks funding for 7 trainees within the Training Program. With admission of its fourth class, it has become clear that the Training Program has gained national recognition and attracted an ever increasing number of applicants, with 27 training grant eligible trainees currently associated with the Program. The trainees are eagerly preparing to take their places as the leaders of the next generations of NIH funded laboratories translating basic scientific principles to address patient needs. The fact that this Program was specifically designed to address the needs for translational research identified in the recent NIH roadmap explains its youth, and emphasizes the importance of promoting its growth and maturation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This proposed research represents a continuation of work on the biochemistry and biology of human Cl, the first complement component and its substrates. Cl is activated by a variety of biological materials and initiates a cascade reaction beginning with the proteolytic cleavage of its natural substrates C4 and C2. Cleavage of C4 and C2 lead to convertase formation with the ability to cleave C3. Generation of capillary permeability, chemotactic, opsonic and lytic activities results and these reactions appear to play fundamental roles in acute and probably in chronic inflammation, irrespective of the method by which the cascade process was initiated. This project has placed an emphasis on isolating and characterizing naturally occurring inhibitors of the early classical components. Current research efforts have been devoted to purifying and characterizing the most recently discovered complement inhibitor - Clq inactivator (ClqINA). We have identified this inactivator as a chondroitin 4-sulfate proteoglycan and research is underway to more specifically define its biochemistry and to assess its role in modulating Clq activity in vivo.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The aims are to investigate the rheological behavior of red cell membranes (RCM), to study the interactions of RCM with the environment, to correlate these functions with the molecular organization of RCM, andto evaluate the role of RCM properties in circulatory regulation, in health and in disease. Rheology of RCM during deformation at constant area and during area stretching is studied on intact cells and on monolayers prepared from RCM components (new Project F), and modeled by theoretical computation. In the supplemental proposal, a similar approach will be used to study the rheology of leukocytes. The rheological data are correlated with the ultrastructure of RCM, with the microviscosity of RCM probed by fluorescence polarization, and with the molecular organization and function of specific membrane components. RCM proteins are examined by covalent labeling, and protein-lipid interactions will be explored by surface chemical and other techniques. The multidisciplinary approach is used to study: changes in RCM during cyclic deoxygenation of sickle cells; the relation of I-i antigens on sickle cell exoface to intracellular Hb F; properties of hexose receptor transport protein; electrochemical forces involved in interactions of RCM exoface with macromolecules; and the micromechanics of cell aggregation. The role of blood cell membrane properties in affecting flow dynamics and metabolic transport will be studied in the macro- and microcirculation (New Project G). Supplemental funds are requested for renovation, for meeting new needs in existing projects, and for adding new projects. The additional support will facilitate greatly our progress toward the ultimate goal of providing the fundamental knowledge needed to improve diagnosis and treatment of cardiovascular and blood diseases.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Primary Care Development Corporation (PCDC) is requesting a grant of $50,000 to underwrite the planning and presentation of a National Summit on Primary Care Emergency Preparedness. This summit will convene primary care providers, representatives from local and state departments of health, elected and government officials, and other stakeholders for a one-day session addressing primary care emergency preparedness. The overarching goal of the Summit is to increase the emergency preparedness level of primary care sites and facilitate their integration into local and regional emergency response planning. To achieve this, the conference aims to serve as a national forum to discuss the preparedness level of primary care centers across the nation, share best practices and innovative strategies to improve the preparedness level and response capabilities, convene stakeholders to identify roles, and set the agenda for preparedness on local, state, and national levels. The summit will be conducted as a one-day conference and will include large group events as well as breakout sessions focused on more specific topic tracks. Expected summit participants include all parties interested in primary care emergency preparedness, such as primary care providers, regional emergency response planners, hospital emergency planners, ambulatory care network administrators, legislators and other government representatives, representatives from local and state departments of health, school-based emergency planners, and others. For planning and recruitment purposes, it is anticipated that the summit will have approximately 300 participants. As a conference focused on the dissemination of research findings and best practices, PCDC will publish a summary of the meeting, outcomes, and lessons learned for public access to ensure that the content and results of the conference are disseminated as widely as possible. PCDC aims to hold the conference on February 7, 2007 at the CUNY Graduate Center in New York City. [unreadable] [unreadable] [unreadable]
|
{
"pile_set_name": "NIH ExPorter"
}
|
The epidermis and skin appendages perform essential barrier functions that protect the body from environmental insults and maintain fluid-electrolyte balance. Defects in the development, differentiation, and proliferation of the cells tht comprise the skin and hair follicles lead to numerous diseases such as skin cancer, psoriasis, bullous skin disease, and alopecia. Transcriptional programs that control skin and hair follicle development have defined processes important for epidermal integrity and hair regeneration. More recently microRNAs have also emerged as important regulators of gene expression in the skin. In contrast, the role of alternative splicing (AS) in skin development and function is essentially unstudied~ there have been no publications focused on alternative splicing factors in the skin or its appendages. Reciprocal epithelial- mesenchymal interactions (EMIs) underlie formation of the epidermis and hair follicles. One important arm involves the interaction of mesenchymal derived Fgf7 and/or Fgf10 with an epithelial-specific splice isoform of fibroblast growth factor receptor 2, Fgfr2-IIIb. Fgf7 and Fgf10 specifically interact with Fgfr2-IIIb, but not mesenchymal Fgfr2-IIIc. Abrogation of this directional signaling pathway results in defects in the epidermis and hair follicles. My lab identified the epithelial-specific splicing factors Esrp1 and Esrp2 that are both necessary and sufficient for the expression of the Fgfr2-IIIb splice variant in epithelial cells. We generated Esrp1 and Esrp2 knockout mice and demonstrate that combined Esrp1/Esrp2 KO is lethal and results in epidermal hypoplasia, reduced follicle numbers, and sparse hair. We hypothesize that the Esrps are required for normal epidermal and follicular development and function by enforcing the expression of epithelial-specific splice isoforms. We will determine the phenotypes of Esrp deletion in specific epithelial cell populations in skin and appendages and identify a comprehensive set of Esrp target transcripts through the following Aims: 1) Determine the phenotypes associated with Esrp ablation in the interfollicular epidermis and in hair follicles. We will define phenotypes associated with conditionally ablation of the Esrps in developing and adult epidermis and hair follicles. 2) Define comprehensive programs of Esrp regulated alternative splicing in the epidermis and hair follicle. We will use high throughput sequencing (RNA-Seq) and splicing sensitive microarrays to define genome-wide programs of alternative splicing in the different cell populations that populate the skin. These technologies will be used in conjunction with conditional deletion strategies to define Esrp regulated targets i the epidermis and hair follicle stem cells. The proposed aims constitute the first comprehensive analysis of alternative splicing in skin development and function, thereby introducing a new paradigm to the field. These studies will reveal novel insights into the processes that impact skin development and function.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Recent work indicates that vomeronasal and taste transduction in vertebrates share similar transduction pathways: G protein-coupled receptors signal through phospholipase C (PLC) to open a member of the TRP family of ion channels, TRPC2 for the vomeronasal organ (VNO) and TRPM5 for taste. The importance of these ion channels for chemosensation is highlighted by the lack of response to pheromones in TRPC2 knockout mice and to bitter, sweet and amino acid tastes in TRPM5 knockout animals. Understanding the mechanisms by which TRP ion channels are gated, has proven difficult in any system, and remains an essential goal for understanding taste and VNO transduction. In the last grant period, we discovered that TRPM5 is activated by intracellular Ca +, suggesting that Ca + may be the second messenger for some forms of taste transduction. In the next grant period we aim to understand the molecular mechanisms that underlie regulation of TRPM5, and how molecular properties of TRPM5 influence taste sensation. We will specifically address the following questions: (1) What are the mechanisms by which TRPM5 channels are activation and inactivated? In these studies we will use patch-clamp recording of expressed TRPM5, together with pharmacological and structural manipulations of the channel. (2) Do blockers of TRPM5 interfere with taste detection? Blockers will be identified by patch-clamp recording from cells expressing TRPM5 and the effects of identified blockers on taste thresholds in mice will be determined (3) Do similar channels to TRPM5 play a role in VNO transduction? TRPM5 and related channels are widely expresed in the body, and mutations in these channels may underlie certain pathological states. By understanding the regulation of these channels, we can understand their contribution to signaling in health and disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Work during the next year will focus on (1) defining the amino acids to which selenocysteine in GSH peroxidase is attached and the sequence of amino acids around it in the polypeptide chain; (2) purifying GSH peroxidase and other Se-containing proteins by HPLC; (3) characterizing Se-GSH peroxidase in rat kidney and lung and determining whether Se is present as selenocysteine; (4) studying GSH peroxidase in a variety of species and tissues; (5) determining whether other Se-containing proteins contain selenium as selenocysteine; (6) studying protection afforded by GSH peroxidase to RBCs exposed to peroxidizing microsomes prepared from Se-deficient rats; and (7) determining whether GSH peroxidase will inhibit lipid peroxidation in vitro in other reaction systems that contain purified liver microsomes.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The mechanism of action of DNA-reactive antitumor drugs such as DNA strand scission agents like neocarzinostatin, macromomycin and bleomycin and intercalation agents like adriamycin and daunorubicin will be studied by examination of their effects on chromatin and nucleosome structure. Chromatin and nucleosomes from drug-treated cells will be mapped for DNA damage sites to determine what regions of the DNA are susceptible to attack by the drugs' nuclease activity. A similar study to determine DNA damage sites will be done using isolated chromatin and nucleosomes. The effect of intercalation drugs on chromatin will be analyzed by spectral techniques and also studies will be done to determine the interaction of these drugs with DNA strand scission drugs. Mechanism of drug action studies will also examine the effects of DNA strand scission drugs using tumor virus SV40 as a model system. A comparison will be made between the ability of these drugs to cut cell-free purified SV40 DNA in vitro and SV40 DNA isolated from drug-treated purified virus. Also the ability of these drugs to alter viral infectivity and propagation will be explored and correlated with the drug's ability to cut viral DNA.
|
{
"pile_set_name": "NIH ExPorter"
}
|
At present, we have little understanding of the key neurobiological mechanisms which result in enhancements in cognition following a single bout of exercise and how these mechanisms accrue to contribute to more permanent changes in cognitive function over time. Such information is vital for the development of physical activity interventions during development to maximize cognitive health and function. Accordingly, this proposal will begin to address this knowledge gap by using an innovative, multi-disciplinary approach cutting across cognitive neuroscience, educational psychology, and kinesiology to establish an empirical basis for two hypothesized neurobiological mechanisms - resting-state cerebral blood flow and functional neural connectivity - proposed to underlie the beneficial effects of single-bouts of physical activity on inhibitory control during preadolescence. Using a within-participant repeated-measures design, state-of-the-art functional neuroimaging measures of resting-state cerebral blood flow and functional neural connectivity will be assessed in a sample of preadolescent children in relation to changes in behavioral task performance resulting from a single bout of exercise relative to a non-exercise condition during two separate, counterbalanced sessions. The significance of the current proposal is its potential to advance our understanding of acute-exercise induced enhancements in cognition during preadolescence by determining the extent to which previously proposed mechanisms explain variance in behavioral changes in cognition associated with exercise. This proposal represents an initial step in an extended program of research aimed at the development of physical activity interventions to maximize their influence on these neurobiological mechanisms to generate the greatest cognitive enhancement and potentially more rapidly incur the more stable facilitations in cognitive processes associated with chronic physical activity participation. Ultimately, this research effort has the potential to influence education policy by providing evidence-based recommendations for exercise programming in schools. That is, despite our growing understanding of the relation between physical health and mental health; educational institutions are de-valuing the importance of physical activity during the school day through the reduction or elimination of physical activity opportunities in school. Thus, a greater understanding of the neurobiological mechanisms associated with single bouts of short-duration physical activity may inform on the functional day-to-day implications of removing physical activity opportunities from the school day for cognitive health. As physical activity trends indicate that children are growing increasingly sedentary and unfit; such an understanding of the potential for single-bouts of activity to contribute to more permanent changes in cognitive function over time and how best to optimize physically active behaviors to maximize cognitive health and function during development may serve to alter the trajectories of cognitive and brain development resulting in enhanced academic achievement in school-aged children.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Abstract Radical prostatectomy (RP) is a commonly used treatment option for localized prostate cancer. Unfortunately, the procedure carries a high risk of erectile dysfunction (ED) which is highly detrimental to the post-surgical well-being of men. The main pathophysiological mechanism behind ED is damage to the cavernous nerves (CN); mechanisms that regenerate or repair CN injury following RP could alleviate or treat ED in these patients. The studies outlined in this proposal will pursue a novel approach to CN repair by harnessing the nerve regenerative powers of Fidgetin-like 2 (FL2), a newly discovered microtubule regulator. In vitro studies indicate that the depletion of FL2 strongly enhances axonal growth in primary cultures of rat peripheral nervous system neurons. Recent work by the investigators involved in this proposal have shown that targeted depletion of FL2 via topical application of FL2-siRNA encapsulated in nanoparticles (FL2-siRNA-np) promotes the closure and regeneration of cutaneous wounds in mouse models and improved erectile function outcomes in a rat model of RP when applied at the time of CN injury. Remarkably, in a CN transection model of RP, at two weeks post treatment with FL2-siRNA there is both improved erectile function outcomes and visible nerve regrowth. These observations lead us to the hypothesis to be tested in this proposal that ?following CN injury, depletion of FL2 activates mechanisms that promote cavernous nerve regeneration, leading to accelerated recovery of erectile function?. This hypothesis will be tested in two specific aims. In the first Specific Aim we will substantiate our preliminary results that depletion of FL2 improves recovery of erectile function following CN injury used as an animal model of RP. We will optimize formulation and treatment regimens for ED. FL2-siRNA will be applied at the site of CN injury both at the time, and one week following injury and recovery of erectile function determined by measuring the intracorporal pressure/blood pressure ratio (ICP/BP) developed after CN stimulation, at various time points after treatment. Following cavernosometry, we will perform pathology studies to provide evidence of CN regeneration and safety of the different formulations. In the second Specific Aim we will conduct experiments to elucidate the mechanism by which the depletion of FL2 promotes functional recovery after CN injury. We will determine the effects of FL2-siRNA on axon growth and regeneration in vitro. Using tissues of controls and FL2-siRNA treated animals we will perform extensive comparative histopathology and gene expression analyses on corporal tissue and from the site of injury of treated versus control rats at various time-points after CN injury. Overall, upon completion of this proposal we will have established the potential of FL2 as a novel target in repairing CN damage following RP, determined treatment regimens, formulation and safety considerations to optimize potential clinical translation and have gained insight into the mechanism of FL2 mediated CN repair.
|
{
"pile_set_name": "NIH ExPorter"
}
|
There is considerable interest in understanding the mechanisms and function of mosquito innate immunity, with recent research in this area identifying important immune genes and metabolic pathways that appear to influence the ability of mosquitoes to transmit malaria. However nearly all such studies have been conducted under highly controlled lab environments, which is not what mosquitoes and Plasmodium parasites experience in the field. Natural malaria transmission occurs across a temperature range of 15-35 0 C, with the potential for considerable variation in conditions across temporal and spatial scales. We wish to test the hypothesis that temperature has significant impacts on mosquito immune function and vector competence, and to explore whether the insights gained under one set of controlled conditions are sufficient to understand variation in mosquito resistance/refractoriness in nature. We argue that mosquito immune function should be affected by environmental temperature because mosquitoes are small-bodied and cold-blooded. In other insect-parasite systems temperature has profound effects on parasite development and overall host resistance. Whether such effects occur with mosquitoes and malaria and if so, whether these effects are due to impacts of temperature on the parasite, on host immunity, or some combination of both, remains unclear. If temperature does affect mosquito immune function, then temperature may be a potent environmental parameter describing variation in mosquito resistance / refractoriness in natural vector populations and might also explain the disparities in immune responses to different Plasmodium parasites (in particular between some rodent malaria parasites and P. falciparum). More fundamentally, if mosquito immune response exhibits thermal sensitivity, the current approach of outlining immunological mechanism under standard lab conditions is insufficient for understanding natural or transgenic vector competence. PUBLIC HEALTH RELEVANCE: Current understanding of mosquito innate immunity derives almost exclusively from studies conducted under standard laboratory conditions. Yet in nature, mosquitoes and parasites interact across a range of environments. We wish to test the hypothesis that environmental temperature significantly shapes mosquito immune function and determine whether notional resistance mechanisms revealed in the lab can explain variation in mosquito resistance to malaria in the field.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Pre-mixed formulations of soluble prandial insulin analogs and NPH micro-crystalline suspensions provide a simplified and cost-effective basal-bolus regimen for patients with Type 2 diabetes mellitus (T2DM) not effectively controlled by basal insulin therapy alone. Such twice-daily products (in vials or pre-filled in pens) account for >$2B in sales in the developed world, and their use is rapidly growing in the developing world. The market leaders are Novo-Nordisk (NovoMix(r) 30 in which insulin aspart is formulated 30% as a soluble zinc hexamer and 70% as an NPH micro-crystalline suspension) and Eli Lilly and Co (Humalog(r) Premixed Insulin in which insulin lispro is formulated 25% as a soluble zinc hexamer and 75% as an NPH micro-crystalline suspension; a 50/50 product is also available). Despite their popularity and clinical utility with appropriate patients, these products are complicated to manufacture, not mixable with other insulin products, fixed in their basal-bolus ratio, and subject to degradation above room temperature. Once opened, pens and vials must be discarded after 10 and 28 days, respectively. We have invented an ultra-stable biphasic insulin analog formulation that would be simpler and less expensive to manufacture, tunable in its pharmacokinetic properties, and ultra-stable at temperatures >100 F. Designated Thermalin-biphasic, our product builds on an innovative structural approach based on a single-chain insulin (SCI) platform. Our SCI analog exhibits a pharmacodynamics profile in an animal model that recapitulates the profile of current pre-mixed insulin products. Its ultra-stability would provide convenience during travel, enable mail-order delivery, and enhance emergency preparedness; in the developing world such a product would circumvent the cost and complexity of the cold chain of insulin delivery. Accordingly, we anticipate that this product would be of broad benefit t patients with Type 2 diabetes mellitus in the developed and developing worlds. We anticipate that Thermalin-biphasic will provide a significant cost savings relative to current insulin NPH- mix products. This savings would accrue from three factors: (a) simplification of transport and delivery through elimination of the need for refrigeration, (b) extended shelf life of vials in the hands of patients, and (c) simplification of the manufacturing process through reduction in post-fermentation processing. This Phase I project will take Thermalin-biphasic through proof-of-concept testing. This novel molecular platform, invented at Case Western Reserve University (Prof. M.A. Weiss) and licensed to Thermalin Diabetes, LLC, confers a dramatic extension of shelf life with retention of potency at high temperatures. The applicant, Dr. B. Frank (PI), was co-inventor of Humalog(r) during his prior career at Eli Lilly. Collaborative studies in will be conducted by Prof. C. T. Roberts, Jr. (cellular signaling and mitogenicity; Oregon Health Sciences University) and Prof. A. Cherrington (PK/PD in dogs; Vanderbilt University School). Thermalin Diabetes, LLC has an exclusive license to the IP, which is owned by CWRU.
|
{
"pile_set_name": "NIH ExPorter"
}
|
It is widely appreciated that reactive oxygen species (ROS) play a major role in the initiation of cancer, and they are also implicated in many cancer therapies, such as ionizing radiation, cisplatin and taxanes. More recently, it has been discovered that cancer cells produce ROS as signaling molecules that promote proliferation. Unfortunately, the molecular details of how redox regulation affects cell signaling events are far from clear. New experimental and computational technologies that we have developed are uniquely suited to identifying the molecular targets that are modified by ROS, either as a result of ROS damage or ROS signaling. With the reagents and methods that we have recently developed, we can now evaluate the "redox profile" of cell populations by targeting uniquely reactive cysteine sulfenic acid (Cys-SOH) groups, the initial intermediates generated following reaction of activated protein thiolate groups with hydrogen peroxide and peroxynitrite (and perhaps other ROS). In this R33 application, our labeling technology will be further developed for quantification and multiplex analysis, so that it will have broad applicability in: 1) the investigation of basic mechanisms of ROS damage and ROS signaling;2) molecular profiling to stratify patients with cancers that are sensitive to ROS-generating therapies;and 3) the development of novel cancer therapies based on the inhibition of ROS-dependent proliferative signaling. The following Specific Aims are proposed: 1) to develop reagents and methods of use for additional new, multicolor fluorescently-labeled Cys-SOH reagents for multiplex analysis of samples;2) to develop quantitative mass spectrometry methods, which have some major advantages over gel-based methods (including direct readout of protein identity and numerous posttranslational modifications);and 3) to use the new quantitative methods to detect and identify Cys-SOH modified proteins generated during ROS-dependent signaling in HEK-293 cells and ovarian cancer cells. Taken together, the approaches developed in Specific Aims 1 and 2 will provide new tools for the research community to use to study the mechanisms of redox regulation and signaling. In Specific Aim 3, these tools will be used to determine the targets of ROS in the regulation of cell proliferation and apoptosis. First we will continue our study of NF-?B regulation in HEK-293 cells in response to cytokine (TNF-?) and tumor promoter (TPA) stimulation. Second, we will use ovarian cancer cells treated with cisplatin or taxane to determine which protein oxidations are critical to regulating survival and apoptosis. Besides providing specific information about the mechanism of redox regulation and signaling, these biological experiments will allow us to further refine our reagents and methods to make them most useful to the cancer biology community. These approaches to detecting functional oxidative modifications to cellular proteins hold promise in identifying specific protein targets that mediate the actions of anticancer drugs, e.g. through their effects on cell cycle arrest, cell division or apoptosis. An outgrowth of these studies could also be the development of new anticancer drugs and the ability to predict efficacy of a given drug in the treatment of individual patients.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Opioids are the cornerstone of treatment for moderate to severe acute and chronic pain, which costs the US economy tens of billions of dollars annually. However, the dose of opioid required to control pain varies up to 10-fold between individuals. In addition, large inter-individual differences exist in the degree to which patients suffer side effects from opioids such as sedation, pruritus, or nausea. Moreover, it is not known why some individuals become addicted to opioids while others do not. The principal hypothesis to be evaluated is that the degree of analgesia provided by opioids in humans displays substantial familial aggregation, and is, in fact, heritable. These studies will use a classical twin paradigm to determine the role of genetics and the environment in influencing analgesia and a range of other opioid effects. Specific Aims: (1) Determine the degree to which opioid analgesic responses show familial aggregation and make preliminary estimates of heritability using both a heat pain model and a model of inflammatory pain with central sensitization. (2) Determine the degree to which non- analgesic opioid responses show familial aggregation and make preliminary estimates of heritability. Side effects such as sedation, nausea, respiratory depression, and pruritus, as well as the positive affective response, a measure of abuse potential, will be monitored. Monozygotic (MZ) and dizygotic (DZ) twin pairs (125 total pairs) will be tested under controlled pain laboratory conditions for their responses to opioid infusion using the complementary pain models while monitoring side effects and additional psychometric indices of mood, sleep, and abuse potential. The selected models provide unique mechanistic information because they involve different peripheral and/or central pain pathways. DNA samples will be collected for zygosity testing and banked for future studies. PUBLIC HEALTH RELEVANCE: Establishing a significant heritable component to human responses to opioids could lead to new analgesic strategies tailored to an individual's genetic makeup, thus reducing patient risk and speeding control of pain. In addition, future large scale candidate gene and genome wide association studies based on these results could uncover key genes involved in pain and analgesic pathways, leading to more rational therapeutic targeting and ultimately more cost- effective treatments.
|
{
"pile_set_name": "NIH ExPorter"
}
|
PROJECT SUMMARY Craniofacial anomalies account for nearly one-third of all birth defects and are a severe cause of morbidity and mortality in infants. In spite of their prevalence, the underlying genetic and molecular mechanisms causing most craniofacial defects remain largely unknown. The long-term goal of the proposed studies is to define the molecular and genetic pathways that control craniofacial development for the purpose of regeneration and repair, tissue engineering, and the diagnosis of and intervention into congenital craniofacial birth defects. Specifically, this application is focused on the identification of the transcriptional pathways and mechanisms involved in craniofacial development. Recent work has identified a novel genetic model for Pierre Robin Sequence (PRS) in mice. PRS occurs in about 1 in every 800 live births and is characterized by small lower jaw, improperly positioned tongue and posterior cleft of the palate, which can result in upper airway obstruction and feeding difficulties. Previously, it was thought that PRS occurred as a result of environmental factors that restrict outgrowth of the mandible, but it is now quite clear that this sequence also has an undefined genetic component(s). Mice that carry one mutant allele of Dlx5/6 locus and one mutant copy of the Mef2c allele die at birth from craniofacial defects resembling PRS. The Dlx5/6 locus encodes two Distal-less related homeobox transcription factors, while the Mef2c locus encodes a MADS box transcription factor. In addition to their genetic interaction, MEF2C and Dlx5 cooperate to induce a robust synergistic transcriptional response. The hypotheses underlying this proposal are that Dlx5/6 and MEF2C form a transcriptional complex downstream of endothelin receptor signaling, that this complex is essential for the activation of a subset of genes that are required for craniofacial development, and that mutations or aberrant expression of Dlx5/6-MEF2C target genes contributes to craniofacial anomalies such as PRS. To address these hypotheses, three specific aims are proposed. Aim 1 will define the physical and functional interaction between Dlx5 and MEF2C and how this results in transcriptional activation. The goal is to define the transcriptional mechanisms that control gene expression during craniofacial development. Aim 2 will analyze the craniofacial defects in Mef2c-Dlx5/6 double heterozygotes in detail. The goal is to identify possible cellular mechanisms underlying the mandible and palate phenotypes in double heterozygotes to understand how these processes control palate closure and jaw growth. Aim 3 will identify upstream regulators of Mef2c transcription in craniofacial mesenchyme and will determine whether Mef2c is a direct target of the endothelin signaling pathway, using a transgenic mouse approach. The goals are to place Mef2c into a transcriptional pathway and to identify the immediate transcriptional effectors of endothelin signaling in craniofacial development.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Candida species are the most important fungal pathogens of humans, and Candida albicans, in particular, is the most common cause of oropharyngeal candidiasis, which is an infection that affects patients suffering from Acquired Immune Deficiency Syndrome (AIDS) among other ailments. Candida are also the fourth most common cause of systemic infections (~30% mortality rate) in intensive care units This fellowship proposal explores the hypothesis that the phospholipids phosphatidylethanolamine (PE) and phosphatidylserine (PS) affect virulence by affecting signal transduction pathways that regulate virulence factors necessary for adhering to and invading host tissues, as well as evasion of the host's immune response. Little is known about the how phospholipid synthesis pathways regulate disease, but some of these pathways differ from those in mammals and may provide new drug targets. Two different pathways synthesize PE in C. albicans. The de novo pathway synthesizes PE from PS, which itself is synthesized by only one enzyme, Cho1p. In addition, the Kennedy pathway makes PE from extracellular ethanolamine. Mutants completely blocked in de novo PS and/or PE synthesis (cho1?/? and psd1?/? psd2?/?, respectively) are avirulent, have a significant drop in PE, expose cell wall ?-glucans (an immune marker for fungal infections) to enhanced detection by the immune system, and adhere to invade host epithelial cells poorly. In addition, these mutants are ethanolamine auxotrophs, and it is possible that the mutants suffer a fitness defect in the host due to insufficient ethanolamine supplies. It s hypothesized that mutations that completely block the de novo PE synthesis pathway affect virulence by compromising cell signaling cascades within the cell, resulting in drastic changes in presentation of cell surface proteins involved in host adherence and immune evasion. These changes in PE synthesis may also decrease fitness of C. albicans in the host due to ethanolamine auxotrophy. This hypothesis will be tested in three aims: 1) Observe differences in cell signaling pathways known to be involved in the regulation of virulence and adherence to host tissues (ie. Rim101 pathway), 2) Determine if PS and PE play a role in evading the immune response 3) Discover if auxotrophy for ethanolamine, a known substrate of PE, affects virulence of the mutants in the host.
|
{
"pile_set_name": "NIH ExPorter"
}
|
DESCRIPTION: (Applicant's Description) The Cooperative Breast Cancer Tissue Resource (CBCTR) represents a collaboration between four institutions that possess archives with large numbers of formalin-fixed breast cancer specimens. These four sites, under Cooperative Agreements with the NCI, have created the CBCTR as a tissue resource for investigators of breast cancer. The CBCTR was originally conceived to provide large numbers of specimens particularly for definitive studies of prognostic markers. The CBCTR has enrolled nearly 9,000 archival breast cancer specimens together with clinical and follow-up information about each case. The material exists as a "virtual bank" with the actual specimens remaining at the individual sites but a Central Database tracking each specimen. The database is mounted on the World Wide Web and may be searched by investigators seeking to ascertain the number and type of material available from this resource. Investigators may then apply to the CBCTR for tissue specimens. Applications are reviewed by a panel of experts, the Research Evaluation Panel, for scientific merit and by the CBCTR Coordinating Committee of Principal Investigator's for feasibility, prior to providing the requested specimens. Kaiser Permanente, Northwest Division has provided over 2,500 specimens to the CBCTR and is reapplying for funding to continue its efforts in this project. This highly integrated and stable medical care system represents an ideal setting for such a project. It has diagnosed a large number of breast cancers since 1970, has archived all such specimens and has complete clinical and follow-up information in its files on all patients. Finally, this organization is committed to research in the public domain and seeks to share its resources with others in the pursuit of breast cancer research.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goal of this project is to comprehensively establish the pathway of T cell activation. We intend to define and order the external signal signals and intracellular messengers responsible for 1 degree and 2 degree T cell stimulation and link those events with sequence of gene activation that governs the activation process. In short, we will identify and sequence the physiologic extracellular stimuli causing T cell activation, identify the intracellular messengers activated by these stimuli, linking the stimuli and intracellular messengers to gene activation, and identify the sequence of T cell activation antigen associated gene activation. The specific aims are: 1. To identify and sequence the physiologic extracellular stimuli yielding T cell activation. 2. To identify the the intracellular messengers activated by each of these extracellular stimuli. 3. To identify the mechanisms by which these stimuli and intracellular messengers cause gene activation. 4. To identify the sequence of T cell activation associated gene activation. These data should provide a comprehensive framework by which T cell immunobiology and immunopharmacology can be examined in health and disease.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Synthetic nucleic acids with antisense sequence complementary to mRNA, and their use for gene activity detection, have advanced our understanding of the molecular mechanisms of diseases in all disciplines of the biological sciences. For in vivo investigations, oligoDNA (ODN) or oligoRNA (ORN) can be modified with phosphorothioate (yielding sODN or sORN) to increase resistance to nucleases. Our hypothesis is that hybrids of sORN with target mRNA is more stable than sODN hybrids. We will compare a modular magnetic resonance (MR) probe comprising supraparamagnetic iron oxide nanoparticles (SPION, a T2 agent) labeled with sODN or sORN. At present, intracerebroventricular (ICV) injection via cortical or lumbar puncture is one of only a few clinically approved methods to deliver drugs to the cerebral spinal fluid (CSF) in humans. We have demonstrated that neural cells of live animals take up SPION-sODN with moderate efficiency and specificity in mRNA targeting in vivo by MR imaging: (1) ICV delivery in mice safely facilitates global distribution of SPION- sODN in mouse brains without lethal effect, (2) specific binding has been shown by in vivo priming of SPION- sODN to target mRNA by reverse transcription (RT), (3) results from electron microscopy (EM) show that iron oxide is located in the end some with a unique association to the endoplasmic reticulum (ER) and nuclei where mRNA is located, (4) changes in SPION-sODN retention above baseline (DR2*) are positively proportional to gene activities (linear regression = 1.0). Our goal is to evaluate the efficiency of SPION-sORN (and SPION-sODN) for targeting astroglia-specific glial fibrillary acidic protein (GFAP) mRNA. Completion of the proposed work provides a platform for novel gene targeting probes as well as a powerful tool for early evaluation of astroglia activation in vivo. Therefore, less SPION-sORN than SPION-sODN is used for gene targeting and reduces accumulation of iron in the brain, leading to longitudinal assessment of neurologic events. We will: Aim 1: Compare in vivo dose and uptake of SPION-sODN or SPION-sORN in mice using ultra-high field MRI. Our hypothesis is that SPION retention (DR2*) will improve when SPION-sORN (SPION-Rgfap) is used to target GFAP mRNA. We will longitudinally compare DR2* of these two probes in the brains of live mice. Aim 2: Validate the correlation between MRI and histological assessments. Our hypothesis is that co- localization of dual-labeled probe (e.g., SPION-Rgfap-Cy3) can be specifically transfected to GFP-expressing glia of transgenic mice in vivo, and can be confirmed under fluorescent, optical and electron microscopes. We will collect brain samples after ICV probe delivery for this correlation study. Aim 3: Validate target binding using primer-free in situ RT to cDNA followed by target specific PCR. The hypothesis is that SPION-Rgfap will bind specifically to GFAP mRNA target in vivo and serve as a primer for in situ RT-PCR. We will collect brain samples, quantify the PCR results, and establish the correlation between MRI DR2* and mRNA copy numbers, using disease model systems. PUBLIC HEALTH RELEVANCE: Glial activation plays an important role in the disease process. However, glial activation can only be detected in postmortem brain samples. The work outlined in this application investigates the mechanism of gene targeting for specific messenger RNA of glia activation in the central nervous system. We will apply a novel nucleic acid- based probe to investigate glial activation in live animals, using magnetic resonance imaging (MRI). The work proposed here, with initial application in live animal models, has important implications for MRI-based analyses of neurophysiologic events at the genetic level. High-resolution MRI of intracellular RNA holds promise for translation to molecular biology in live subjects;to permit real-time longitudinal MRI in future applications for neuroscience and preclinical evaluation in medical research.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Oxygenation plays a critical role in health and medicine. Deficits or excesses in cells, tissues and organisms are associated with disease, damage, poor healing, and even death. For example, hypoxia is implicated in cancer progression, metastasis, and resistance to therapies. Hypoxia inducible factor 1 (HIF-1) expression in low oxygen environments accelerates angiogenesis and promotes tumor survival, and radiation and drug treatments can be less effective when reactive oxygen is not present to enhance the tumor damage or when hypoxia-induced changes in the tumor microenvironment otherwise affect drug delivery and action. Despite the importance of oxygen partial pressure (pO2) to biomedicine, its assessment is often limited to oxygen consumption rate (OCR) measurements, invasive probes, or expensive imaging modalities. Often methods provide only average values for whole populations of cells, or are restricted to single point measurements in space or time. The long-term goal of this project is to develop a versatile material set and platform of pO2 imaging technologies that can enhance preclinical studies and aid in diagnosis, treatment and surgery in many medical contexts. Building upon success in proof of concept studies and the strengths of our team in materials synthesis, imaging, and cancer biology, the objective of this application is to develop dualemissive difluoroboron ?-diketonate-poly(lactic acid) (BF2bdkPLA) dye-polymer nanoparticles (BNPs) in conjunction with imaging methodologies for luminescence detection and ratiometric O2 sensing in breast cancer in vitro and in vivo mouse models. This objective will be accomplished by pursuing the following specific aims: 1) BNP dyes will be chemically modified for broad range emission color tuning and oxygen sensitivity modulation; imaging methods will be developed for their use in in vitro ratiometric O2 sensing (i.e. both fluorescence (F) and phosphorescence (P) detection). 2) BNP polymers will be adapted for passive and active targeting for pO2 monitoring with improved spatial specificity; targeted BNPs will be investigated in vitro and in vivo. 3) BNP hypoxia imaging agents will be tested for their ability to detect tumors, and to monitor tumor progression and radiation and chemotherapy response over time in a mouse mammary window model. The proposed cost- effective technology is innovative because it enables dynamic hypoxia imaging with improved combined spatial and temporal resolution compared to existing approaches with a modular, tunable, synthetically accessible materials platform. The expected outcome is a versatile oxygen nanosensor technology in conjunction with common optical imaging modalities to quantify pO2 in cells, tissues and in vivo. BNPs with greater specificity, tissue penetration of light, and multiplexing capability will result. This work will have a positive impact on cancer car because it will better illuminate the relationships between hypoxia, cancer progression, and treatment protocols. Ultimately, BNPs will shed light on many medical challenges by helping to map relationships between oxygenation and biological function. This is already being realized.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The beta thalassemias are characterized by deficiency of adult (b) globin chains of adult hemoglobin (Hb), an excess of toxic, unmatched a globin chains, and intramedullary hemolysis. The resulting anemia only after fetal (g) globin synthesis and Hb F is suppressed in infancy. Induction of fetal (g) globin synthesis and Hb F is suppressed in infancy. Induction of fetal (g) globin to levels which improve globin chain balance by even 10% can prolong red blood cell survival and diminish clinical morbidity. Arginine Butyrate, a prototype fatty acid, has been effective when given intermittently or Pulsed, inducing Hb F and increasing total hemoglobin by 3 gm/dl over baseline levels in 5/6 beta thalassemia patients. EPO-rhu can prolong red cell survival but does not correct underlying globin chain imbalance. We hypothesize that these two agents should have additive therapeutic benefit. A clinical pilot study is proposed to test the hypothesis that therapy with Pulsed Butyrate, or rhu-EOP + Pulsed Butyrate, will induce g globin chain synthesis sufficiently to improve non a: a globin chain balance and red blood cell survival, and increase total Hb in a significant proportion of patients with beta thalassemia intermedia. Baseline hematologic levels will be assayed four times over a two-month period. Butyrate will then be administered during an Induction Phase, to determine a patient's optimal dose, followed by a "Maintenance Phase" of therapy for 3 months. Pulsed Butyrate will also be tested with rhu-EPO. The proportions of patients on each arm of the study in whom the following endpoints are achieved, compared to baseline levels, will be analyzed: 1) an increase in total of Hb in at least 2.0 grams/dl, 2) an increase in hematocrit of at least 5%, 3) a decrease in hemolysis, measured by LDH and bilirubin, 4) improvement in globin chain synthesis by 10%. Whether specific genotypes and in vitro response to Butyrate correlate with clinical responses will also be analyzed. These studies should determine he proportion and some genotypes of beta thalassemia patients which can benefit from Pulsed Butyrate +/- rhu-EPO therapy.
|
{
"pile_set_name": "NIH ExPorter"
}
|
We have demonstrated in 10 normal subjects that breathing a He (80%) O2 (20%) mixture has no significant effect on static lung pressure volume behavior. Partial flow volume (FV) curves were found to be superior to full FV curves in detecting response to bronchodilator in 49 subjects with respiratory symptoms. In 15 of 30 patients with significant reduction in expiratory flow we found normal density dependence comparing air and HeO2 FV curves at 50% VC. No subject had asthma. The response to bronchodilator appears predictable based on control level of density dependence. In autopsy human lungs we have been moderately successful in predicting the FV curve from measures of airway pressure-area behavior and static pressure volume curves. Normal values for total respiratory resistance at an oscillation frequency of 3 cps have been established. BIBLIOGRAPHIC REFERENCES: Rodarte, J.R., Hyatt, R.E. and Westbrook, P.R.: Determination of Lung Volume by Single and Multiple Breath Nitrogen Washout. Amer. Rev. Resp. Dis. 114:131, 1976. Frazier, A.R., Rehder, K., Sessler, A.D., Rodarte, J.R. and Hyatt, R.E.: Single Breath Oxygen Tests for Individual Lungs in Awake Man. J. Appl. Physiol. 40:305, 1976.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The most frequent cause of visual loss in childhood is functional amblyopia. Recent studies show that a high spatial frequencies and under conditions of crowding, qualitatively different acuity losses are found in different amblyopes. The present proposal will capitalize on these differences as a means of clarifying the specific losses in peripheral vision and amblyopic vision compared to normal foveal vision. Just as color deficient individuals had contributed much to our knowledge of color vision, we expect amblyopes will contribute much to our knowledge of spatial vision. Several new psychophysics methodologies will be used to help unravel the comples story: 1. There is a golden opportunity to learn about the amblyopic distortion by comparing the amblyopic eye to the nonamblyopic eye. Under the proper conditions, amblyopes (even those with anomalous correspondence) can not determine the eye-of-origin of a stimulus. By using a double judgment methodology, eye-of-origin confusions will be used to convert subjective interocular comparisons into objective signal detection measurements. New methods for analyzing double judgment tasks with the effects of response bias and response correlation minimized will be used to extend signal detection methodology to the suprathreshold regime. The proposed experiments will focus upon the interactions between local features in peripheral as well as amblyopic vision: 1-Repetitive arrays of features will allow apatial frequency analysis to be used. Spatial frequencies within 3/2 octave of the cutoff spatial frequency will be exhaustively investigated. Effects of phase and background contrast will be explored. 2-A hierarchy of localized luminance distributions will be used to measure contrast sensitivity and phase sensitivity will be studied in amblyopes and peripheral vision. 3. After the first year, nonlocal phase effects, effects of crowding and two dimensional stimuli (ie vernier acuity) will be studies. 4. Quantitative modelling will be used to explain amblyopic and peripheral losses in terms of underlying mechanisms.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objective of this proposal is to update, revise and republish a second edition of the "Registry of Gallus domesticus genetic stocks" in 1975. The scope of the second edition will be enlarged to include the Japanese quail and the turkey, as well as the chicken, and it will be covering research and teaching institutions in Canada as well as all of the United States. The Registry will include specialized lines, stocks carrying mutant traits and rare and unusual breeds and varieties. The compiled data will include a brief description of the trait, line or breed and the names and addresses of those people who have these stocks in their possession and their availability. This Registry will be made available to all biologists interested in it.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Investigators in this project will continue to benefit from a state of the art facility, with advanced nstrumentation and highly trained technicians at their disposal. The personnel have unique experience in sorting extremely rare cells to high purity and in obtaining nine color data with small sample sizes. They routinely handle human specimens with biohazard precautions, and can work with spores, live bacteria and bacterial products. There is no equivalent expertise in the State of Oklahoma. Costs are contained in part by OMRF institutional support. Training is another important activity of this core facility. While investigators can simply hand samples to the technicians and receive formatted data, they are encouraged to learn at least how to analyze their own results. Most students and postdoctoral fellows prefer this, and a few have even earned how to sort on the FACSAria with close supervision. Advice on related procedures, such as reagent selection and magnetic bead enrichments is also readily available. Many appropriately labeled antibodies are available to facilitate pilot experiments and encourage innovation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Approximately 25% of current pharmaceuticals are either directly obtained from nature or are structurally based upon natural products that include important anti-cancer drugs and painkillers from plants. The study of selected medicinal compounds from plants requires detailed investigation of exactly that plant species that contains the entire biosynthetic pathway to the drug. Detailed genetic and biochemical information on these highly specialized plants is often missing. It is proposed to fill this knowledge gap and advance research in the field of plant-derived pharmaceuticals discovery and production in the poppy family (Papaveraceae) by identifying genes expressed in plant organs distinct in their alkaloid spectrum in opium poppy Papaver somniferum and the California poppy Eschscholzia californica through comparative transcriptomics using a combination of EST, 454 FLX and Solexa sequencing. Microarrays that represent the transcriptome will be developed for each species so that the temporal and spatial expression patterns during normal development in all three poppy species and in transgenic plants of P. somniferum in which six of the known alkaloid synthesis genes are systematically over- or under-expressed can be determined. BAC libraries will be prepared so that physical maps can be constructed of genomic regions surrounding alkaloid biosynthesis genes in these two members of the poppy family. Molecular evolutionary analysis of sequence data will be carried out within the context of gene family phylogenies, gene expression patterns and alkaloid content in order to identify novel genes that may encode enzymes or transcription factors involved in alkaloid biosynthesis. Functional analyses will be performed with the genes hypothesized to encode enzymes involved in the biosynthesis of medicinal alkaloids. This genetic and biochemical information will be made publicly available in the form of a user-friendly database (PoppyDB) and viewer (PhyloPathway) for metabolic engineering, biomimetic synthesis and directed evolution for the improved production of drugs and for the development of novel drugs. Project Narrative: Approximately 25% of current pharmaceuticals are either directly obtained from nature or are structurally based upon natural products that include important anti-cancer drugs and painkillers from plants. The study of selected medicinal compounds from plants requires detailed investigation of exactly that plant species that contains the synthetic pathway to the drug. Detailed genetic and biochemical information on these highly specialized plants is often missing;it is proposed to fill this knowledge gap and advance research in the field of plant-derived pharmaceuticals discovery and production in the poppy family.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Cell surface molecules play important roles in regulation, differentiation and development. They can also be used to manipulate the human chromosomal content of human X Chinese hamster somatic cell hybrids. Immunological analysis of cell surface molecules influenced by genes on chromosome 21 will be identified and characterized using monoclonal antibodies. Deletion and irradiation hybrids will be isolated and used foro regional gene mapping of chromosome 21. Further studies will be undertaken on the analysis of the interferon system and the interaction of genes on chromosome 21. Comparative mapping provides a means for studying genome organization and chromosomal evolution. A gene map for chimpanzee chromosome 22 will be established and compared with its homologous human chromosome 21. A long term goal of these studies is to identify chromosome 21 encoded cell surface molecules which may plan a role in the pathogenesis of Down Syndrome.
|
{
"pile_set_name": "NIH ExPorter"
}
|
PROJECT SUMMARY (See instnjctions): Core A The administrative core is responsible for the operation of the program and supervision of the conduct ofthe proposed grant. The core facilitates communication between invesfigators and visits of outside investigators to MD Anderson Cancer Center. The core maintains the central databases for the clinical and translational research described in the projects as well as the GMP - Immune Assessment Core (Core E). The core supports the FDA IND submissions and reporting required for protocols in the POI. The core manages program personnel and budgets for each section ofthe grant. The core is responsible for preparing reports forthe research projects and publicafions. The CML group meets multiple times per week in meeting related to leukemias, stem cell transplantafion and specific laboratory research related to this program. We discuss the design, status and results of clinical and laboratory research, sample acquisition, status of patients participating on clinical research studies, pathology, cytogenefics, molecular analysis review. This includes a weekly CML POI program meetings and broader meefings involving the Leukemia, Stem Cell Transplant and CellularTherapy (SCTCT) and Molecular Pathology programs. Specifically, the Administrative Core has the following objectives to accomplish: Provide leadership to and oversight of the cores and research projects ofthe Program. Convene all necessary meetings, including meetings ofthe External Scienfific Advisory Board, regulariy scheduled meetings of the investigators, and discussion of all the clinical trials in the program. Prepare grant continuafion submissions for the NCI and to comply with internal reporting requirements. Coordinate data quality control and quality assurance in conjuncfion with the Biostatistics and other projects and cores. Monitor and control expenditures and maintain budget information. Track and maintain record of all publications, including abstracts and manuscripts, resulfing from the program project.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Adhering to the original philosophy that has guided the Alcohol Research Center from its inception, we bring together researchers with backgrounds in biophysics, biochemistry, cell biology, molecular biology, pharmacology and pathology around the central theme of "Alcohol and the Cell". The common focus of the different research components remains the elucidation of (1) the relationship between the structural and functional effects of ethanol on cells and subcellular structures, (2) the adaptations occurring during long-term exposure of the organism to ethanol, and (3) the molecular basis for irreversible cell injury associated with long-term ethanol intake. Studies focus on a variety of different tissues, including liver, heart, brain, cell lines of different origins and model membrane systems. Techniques to be used include electron paramagnetic resonance, nuclear magnetic resonance, fluorescence lifetime and energy transfer measurements, real-time fluorescence microscopic imaging of single cells, laser confocal microscopy of signal cells and perfused organs, electron microscopy, HPLC separation of lipids (both on an analytical and preparative scale), capillary gas chromatography, single cell electrophysiological measurements (voltage clamp and patch clamp) and molecular biological techniques. Individual projects focus on the effects of ethanol on the following parameters: (1) Cell Membrane Constituents, including (a) membrane lipids, (b)protein kinase C, and (c) phospholipid pools; (2) Cellular Function, including (a) Ca2+-dependent signal transduction, (b)E-C coupling in cardiac myocytes, and (c) cardiac Ca2+ channels; (3) Cell and Tissue Injury, including (a) apoptosis, and (b) aldehyde dehydrogenase gene expression. Pilot projects for the first two years focus on (1) ethanol effects on neuronal survival and (2) ethanol effects on connexin-43 expression. We expect that these studies will contribute to elucidating the relationship between ethanol-induced alterations at the molecular and cellular levels and the development of alcohol-related diseases.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This is a renewal application by the University of California, San Francisco (UCSF) to participate in the scientific activities of the Cancer and Leukemia Group B. UCSF joined the group as an affiliate of the University of California, San Diego, in 1990 and became a main member institution in 1992. The Principal Investigator (PI) on that application was Dr. Henderson, who was succeeded as PI by Dr. Venook upon Dr. Henderson's resignation in 1995. Dr. Venook has been active in CALGB since 1989. The UCSF faculty is very well represented in the scientific endeavors of the group. Eight members serve on core committees: Dr. Venook on the Gastrointestinal and Pharmacology and Experimental Therapeutics Committees; Dr. Small as Vice-Chair of Prostate and Chair of its Correlative Sciences subcommittee; Dr. Warren on Surgery, Gastrointestinal and Solid tumor Correlative Sciences; Dr. Henderson on Breast (former chairman); Dr. Linker on Leukemia; Dr. Triphathy on Solid Tumor Correlative Sciences; Dr. Davoren on the clinical Economics Working Group; and Dr. Kaplan as Vice-Chair of the AIDS Malignancy Working Group. Drs. Tripathy and Esserman head the current CALGB efforts to establish a high-risk breast and ovarian cancer registry. Drs. Venook, Small, Linker, Tripathy and Henderson are PIs on eight current studies; Dr. Warren is a co-PI on two studies and his laboratory is coordinating the correlative science components of each. Dr. Venook directed and has submitted for publication CALGB's study of paclitaxel in patients with liver dysfunction and Dr. Roach presented to the group a first-authored paper on racial issues in breast cancer using CALGB data. Drs. Kaplan and Gray have given plenary session scientific presentations at the group meetings. While accrual in the first three years of this past grant fell below predictions, accrual has improved dramatically and UCSF has met its target for the final year of the grant, with 138 patients accrued, 38 of whom are minorities. The enthusiastic participation of four UCSF hospitals (Moffitt-Long, Mt. Zion, San Francisco General and the Veterans Administration), the addition of an affiliate (St. Joseph's Hospital, Eureka, CA) and the continued intellectual involvement of committed clinical investigators will enable UCSF to maintain and improve upon its performance in the coming years.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Acute inflammation is characterized by increased microvascular permeability to plasma proteins and leukocyte recruitment into inflammatory sites. A large increase in permeability of the microvessel wall is a critical event resulting in edema formation and organ dysfunction. The long-term goal of our research is to investigate the mechanisms that regulate microvessel permeability under inflammatory conditions. The objective of this proposal is to investigate the direct correlation between vascular structural changes, transport pathway formations, signal transduction pathways, and the magnitude and time course of the permeability changes in intact microvessels in response to different stimuli. Three specific aims are proposed: 1) Investigate the cellular mechanisms of endothelial gap formation and inflammatory mediator-induced permeability increases in intact microvessels under acute and chronic inflammatory conditions; 2) Investigate the role of pericytes and the basement membranes in the regulation of microvessel permeability under acute and chronic inflammatory conditions; and 3) Identify the cellular mechanisms responsible for ROS-induced permeability increases under acute and chronic inflammatory conditions. These aims will be accomplished using combined confocal microscopy, electron microscopy, with quantitative assessments of microvessel permeability in intact microvessels. Our newly established methods enable us to three- dimensionally visualize and quantify inflammatory mediator-induced gap formation, characterize the changes in endothelial adhesion proteins, as well as to detect changes in actin cytoskeleton in endothelial cells and pericytes in individually perfused microvessels. The electron microscopy study allows ultrastructural changes to be correlated with confocal image findings. The proposed research will provide new information and possibly new concept for a better understanding of the mechanisms that regulate fluid and solute transport when permeability is increased under inflammatory conditions. The insight gained will be directly applied to the identification of an effective target to prevent the permeability increase and contribute to the development of targeted and clinically applicable anti-inflammatory therapies.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), Division of Intramural Population Health Research (DIPHR) requires data support for the baseline/child aged 3 years assessment for an observational cohort study titled Sprouts: Development of eating behaviors in early childhood. The study will recruit approximately 200- 250 mother-child dyads from among participants of a DIPHR-funded pregnancy/infancy cohort study. The contractor will monitor and manage data including but not limited to laboratory cognitive and behavioral assessments, eye-tracking, dietary assessment, self-administered questionnaires, biospecimens, clinical data, and anthropometrics. Most data will be collected electronically through the NICHD web-based data collection system called the Clinical Trials Database (CTDB, https://ctdb.nichd.nih.gov/ctdb/). The clinical site will collect dietary intake data using either the National Cancer Institute Automated Self-Administered diet recall (ASA24, https://asa24.nci.nih.gov/researchersite/) or the Nutrition Data System for Research (NDSR) (http://www.ncc.umn.edu/products/), to be determined. The child age 4-years assessment is anticipated to begin in fall of 2019 and to be complete in the fall of 2021, to be followed by data management, cleaning and analysis, culminating in delivery of the final datasets and documentation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Many species of protozoa are obligate intracellular parasites, including some, such as malaria, that are important agents of disease. Little is known as to why these eukaryotic cells can grow and reproduce only within another species of eukaryotic cell. Do they parasitize directly the energy-forming mechanisms of the host cell, or do they gain an energetic advantage by parasitizing the biosynthetic capabilities of the host cell? How are interactions with the host cell regulated? Extracellular maintenance of the parasites provides an approach toward answering these and other basic questions in the biology of parasitism. Only one kind of intracellular parasite has been kept alive and developing extracellularly. This is the bird malaria Plasmodium lophurae. The erythrocytic trophozoites of this organism, when removed from their host erythrocytes, will develop into schizonts in a medium of red cell extract. Exogenous ATP is a requirement for this development. We have now observed an initial extracellular development of merozoites of P. falciparum for which ATP and pyruvate are essential. We propose to investigate in detail this phenomenon both with regard to the constituents of the medium and to the fine structure of the developing merozoites. Special attention will be given to determining whether ATP can be replaced by ADP or AMP or by other nucleotides and to attempting to specify the role of the ATP. We will also investigate the possible effects of red cell membrane proteins on this initial extracellular development of falciparum merozoites. The information to be obtained will contribute specifically to knowledge of the physiology of the pathogenic erythrocytic stages of malaria parasites, with potential consequences for treatment and control of malaria. It will also bear on intracellular parasitism in general and on intracellular regulatory mechanisms.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Excess thyroid hormone induces a unique form of cardiac hypertrophy characterized by enhanced myocardial performance which may be related to switching from a low ATPase form of myosin to one with higher ATPase activity. At present our laboratory is attempting to define the mechanism(s) by which thyroid hormone regulates myosin isoenzyme expression. Although we mainly are concerned with the process of myosin isoform switching, we are impressed with the lack of information about nuclear thyroid hormone binding proteins themselves. Accordingly, the major aim of the proposed research is to investigate the role of nuclear T3 receptor in the initiation of cardiac hypertrophy and in the control of myosin isoenzymes expression. To accomplish these objectives, we plan, first, to clone the cDNA coding for the T3 receptor protein by using polyclonal and monoclonal antibodies to identify cDNAs coding for this protein in a specially constructed Lambdagt11 expression clone bank. Secondly, we propose to identify specific nuclear DNA binding sites for the T3 receptor. We have developed a strategy for isolation of the T3 receptor with its DNA fragment attached and support is requested to clone and sequence these fragments. These results should provide significant new information related to the structure of the T3 receptor protein and its DNA binding sites. These investigations will be carried out by a collaboration between Dr. Morkin's group in Tucson and Dr. Keith Latham's laboratory at the Uniformed Services University of the Health Sciences in Washington, D.C. Dr. Latham recently has developed polyclonal and monoclonal antibodies to the receptor protein. We believe the team we have assembled can make significant progress on elucidation of the mechanism of thyroid hormone actions on the heart and in other tissues.
|
{
"pile_set_name": "NIH ExPorter"
}
|
A peptide containing immunoreactivity to the amphibian peptide bombesin was isolated from bovine milk. The isolation procedure, however, gave relatively poor yields in spite of the precautions taken against loss due to absorptivity on the walls of various containers. New procedures are being developed to increase the yield of peptide recovered and decrease the number of steps in the scheme. Amphibian and mammalian bombesin-like peptides are known to affect gastric secretions either by i.p. infusion of the peptide or by intracerebral ventricular administration; a bombesin-like peptide in milk may have a similar mode of action. Thus, a preliminary assessment was under taken on a large number of potential analogues of milk bombesin on their neuromodulatory role of gastric secretion in rats. The data can be summarized as follows: (i) Trp in position 8 is required for biological activity; (ii) Gln and His at positions 6 and 12, respectively, are also important; and (iii) and unknown extension of sequences preceeding Gln6 are involved since neuromedin C (GRP 18-27) appears inactive.
|
{
"pile_set_name": "NIH ExPorter"
}
|
LHRH neurons, critical for reproduction, are derived from the nasal placode and migrate into the brain where they become integral members of the hypothalamic-pituitary-gonadal axis. We study mechanism(s) underlying LHRH neuronal differentiation, migration and axonal targeting in normal/transgenic animals, and nasal explants. Using these same models, our work also addresses the mechanisms regulating (intrinsic and trans-synaptic) LHRH gene expression, peptide synthesis and secretion in LHRH neurons. Previous work in our lab 1) showed GABA was an important factor in the migration of LHRH neurons in nasal regions, 2) identified a novel gene termed NELF which we hypothesize acts via a homophilic interaction to influence LHRH neuronal migration on olfactory axons and 3) demonstrated that biosynthesis and secretion of LHRH in vitro mimics that seen in vivo. Over the past year, we have characterized the expression of GABAA receptor subunits in LHRH neurons and found two subunits, alpha 2 and alpha 6, show inverse changes over development ? alpha 2 increases while alpha 6 decreases. In addition we have documented LHRH expression in a novel location, the developing incisor and are determining lineage relation between these LHRH cells and neuroendocrine LHRH cells derived from the nasal placode. We have also performed a differential screen of LHRH neurons after GABAergic treatment and have begun to examine the role of these genes both in vivo and in vitro. One gene that was differentially expressed was the peptide CCK. To date we have shown that CCK is co-expressed in LHRH neurons during development and influences both movement and maturation of LHRH neurons. We also examined the expression pattern of calcium channels in LHRH cells as a function of development and the functional consequences of changes in these expression patterns with respect to LHRH cell migration and/or regulation. We have found that although N and L type channels are present, disruption of neither alters LHRH movement. Finally, based on our previous work which showed estrogen receptor beta subtypes in LHRH neurons, we have examined LHRH neuronal activity and the effects of estrogen on this parameter. We found that estrogen has a direct effect of LHRH neuronal activity and increases the number of LHRH cells which participate in a synchronized calcium pulse. We hypothesize that this phenomenon is related to the positive feedback that occurs in vivo during the preovulatory surge. To other studies in progress examine the role of anosmin-1 in olfactory receptor/LHRH development and the electrical properties associated with LHRH neuronal activity. Future studies are directed at the molecules and cues important for development of the olfactory and LHRH neuronal systems as well as the mechanisms regulating LHRH neuronal activity. Specific studies in progress focus on: 1) isolation of midline cues which influence olfactory axon outgrowth; 2) the role of NELF and other molecules in LHRH migration, 3) identifying pacemaker molecules in LHRH neurons that participate in establishment/maintenance of rhythmic activity, 4) genes differentially expressed in LHRH neurons as a function of GABAergic signals and 5) the mechanisms by which estrogen alters LHRH neuronal activity.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Funds are requested to establish a state-of-the-art, high-resolution NMR facility for the study of biological systems. The facility which is to occupy the first floor of the new wing of the Biochemistry Building, Department of Biochemistry, University of Wisconsin-Madison (under construction), is designed to handle the severe problems of resolution and sensitivity encountered with biological macromolecules and in vivo preparations. The facility is to be based on two commercial multinuclear spectrometers (a 9.4 T wide bore spectrometer and an 11.8-16.5 T) narrow bore spectrometer) and an off-line data station for analyzing data. The instrumentation has been chosen to optimize the collection and analysis of two-dimensional NMR data sets and for carrying out biomedical NMR experiments requiring the ultimate in spectrometer sensitivity. The center will endeavor to provide the latest equipment and techniques in high-resolution, multinuclear magnetic resonance experimentation for collaborative and service projects covering several biomedical areas: structure and function of proteins, nucleic acids, and natural products; active sites and mechanisms of enzymes; modes of action of drugs; and physiological levels of metabolites in living cells and organisms. In addition to the primary function of providing effective research tools to these users, the facility will engage in instrumental development and will explore novel biomedical applications of NMR spectroscopy. Core research projects include homonuclear and heteronuclear two-dimensional NMR spectroscopy of macromolecules, isotopic labeling of biomolecules for NMR analysis, and multinuclear magnetic resonance studies of viruses, living cells, and intact tissues. The progress and administration of the facility will be reviewed regularly by an Advisory Committee of experts in the field of biological NMR, who will also evaluate the research proposals of long-term users. An Instrumentation Consultant Board will provide advice on development of equipment and techniques. A local Steering Committee, consisting of facility staff and representatives from the major user departments on campus, will deal with short range concerns.
|
{
"pile_set_name": "NIH ExPorter"
}
|
We use bacterial toxins as killing agents to eliminate cancer cells. To accomplish this, we modify the toxin so it no longer binds via its own cell-binding domain and substitute in place of the binding domain a monoclonal antibody. The antibody is chosen to bind cancer cells preferentially over normal cells. These toxin-antibody molecules are called immunotoxins. Immunotoxins are promising but imperfect anticancer agents. Our goal is to understand the interaction of various toxins with eukaryotic cells and use this information to design better agents for treating cancer. To study interactions, we add toxins to mammalian cells and study the pathway of death. In tracking the killing of cancer cells by immunotoxins, we made the observation that cells grown to high density are resistant to killing. We wish to understand this phenomenon and determine its relevance for cancer therapy in general. A convenient and potentially useful way to study cell-killing pathways is to use RNA interference to identify pathways that participate in toxin delivery to the cytosol - where it acts. Recently, we initiated studies with a newly described toxin from V cholera, called Vibrio Cholera Exotoxin (CET). This toxin is related to the exotoxin from Pseudomonas, exhibiting about 50% identity in selected domains (domains II and III). However, antibodies that neutralize the exotoxin from Pseudomonas do not neutralize CET, despite the close similarity.Truncated versions of Pseudomonas Exotoxin (PE) have been fused with antibody fragments to produce potent cytotoxic agents termed recombinant immunotoxins. These agents are targeted to kill cancer cells based on the binding specificity of the antibody fragment. Potency is derived from the enzymatic nature of the toxin as it translocates to the cytosol, ADP-ribosylates elongation factor 2 and terminates the synthesis of new cellular protein. Most prior investigations reported that PE and PE-immunotoxins kill cells via apoptosis. Here we report that PE and PE immunotoxin inhibit protein synthesis and cell growth of colon cancer cell lines but do not provoke an apoptotic response. However, the addition of the BH3-only mimetic, ABT-737 in combination with the immunotoxin produces a profound apoptotic response in these cells that neither agent alone can achieve. Tissue culture data for ABT-737 activity has now been confirmed in xenograft models confirming this approach as a viable approach for overcoming resistance to immunotoxin action.We are conducting whole genome screens using RNAi agents to silence all human genes. Immunotoxin is then added to RNAi-treated cells with the goal of identifying genes that inhibit immunotoxin action. These inhibitory genes are identified because, when silenced, cells display greater sensitivity to immunotoxin action.Likewise we conduct large scale drug screens: again to identify and inhibit gene products that reduce the effectiveness of immunotoxin action. These are likely to include gene products that interfere with cell killing and might also be useful in reversing some forms of drug resistance to chemotherapy. Our overarching goal is to make our immunotoxin program more effective by eliminating the cellular barriers to targeted therapy.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Toxoplasma gondii is a common opportunistic pathogen in HIV+ individuals. Although the immunocompromised state induced by HIV infection clearly promotes T. gondii expansion, it is unclear whether T.gondii itself might influence HIV growth either positively or negatively. Previous studies in our lab have provided evidence for both effects with T.gondii stimulating HIV LTR activity in transgenic mice and on the other hand the parasite inhibiting R5 HIV infection of human T cells through its production of a chemokine mimic, cyclophilin, that binds to the viral co-receptor CCR5. In collaborative work with Leonid Margolis (NICHD) recently accepted for publication, we extended this analysis of HIV-T.gondii interactions to a more physiologic model which employs infection of explanted human lymphoid tissue. We demonstrated that both pathogens readily replicate in ex vivo infected blocks of human tonsillar tissue. Importantly, we found that live T. gondii preferentially inhibits replication of R5 versus X4 HIV-1 isolates in the coinfected tissue blocks, an outcome that was not attributable to parasite induced depletion of the target CD4+ T lymphocytes. This inhibition of R5 viral replication could be reproduced by treatment of the tissue blocks with recombinant T. gondii cyclophilin suggesting blockade of CCR5 by this parasite protein as a possible mechanism. These ex vivo findings demonstrate that in human lymphoid tissues T.gondii has an inhibitory rather than stimulatory influence on HIV replication and raise the possibility that in co-infected individuals, T. gondii may influence the overall outcome of viral infection by preferentially suppressing R5 variants. In a second major project carried out this year, we have studied the stimulation of dendritic cells (DC) by T. gondii infection in vivo. DC provide a major source of IL-12, a cytokine critical for host resistance to this protozoan pathogen and thus this response is a key element in determining innate immunity to the parasite. Earlier work from the lab involving injection of a soluble tachyzoite extract, STAg, implicated CD8 alpha + DC as the major cell producing IL-12p40 in response to the parasite and moreover argued that the response does not require priming of these cells by IFN-gamma or other T cell/NK cell products. In contrast when we analyzed the DC responding to live T. gondii infection using IL-12 p40 reporter mice and an avirulent parasite strain, only CD8 alpha- CD11b high DC were observed to produce p40 and this response was greatly reduced when IFN-gamma or NK cells were depleted in vivo by antibody treatment. These findings suggest that IL-12 production by live infection (in contrast to STAg injection) may involve a different DC population that requires prior priming from another sentinel cell responding to the invading parasite.
|
{
"pile_set_name": "NIH ExPorter"
}
|
It is estimated that, globally, over 1.1 billion people lack access to safe drinking water. Even in the United States (with the safest drinking water in the world), over 4000 Americans fell ill to drinking water and recreational water outbreaks between 1997 and 1998. Last July, the Environmental Protection Agency proposed to increase protection against Cryptosporidium by proposing its Long Term 2 Enhanced Surface Water Treatment Rule. Health education is essential for public awareness of this far-reaching issue. A previous market survey of educators indicated that current water classroom activities do not include DNA-based experiments. Those same educators also communicated that such experiments were often are prohibitively expensive for most institutions of learning. To address these needs, Edvotek, Inc. the applicant organization, will research and in Phase III market several inquiry-based educational products for microbial detection in water. Two of these are classroom experiments illustrating detection of coliforms by chromagenic assays and techniques using the polymerase chain reaction (PCR). During the phase II proposal, two additional educational products, extensions to the PCR experiment, will be researched. To address the issue of affordability, Edvotek(R) will complete development of two prototype low-cost thermal cyclers for the performance of classroom PCR experiments. To allow shared, long-distance learning from various U.S. locations, a Web-based data entry site will be developed. Finally, the educational quality of such experiments will be determined by educator input assessing the ability of these products to meet National Science Standards and their impact to classroom science. The development of these products will address a strong market need in the educational community. This will allow for a greater public awareness of this important health topic and provide the foundation for a better workforce in the future.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The epidermal growth factor receptor (EGFR) is a transmembrane receptor involved in the transduction of proliferative and survival signals which is overexpressed in up to 50 % of patients with colon cancer. Preclinical data suggest that pharmacological inhibition of the EGFR results in tumor growth inhibition and, therefore, the EGFR represents a potential target for cancer therapy. ZD1839 is a novel, orally available, small molecule inhibitor of the EGFR in clinical development. Important questions in the development of this agent, as well as other mechanistic based compounds are the definition of patient populations more likely to respond to the drug based on the presence and functionality of the target pathway in tumor tissues and the assessment of target inhibition in clinical trials. This proposal outlines a series of clinical, pharmacological and biological studies of ZD1839 in patients with colon cancer. Our long term goal is to develop ZD1839 for the treatment of patients with colon cancer, as well as with other EGFR-driven tumors, utilizing rationally derived biological concepts based on the putative mechanism of action of the agent. The specific aims of this proposal are: Specific Aim # 1: to determine the relationship between expression and activation of the EGFR receptor in malignant tissues and the activity of ZD1839 in patients with colon cancer, Specific Aim # 2: to characterize the effects of ZD1839 on the activation and signaling of the EGFR in tumor and normal tissues of patients with colon cancer who are treated with ZD1839; and, Specific Aim # 3: to relate the pharmacodynamic effects of ZD 1839 to relevant indices of clinical activity. Patients with advanced, measurable, colon cancer will be treated with ZD1839 at a dose of 500 mg/day on a protracted oral dosing schedule. Tumor tissues, normal skin tissues, and plasma will be collected at baseline and 28 days post-treatment. The expression and activation of the EGFR will be determined using validated immunohistochemical methods developed at our institutions. The result from the biological studies will be correlated with indices of drug activity. These results will provide hypothesis-generating data regarding the subset of colon cancer patients more likely to benefit from treatment with this agent as well as information regarding the biological activity of ZD1839 in colon cancer. Collectively, these data will impact in the future development of ZD1839 in patients with colon cancer as well as other tumor types.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The mechanisms involved in the storage and periodic elimination of urine exhibit marked changes during prenatal and postnatal development. In the young fetus, prior to maturation of the nervous system, urine is presumably eliminated from the urinary bladder by non-neural mechanisms. At later stages of development, micturition is regulated by spinal reflex pathways. As the central nervous system continues to mature during the postnatal period, reflex voiding is brought under voluntary control involving higher brain centers. In adults, injuries or diseases of the nervous system can lead to the reemergence of primitive functions that were prominent early in development but then were suppressed during neural maturation. Therefore, developmental studies of micturition reflex pathways are likely to provide key insights into the mechanisms underlying neurogenic disorders of urinary bladder function in adults. This research proposal will focus on alterations in spinal reflex voiding mechanisms that underlie the maturation of voiding function during the early postnatal period. Aim I). To determine the organization of urinary bladder interneurons and parasympathetic preganglionic neurons in the lumbosacral spinal cord. A combination of transneuronal tracing with pseudorabies virus (Bartha strain) and conventional retrograde dye mapping techniques (Fluorogold) will be utilized. Changes in connectivity between the various spinal elements will be assessed in postnatal rats prior to (P1, P7, P14) and following (P21, P28, P36) the emergence of the spinobulbospinal micturition reflex using design-based stereological techniques. Aim 2). To examine changes in central processing of afferent (A-s and/or C-fiber) information from the lower urinary tract of postnatal rats prior to and following the emergence of the spinobulbospinal micturition reflex. A combination of axonal tracing techniques and Fos protein expression will be utilized. These experiments will determine if A- and/or C-fiber bladder afferents are active during early postnatal development and how afferent information is processed by different neurons in different regions of the spinal cord. Aim 3). To examine the termination pattern, area occupied and morphology of perineal afferent nerve fibers and bulbospinal projections in close apposition to interneurons and/or preganglionic neurons in the lumbosacral spinal cord in postnatal rats prior to and following the emergence of the spinobulbosninal micturition reflex. These studies will determine if perineal afferent projections (Dil or WGA-HRP-labeled) and/or bulbospinal (CRF-IR) projections and varicosities are prominent in the lumbosacral spinal cord and which neuronal populations are in close apposition to afferent or bulbospinal efferent projections. Anatomical studies will be combined with immunoassay for CRF in the lumbosacral spinal cord.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The mission of the West Virginia University Injury Control Research Center (WVU ICRC) is to advance the science and practice of injury prevention and control through research, education, outreach and service. The long-term goal of the Center is to reduce the societal burden imposed by injury. We aspire to prevent injury-related mortality and morbidity, and reduce the adverse health and economic consequences associated with injury. The WVU ICRC addresses national priorities for injury control while simultaneously emphasizing topics that are most relevant to West Virginia and the surrounding Appalachian region, a region that is economically depressed and exhibits disproportionately high injury rates. As a center of excellence, the WVU ICRC is much more than a collection of individual research projects. Rather, the Center is an interdisciplinary organization that applies comprehensive approach designed to advance the field of injury prevention and control. Accordingly, the activities proposed in this center grant application are intended to accomplish the following overarching aims: 1. To advance scientific discovery in the field of injury prevention and control, including the development of new and effective injury prevention and control strategies; 2. To improve injury prevention and control capacity at the local, regional, and national levels; 3. To increase public awareness of the burden of injury and public support for injury prevention and control efforts; and 4. To strengthen injury prevention and control programs and policies. To fulfill these aims, the WVU ICRC has developed four Core areas (Administrative, Outreach, Education, and Research). Each Core has established specific goals, objectives, and methods, which, collectively, comprise the cross-cutting activities of the Center. In this application we propose four new research investigations focusing on national and regional injury priorities including motor vehicle injuries, falls in the elderly, prescription dru overdoses, and suicide. The proposed research spans the spectrum of injury control including primary prevention, acute care, and public health translation. Further, we propose an expansion of our educational program and the establishment of a variety of innovative training opportunities for our students in collaboration with our many external partners. Finally, we plan to expand the impact of our Center through a series of activities designed to actively disseminate research findings, promote enhanced regional collaborations, and support community-based injury control practitioners and policymakers.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Danya International, Inc., proposes to develop a highly engaging, animated video series presenting obesity prevention information in a developmentally appropriate, stylistically captivating adventure format to children in kindergarten through second grade (ages 5 to 8). The video series, Adventures in Nutropolis, will contain 10 animated, interactive episodes, each approximately 10 minutes long, accompanied by teacher's guides designed to facilitate the use of the materials in early elementary school classrooms and parent information sheets to connect school-based learning to the home environment. Specifically, the Adventures in Nutropolis materials will teach children about the main contributing factors to obesity and model effective, age-appropriate obesity prevention behaviors as a means of encouraging early elementary school children to identify with and incorporate healthy behaviors into their lives. Each episode will contain interactive activities to allow children to put what they have learned about healthy behaviors into practice. During Phase I, we will fully develop the animated pilot episode and corresponding teacher's guide, as well as conduct a small evaluation to determine feasibility. Storyboards and script treatments for the remaining episodes and outlines for the associated teacher's guides and parent newsletters will also be drafted. Materials will be developed with input and feedback from a focus group of children in the target age group and an Advisory Panel of experts in the fields of pediatric nutrition, youth health promotion, pediatric physical activity, and elementary school curriculum development. In later phases of the project, we will develop the remaining episodes of the video and teacher's guides, and conduct a full-scale evaluation of the entire product. Additional materials to enhance student learning, such as an interactive Web site with game-based lessons, will also be created.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Increasing evidence points to a causative role for amyloid beta-peptide (Abeta) in the pathogenesis of Alzheimer's disease (AD). Thus, mechanisms through which Abeta affects cellular properties have come under intensive study. We have identified a novel Abeta binding protein Alcohol Dehydrogenase termed ABAD, with properties of a beta- hydroxyacyl-Co-enzyme A dehydrogenase and a generalized alcohol dehydrogenases, which is expressed at high levels in AD-affected brain. Recombinant ABAD binds Abeta (1-40/42) specifically, via the N- terminus (residues 1-20). ABAD, associated with endoplasmic reticulum (ER) and mitochondria functions as a co-factor in Abeta-induced cell stress; co-transfection of cells with constructs driving over-expression of betaAPP(V717G) and wild-type (wt) ABAD caused generation of reactive aldehydes and induction of DNA fragmentation in COS and neuroblastoma cells. In contrast, similar co-transfection studies with constructs encoding betaAPP(V717G) and mutationally-inactivated ABAD(Y168G/K172G) did not result in cytotoxicity. We propose that under quiescent conditions ABAD contributes to neuronal homeostasis, but in an Abeta-rich environment, the intact enzyme acquires pathogenic properties, exacerbating cell stress and cytotoxicity. In view of increased levels of ABAD in neurons in AD brain, we will prepare transgenic (Tg) mice with targeted over-expression of ABAD in neurons, using the thy-1 promoter, and analyze their response to an Abeta-rich environment provided by cross-breeding with Tb mice over-expressing variant betaAPP. The biologic implications of ABAD for the neuronal stress response will be further probed by over-expressing a dominant-negative form of ABAD in Tg mice, thereby selectively deleting ABAD function in neurons. In each case, mice will be analyzed for neuropathologic, electrophysiologic, biochemical and behavioral endpoints, as well as in terms of their response to ischemic stress.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The aim of this research is to describe the relationship between the structure and function of antibody receptors and of those molecules that interact with these receptors in an immune response. Our appproach is to analyze the structures of a variety of cell-surface molecules of thymus-derived lymphocytes (T-cells) and to define their function and interactions in terms of the molecular mechanisms of cell recognition phenomena in the immune response. Microchemical methods for structural analysis are being developed and used in conjunction with functional cellular assays. The specific goals are: 1) to develop assays and chemical methods for structural characterization of the T-cell receptor; 2) to analyze the structure of the urinary protein alpha1-microglobulin, counterparts of which are found on T-cells; 3) to determine the structure of TL antigens and compare it with that of H-2 and other lymphocyte antigens; 4) to develop an extensive program of microchemical methods for the complete structural analysis of cell surface proteins; 5) to compare fetal and tumor-specific antigens with other cell-surface molecules such as H-2 antigens and with immunoglobulins in order to determine whether their variability is based on similar principles. The overall goal of these studies is to calrify our understanding of the ontogeny, physiology and genetic mechanism of the immune response.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The objective of the Computer Resource is to aid the investigation of the structure, dynamics and function of biomolecules by providing a unified, powerful and highly interactive computer system for the collection, processing, storage, analysis and display of biomolecular data and for the construction, analysis and evaluation of models of biomolecular phenomena. Major areas of biomedical research of the Resource include: protein crystallography; relationships between structure and biological function for peptides and proteins; structural studies of polynucleotides; large system molecular dynamics and electropherogram analysis of proteins.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Many pathogens use the intestinal mucosa as the primary site of entry. The intestinal mucosa must induce a rapid and strong immune response to defend against such pathogenic invasion, yet an excessive immune response to commensal flora leads to chronic inflammation like inflammatory bowel disease. While it is evident that mucosal antigen-presenting cells (APCs) play a key role in orchestrating the effector response to pathogens, it is unclear how APCs maintain homeostasis with commensals. At the initiation of an immune response, mucosal APCs sense pathogens by using toll- like receptors (TLRs) that in turn signal through the adapter molecules MyD88 (myeloid differentiation factor 88) or TRIF (Toll/interleukin-1 receptor domain-containing adapter inducing IFN-b). Most research has focused on the MyD88 pathway because it is used by almost all TLRs and it leads to a robust NF-kB response. However, strong NF-kB activation may also destroy host tissues. By contrast, the TRIF pathway is only induced by TLR4 or TLR3 and supports regulatory responses through induction of IFNb. Therefore, there must be unique roles of TRIF pathway in intestinal immune regulations, but how this pathway contributes to host defense against intestinal bacterial infection has not been studied. Our ultimate goal is to protect the intestinal mucosa from infection and chronic inflammation through the future development of vaccines and immunotherapies. But fundamental research is needed to understand how to capitalize on specific aspects of TLR signaling in the intestinal mucosa to achieve this goal. The objective of this application is to understand how APCs regulate intestinal immune responses through TRIF signaling. We wish to test the hypothesis that TRIF signaling in APCs regulates protective immune responses in the intestine. Using Yersinia enterocolitica as a model enteric pathogen, we found that TRIF-deficient mice had a significant defect in macrophage bactericidal activity. They succumb to infection due to systemic Y. enterocolitica dissemination. These findings lead us to examine how TRIF signaling in APCs regulates intestinal immune responses during bacterial infection. Once the mechanism of TRIF-mediated immune defense is elucidated, this signaling can be pharmacologically manipulated leading to innovative approaches in the management of inflammatory and infectious diseases affecting the intestine. Targeting TRIF is advantageous because clinically approved TLR3 ligand is available.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Neural tube defects (NTD), resulting from a failure in the transformation of a flat neural plate into a closed neural tube, occur in approximately one in every thousand births. When the anterior neural tube fails to close, it results in a fatal birth defect called anencephaly in which the brain and skull fail to develop. Our long-term goal is to use zebrafish to define the genetic and cellular mechanisms that control closure of the anterior neural tube. We found that zebrafish embryos with reduced Nodal signaling have a phenotype analogous to anencephaly. The role of Nodal signaling is not within the neural tube itself, but within the head mesendoderm, a combined mesodermal and endodermal tissue that underlies the anterior neural tube. Rescue of head mesendoderm in Nodal signaling mutants effectively corrects their NTD. Head mesendoderm/mesoderm has a conserved role in anterior neurulation in vertebrates, but this role is poorly understood. This proposal will test the hypothesis that hea mesendoderm in zebrafish is required during late blastula to early gastrulation stages to initiate organization of neural tube precursors into a polarized neuroepithelium. Our objectives are to: (1) identify the sequential biochemical changes that establish adherens junctions, tight junctions, and asymmetric protein localization in the developing neuroepithelium, (2) define which of these biochemical changes depend upon head mesendoderm, (3) map when and where during development mesendoderm functions to promote anterior neurulation, and (4) test candidate proteins for essential roles in mediating mesendoderm function. These experiments will identify new roles for head mesendoderm in the development of the neural tube. Further, as the processes that drive neurulation are largely conserved between species, this research will increase our understanding of normal development and give us insight into potential causes of anencephaly in humans. Importantly, the hypothesis-driven experiments in this proposal are designed to be carried out by undergraduate students. They rely on well established techniques that have been mastered many undergraduate students in my laboratory. Further, the experiments are divided into specific sections that each rely on one molecular technique. This will enable students to take a project from beginning to publication within the time frame of their undergraduate studies. It is expected that this project will give undergraduate students valuable experience with the field of developmental genetics that will provide the basis for their future careers and studies while making important contributions to the science in my laboratory and to the understanding of the mechanisms underlying neurulation. PUBLIC HEALTH RELEVANCE: Failures in neural tube closure are some of the most common birth defects in humans, but the causes are not well understood. We will use the genetic and embryological techniques available in zebrafish to uncover new mechanisms required for neural tube closure. As the cellular mechanisms that drive formation of the neural tube are largely conserved among vertebrate species, this work will provide insights into new ways to diagnose and treat neural tube defects in humans.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The Northwest Clinical Community Oncology Program (NW CCOP) has been in existence since the inception of the National Cancer Institute's Clinical Community Oncology Program. As one of the founding CCOPs, the NW CCOP plays an integral role in achieving the National Cancer Institute's (NCI) objective of connecting community cancer specialists and primary care providers with NCI sponsored clinical trials. The NW CCOP is an experienced consortium of hospitals and independent oncology practices spanning three states serving local communities within western Washington, northern Oregon and northern Idaho. The overall purpose of the NW CCOP is to bring clinical advances through the most cutting edge research protocols to local communities. NW CCOP utilizes the Clinical Trials Support Unit (CTSU) as well as the Southwest Oncology Group (SWOG), National Surgical Adjuvant Breast and Bowel Project (NSABP), University of Rochester Cancer Center (URCC), M.D. Anderson Cancer Center (MDACC), and the Sun Coast CCOP Research Bases. The NW CCOP provides community oncologists with the opportunity to participate in a variety of treatment and cancer prevention and control research trials provided by these research bases. The specific aims of the NW CCOP are to: increase patients' access within Washington, Idaho, and Oregon to advances in treatment and cancer control and prevention clinical trials within their local communities; Expand outreach efforts to minority and underserved populations; Increase the knowledge, awareness, and involvement of the primary healthcare providers to promote increased participation in cancer treatment, and cancer control and prevention trials; and Provide enhanced systems supporting the consortium through communication and training among providers and coordinators thereby promoting quality and timely data collection while maintaining a level of excellence in protection of study participants.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This research proposal is designed to illuminate the developmental mechanisms underlying the formation of specific connections between motoneurons and muscles. These studies address the questions of how growing motor axons in the axolotl (Ambystoma mexicanum) arrive at their appropriate targets and whether they can distinguish between their own targets and those appropriate to different neurons. Experiments will involve microsurgury, electrophysiology, electron microscopy and several histological methods including fluorescence microscopy. The project has the following aims: 1. to describe the segmental innervation of axolotl hindlimb extensor muscles. This will be done by recording from muscles and labeling motoneuron pools; 2. To describe the development of the adult innervation pattern, in particular the contribution of synapse elimination to refining the embryonic pattern; 3. to study the behavior of motor axons after microsurgery on the developing spinal cord or lumbar plexus in axolotl embryos; and 4. to study the segmental innervation of adult muscles tranplanted to new positions. The axolotl nerve-muscles system has been chosen because a) it is accessible both for recording from muscles and stimulation of their nerves; b) mature axololts recover normal limb function after damage to peripheral nerves, suggesting that some of the mechanisms responsible for establishing motor projections during embryogenesis persist and might be amenable to study in adult life; c) axolotl embryos survive microsurgery on both developing spinal cord and muscle tissue, permitting experiments so far impossible with mammalian embryos; and d) there is an expanding library of mutants useful for future experiments. Through this work I hope to learn how synaptic connections are made selectively between neurons and certain muscles but not others. Similar selectivity probably underlies the orderly assembly of the nervous system during embryogenesis. I hope further that results from these experiments will help identify the location and nature of the molecular cues that are probably involved in axon guidance and selective synapse formation.
|
{
"pile_set_name": "NIH ExPorter"
}
|
We propose to expand on the findings of two recent studies that suggest the organization of nursing care and collaboration between nurses and physicians are as important as the organization of medical care and technology in determining compelling clinical outcomes such as excess mortality. The primary objective of the research is to extend the findings of the American Association of Critical Care Nurses (AACN) Demonstration Project by delineating combination of organizational structure and process factors at the work unit level that are the best predictors of desired organizational and patient care outcomes, including outcomes as nursing retention and patient mortality. This regionally based work is sen as a foundation for examining such determinants in a large scale national probability sample of critical care nursing units. The study will use an hypothesis-testing correlational design with the critical care unit as the primary unit of analysis. The degree to which a unit departs from an "ideal-type" discretionary pattern of work design and operation is hypothesized to be correlated with various measures of organizational and clinical performance. We expect that the closer a unit is to the "ideal- type" the better will be its performance in terms of organizational outcomes (retaining nurses and shorter length of stay) and clinical outcomes (quality of nursing care, less excess mortality, and more satisfied patients). We will survey and observe patients, nurses and physicians in each of 27 critical care units in 14 hospitals in Washington State regarding a variety of organizational and clinical factors believed to be associated with organizational outcomes, such as nurse retention and clinical outcomes such as quality of care, patient satisfaction, and excess mortality.
|
{
"pile_set_name": "NIH ExPorter"
}
|
This is a phase I/II trial of an oral agent known as DFMO for treatment of high-grade squamous intraepithelial lesion. Since the only current treatment option for HSIL is surgical, the availability of an oral agent would represent an advance in therapy if effective. Also, since the lesions are often multi-focal, systemic therapy offers a 2nd major advantage. We propose a study design in which we will enroll 14 subjects initially, study them on DFMO for 6 mos & perform anoscopy at defined intervals to determine if they have shown signs of disease progression or regression. After 6 months, therapy will be discontinued and they will be followed for another period of 12 months off therapy. If anyone of the initial 14 subjects shows signs of disease remission, then another 11 subjects will be enrolled for a total of 25 subjects. The availability of the GCRC will be essential to the successful completion of these studies since all the necessary equipment is already there and subjects are accustomed to using this facility as part of the follow-up in one of the other two studies.
|
{
"pile_set_name": "NIH ExPorter"
}
|
Universal newborn screening and identification of hearing loss is an important national goal that is achievable with today's technologies. Without appropriate intervention, children with hearing impairments will fall behind their peers in achieving developmental milestones with lifelong effects. Appropriate early hearing detection and intervention (EHDI) programs should be family-centered, interdisciplinary; culturally competent, and built upon informed choice for families. Although a large amount of information related to EHDI is available in bits and pieces from various sources, obtaining and using the information for decision making represents one of the serious challenges facing families and professionals today. Since, EHDI is a relatively new field, many professionals are not prepared to adequately assist families in making important decisions about their children's future. Comprehensive, easily accessible, and culturally sensitive EHDI informational resources for families and professionals would be important new tools to facilitate successful child and family development. The specific aims of this project are: 1) To identify and prioritize content and presentation formats for family and professional resources in EHDI; 2) To develop and test an accessible digital format for EHDI Resources; 3) To evaluate the effectiveness of EHDI Resources through systematic field testing; and 4) To disseminate and distribute EHDI Resources nationally. The hypothesis that will be tested is that after using the EHDI Resources, parents, siblings and professionals would demonstrate an increase in specific knowledge and self-efficacy related to EHDI. In addition, user satisfaction with the EHDI Resources will be evaluated. Evaluation will include formative, process, and outcome procedures. A multi-site randomized controlled study will be conducted as part of outcome evaluation. The EHDI Resources will include English and Spanish narration, incorporate a wide range of accessibility features, and be distributed in three formats, CD-ROM, DVD-ROM, and via the web. When completed, this project will have successfully developed, evaluated and distributed an important new set of EHDI informational resources for families and professionals.
|
{
"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. Eight years ago, Aurora kinase A was classified as an oncogene. This protein has been found at high levels in many forms of cancer, including gastric, colorectal, breast, esophageal and lung cancers [unreadable]indeed, as many as 86% of lung cancers exhibit inappropriate expression of Aurora A. Simply overexpressing this kinase can transform certain normal cells into tumors. Importantly, tumors generated by high Aurora A kinase activity demonstrate increased resistance to drugs that target microtubule formation, including Taxol. In fact, clinical trials are currently underway for small molecule inhibitors of Aurora kinase activity in treating cancer. However, very little is known about the molecular targets of this kinase. How does this transformation occur? Are proteins outside of the normal function of Aurora A involved in this process? It is the goal of the current proposal to tap into the combined power of high performance mass spectrometry, quantitative strategies in proteomics, and selective phosphopeptide enrichment methods to comprehensively characterize substrates of Aurora A kinase, on a proteome-wide scale. This information is a critical foundation from which we can begin to understand how unregulated Aurora A kinase activity causes lung cancer, and how to treat it.
|
{
"pile_set_name": "NIH ExPorter"
}
|
The goal of this research program is to analyze cell growth and development. Each project in the program uses defined cell populations to permit an integrated genetic and biochemical approach. Within this common framework we are addressing several problems. The prokaryote Caulobacter expresses different sets of genes at different stages of its cycle. The mechanism of temporal control of gene expression in this organism is being studied by examining the organization of the genome as a function of the cell cycle. Both Caulobacter and budding yeast couple the cell division cycle to membrane biogenesis. The coupling mechanisms in both cells will be analyzed by studying mutants conditionally defective in the biosynthesis of mebrane lipids. Chromosomal mutations of E. coli that prevent the expression of conjugative plasmid functions will be used to study the relation between gene expression and envelope biogenesis. The genes altered by these mutations will be cloned, their gene product identified and the role of these gene products in the synthesis and processing of envelope proteins determined. The role of specific E. coli DNA binding proteins in DNA replication and repair will be studied by analyzing strains carrying mutations in one of these proteins as well as by purifying the protein and characterizing its functions in DNA replication and repair in vitro. The relation between positional information and patterns of differentiation in multicellular aggregates will be studied in Hydra and Dictyostelium. The stem cell system in Hydra will be studied in order to elucidate the effect of position on the probability of stem cell self-renewal. Dictyostelium will be used to identify specific morphogenetic factors and to determine their mechanism of action.
|
{
"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.