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Intra-atrial reentrant tachycardia is a well recognized sequelae of the Fontan operation. The incidence of both early and late intra-atrial reentrant tachycardia is 10-35%. (1-5) Data from animal models as well as studies in humans in the clinical electrophysiology laboratory suggest that the suture line of the lateral cavopulmonary connection and the natural anatomical right atrial boundaries (vena cava orifices, coronary sinus orifice, tricuspid valve, and possibly the crista terminalis) can serve as barriers creating a pathway which, along with changes in intra-atrial conduction, provide the substrate for an intra-atrial reentrant tachycardia. (6-12) The objective of this study is to investigate if a strategically placed linear incision, incorporated at the completion of the fenestrated Fontan, will prevent the postoperative intra-atrial arrhythmia. The study is a randomized prospective design in which candidate patients will receive either the conventional fenestrated Fontan repair (control group), or a fenestrated Fontan repair with a cryosurgery lesion or surgical incision (test group). The primary endpoint will be based upon the failure to induce the intra-atrial reentrant tachycardia in the test group compared to the control group both during the immediate post operative period (prior to hospital discharge) and at the time of fenestration closure (6-12 months after surgery). The patient groups will also be followed for a five year period to determine the effect of the surgical incisional lesion on the prevention of late clinical reentrant intra-atrial tachycardia.
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{
"pile_set_name": "NIH ExPorter"
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The availability of functionalized molecules is of preeminent importance in the fields of organic and medicinal chemistry and to the pharmaceutical and related industries. Not only should compounds be accessible, but also the means to readily and predictably modify them to change their properties and/or eliminate side effects is of tremendous significance. The chemistry that is proposed in this grant application involves the invention or accentuation of techniques to access molecules that contain basic nitrogen centers as key structural features. Our prior work in this field has been incorporated into the everyday repertoire of synthetic organic chemists and has greatly impacted the field of organic synthesis, particularly in drug discovery in the pharmaceutical industry. In addition, we are proposing work on the synthesis in enantiomerically pure (single handed) form of a wide variety of amines from extremely simple (alkene) precursors. Essentially all modern small molecule medicines must now be sold in single-handed forms and the successful accomplishment of this feat can add to the cost of the medicinal agent. Also included are new methods for the preparation of nitrogen heterocycles. In many ways, compounds of this type form the backbone of the pharmaceutical industry. We also propose to carry out mechanistic studies on all of the chemistry included. Further, information gained from this work will help to understand the mechanism of the processes that are being developed in order to increase the efficiency of the methods that we are developing. Overall, the successful realization of the chemistry proposed in this grant application will have a major impact on the ability of chemists and those in auxiliary fields to develop new medicinally important, sensor or biologically relevant compounds and materials that will move science forward.
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{
"pile_set_name": "NIH ExPorter"
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This is a phase I/II open label combination therapy study in two parts separated by at least eight weeks. In Part 1, HIV-infected infants and children will receive therapy with saquinavir plus two nucleoside antiretroviral agents chosen by the investigator and patient (or guardian). In Part 2, HIV-infected infants and children will receive therapy with saquinavir, nelfinavir and nucleoside antiretroviral agent(s) as chosen by the investigator and patient (or guardian). At least four patients (two each in Parts 1 and 2) will be enrolled in each of three age strata: six months to two years; greater than two years to six years; and greater than six years to 13 years. Each patient will receive therapy for at least 24 weeks. Clinical and laboratory pharmacokinetic, safety, virologic, and immunologic evaluations will be performed every four weeks for the first 16 weeks and then every eight weeks until week 72. Pharmacokinetic studies will be done at day 0, day 1, every four weeks until week 24, and at week 72 in patients who are willing and able to take both the adult and pediatric formulations, and patients who take only one formulation will have pharmacokinetic studies done at day 0, every four weeks until week 24, and at week 72. Saquinavir dosing may be adjusted based on the results of initial pharmacokinetic studies. Any patient in Part 1 who does not achieve acceptable saquinavir levels after dose adjustment will be given the opportunity to further modify dose and/or dose interval, or add nelfinavir to the original regimen at week eight.
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{
"pile_set_name": "NIH ExPorter"
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Fanconi Anemia (FA) is an inherited DNA repair disorder characterized by congenital abnormalities, cancer predisposition, and progressive bone marrow failure. FA is caused by biallelic mutations in one of sixteen FANC genes, the products of which cooperate in the FA/BRCA DNA repair pathway. Although the precise biochemical functions of the FA/BRCA pathway remain unclear, the pathway promotes homologous recombination (HR) repair. Due to the underlying DNA repair defect, FA cells are hypersensitive to genotoxic DNA crosslinking agents. The mechanism of the bone marrow failure (BMF) in FA remains elusive. Our recent studies suggest that BMF results, at least in part, from increased p53 expression in hematopoietic stem and progenitor cells (HSCPs), leading to progressive cell cycle delay and apoptosis. BMF may also result from the accumulation of DNA damage from the endogenous crosslinking agent, acetaldehyde, and the selective toxicity of this agent to hematopoietic stem cells. Recently, we identified hyperactive TGF? signaling as a mechanism of bone marrow suppression in FA. Disruption of TGF? signaling, through the use of shRNAs, sgRNAs, and small molecule inhibitors confirmed the suppressive role of the pathway on FA cell growth. We hypothesize that an upstream inhibitor of the TGF? pathway (i.e., a monoclonal antibody to TGF? itself) will inhibit this pathway and rescue the function of the HSPCs and FA stromal fibroblasts, resulting in an increased probability of rescuing bone marrow function in FA patients. The specific aims of this R01 are: 1) To determine whether hyperactivation of the TGF? pathway suppresses growth of primary FA cells and FA cell lines, 2) To determine the mechanism by which TGF? inhibitors promote FA cellular growth and enhanced resistance to crosslinkers, and 3) To determine the mechanism of TGF?-mediated suppression of bone marrow in vivo in mouse models.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of this research program is to develop safe and effective treatments for hereditary neurological disorders. Specific research accomplishments in the past year include the following: (1) a collaborative study to establish reliable biomarkers for spinal muscular atrophy, (2) completion of a phase 2 study of high dose idebenone treatment for Friedreich's ataxia, and (3) development of a phase 2 clinical trial to test the feasibility of dutasteride treatment for spinal and bulbar muscular atrophy (Kennedy's disease).
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{
"pile_set_name": "NIH ExPorter"
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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. 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. The objective of this research is to understand the mechanisms whereby hormonal and local factors control the structure-function of the ovulatory follicle and corpus luteum in the ovary, and hence fertility, during the menstrual cycle in primates. Ongoing research is testing the hypothesis that the vital actions of the pituitary hormone, luteinizing hormone (LH) in promoting ovulation of the mature follicle and its subsequent conversion into the corpus luteum, and in controlling the functional lifespan of the corpus luteum, is either direct or indirect via local steroid action (e.g., via stimulation of progesterone [P] synthesis and P receptor expression). Because of similarities to ovarian function in women, monkeys are used in experiments where either LH and steroid hormones are depleted or steroids alone are depleted and replaced. The ovulatory follicle or corpus luteum is then removed to analyze molecular and cellular processes that are LH-dependent and either steroid/P-independent or -dependent. Gene and protein expression are analyzed for various processes, including: (1) protease enzymes that may be critical for tissue remodeling during ovulation, luteal development and luteal regression, and (2) local regulatory systems, e.g., the corticotropin-releasing hormone (CRH)/urocortins (UCN)-receptor-binding protein system. Current genome-wide analyses are identifying processes that are regulated by LH and/or steroid hormones in the follicle or corpus luteum. The activity of these processes is being manipulated, via gene knockdown or protein antagonist approaches, to elucidate their biologic relevance to primate ovarian function and cyclicity.
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{
"pile_set_name": "NIH ExPorter"
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The proposed research into the income and life styles of middle class Black Americans is the first stage of more extensive research into the economic conditions, attitudes, values, and mobility of middle class Blacks. The present study is a preliminary investigation into (1) the factors related to the achievement of middle and upper income among black and white families, (2) life styles among middle and upper income black and white families, and (3) a profile of Black wives in middle and upper income families. While future research will involve the collection of original data, the present project will make use of the Bureau of Census' Public Use Samples of the 1970 decennial census. Both contingency analysis using chi square and regression analysis will be the principle technique, and will involve two regression equations (one for the Black sample and one for the white sample) using the same independent variables. The second problem will be investigated with the aid of both contingency analysis and partial correlation using dummy variables, and the third primarily with contingency analysis.
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{
"pile_set_name": "NIH ExPorter"
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The long-term objective of these studies is to generate novel methodologies for genetic manipulation of vertebrate embryos for the ultimate purpose of understanding gene function during development. The investigators propose experiments to test the hypothesis that bacteriophage protein pairs can stimulate homologous recombination (HR) in vertebrate Xenopus and Zebrafish embryos. The bacteriophage protein pairs include Red__ and Red__ from phage lambda, and RecE and RecT from the lambdoid prophage Rac. The protein pairs have been employed for both efficient gene conversion and gap repair using short homology regions. Recombination involves co-operation between a 5'-3' exonuclease (RecE or Red-alpha with a single strand binding protein (RecT or Red-Beta). Our collaborator, A.F. Stewart, pioneered the use of these proteins to initiate homologous recombination in E. coli. Recently Dr. Stewart's lab has demonstrated that these proteins can promote homologous recombination in mouse ES cells using single stranded oligonucleotides to target specific sequences. This has been termed ssOR for single-strand oligo repair. This exploratory grant will investigate several approaches in early Xenopus or Zebrafish embryos to target specific chromosomal sequences using the phage protein pairs to promote ssOR. The investigators propose to (1) develop assays for ssOR in cytoplasmic extracts of Xenopus eggs to explore the specific requirements for HR in the Xenopus embryonic cytoplasm. The assay will examine requirements for repair of antibiotic resistance genes on extrachromosomal plasmids and will use an E.coli colony readout. (2) The investigators will target a chromosomal Green Fluorescent Protein (GFP) transgene in Xenopus tropicalis embryos to generate visually screenable loss of function phenotypes. Purified phage proteins and single strand oligos will be used target a GFP transgene to alter the expression of GFP expression through coding changes, frameshift mutation, or stop codon insertion. (3) The investigators will explore conditions to target several zebrafish chromosomal loci using ssOR to generate well-characterized point mutations that give scorable phenotypes.
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{
"pile_set_name": "NIH ExPorter"
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This study is designed to further study the safety and efficacy of orally administered SP-303 in patients with AIDS. Prior studies withis agent have shown some reduction in stool volume in patients with travellerUs diarrhea. This study will include HIV positive men and women 18 years and older with a confirmed stool volume at screening of 300 grams. Patients will be hospitalized in the GCRC for one day of screening and six days of treatment. Responders to treatment (identified as having a 50% reduction in stool volume from screening) will continue on their initial therapy for three additional weeks.
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{
"pile_set_name": "NIH ExPorter"
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Dr Turco is studying the lipophosphoglycan (LPG) of Leishmania, a parasite causing significant health problems to humans. The structural characterization of the LPG is being pursued as is the understanding of the enzymatic synthesis of the compounds. LPG fractions are being isolated and analyzed by MALDI and negative ion electrospray mass spectrometry using partial degradation approaches. A particular focus is to understand how the different LPG structures are responsible for interspecies differences and how intraspecies developmental modifications affect the LPG structures. Therapeutic advances can be expected to follow detailed understanding of these glycoconjugate systems.
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{
"pile_set_name": "NIH ExPorter"
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The primary aim of the study is to test the impact of real-time TIPI probabilistic outcome information on patient care. Specifically, this investigation aims to test, in a controlled prospective clinical trial, the impact of the TIPI computerized electrocardiograph on physician ED triage of patients with symptoms suggestive of ACI.
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{
"pile_set_name": "NIH ExPorter"
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The present project combines cellular and computational techniques to explore the role of specific forms of synaptic plasticity in the modulation of information processing in a behaviorally relevant neural network. This project will include an in-depth analysis of short-term synaptic plasticity (STSP) at an identified class of inhibitory interneurons (L3Os) in the siphon-withdrawal reflex (SWR) in Aplysia, and the long-term modulation of this synaptic plasticity. Although STSP is present at synapses in both invertebrates and vertebrates, its behavioral role is not well understood and difficult to address in many systems. However, recent studies in our laboratory have begun to link short- and long-term plasticity at the L3O synapse to behavioral plasticity in the SWR. Thus, the SWR circuit is well suited for a functional examination of STSP and will provide insights into the role of STSP in other systems. Furthermore, the knowledge gained with this project will provide important predictions to expand our understanding of the neural circuitry that underlies adaptive behavior, a connection that is often difficult to make in more complex nervous systems but is essential to our understanding of the human brain.
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{
"pile_set_name": "NIH ExPorter"
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Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The broad goal of this study is to investigate the chromosomal consequences of cell's inability to maintain the three critical aspects of the DNA precursors pools: their quantity, their balance and their purity. Quality of DNA precursor pools has been long recognized for its important role in avoidance of mutagenesis, which is significant for evolution of any species. Recently, we have argued for a more immediate consequence of limited, imbalanced or contaminated DNA precursor pools, which is formation of chromosomal lesions that threaten cell's survival and that require complex DNA mending system called recombinational repair. We have proposed two general models of how base analog incorporation into DNA could lead to chromosomal fragmentation that requires recombinational repair. In our previous work, we identified uracil and hypoxanthine as the major base analogs contaminating DNA precursor pools in E. coli. At the same time, the identity and the sources of other natural base analogs contaminating DNA precursor pools remain unknown. Moreover, even though DNA precursor pool imbalance at first leads to reparable chromosomal lesions, it eventually generates irreparable chromosomal lesions of unknown nature. Our recent observations provide insights into the possible scenarios leading to irreparable chromosomal lesions and into the nature of these lesions, while introduction of genome analysis presents an opportunity to identify the most affected parts of the chromosome. Aim 1 of this study addresses specific mechanisms of chromosomal fragmentation, induced by hypoxanthine, uracil or fluorouracil. Aim 2 focuses on characterization of the mysterious origin DNA disappearance during thymine starvation, coinciding with thymineless death. Aim 3 tests our recently-proposed multi-stage model for thymineless death, which begins with stalling of the existing replication forks, proceeds through unknown number of intermediate stages and ends with the replication origin destruction. Collectively, this work will emphasize the importance to keep DNA precursor pools plentiful, balanced and sanitized by characterizing various sources of imbalance and contamination of the pools, as well as the chromosomal consequences, including irreparable lesions, that result from pool contamination and imbalance.
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{
"pile_set_name": "NIH ExPorter"
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The 10th annual WORLD Symposium will be held February 11-13, 2014 (San Diego, CA, USA). This research meeting is a multidisciplinary forum presenting the latest advances in basic science, translational research, and clinical treatments for lysosomal diseases. The theme of the meeting emphasizes the primary aim, which is to assess the mechanisms, and obstacles, for taking bench research into human therapy. Additional aims build upon this, and implement the goals of the Lysosomal Disease Network (LDN) to: 1) further develop the mechanisms and mission of the LDN; 2) foster interdisciplinary collaboration between scientists, leading to improved knowledge regarding the biochemical, immunologic, genetic, and clinical manifestations of these diseases; 3) identify and discuss the latest findings in diagnostic testing, screening, and treatment; and 4) identify areas that need more basic/clinical research, public policy, and regulatory attention. Because many LDN constituents investigate and/or treat specific aspects of these diseases, they may have little exposure to work done in areas outside their current area of interest. This meeting allows for the sharing of knowledge and advances across lysosomal diseases, and provides an opportunity to discuss treatment outcomes. Evidence of the synergistic nature of the WORLD Symposium can be seen in the growth of the multi-center consortium Lysosomal Disease Network (U54 NS065768, a member of the Rare Diseases Clinical Research Network). In addition to researchers and clinicians, the WORLD Symposium welcomes patients, parents, caregivers, and all patient advocates who wish to attend. This unique feature allows a small, geographically-divergent patient population access to information on the latest scientific advances in lysosomal diseases and gives scientists and clinician's unprecedented access to patients outside the clinic setting; providing a forum for sharing information that otherwise does not exist. The program is organized into six sessions, extending from basic research through translational research to clinical research. For each session, the Program Committee will select two invited speakers and will fill the remainder of the program from submitted abstracts (the submission deadline is October 1, 2013). For the third consecutive year, the Symposium will include a didactic component, Lysosomes 101, Fundamentals of Lysosomal Biology and Disease, on the Monday preceding the research meeting (February 10, 2014). Building on-but not supplanting-the curriculum of the previous years, this educational session will offer attendees the basic concepts of the biochemistry, immunology, molecular biology, and genetics of lysosomal disease. In 2014, Lysosomes 101 will expand from 4 hours to a full day of instruction, providing a common understanding for the cutting-edge research presentations that constitute the Symposium content. In order to foster relationships between senior and young investigators, we will once again sponsor travel for young investigators, facilitating their attendance of Lysosomes 101 and the WORLD conference. The mentoring of new scientists is a major goal of the LDN and the RDCRN.
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{
"pile_set_name": "NIH ExPorter"
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Continuation is planned of a long-term study of plasmid molecular biology, focusing on the model rolling circle plasmid, pTl8l. Studies are proposed of the mechanisms of initiation and termination of leading strand replication, with special attention to the possible involvement of a type C cruciform structure containing the specific nick site. A comprehensive mutational analysis of the plasmid-coded initiator (Rep) protein is proposed, with the aim of isolating and localizing mutations affecting each of the protein's nine known functions. Attention will be devoted to the effects of local and distal secondary structures and other special sequence elements on the function of the pTl8l leading strand origin, with special reference to cis-inhibitory sequences. We have predicted and verified the replication-specific inactivation of the Rep protein and have demonstrated that the inactive form of the protein is modified. The nature of the modification and the mechanism of its formation will be determined.
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{
"pile_set_name": "NIH ExPorter"
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This project is designed to determine the mechanism responsible for increased orotic acid excretion in animals consuming diets that are marginal or deficient in arginine or other urea cycle intermediates. The present studies will determine the involvement of the urea cycle and the shunting of excess nitrogen in pyrimidine biosynthesis. Studies will examine the influence of arginine deficiency and amino acid excess on the shunting of nitrogen from urea biosynthesis into pyrimidine biosynthesis. Additionally, the capacity of the urea cycle to detoxify ammonia as indicated by the flux of nitrogen through the pathway and enzymatic activities will be determined. These studies will also examine some of the long term complications of consumption of a diet that is deficient in arginine or other urea cycle intermediates in the rat.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary/Abstract Neurodevelopmental disorders currently affect 1 in 6 children but males are disproportionately affected. The cause of this robust sex difference is not known. Neurodevelopmental disorders involve disruptions in approach and avoidance behaviors, which rely heavily on amygdala function. Our transgenic mice missing one copy of the cell-adhesion molecule gene, PCDH10, exhibit male-specific behavioral and amygdalar impairments, including social approach and threat conditioning deficits, decreased connectivity between the lateral and basal nuclei of the amygdala, decreased NMDAR expression in the basolateral amygdala (BLA), and increased filopodial spine density in the BLA; impairments in behavior, structure, and function that are relevant to neurodevelopmental disorders. With this experimental system, we will study the mechanisms of male-specific deficits in approach and avoidance behaviors and the mechanisms underlying behavioral rescue on behavioral, cellular, molecular, and circuit levels. We will manipulate neonatal levels of testosterone and measure social approach, threat conditioning, NMDAR expression, synaptic properties, and calcium activity in excitatory BLA cells in vivo, using Western blots, whole-cell patch-clamp electrophysiology, and fiber photometry. We will also study the effects of d-cycloserine, shown to rescue social approach, on ex vivo and in vivo BLA activity. We will study main effects of sex, genotype, and treatment to help uncover mechanisms that underlie the male preponderance of neurodevelopmental disorders in order to develop novel treatment strategies.
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{
"pile_set_name": "NIH ExPorter"
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Candela Laser Corporation, in cooperation with The Johns Hopkins University, proposes to develop a new procedure for treatment and closure of choroidal vessels and neovascular membranes with reduced damage to the retina and better preservation of vision. Choroidal neovascularization is the most frequent cause of blindness in patients over the age of 65. The success of current argon/krypton laser treatments is severely limited by patient eligibility criteria, a high recurrence rate, and loss of vision due to the treatment itself. The treatment procedure to be evaluated in this study uses small doses of Indocyanine Green injected prior to each laser burn to selectively enhance absorption of laser light in the designated target area during laser irradiation. In contrast to previous studies, dye concentration is monitored precisely. Laser energy is delivered during the transit of a dye bolus or when the dye concentration reaches its maximum in the target area. A diode laser at 805 nm, matched to the absorption maximum of ICG will be coupled to an infrared video fundus camera delivery system. Optimum parameters for dynamic dye-enhanced laser treatment will be evaluated in a rabbit model in the Phase I study.
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{
"pile_set_name": "NIH ExPorter"
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The long term goal of this project is to assess the potential of antisense oligonucleotides in treatment of genetic defects. A novel application of antisense oligonucleotides is proposed, in which these compounds are used to restore correct functioning of a defective gene rather than for inhibition of expression of an undesirable gene, as is commonly used. Specifically, they are used to reverse aberrant splicing caused by mutations seen in a number of genetic diseases and in certain cancers. In this granting period the specific experimental system to be studied will be a subset of mutations in human beta-globin gene which cause thalassemia via-aberrant splicing of beta-globin pre-mRNA. The proposed approach stems from our recent observations that antisense 2'-O-methyl- ribooligonucleotides are able to inhibit aberrant splicing induced by thalassemic mutations and restore correct splicing in cell free splicing extracts. The specific aims are intended to extend these results towards the long term goal of the project: 1) To optimize the effects of antisense oligonucleotides on aberrant splicing in vitro, in cell free extracts. The exact position of the targeted oligonucleotides as well as their length will be varied to obtain the highest efficacy of restoration of correct splicing. The targeted thalassemic mutations will include those studied recently i.e., IVS1-110 and IVS2-654 as well as others, such as those causing aberrant splicing at the 5' splice site (IVS1-5, IVS1-6). 2) To test the most effective antisense oligonucleotides identified in Specific Aim 1 in tissue culture cells. These will include HeLa or CHO cells stably transfected with thalassemic globin genes cloned under the CMV promoter and human x mouse hybrids expressing appropriate thalassemic beta-globin pre-mRNA. The oligonucleotides that will be most effective in reversal of aberrant splicing (presumably due to their efficient uptake by the cells and/or optimal targets) will be identified. 3) To test whether oligonucleotides other than 2'-O-methyl-oligonucleotides have better ability to restore correct splicing in vitro and in vivo. Initially 2'-O- methyl-phosphorothioates and methylphosphonates will be used. These modifications may improve the stability and/or the efficiency of cellular uptake of the oligonucleotides and will result in their increased activity.
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{
"pile_set_name": "NIH ExPorter"
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Type-1 diabetes (T1D) is hallmarked by autoimmune attack on the insulin-producing p cells of the pancreatic slets. Infiltration of the islets by autoimmune/inflammatory cells, or insulitis, occurs in a percentage of genetically susceptible individuals, and a fraction of these subjects progress to clinically overt diabetes. Our view of T1D pathogenesis, especially in humans, remains clouded and controversial in many regards. The main reasons for :his unsatisfying picture are the complexity and heterogeneity of T1D, and the fact that it is not usually diagnosed until very late, when most initiating and propagating autoimmune phenomena have already played out. One approach to surmounting the latter problem would be to develop a method to non-invasively image the pancreatic slets in vivo in real time, which has been one of the prime objectives of the collaborative JDC/MGH imaging program. The overall goals of this component of the program, Project 2, has been and remains first, to help evaluate the applicability of novel imaging methodologies (developed within Project 1 and the Core) to mouse models of T1D; second, to exploit successful visualization strategies to answer currently unresolved questions about disease pathogenesis in these models (certain in concert with Project 3); and third, to perform proof-of-principle experiments for translation of successful strategies to human T1D patients (under the auspices of Project 4). During the past funding cycle, Project 2 successfully developed a method to non-invasively image islet inflammation in murine T1D. This technique is based on visualizing, via magnetic resonance imaging (MRI) of magnetic nanoparticles (MNP), the microvascular changes (vessel dilation, vascular leakage) that routinely accompany inflammation, coupled with uptake by neighboring macrophages. A mock clinical trial on recent-onset diabetic mice clearly established the power of the MNP-MRI technique to distinguish between individuals that were or were not responding to anti-CD3 monoclonal antibody therapy. These promising results prompted initiation of a collaborative clinical trial to assess the ability of an analogous MNP, Combidex, to reflect vascular changes in association with human T1D (Project 4). For the next funding cycle, Project 2 proposes to apply the MNP-MRI technique to additional questions concerning the pathophysiology of T1D, but mostly to capitalize on exciting recent advances in fluorescent protein tomography (FTP) made by Project 1 and The Core. At the cutting-edge of whole-body live-rodent imaging, this technique permits non-invasive visualization of proteins and cells of interest in deep tissues (eg the pancreas) of living mice. The Specific Aims are: 1. To develop a system to co-image islet p-cell mass and p-cell activity non-invasively in vivo. A triple- transgenic mouse approach will be taken, exploiting the newly engineered red fluorescent proteins, mRaspberry and dKeima, in conjunction with FTP. Also under this Aim, a series of calibration and validation experiments will put the systems utility to the test. 2. To exploit the p-cell mass/activity co-imaging system to tackle outstanding issues concerning type-1 diabetes in mouse models. Questions to be tackled include: a) How does p-cell mass/activity change during the natural history of T1D in the NOD mouse model? b) How do genetic factors impinge on the mass/activity curves? c) Can the residual level of p-cell mass or activity predict onset of overt diabetes? d) Do different therapeutic regimens influence p-cell mass/activity? 3. To exploit MNP-MRI to guide dissection of the cellular and molecular elements predicting development of clinical diabetes in individual NOD mice. The novel capability of predicting if and approximately when a given mouse will convert from insulitis to diabetes will be used to address the influence of a NK cells b T helper cell subsets and c CD4+25+Foxp3+Treg cells within the infiltrate. Successful achievement of these Aims should provide novel insights into the pathogenesis of T1D, as well as furnish proof-of-principle for human imaging endeavors. Joslin Diabetes Center, Boston, MA PHS 398 (Rev. 04/06) PageM Form Page 2 Project 2: Imaging p-cell attack in mouse models of type-1 diabetes PI: MathiS, Diane PD: Mathis, Diane KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator(s). List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Role on Project Mathis, Diane DMATHIS JDC PI OTHER SIGNIFICANT CONTRIBUTORS Name Organization Role on Project Kulkarni, Rohit JDC Consultant Weir, Gordon JDC Consultant Human Embryonic Stem Cells ^ No CH Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://StemcellS.nih.gov/reqistrv/indeX.aSP. Usecontinuation pages as needed. If a specific line cannotbe referencedat this time, includea statement that one from the Registry will be used. Cell Line PHS 398 (Rev. 04/06) Page^O Form Page 2-continued Number the following pages consecutively throughout the application. Do not use suffixes such as 4a, 4b. Project 2: Imaging p-cell attack in mouse models of type-1 diabetes PI: Mathis, Diane PD: Mathis, Diane The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. PROJECT 2 TABLE OF CONTENTS Page Numbers Cover Page 93 Description,
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Cooperativity plays an important role in metabolic regulation by controlling the rates of enzyme catalyzed reactions. Two types of allosteric interactions have been proposed to explain cooperative ligand binding equilibria, and coupled conformational changes, in oligomeric enzymes. However, the full range of kinetic cooperative behavior which may result from these allosteric interactions has not been examined rigorously. As a result, certain cooperative phenomena, particularly the class of effects known as half-of-the-sites reactivity, may have been interpreted too narrowly. In particular, the possibility that positive equilibrium cooperativity may be accompanied by negative kinetic cooperativity and metastable behavior seems to have been generally overlooked. Our objective in the research proposed here is to clarify the types of kinetic cooperative behavior that may arise from various allosteric interactions.
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{
"pile_set_name": "NIH ExPorter"
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We wish to define the entire domain of the bacteriophage T4 rIIB mRNA that is scanned by the ribosome during the initiation of protein synthesis. We will use exhaustive genetics to select mutations that diminish translation of the rIIB mRNA and biochemistry to determine contacts between ribosomes and the mRNA. We will ask the congruence between the genetic and biochemical answers. Additionally, we will study a translational repressor that interacts with the rIIB mRNA. We will use genetics and biochemistry here as well. Our goal is to understand in molecular terms a set of interactions that control gene expression in prokaryotes at the post-transcriptional level.
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{
"pile_set_name": "NIH ExPorter"
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The purpose of the 2nd World Parkinson Congress (WPC 2010) is to address a need for worldwide dialogue to address the multifaceted problems of PD and propose solutions including new approaches to research and better treatment options for people with PD. The WPC 2010, to be held from September 28 - October 1, 2010 in Glasgow, Scotland is the only international meeting in the field of Parkinson's disease (PD) responding to a need to bring the whole PD community together for high-level scientific and lay sessions. Delegates at the meeting will represent neuroscientists, physicians, people with PD, caregivers, nurses, allied health professionals, government representatives, and other members of the advocacy community from more than 50 countries. The four-day program highlights current work being done to advance the science of PD and to improve care for people living with the disease. Exposing researchers from the US to the scientific advancements made in other parts of the world will help them explore new ideas and build collaborations they might otherwise not build. Broad support and involvement from the PD community will allow the WPC to create a venue for cross pollination of the community to build collaborations and interactions between health professionals, researchers, and patients who might not otherwise meet. These interactions will spur innovative research, identify solutions to unmet needs and advance therapies for people with PD. Thirty-eight, of a total of 75 committee members, including people living with PD, designed an elaborate program to raise awareness among scientists and clinicians about unique aspects of PD to deepen their commitment to PD research and possibly foster research on less studied aspects of PD. At the same time, the program is expected to expose patients and caregivers to research and ideally increase willingness and interest in participating in clinical trials, which is a major challenge for researchers today. Speakers are being invited from all areas of the PD community with sessions formatted in a variety of ways to maximize the learning potential and dialogue, including plenaries with up to 1,000 delegates, parallel sessions with 200 - 550 delegates and more intimate workshops, of around 100 delegates. The morning plenaries are designed around a theme and used as a meeting ground for all attendees to start each day with a topic that sets the stage for following sessions and stimulates further discussion later in the day. Parallel sessions and workshops will cover 1) new developments in science, 2) concepts on clinical care, and 3) issues related to quality of life;providing day-long discussions to hold the diverse interest of scientists, clinicians, and patients. There will be both Scientific and Living with Parkinson's posters each day of the Congress that were reviewed and accepted by the Program Committee. This broad range of posters sessions is designed to maximize interaction among researchers, clinicians, patients, and careers. The three most outstanding posters each day will be presented in a 'Hot Topics'session at the beginning of each day. Poster tours will be held paying keen attention to the most promising work in the field and to increase dialogue and collaboration between the stakeholders in the PD community. PUBLIC HEALTH RELEVANCE: The purpose of the 2nd World Parkinson Congress is to create a worldwide dialogue to discuss the multifaceted problems of PD, propose solutions including new approaches to research, build innovative collaborations, and create better treatment options for people with PD. It is the only meeting in the field of Parkinson's disease (PD) that addresses a need to bring the whole PD community together for high-level scientific sessions and discussions on current work being done to advance science, improve care for people with PD and help sensitize researchers to the needs of people with PD and conversely help patients understand the challenges researchers and health professionals face when studying PD. Uniquely, the delegates at the meeting represent neuroscientists, physicians, people with PD, caregivers, nurses, allied health professionals, government representatives, and other members of the advocacy community.
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{
"pile_set_name": "NIH ExPorter"
}
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The tularemia pathogen, Francisella tularensis, is a highly virulent human pathogen and is easily transmitted through aerosols, thus its potential for use as a weapon of bioterrorism is high. The bacterium is a facultative intracellular pathogen, suggesting a crucial role for T cells in protective immunity against the organism as has been confirmed by studies in animal models. T cells contribute to protection against microbial infection by recognizing peptides presented by major histocompatibility complex (MHC) molecules and lipids presented by the MHC-like CD1 proteins. T cells reactive with peptide antigens from F. tularensis have been studied in both animal models and humans. However, T cell responses to the lipid antigens of F. tularensis have not been investigated. Lipids within the cellular envelope of Francisella species contain unusual hydroxy and long chain fatty acids, two components characteristic of lipid antigens from Mycobacterium species presented by CD1. In addition to its lipid chemistry, F. tularensis displays a number of other similarities to Mycobacterium tuberculosis in its site of intracellular growth, formation of granulomas by the host, and requirement for T cell function and IFN-gamma, production for protective immunity. Our overall hypothesis is that Francisella tularensis contains lipid antigens for the CD1 antigen presentation pathway and that F. tularensis-reactive CDl-restricted T cells contribute to the control of tularemia infection. We first propose to establish human CDl-restricted T cells that recognize lipid antigens of F. tularensis and identify the structure of lipids that stimulate CDl-restricted T cells. Second, we will determine the function of F. tularensis-reactive human CDl-restricted T cells. Protective immunity against F. tularensis requires IFN-gamma, production and individuals vaccinated against F. tularensis infection generate antigen-specific cytotoxic T cells (CTLs). CDl-restricted T cells against microbial lipids produce IFN-gamma and function as CTLs against infected macrophages. We propose to determine the cytokine pattern of F. tularensis-reactive CD1- restricted T cells and whether F. tularensis-reactive CDl-restricted T cells lyse bacteria-infected macrophages to reduce the bacterial load. From these studies we hope to expand the diversity of lipid antigens known to activate CDl-restricted T cells with the future goal to design lipid-based vaccines against tularemia infection.
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{
"pile_set_name": "NIH ExPorter"
}
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DNA is a primary target for many chemical agents which have been implicated as environmental hazards. Unless repaired prior to replication, DNA-damaging events may lead to mutation or other genetic damage. We propose to establish a Specialized Center of research in Genetic Toxicology that will focus on the chemistry and molecular biology of DNA adducts in mammalian cells. Research conducted in this Center will explore the molecular toxicology of adducts derived from important classes of environmental mutagens. Chemicals to be studied initially include benzo(a) pyrene, aromatic amines and several potent mutagens isolated from fried foods. The mutagenic properties of pyrimidine dimers, modified nucleosides and DNA containing apurinic/apyridinic sites will also be examined. Research projects will utilize oligodeoxy-nucleotides that have been selectively modified with chemical mutations or their active metabolites. These modified oligonucleotides, prepared by organic synthesis, will be used to explore short-and long-term consequences of adducts bound to a defined fragment of DNA. The interaction of DNA and RNA polymerases with chemically-modified DNA primers and templates will be studied and the ultimate effects of these changes on replication and transcription will be determined. Longer-term effects of chemical damage to DNA will be studied by examining patterns of mutations produced during repair and replication of modified DNA. Finally, monoclonal antibodies will be raised against selected DNA adducts. These antisera will be characterized and used to develop sensitive and specific immunological assays for the detection and quantitation of mutagens and adducts in biological samples.
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{
"pile_set_name": "NIH ExPorter"
}
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The specifipcroteolytiactivatiosntepsof bloodcoagulationarccatalyzedby scrineprotcinasetshatarehomologous toeach otherand tothearchetypalserincprotcinascosf digestionI.n many cases,explanationfsorthemolecularbasis of thedistinctimvaecromolccularsubstratscpecificitoyf thecoagulationreactionrsemain incomplete.Evidence suggeststhatprothrombinactivatiobny theprothrombinascornplcxr,esultfsrom interactionastan extended macromolccularrecognitiosnite(Exositeso)n theenzyme followedby activcsiteinteractionpsriorto cleavageand productrcleascW.c willuseprothrombinactivatiocnatalyz_ by theprothrombinasecomplex as a paradigm for specifimcacromolccularsubstratrcecognitioannd cleavageby coagulationcomplexes to investigatehebasisforits substratscpecificiatnyd functionU.sing fluorescenstubstratacnalogs,bindingand stoppedflow kineticmeasurements by resonanceenergytransfewrillbc used toinvestigatcexositcbindingand providea completekineticand thermodynamic descriptionf thestcpwiscinteractiontshatleadto substratrcecognitionand cleavage.Wc willtesthe hypothesisthatextendedsurfacesinfactorXa playa major roleinexositc-dcpcndcnstubstratreecognitionby prothrombinascT.hese structurafleatureswillbc exploredwithexositcprobesthatbindXa and selectiveliynhibit proteinsubstratbcinding.Using thebindingfootprinotf cxositc-directperdobesas a guidelinew,c willprovidefull biochemicalprooffortheseideasby preparingrecombinantXa derivativewsith a selectivdefectin cxositc-depcndcnt substratrcecognitiownithinprothrombinascW.c willinvestigatehehypothesisthatheorderedaccessibiliotfythetwo bonds inprothrombinby prothrombinasearisesfrom thecombined constraininegffectosf cxositctetherinagnd membrane bindingby thesubstratcT.hese bindinginteractionasrcproposedtoboth facilitatned constraibnond presentatioand thespecifiactionofprothrombinascon prothrombin.Inthelastseriesof studiesw,c willinvestigate thehypothesesthatheinitiaelxositetetherinignteractiowniththesubstratoevercomes both cnthalpiacnd cntropic limitationosf activesitedockinginteractiontsherebyieldingvastimprovements incatalytircateand leadingtothe orderedcleavageofprothrombin.We believethatheapproachescontainedinthisproposalwillprovideunanticipated insightsintothefunctionof thecoagulationenzymes and a new perspectivoen themechanism of actionof prothrombinascon itsbiologicaslubstrateT.he delineationf thedeterminantosf macromolecularsubstratscpecificity willlikelysuggestnovelapproachesfortherapeutitcargetinogf thesereactionisnthromboticand vasculardisease states.
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{
"pile_set_name": "NIH ExPorter"
}
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This revised application for a Clinical and Translafional Science Award (CTSA) would support the confinued development of the Los Angeles Basin Clinical and Translafional Science Institute (CTSI). Our vision is to improve health of the diverse and underserved populafion of urban Los Angeles and, in doing so, to gain knowledge that we can share with others to improve health in urban settings and megacities across the globe. In pursuit of that vision, we have established a unique partnership of leading academic, clinical and community health organizations in the urban heart of Los Angeles. We are transforming this partnership into a preeminent clinical and translational science institute focused on better heath for diverse, urban communities. We have established four main goals for the CTSI: abuild on many independent strengths to create an integrated academic home that places high priority on clinical and translational science;acreate new translational research teams and conduct projects that leverage our unique populations and partnerships to address the best scientific opportunities and most important health priorifies of our local communities;atransform education and training to create a cadre of people with a major career focus on clinical and translational research;and aimplement and disseminate what we learn to improve health in our local communities and advance translational research through regional and national networks and collaborations. To achieve these goals, we have assembled an exceptional leadership team that is creating through the CTSI: aoutstanding resources for active development of new research projects and teams, aa robust infrastructure to promote and support basic, clinical and community research, acutting edge methodologies for data acquisifion, integration, management and analysis;aintegrated training of basic, clinical and community researchers using distance education and a focus on research for the diverse communities;aa novel approach to dissemination of our findings using professional communications expertise;and aa professional evaluation group to track and evaluate our progress and impact. Using $3.3M of institutional support, we have already made significant progress in transforming our local environment to a home that highly values clinical and translafional research. We demonstrate strong ability to generate and support exciting new research, from the bench to our communities. With a CTSA and a pledge of $18.7M of institutional support over five years, we will expand to a fully functional institute that will have a have a major impact to improve urban health and advance the discipline of clinical and translational research.
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{
"pile_set_name": "NIH ExPorter"
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This proposal requests SBIR Phase II support to complete and field test an Internet-based language intervention system designed to promote language development in preschool children with language disorders. Language delays are pervasive among school-age children. Early intervention is crucial since the consequences of language delay can be serious and cumulative. Yet research shows that most children with language delays never receive language intervention services during their preschool years. We propose to complete an Internet-based early language intervention system designed specifically for the remediation of language disorders in 3-5 year old children with language function in the range of 24-42 months. The curricular design will be guided by contemporary linguistic research. Online delivery of the intervention curriculum will be individualized and controlled by an artificial intelligence system that tracks performance and adjusts instructional support. By combining easy access via the Internet with centralized data collection and curricular control, it will become possible to implement and easily manage coordinated school and home language intervention strategies, and to provide services to more children without further taxing limited professional resources. In Phase I we tested the feasibility and technical merit of this objective. A prototype program module was developed and tested with 23 students who were enrolled in an Early Essential Education program, including 19 preschoolers with language delays. Data were collected over the Internet while testing six of these children. In Phase II we plan to fully develop and field-test this Internet-based language intervention system. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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The present project investigates the structural aspect of interactions between myosin and actin (crossbridges) in muscle cells. (i) We characterized the binding of myosin to actin, i.e. formation of crossbridges, in the presence of ATPgS, a slowly hydrolyzable ATP analogue. The binding characteristics indicate that the ATPgS- myosin binds to actin strongly and the binding is cooperative in the absence of calcium. Thus the ATPgS-crossbridge state resembles those states accompanying or following the release of phosphate in the actomyosin ATPase cycle. This ATP analogue binds to myosin at nearly full saturation level, resulting in a pure ATPgS-crossbridge state, which could considerably facilitate studies of the strong binding states. (ii) Calculations showed that some of the features in the density maps reconstructed from five or seven equatorial reflections could be due to effects of insufficient terms used in the Fourier synthesis. However, through model calculations, it is possible to estimate, albeit approximately, the underlying density distributions derived from experimental data without the spurious features. (iii) Equatorial X-ray diffraction studies on intact and skinned cardiac muscle have been initiated. Preliminary results indicate that the interfilament spacing is inversely proportional to the amount of stretch beyond rest length, even in the skinned preparation.
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{
"pile_set_name": "NIH ExPorter"
}
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Activated protein C (APC) and other components of the protein C anticoagulant pathway can protect against death from severe sepsis by inhibiting both thrombosis and inflammation. The role of the endothelial cell protein C receptor (EPCR) and its potential interactions with protease activated receptors are incompletely understood, but preliminary data suggest a major role of EPCR in protection of the lung from bleomycin induced injury and hypoxia. We propose to study the impact of either over or under expression of EPCR on the development of acute lung injury induced by bacterial inhalation, hypoxia, hyperoxia and acid aspiration. The impact of altering EPCR expression on survival, cytokine elaboration, lung fibrosis, and thrombosis will be determined. The ability of APC to alter these pathological responses in animals over or under-expressing EPCR will be determined. EPCR expression appears to be induced on lung epithelial cells in response to bleomycin. We will determine the potential role of this induction by comparing the responses of EPCR null mice to mice selectively deficient in endothelial cell EPCR. Finally, it has been proposed that APC provides protection against acute inflammatory injury by activating PAR 1 in an EPCR dependent fashion. This possibility will be examined by determining the ability of APC to protect PAR 1 deficient mice from endotoxin inhalation with special attention to the lung pathology. We will attempt to generate mice deficient in both PAR 1 and EPCR to examine interactions between these receptors. These studies will provide insights into the mechanisms by which the protein C anticoagulant pathway protects the lung from injury.
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{
"pile_set_name": "NIH ExPorter"
}
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DESCRIPTION: GVHD continues to limit the effectiveness of both related and unrelated allogeneic hematopoietic stem cell transplantation (HSCT). Yet the mechanism of initiation of GVHD remains unclear. The hypothesis driving this proposal is that a CC chemokine is a major regulator of the initiation of tissue damage in GVHD. Chemokines are a family of related proteins that regulate leukocyte migration. Recently the applicants and other groups cloned and characterized a novel subfamily of CC chemokines they termed Exodus-1, 2 and 3 for their extraordinary T-cell chemotactic ability. These three chemokines share a similar sequence around their amino terminal that no other chemokines have, asp-cys--cys-leu (DCCL). Exodus-2 and 3 are both ligands for the CC chemokine CCR7. These CCR7 chemokines are distantly related to other chemokines (twenty-eight percent homology with MIP-1a) but share among themselves several biological activities, including 1) preferential stimulation of T-lymphocyte and NK cell chemotaxis and 2) inhibition of hematopoiesis and regulation of hematopoietic progenitor migration. Recent data indicate that Exodus-2and 3 mediate T-cell adhesion to the endothelium, stimulating T-cells to stop rolling and adhere to the endothelial region expressing chemokines. Exodus-2 has been also reported as 6Cline, SLC, and TCA4. Exodus-3 has also been described as MIP-3b and ELC. They observed that Exodus-2 but not 3 is highly expressed in several target organs in human and murine GVHD. Therefore, it is hypothesized that Exodus-2 is an important mediator of T-cell tissue infiltration during initiation and/or progression of GVHD organ toxicity. This study will test this hypothesis in four aims focusing on Exodus-2 because its expression pattern is most compatible with a role in GVHD. Aim 1 - the timing and location of Exodus-2 expression during the initiation in models of murine GVHD will be analyzed. Aim 2 - the effect of anti-sera or competitive inhibitor neutralization of Exodus-2 in murine GVHD will be analyzed. Aim 3 - the effect of homozygous deletion of Exodus-2 in a knock-out mouse on GVHD pathology will be investigated. Aim 4 - the role of Exodus-2 expression in specific GVHD target organs will be studied using transgenic mice that over-express Exodus-2 selectively in liver or skin or gut. At the end of this proposal, the mechanisms by which Exodus-2 initiates GVHD should be better understood, and the possibility whether or not it might lead to effective therapeutic agents would be better defined.
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{
"pile_set_name": "NIH ExPorter"
}
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Chemically-induced neoplasia is a major public health concern and is the driving force behind much of the research conducted by the NTP. Present work has focused on mechanisms of chemically-induced forestomach carcinogenesis. One area of interest has been the relationship between chemical-induced early cell proliferation and carcinogenicity. Ethyl acrylate (EA) was selected as a model chemical for these studies because chronic gavage administration of EA resulted in a dose- and concentration- dependent increase in the incidence of forestomach (FS) neoplastic lesions in both sexes of F344 rats and B6C3F1 mice. No neoplastic lesions were found at any other site. The current work investigated the correlation between the induction of cell proliferation in the male rat FS (target) and liver (nontarget) and the carcinogenicity of EA. Cell proliferation was measured by assessing bromodeoxyuridine (BrDU) incorporation into DNA administered by osmotic minipump concurrent with gavage treatment with EA (50, 100 or 200 mg/kg/day) for 2, 4, or 8 days. BrDU incorporation was detected immunohistochemically. Results of these studies indicated that EA induced epithelial cell proliferation in the FS was dose- and time- dependent. Minimal or no cell proliferation was detected in the livers of EA-treated rats. These results suggest a positive correlation between cell proliferation in the target tissue and carcinogenicity.
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{
"pile_set_name": "NIH ExPorter"
}
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PROJECT SUMMARY CYP2D6, a key drug-metabolizing enzyme, is involved in the metabolism of ~20% of all drugs. We hypothesize that natural variation in retinoic acid concentrations and retinoid signaling in the liver regulate CYP2D6 expression and activity in humans, which in turn contributes to CYP2D6 pharmacokinetic variability and CYP2D6 induction during pregnancy. We will take a mechanistic approach and test the relationship between retinoid concentrations and CYP2D6 activity in 2 studies evaluating from the perspective of CYP2D6 induction during pregnancy, repression of induction with vitamin A administration and repression of normal CYP2D6 activity with 13-cis-retinoic acid (isotretinoin) in non- pregnant adolescent patients. Pregnant and adolescent patients often require treatment for chronic conditions, which can include CYP2D6 substrates. After accounting for genetic variation, there is still a great deal of unaccounted variability in CYP2D6 activity in these special populations making clinical management challenging. Basic science studies suggest that retinoids play an important role in CYP2D6 regulation. Our objective is to understand the role of retinoids in CYP2D6 activity in pregnant, postpartum and adolescent patients and translate new laboratory findings into humans. Our Specific Aims are: Specific Aim 1. To determine if vitamin A administration decreases CYP2D6 activity during pregnancy. In this aim, we will evaluate the effect of vitamin A administration on pregnancy-induced CYP2D6 activity. This study will provide mechanistic understanding of CYP2D6 induction and variability during pregnancy as well as provide a potential clinical strategy for management of pregnant women that require CYP2D6 substrates. Specific Aim 2. To investigate if isotretinoin (13-cis-retinoic acid) administration decreases CYP2D6 activity in adolescent patients. In this aim, we will conduct a drug-drug interaction study evaluating the effects of 13-cis-retinoic acid on non-induced CYP2D6 activity in adolescent patients. Secondary analysis will evaluate the relationship between retinoid concentrations and CYP2D6 activity in these special populations. In both Specific Aims we will utilize dextromethorphan as our CYP2D6 probe substrate. Through these complementary aims, we will be the first to conduct mechanistic testing of endogenous regulation of CYP2D6 activity in humans. The results could overcome a critical barrier to safe and effective administration of CYP2D6 substrates in adolescent and pregnant patients. Based on our compelling preliminary data, we expect to identify a novel mechanism of CYP2D6 regulation in humans and a potential management strategy for CYP2D6 induction and variability during pregnancy, with the intention of improving safety and efficacy of medications for these vulnerable patients. This work is critical for the provision of individualized therapy and along with genetics, the first step in development of an endogenous biomarker for CYP2D6 activity.
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{
"pile_set_name": "NIH ExPorter"
}
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Syntheses of the following six classes of compounds are proposed: a) 2,4-Diamino-5-(1-adamantyl)-6-substitute pyrimidines. b) 2,4- Diamino-5-substituted-6-methylpyrimidines with either -(CH2)n-Adamantyl or CH2-NH-(CH2)2-Adamantyl at carbon 5. c) 2,4-Diamino-6-(or 7) substituted pteridines with either adamantyl or -CO-Adamantyl groups at carbon 6 or 7, respectively. d) 2,4-Diamino- or 2-amino-4- hydroxylpteridines with -CH2NR-Adamantyl side chain at C-6. e) 5- Adamantyl-6-methylpyrimidines having an SH-group at C-2 and C-4 or NH2 group at C-2 and SH group at C-4. f) 2,4-Substituted purines having Adamantyl or -CH2 admantyl groups at N-9. Compounds in classes a-e are designed as inhibitors of dihydrofolate reductase. Class d in addition contains potential blocking agents of biochemical pathways involving tetrahydrofolate cofactors. Compounds in class e are designed as potential inhibitors involved in the interconversion of purines nucleosides or in the biosynthesis of nucleic acids. All compounds synthesized will undergo preliminary screening as growth inhibitors of mammalian cells in culture. Compounds having growth inhibitory activity with ID 5 equals less than 10 minus to the 6th power will be investigated in cell culture for their mode of action and as inibitors of isolated target enzymes. (The most powerful growth inhibitors ID 50 equals less than 10 minus to the 8th power) will undergo testing as antineoplastic agents in vivo in the following systems: L-1210 leukemia, P-788 leukemia, S-180 ascites, S-180 solid tumor. TA3 ascites and spontaneous mammary tumors, all above in mice and Walker carcinoma 256 and Murphy-Sturm Lymphosarcoma in rats. The relationship between antiproliferating activity and the inhibition of target enzymes in vivo will be further examined. C14 and H3 labeled compounds will be prepared for tissue distribution and cellular uptake study. The relationship between hydrophobicity of the C-5 substituent of diaminopyrimidines and their binding to dihydrofolate reductase as well as their cellular uptake will be studied.
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{
"pile_set_name": "NIH ExPorter"
}
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The major focus of this proposal is the elucidation of the mechanisms of action of cytosine arabinoside, ara C. This drug, the single most effective agent in the treatment of AML, appears to exert most of its actions through fraudulent incorporation into DNA. We will attempt to define in precise molecular terms the effects of this misincorporation on the functions of DNA; better define the nature and locus of ara C misincorporation; further explore the intermediary metabolism of the principal metabolite uracil arabinoside (ara U); and finally study which of several biochemical parameters may be predictors of clinical response to the drug. The role of DNA in controlling the life and growth of cells is so central that any derangement is likely to be damaging or lethal. The complex, interrelated functions of DNA make assessment of the impact of misincorporated ara C on any one function of DNA in living cells highly problematic. We have thus taken the approach of using synthetic DNA fragments as simplified in vitro models. We have developed methodology for the chemical synthesis of DNA oligomers containing ara C. We will utilize these oligomers for in vitro assays of DNA functions including chain elongation, replication fidelity, chemical stability, fragment ligation, and as a substrate for 3' 5' exonuclease activity. We will look for evidence of sequence specificity at the nearest neighbor and short sequence levels of DNA organization. We will also look for evidence of removal of ara C from DNA by excision repair mechanisms and for its spontaneous deamination to ara U. Although ara U is usually considered an inactive metabolite of ara C, reports indicate that it may have some activity and that it may modulate the activity of ara C. We will examine whether ara UTP cn serve as a substrate for dUTPase, and whether ara U can be excised from DNA by the uracil DNA glycosylase system. During our clinical treatment program for childhood acute myelogenous leukemia we will study the efficacy of ara C as a single agent in producing early cell kill. We will measure biochemical parameters related to the metabolism of ara C and subsequently look for correlations between these parameters in each patient and the degree of early cell kill produced in that patient.
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{
"pile_set_name": "NIH ExPorter"
}
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This Collaborative is between the University of California, San Francisco Cancer Center (UCSFCC) and San Francisco State University (SFSU). It will initiate two pilot Research Projects and two Planning and Development activities for collaborative undergraduate and/or pre- doctoral research training in cancer biology. The collaborative pilot Research Projects will involve two teams of SFSU/UCSFCC investigators; both collaborations will consist of a newly hired SFSU. Assistant Professor of Biology and a senior UCSF faculty member who is also a member of the UCSFCC. UCSF Professor Elizabeth Blackburn, Ph.D. will serve as mentor and co-investigator with Sally Pasion, Ph.D. to characterize telomere ends in fission yeast DNA replication and repair mutant. Wilfred Denetclaw, Ph.D. (appointment as a SFSU Assistant Professor of Biology effective September 2000) will be mentored by his former UCSF post-doctoral supervisor Professor Charles Ordahl. Their collaborative Research Project will focus on the biological markers for dermomyotomal myotome progenitor cells and their epithelial-to-mesenchymal transition into the myotome layer. The research questions and mentoring commitments built into each collaborative pilot Research project are designed to result in the submission of RO1 proposals during the third year of this P-20 planning grant. Student research training will be the focus for the Planning and Development section of this proposal. One pilot Planning and Development activity will use a joint SFSU and UCSFCC faculty committee to develop a more aggressive and concerted placement process/program to recruit SFSU undergraduate (MARC and MBRS- RISE) and SFSU MA/MS degree (NIH Bridge and MBRS-RISE) students into UCSFCC laboratories specifically will review and evaluate the past involvement of, 1) SFSU undergraduates (MARC and MBRS- RISE) in UCSF summer research experiences and 2) the academic year thesis research experiences of SFSU RISE) in UCSF summer research experiences and 2) the academic year thesis research experiences of SFSU MA/MS degree students whose research was executed in UCSF faculty laboratories as a basis to develop an action plan that will provide an excellent training experience in an environment permeated with a cancer biology focus. The second pilot Planning and Development activity will be directed at doing the ground work necessary to develop a joint R-25E or R-25T or T- 32 pre-doctoral training proposal during the third year of this P-20 project. Realization of this goal will require serious planning through joint UCSFCC-SFSU faculty meetings and retreats to 1) identify and prioritize short and long-term pre-doctoral student cancer research training goals and 2) develop a mutually acceptable collaborative pre- doctoral cancer research training program within the guidelines of a R- 25E or R-25T or T-32 funding mechanism. A core element of any final proposal will be the application of recruitment and retention strategies that will result in an increased enrollment of under-represented strategies that will result in an increased enrollment of under-represented minorities into and graduation from Ph.D. programs.
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{
"pile_set_name": "NIH ExPorter"
}
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In this program project, we propose a Personalized Medicine approach to the study of advanced lung disease. We hypothesize that sub-phenotypes of common diseases, including pulmonary arterial hypertension (PAH), have a profound influence on outcome and responsiveness to therapy. The overarching translatlonal goal of this program is to define common mechanistic and therapeutic pathways for PAH in the context of major lung and systemic diseases, such as COPD and HIV. Our proposed three major projects and two cores are designed to integrate and synergize fundamental translatlonal research addressing major current and high impact problems in the PAH and advanced lung disease field. Because translational medicine requires a bench-to-bedside-to-bench integrated approach, we developed a full translatlonal continuum from preclinical models in three species, including a novel primate PAH model, screening and clinical drug development programs, human hemodynamic phenomic assessments, genetics, and clinical epidemiological trials focused initially on two major disease targets, COPD and HIV, which represent two prototypic models of the pulmonary hypertension sub-phenotype. Our three proposed projects and cores all individually and collaboratively align across this translational spectrum, driving an effort to understand fundamental mechanisms of disease, identify small molecule therapeutic agents, develop screening biomarkers for vascular sub-phenotypes of lung disease, and to set the stage for phase II and III clinical trials. Successes in the first five years of this project are expected to develop into phase ll-lll clinical trials and extension to other advanced lung diseases, such as interstitial lung disease and obstructive sleep apnea, in years 6 to 10. Project 1: Pulmonary hypertension in COPD: Genetic and Environmental Determinants Project 2: ROS signaling and NOS uncoupling in pulmonary vascular disease Project 3: Pulmonary vascular-targeted NO therapeutic strategies Core A: Administrative core Core B: Pre-Clinical Models of PAH Core C: Translational Vascular Phenomics, Genomics and Epidemiology Core
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{
"pile_set_name": "NIH ExPorter"
}
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The purpose of this project is to implement and to further develop the methods of molecular dynamics, stochastic dynamics, Monte Carlo, hydrodynamics and computer graphics to investigate the solution structures of peptides, proteins, nucleic acids and lipid bilayers. The project thus involves questions of both methodology and modeling. This past year the methodological part of the research principally involved: (i) statistical analysis of the accuracy of order parameters and diffusion constants computed from dynamics simulations; (ii) statistical analysis of concerted dihedral transitions in polymers, with application to "kink" motions in membrane chains; (iii) an exhaustive comparison of Monte Carlo and dynamics algorithms for conformational searching using the bistable oscillator as a paradigm; (iv) an analysis of systematic bias in rates calculated from short trajectories. Simulations pertaining more to modeling included: (i) MD simulations of the series butane, octane, dodecane, hexadecane, and eicosane, with extensive analysis of the viscosity dependence of translational and rotational diffusion, and isomerization; (ii) MD simulation of a fluid phase dipalmitoyl phosphatidylcholine lipid bilayer in water, including an analysis showing that on the 100 ps timescale there is effectively no difference in the reorientational dynamics of the carbons in the membrane interior and in pure hexadecane. The result strongly implies that the apparently high viscosity of the membrane is more closely related to molecular interactions on the surface rather than in the interior.
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{
"pile_set_name": "NIH ExPorter"
}
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The long term objective of this work is to understand the function of the intraflagellar transport (IFT) proteins in mammals. These proteins form a complex composed of -17 polypeptides that is transported along ciliary and flagellar microtubules. IFT proteins are required for most types of ciliary assembly and are thought to function by transporting ciliary proteins from their site of synthesis in the cell body to their site of assembly in the cilium. Cilia and flagella are very prevalent in mammals and play important motility and sensory functions. The importance of these organelles is highlighted by analysis of the Tg737orpk mouse, which has a mutation in the IFT88 subunit of the IFT particle. This leads to ciliary assembly defects in many organs and causes a pleiotropic phenotype that includes blindness (retinal degeneration) and cystic kidney disease. The retinal degeneration and blindness are thought to result from an inability to properly form rod and cone outer segments in the retina. These structures, which are required to detect light in the eye, are developmentally derived from cilia. The cystic kidney disease is thought to result from defects in primary cilia, which are solitary non-motile cilia, that project from the surface of the epithelial cells that line the ducts and tubules of the nephron. These cilia are thought to be sensory organelles that play important roles in monitoring the need for cell division and controlling proliferation and differentiation. Very little is known about the functions of individual IFT particle proteins. In this proposal we will identify binding partners of IFT20, a Golgi-associated IFT particle protein. We will also address the function of IFT27, which is small G protein, in a mammalian cell culture system using shRNAs to reduce the level of the protein. Lastly, we will address the function of IFT proteins in mouse by analyzing conditional alleles of IFT genes. The work proposed here will provide new information on how cilia are assembled and how defects in these organelles cause blindness and cystic kidney disease in humans.
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{
"pile_set_name": "NIH ExPorter"
}
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Pancreatic cancer remains one of the most deadly human malignancies. During the past decade, unprecedented progress has been made identifying the genetic basis for this disease, including the discovery of a number of common somatic mutations now confirmed to play important pathogenic roles. However, the recent application of whole genome deletion analysis and high throughput DNA sequencing has accelerated the rate of novel mutation detection well beyond any ability to functionally evaluate identified candidate genes. This mismatch between gene discovery and functional annotation will only increase with the completion of the already in-progress sequencing of the pancreatic cancer genome, an effort currently being pursued by investigators here at Johns Hopkins. In order to alleviate this bottleneck, and provide a system for higher throughput annotation of the pancreatic cancer genome, we have generated the first zebrafish model of exocrine pancreatic cancer. Based on the low costs and modest floorspace required to maintain adult zebrafish, as well as the ability to rapidly generate large numbers of transgenic lines, this organism offers many advantages in evaluating the molecular basis of human cancer. When an oncogenic version of human KRAS is expressed in developing zebrafish pancreas, pancreatic progenitor cells fail to undergo normal exocrine differentiation, leading to the subsequent formation of invasive pancreatic cancer. Zebrafish pancreatic cancers invade and metastasize, and exhibit many features in common with the human form of the disease, including abnormal activation of hedgehog signaling. In addition creating the first zebrafish model of exocrine pancreatic cancer, we have successfully generated transgenic lines in which a modified Gal4 transcriptional activator is expressed in pancreatic progenitor cells. Using transposon technology to insert UAS-regulated transgenes into the zebrafish genome, we now have the opportunity to functionally evaluate a wide variety of genetic lesions for their ability to modify pancreatic cancer initiation and/or progression, achieving a level of throughput not technically feasible in the mouse. Using these techniques, we now plan to pursue the following Specific Aims: First, to functionally annotate candidate dominant mutations identified in the pancreatic cancer genome, through their modular introduction into the zebrafish tumorigenesis model;second, to study the effects of graded changes in hMYC expression in pancreatic tumorigenesis, using an inducible Gal4/UAS system targeting progenitor cells in zebrafish exocrine pancreas;and third, to develop Cre-based models of KRAS-mediated pancreatic neoplasia in zebrafish. Together, these studies will provide important new information regarding the genetic basis for pancreatic cancer, allowing for the more rapid development of effective targeted therapies. RELEVANCE (Seeinstructions): Pancreatic cancer represents one of the most deadly human malignancies, with five year survival rates of less than 5% and no change in this figure over the past four decades. By determining the genetic basis for this disease, our program will generate clinically relevant information that is likely to directly impact on strategies for chemoprevention, early detection and treatment.
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{
"pile_set_name": "NIH ExPorter"
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This proposal addresses the molecular control of neuronal cytoskeletal proteins by extracellular signals. Understanding how the neuronal cytoskeleton is regulated is important because several disease processes, including Alzheimer's disease and schizophrenia are thought to affect the cytoskeleton and thereby alter neuronal function. During outgrowth, development, and synaptogenesis, the morphology of neurons is controlled by local environmental cues like growth factors and neurotransmitters. The microstructure of adult neurons is also thought to be modified in response to synaptic or neurohumoral activity. We propose to examine the cellular basis for morphological plasticity in neurons by focusing on the regulation of the phosphorylation state and function of the microtubule- associated protein MAP2. MAP2 is thought to play an important role in the development and maintenance of dendritic morphology via its interaction with microtubules and other cytoskeletal elements. Multiple protein kinases and protein phosphatases are known to modify MAP2 phosphorylation and MAP2 function in vitro, but little is known about the properties of MAP2 phosphorylation in vivo. Our previous studies revealed that one signal which alters MAP2 phosphorylation in intact neurons is activation of excitatory amino acid receptors. MAP2 becomes rapidly and selectively dephosphorylated as a result of N-methyl-D- aspartate receptor activation. This effect appears to involve a novel signal transduction pathway for NMDA receptors. In the proposed studies, we will define the signal transduction pathways which modulate the phosphorylation state of MAP2 in hippocampal slices and cultured hippocampal neurons. Hippocampal cells metabolically labeled with 32P- orthophosphate will be used to determine the mechanism, receptor specificity and temporal properties of excitatory amino acid-induced dephosphorylation of MAP2. The stimulation of MAP2 phosphorylation by neurotransmitter and growth factor-dependent protein kinase activity will also be examined. Two forms of MAP2 will be studied: conventional MAP2 found in adult neurons, and an alternatively spliced variant unique to immature neurons called MAP2c. This variant lacks a large middle portion of adult MAP2, and thus contains a relatively small number of phosphorylation sites. The immature form of MAP2 will help us to define which phosphorylation sites are under the control of NMDA receptors. Once we identify the kinase and phosphatase pathways for MAP2 and MAP2c regulation, we will determine the precise location of phosphorylation sites on the smaller protein MAP2c. Such information will be important in eventually determining the role of phosphorylation of MAP2 and MAP2c in regulating neuronal morphology. These results will contribute to an understanding of how extracellular signals regulate MAP2 function in vivo. More generally, these experiments will allow formulation of a molecular model for transmembrane regulation of the neuronal cytoskeleton.
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{
"pile_set_name": "NIH ExPorter"
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Previous research on aging and memory has typically demonstrated that performance on memory tasks tends to decrease with increasing chronological age after a peak in early adulthood. However, this work has been based almost exclusively on episodic memory tasks which require the exact reproduction of input. On the other hand, there is almost no research which examines age/cohort differences in semantic memory involving comprehension, integration, and memory for the gist of the input. The proposed research is designed to investigate adult age-cohort differences in comprehension and memory for meaningful text materials from a contextual perspective. More specifically, the proposed research is designed to examine these processes as a function of the context defined by the input and output conditions of the task, macrostructure and microstructure of the text, goals of the task, and knowledge schema, intellectual abilities and personality states of the reader. A set of experiments involving participants from each of three age/cohort groups (25-35 years, 1944-1954; 45-55 years, 1924-1935, 65-75 years, 1904-1914) are planned over a two year period. The separate experiments are designed to investigate the role of the variables noted above. The proposed research will provide a contrast to previous work based on associative or information processing views of learning and memory. In particular, the proposed research should begin to lay the groundwork for an understanding of how adults and the aged process meaningful material. Such understanding is crucial to an ecologically valid description and explanation of adult development across the life span.
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{
"pile_set_name": "NIH ExPorter"
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Radon progeny in indoor air present a greater radiological health hazard to the greatest population than any other radiation source. The objective is to develop a very low-cost, featherweight, mailable alpha particle dosimeter medium that can be deployed by the general population at any site. The proposed medium is a thin material which maintains trapped electrons at alpha particle impact sites for extremely long times, but will scintillate visible light when illuminated by near-infrared (common GaAs LED's, etc.). The proposed effort would generate and quantify the performance of experimental samples of Quantex's unique electron trapping (ET) materials in appropriate alpha-transmissive thin-film envelopes to demonstrate feasibility. Sample fabrication experiments will explore the optimum ET material composition for alpha particle selectivity, photomultiplier and infrared LED configurations, and "erasing" requirements. Initial cost analysis and amenability to mass manufacturing will be studied and summarized. The proposed approach would revolutionize radon progeny site sampling, and require only relatively inexpensive apparatus at central counting stations. Commercial application would be widespread for dwellings and offices, and could also be very significant for mining personnel.
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{
"pile_set_name": "NIH ExPorter"
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This K01 mentored research scientist development award seeks funding to support the broad, long term objective of developing a program of research in HIV/AIDS intervention for Afro-Caribbean American youth. The specific aims of the proposed study and related training activities are: 1) to advance knowledge regarding HIV/AIDS prevention focusing on mother-daughter relationships; 2) to gain expertise in appropriate research approaches including community-based participatory research for designing and testing an HIV/AIDS prevention intervention; 3) to expand understanding of statistics appropriate for use with intervention designs ; 4) to establish and maintain relationships with faith-based communities as a foundation for ongoing research; and 5) to develop and submit an independent externally-funded research grant. Training activities will complement the research objectives by providing mentoring, academic training, practical experience, and consultation. This proposal is health-related because Afro-Caribbean American youth are at risk for sexually transmitted disease including HIV/AIDS. The safe choice intervention emphasizes abstinence but also teaches them how to make responsible decisions should they decide to be sexually active. In this way, adolescents can eliminate the risks for sexually-transmitted HIV/AIDS. Churches have been a powerful source of influence for many Caribbean families and have been underutilized by researchers to effect changes in sexual behavior. This descriptive-comparative research design will develop and test a church-based, theory-driven, and culture-specific intervention from an existing intervention that works. The study will utilize a community based participatory approach using convenience samples. Two separate focus groups (10 mothers and 10 daughters) will be conducted to gain culture-specific information which will be used to tailor the existing intervention. In the "Act" phase, the tailored intervention will be implemented and feasibility and acceptability will be evaluated. The premise of the study is to identify culture-specific determinants of risk behaviors comprehensively using quantitative and qualitative methods to generate knowledge that can be directly applied to practice and develop a program of research focusing on safe choice as a means of HIV/AIDS reduction. The goal for future research is to identify and implement culturally-appropriate HIV/AIDS intervention strategies for Afro-Caribbean Americans. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Reduced cerebrospinal fluid levels of the serotonin metabolite 5- hydroxyindoleacetic acid have been reported to be commonly associated with suicidal behavior. Alcoholics are known to often manifest suicidal behavior. Therefore, we compared cerebrospinal fluid levels of monoamine metabolites among alcoholics who had (N=18) or had not (N=132) attempted suicide, and controls (N=29). There were no significant differences among the three groups for cerebrospinal fluid concentrations of either 5-- hydroxyindoleacetic acid, the dopamine metabolite homovanillic acid, norepinephrine, or the norepinephrine metabolite 3-methoxy-4-- hydroxpenylglycol. However, in an expanded data set of almost 300 alcoholics there were significant differences for age of onset alcoholics who attempted suicide had an early age of onset of heavy drinking. They also had significantly more lifetime psychiatric diagnoses of major depression, antisocial personality disorder, panic, phobic disorder and more family history of alcoholism.
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{
"pile_set_name": "NIH ExPorter"
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The proposed studies will define the various molecular forms of human and murine type I and type II interferons in terms of the extent of physicochemical and biologically detectable variations. By correlating these variations with their abilities to exert diverse antiviral, antitumor and non-antiviral activities in homologous and heterologous cells, we proposed to define structure-function relationships of the interferons. By isolation of interferon messenger RNAs from the various induced cells producing distinguishable human interferons or distinguishable mouse interferons and by translating these mRNAs in various cellular and cell-free systems under conditions either promoting or restricting posttranslational modifications, we propose to isolate modified interferons whose physicochemical and biological (antiviral : non-antiviral activity ratios and host-specificities) properties can be compared to native interferons, potentially enabling production of interferons with selected properties. We propose also to chemically or enzymatically degrade native or modified interferons to produce small "active-core" fragments which can themselves be characterized, to detect differences from native interferons in terms of physicochemical and biological properties. Studies on the affinities of the distinct forms of native interferons and of modified or fragmented interferons for various tissues are proposed to determine the physiological significance of distinct interferon forms.
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{
"pile_set_name": "NIH ExPorter"
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This program project examines the structure, biosynthesis, and roles of selectin ligands in cell-cell communication. It also aims at elucidating the roles of carbohydrate binding protein-carbohydrate interactions in tumor cell dissemination and inflammation. This program project is a close collaboration among investigations whose expertise ranges from carbohydrate synthetic chemistry, carbohydrate chemistry, carbohydrate biosynthesis, to cellular and molecular biology. This work centers around the sialyl Le/x and related oligosaccharide structures originally discovered by the participants in this grant as E- selectin ligands. The structure and biosynthesis of E-, P-, and L-selectin ligands in humans and mice will be investigated. In particular, the roles of specific fucosyltransferase(s) and sulfotransferase(s) in the formation of selectin ligands will be determined by using knockout mice. Interactions of selectin or selectin-related molecules with sialyl Le/x on tumor cells will be characterized. In particular, the roles of tumor cell carbohydrates in nature killer (NK) cell-mediated cytolysis will be elucidated. Concurrently, the roles of NK cell carbohydrates and adhesion molecules will be determined in tumor cell targeting. These studies will provide new information on selectin-carbohydrate and selectin-related molecule-carbohydrate interactions under normal and pathological conditions. The knowledge obtained in these studies may help in the design of improved therapeutic interventions against malignancies and other diseases.
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{
"pile_set_name": "NIH ExPorter"
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The overall goal of this proposal is develop a novel method for HIV vaccine development. The hypothesis which is set forth is as follows: that bacteriophage vectors can be adapted for delivery of exogenous genes to mammalian cells, including dendritic cells, and that these modified bacteriophage vectors can be used to elicit specific and potent antiviral cytotoxiclymphocyte (CTL) responses to an encoded HIV-1 antigen. This hypothesis will be tested as follows: genetically modified phage vectors will be constructed, and their ability to elicit HIV-specific CTL responses will be examined using a small animal model system. Overall, the work is expected to contribute to the development of phage vectors suitable for use in HIV vaccine delivery. Such vectors would be simple to use, cheap to manufacture and considerably safer than vectors based on potentially pathogenic human or animal viruses. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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This proposal outlines an intensive 5 year training program designed to prepare the principal investigator for a career as a physician scientist in tumor immunology. The candidate is currently a third year hematology fellow with 1.5 years of basic science experience in the lab of Dr. Jenny Ting at the University of North Carolina Chapel Hill. This award will enable the investigator to achieve scientific independence by providing a dedicated period of bench research in a comprehensive educational environment. At the end of the term, he will have acquired the necessary technical skills as well as scientific rigor to contribute to the fields of tumor immunology as an academic physician. The applicant's research in Dr. Ting's lab focuses on the migration and function of murine dendritic cells (DCs). The aims of this work are (1) to compare the migration of immature and mature DCs after subcutaneous and intravenous injection, (2) to describe the impact of cancer on DC migration, and (3) to explore the potential therapeutic benefit of increasing DC migration to the tumor by genetic manipulation. This will be done by injecting DCs cultured from the bone marrow of transgenic mice expressing enhanced green fluorescent protein (EGFP). These cells will be identified in vivo using micro-fluoroscopy and flow cytometry and isolated with cell sorting. The functional changes induced by migration from injection site will then be assessed. This inventive strategy will lead to a better understanding of activation and maturation of DCs in vivo and novel approaches to immunotherapy. The applicant's work will be reviewed regularly by Drs. Jenny Ting, Jon Serody, and Beverly Mitchell. Dr. Ting will serve as the primary laboratory mentor. Her lab is well funded and committed to applying cellular and molecular approaches to cancer and transplantation. Dr. Serody has extensive basic science experience with DCs and is currently leading UNC's effort to develop DC vaccines. Dr. Mitchell is chief of the hematology/oncology division at UNC and is nationally recognized in the field of hematology as a clinician scientist. All have extensive experience in developing the careers o f young scientists. This work has the support of the Lineberger Comprehensive Cancer Center which has made accessible its core facilities.
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{
"pile_set_name": "NIH ExPorter"
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Group B Streptococcus type III-tetanus toxoid conjugate vaccine will be administered to healthy, nonpregnant adult volunteers. A single dose of one or two vaccine concentrations will be given intramuscularly with blood drawn before and 28 days after immunization.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary Most immigrants begin their labor market trajectory in the US with lower earnings than natives with comparable levels of skills and work experience. Whether and how fast immigrants catch up to natives is critical for their own wellbeing and that of their children as numerous studies have found a strong relation between low parental income and a variety of negative outcomes for children, including worse health, greater likelihood of dropping out of school, higher risk of teenage pregnancy, and lower emotional and cognitive development. Our knowledge of the economic progress of immigrants comes mostly from studies using a synthetic cohort approach whereby arrival cohorts are followed across decennial censuses. This approach suffers from well- known estimation problems. Chief among them is that repeated cross sections necessarily exclude individuals who return to their countries of origin between censuses, and also undercount circular migrants. The exclusion of circular and return migrants could lead to an overestimation of immigrants? ability to narrow the earnings gap with natives over time in the repeated cross-sectional approach. Our knowledge of the earnings assimilation of immigrants is also severely outdated as it is largely based on information from immigrants arriving in the 1980s or earlier. In this study we will examine immigrant men?s earnings trajectories, and measure the extent to which the earnings gap with natives narrows over time, using a unique dataset that links respondents of the Survey of Income and Program Participation (SIPP) to their longitudinal earnings from individual tax records. We will follow immigrants? earnings for up to 20 years after arrival and compare their earnings trajectory to that of similarly-qualified natives. This longitudinal approach offers better estimates of the economic assimilation of foreign-born individuals over time than an approach using synthetic cohorts based on repeated cross sections. We will explore differences in the earnings growth trajectories of immigrants according to their level of education, country of origin, and racial and ethnic identification. Examining differences in the earnings assimilation of immigrants of different race and ethnicity is important because previous research has suggested that minority immigrants face greater obstacles in assimilating into the US labor market. We will also compare the earnings trajectories of different immigrant arrival cohorts to test the argument that immigrants are declining in their labor market ?quality? over time. Finally, we will use fixed-effects models to control for unobserved differences in immigrant selectivity, thereby isolating insofar as possible the effect of the context of reception on immigrants? earnings growth.
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{
"pile_set_name": "NIH ExPorter"
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Immunization of tumor-bearing hosts with Vibrio cholerae neuraminidase (NANAase)-treated tumor cells alone or in combination with chemotherapy slows the growth or causes regression of established tumors. Although this type of immunotherapy has been explored in a number of animal models and has been used in man, information is lacking on why NANAase treatment enhances immunogenicity. We have developed a chemoimmunotherapy model for treating L1210 ascites tumors in DBA/2J mice by combining 1-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosurea (MeCCNU) with NANAase-treated purified L1210 plasma membranes. Because the immunogen is a purifed subcellular particle of known protein and glycoprotien composition, we can analyze at the molecular level the NANAase modifications of the membranes and how these affect the immune response to the tumor. We will use two-dimensional immunoelectrophoresis to analyze the immune response of DBA/2J mice to NANAase-treated L1210 plasma membranes to determine the relationship between glycoproteins modified by NANAase and the antigens to which the host responds. We will also determine whether the plasma membranes of tumor cells surviving chemotherapy are more closely related to NANAase-treated membranes prepared from the parent tumor than to untreated membranes from that tumor. This will be done by producing an L1210 tumor line resistant to MeCCNU chemotherapy and by using biochemical and immunochemical techniques to compare its plasma membranes with those of NANAase-treated and untreated membranes of the parent tumor. In addition, we will study the role in the immune response to NANAase-modified membranes of naturally occurring serum factors that recognize determinants on NANAase-treated cells; we will determine whether modified membranes are more effective in animals preimmunized to increase the level of these factors or whether modified membranes preincubated with serum containing these factors are more effective than those that have been preincubated.
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{
"pile_set_name": "NIH ExPorter"
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The proposed research will examine the long-term retention of paired-associates by four- and seven-year old children. The question of development differences in memory has been attacked in many previous studies. Unfortunately, only a few of such studies have been methodologically sound. By far the most common methodological deficiency has been the failure to take into account variations in amount of original learning when retention differences are examined, i.e., degree of original learning has been confounded with rate of forgetting. Moreover, in those few studies that were methodologically sound, none examined age differences with retention intervals of more than a few minutes and with children ranging down to the preschool years. That such an experiment may yield important results concerning human memory development is suggested by a recent review by Campbell and Spear (1972) of theory and of relevant animal research. The proposed research is designed with these considerations in mind. The Ss will be 60 preschoolers (mean age about 4 years, 2 months) and 60 second graders (mean age about 7 years, 6 months) enrolled in schools in Evanston. At each age level the Ss will be randomly assigned into one of three retention interval (RI) conditions: 10 minutes, 48 hours, or 10 weeks. Initial paired associate (PA) learning will ensure a high degree of learning for all Ss and one that is approximately equal for the two age levels. PA retention testing will be carried out after the intervals specified above. Major interest will be in main effects of age on retention test performance and, particularly, in the interaction between age and RI.
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{
"pile_set_name": "NIH ExPorter"
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The long term goals of this study are to define the changes in ion channel activity in colonic inflammation. Ulcerative colitis is an inflammatory bowel disease characterized by recurrent episodes of colonic inflammation and tissue degeneration. The overall hypothesis is that inflammation induces specific changes in expression, regulation and kinetics of ion channels in smooth muscle cells. These changes affect gastrointestinal motility contributing to the clinical expression of altered contractions. In this proposal, we will test the hypothesis that reactive nitrogen species produced during colonic inflammation cause nitration of the tyrosine residues within the c-terminus of the calcium channel and result in down-regulation of channel function. Based on our preliminary findings, our working model is that nitration of tyrosine residues within the c-terminus of the Ca2+ channel prevents phosphorylation by c-src kinase and alters channel gating, downstream intracellular signaling and gene transcription. Specific aim 1 is to determine the mechanisms by which c-src kinase alters the gating properties of the smooth muscle calcium channel. In this aim, we will determine the biophysical properties of the calcium channel and examine the effect of tyrosine phosphorylation and nitration on voltage-dependent inactivation using single channel and whole-cell patch clamp techniques. We will also investigate the interaction of src kinase with the calcium channel using fluorescence resonance energy transfer (FRET) and siRNA approaches to delineate the functional effect of src-Ca2+ channel interaction in muscle contraction. Preliminary data show that mutation of the terminal tyrosine residue within the c-terminus of the Ca2+ channel prevents the docking of the src homology 2 (SH2) domain. In specific aim 2 we will examine the role of nitration of the Ca2+ channel in intracellular signaling and gene expression following inflammation. In this aim we will measure phosphorylation of the transcription factor CREB, and the downstream activation of cyclic AMP response element (CRE). We will test the effect of Ca2+ channel nitration on these processes and define the role of the terminal tyrosine residues using mutants of the Ca2+ channel. Analogous to protein phosphorylation-dephosphorylation, we will test whether denitration is a physiological process in specific aim 3. Our preliminary findings indicate that Ca2+ channel are substrate(s) for denitration. We will test whether denitration reverses the down-regulation of calcium influx in whole tissue segments, and determine the source of denitrating enzymes within the colonic wall. The information obtained from these studies will increase our understanding of the potential changes in ion channel activity with inflammation and help identify novel therapeutic agents in the treatment of motility disturbances in the pathophysiology of the colon.
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{
"pile_set_name": "NIH ExPorter"
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Facility Maintenance and Operations: Summary The NEIDL Facilities Maintenance and Operations Core is comprised of three functional areas, including Facilities (FAC), Information Technology (IT) and Community Relations (CR). Each is tasked with closely related but distinct responsibilities that collectively maintain the function, safety, and standing of the NEIDL BSL-4 operation. The Facilities Core is integrated with its equivalent units at the institutional level. The aims of the Facilities functional area are to 1) Support the operations and maintenance of the laboratory and support systems; 2) Improve the laboratory infrastructure for function, safety, and finance; 3) Maintain training, documentation and continue refinement of standard operating procedures. The aims Information Technology functional area are to 1) Continue the development, implementation, and staff training of the BSL-4 ?in-suit?, hands free voice over internet protocol (VoIP) communications systems; 2) Implement, modify as need, and train staff on the select agent inventory software and its radio frequency identification (RFID) tracking systems in the BSL-4 laboratories; 3) Continue the development, implementation, modification, and staff training of an online library of ?video-enhanced SOPs? of BSL-4 operations for in-lab reference, training, and safety review. Finally, the aims of the Community Relations functional area are to: 1) Broaden community outreach and expand efforts to inform and educate the community as well as build and sustain community trust about the NEIDL and its mission, in partnership with the Community Liaison Committee as well as engage other tactics; and 2) Use the NEIDL and its scientific and support personnel as an ongoing educational training resource for students, their parents and the lay public in the surrounding community.
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{
"pile_set_name": "NIH ExPorter"
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Maintaining functioning among older men and women is an important public health goal as the U.S. population lives longer with chronic disease. Lower extremity peripheral arterial disease (PAD) affects 12% of men and women age 65 and older. This project will study the independent impact of PAD on lower extremity functioning and social independence among men and women age 55 and over. We well determine whether the ankle brachial index (ABI), an accurate, reliable measure of lower extremity arterial perfusion, predicts baseline and subsequent lower extremity function. Our primary specific aims are 1)To assemble a cohort of 470 men and women with PAD and 150 men and women without PAD, all age 55 or older, and follow them prospectively for up to 38 months; 2)To define the relationship between baseline lower extremity functioning and baseline lower extremity arterial perfusion as measured by the ABI; 3)To determine the relationship between the rate of decline in lower extremity functioning over 24 months follow up and baseline lower extremity arterial perfusion as measured by the ABI; 4)To determine whether current cigarette smoking, lack of regular exercise, diabetes mellitus, and lower physical activity levels are associated with greater functional decline among PAD subjects, independent of the ABI level. We well test the hypotheses that 1)Lower extremity arterial perfusion correlates directly and independently with lower extremity function; 2)Lower extremity function among PAD subjects deteriorates at a predictable rate over time; 3)PAD subjects with a lower ABI level experience greater declines in lower extremity function over time; 4)PAD subjects who smoke, are diabetic, do not regularly exercise, or have lower physical activity levels experience greater decrements in lower extremity function over time, independently of their ABI level. In an ancillary study, we will obtain blood samples and determine whether specific hemostatic factors and genetic polymorphisms are associated with higher rates of cardiovascular events and progression of PAD. Results of this study will be used to develop subsequent intervention studies designed to improve quality of life and prevent functional decline among men and women with PAD.
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{
"pile_set_name": "NIH ExPorter"
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It remains largely unknown how energy metabolism coordinates hematopoietic stem cell (HSC) maintenance and lineage differentiation. The hypoxic microenvironment in stem cell niches limits mitochondrial aerobic metabolism (respiration/oxidative phosphorylation) in HSCs, which preserves this essential cell reservoir from oxidative damage by attenuating the production of reactive oxygen species (ROS), a byproduct of mitochondrial respiration. However, cell intrinsic mechanisms regulating HSC metabolic activities are poorly defined. Furthermore, how energy metabolism orchestrates HSC differentiation in concert with other regulatory networks has not been characterized. Lack of such knowledge impedes the understanding of the pathogenesis of blood diseases associated with mitochondrial dysfunction and metabolic conditions. PTPMT1, an evolutionarily conserved PTEN-like phosphatidylinositol phosphate (PIP) phosphatase, is localized to the mitochondrial inner membrane where ion channels and transporters, important for mitochondrial ion homeostasis and thus metabolism, reside. Our preliminary studies have shown that targeted disruption of PTPMT1 in mice results in post-implantation embryonic lethality. Deletion of PTPMT1 from adult bone marrow (BM) cells of inducible knockout (PTPMT1fl/fl/Mx1-Cre+) mice impairs myeloid and lymphoid cell development. Moreover, postnatal hematopoiesis in hematopoietic cell-specific knockout (PTPMT1fl/fl/Vav1-Cre+) mice is completely blocked. These mice succumb to pancytopenia within 3-6 days of birth. Strikingly, HSCs in the BM are increased by ~30-fold and ~10-fold in PTPMT1fl/fl/Mx1-Cre+ mice and PTPMT1fl/fl/Vav1-Cre+ neonates, respectively. Preliminary mechanistic studies show that cellular respiration of PTPMT1-depleted cells is decreased while glycolysis is enhanced. The objective of this application is to extend these studies to further determine the role and signaling mechanism of mitochondrial phosphatase PTPMT1 in HSCs. We hypothesize that PTPMT1 coordinates HSC homeostasis and lineage commitment by modulating mitochondrial metabolism. We plan to test our hypothesis and accomplish the objective of this application by pursuing the following two aims. (i) To define the role of PTPMT1 in hematopoiesis. (ii) To determine the mechanisms by which PTPMT1 modulates HSC function. The proposed work is innovative, because it explores the significance of finely controlled mitochondrial metabolism for HSC maintenance and lineage differentiation. The combination of the work proposed is collectively expected to yield novel insights into the bioenergetic regulation of hematopoietic cell development. In addition, the information gathered will lead to a better understanding of the pathophysiology of blood disorders resulting from mitochondrial dysfunction and metabolic conditions.
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{
"pile_set_name": "NIH ExPorter"
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Our previous funding round of this project was devoted to creating the underlying data standards and infrastructure to support sharing microarray data. The availability of the MGED (Microarray and Gene Expression Data Society; www.mged.org) standards and infrastructure and our accumulated experience using and evaluating them puts us in an excellent position to deliver tools and resources directly to the bench biologists who are generating high-throughput gene expression data: We now propose to shift our main efforts away from building computational infrastructure - the over-arching purpose of this new proposal is to facilitate scientific discovery by providing bench biologists with the tools they need to effectively share gene expression data and to take advantage of well-annotated gene expression data in their research. Our work to create and promote data sharing standards and resources will have greatest impact when those standards and resources are in common use by biomedical researchers. [unreadable] [unreadable] The need for and potential benefits of standards for microarray and other high-throughput technologies is clear, yet the positive impact of the standards thus far developed has not yet been fully realized. This is in large part because the standards and tools we have developed still require expert knowledge, yet the targeted users are bench biologists who are not experts in this domain, and should not be expected to become so. In this application, in addition to building the next generation of data exchange standards, we are proposing to use the infrastructure we have built in the previous round of funding as the basis for data exchange resources that are useful and usable by bench biologists. Therefore, our aims are to: [unreadable] 1. Develop tools to help researchers easily annotate microarray experiments. [unreadable] 2. Extend popular data analysis and visualization tools (BioConductor, MeV, GenePattern, Java TreeView) to use MAGE-TAB-encoded experimental annotations. [unreadable] 3. Generalize the MAGE-TAB data exchange standard to work with other high-throughput biomedical data, such as ultra-high-throughput sequencing. [unreadable] 4. Provide biologist-friendly ontology terms that can be used to annotate microarray data as well as serve as meaningful terms in computational analyses. [unreadable] 5. Participate in outreach, education and information gathering efforts to engage the community in the development of standards and to ensure widespread use and critiques. [unreadable] [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Polyoma virus functions important in the initiation and maintenance of cell transformation, and in turmorigenesis, will be studied. Altered polyoma viruses lacking restriction endonuclease sites in the "early" and "late" portions of the genome have been isolated, and partially characterized with respect to the extent of the genomic alteration that has occurred. The following biological properties of these viruses will be studied, a) ability to induce host DNA synthesis; b) ability to induce cell transformation, and c) ability to induce tumors. BIBLIOGRAPHIC REFERENCES: Anderson, D.M., and Folk, W.R. (1976) Iodination of DNA. Studies of the Reaction and Iodination of Papovavirus DNA, Biochemistry 15, 1022. Folk, W.R., and Bancuk, J.A. (1976) The Polyoma Genome in Hamster BHK 21-C13 Cells: Integration into Cellular DNA and Induction of the Viral Replicon, J. Virol. in press (October).
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{
"pile_set_name": "NIH ExPorter"
}
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The purpose of this application is to: a) Continue the experimental intercomparison of clinically active fast neutron beam radiotherapy projects in regard to fast neutron dosimetry, b) include new clinical projects in this intercomparison as they become active, c) assist the cancer therapy effort in general and the Radiological Physics Center-Houston in its role as the dosimetry quality control arm of RTOG by development of a fast neutron dosimetry protocol based on the best experience and advice currently available. This protocol would then be subject to review by a suitable scientific body. The field of neutron therapy is in a period of rapid development in facilities of varied backgrounds. This enriches the base of the physics advice and dosimetry methods available but accentuates the need for a mutually agreeable aproach to the dosimetric basis for national clinical trials of fast neutrons at these centers. This task group will ensure the compatibility of current practices at the various facilities by experimental intercomparison of dosimetry at new facilities as they become active and/or at one established facility so that such experimental exchanges occur at least once per year. Subsequent to acquisition of data and dependent upon opportunity for review and evaluation, meetings will be held to compare the data, plan experimental resolution of differences and develop a protocol of significance to the fast neutron beam therapy trials and to the radiological community, irrespective of the centers' relation to any such proposed national clinical trial.
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{
"pile_set_name": "NIH ExPorter"
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The goals of this research are to define the etiologies and pathogeneses of otitis media with effusion. Our past studies have documented several pathogenic pathways whereby nasal inflammation provoked by allergy or viral infection are manifest as eustachian tube obstruction, a recognized cause of otitis media with effusion. The purpose of this ongoing research is to define the role and interaction of allergy and viral infection in promoting nasal congestion, tubal obstruction and middle ear pathophysiology. These relationships will be explored with an emphasis on amplification, modulation and prolongation of primary responses. Specifically, we will utilize human models for challenges with inflammatory mediators, allergens and viruses, and animal models of IgE hypersensitivity to: 1) define the role of inflammatory mediators in causing tubal obstruction and middle ear disease; 2) document the potential of allergen and viral priming and late allergic reactions to enhance and sustain the nasal and middle ear pathophysiologies; and 3) determine if allergic rhinitis subjects are hyperresponsive to upper respiratory virus infection. Age related response differences between children and adults will be further defined. Children who are indentified in prospective studies as "otitis prone" will be evaluated for immune defects to explain their recurrent ear infections. Middle ear effusions (MEE) from children with acute and chronic OME will be assayed for histamine, prostaglandins, tryptase and eosinophil derived major basic protein and neurotoxin. Enhanced basophil histamine release and production of histamine releasing factors will be explored as possible predisposing factors in these children. This collaborative research effort by clinical immunologists, otolaryngologists, microbiologists, physiologists and biochemists will better define the role of allergy and viral infection in the pathogenesis of OME and should result in improved diagnosis and management of this most common pediatric disease.
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{
"pile_set_name": "NIH ExPorter"
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Chronic kidney disease affects approximately 26 to 30 million adults in the United States and is associated with substantial morbidity and mortality. The underlying causes of CKD and risk for progression in an individual patient are frequently not known in clinical practice. Nevertheless, several pathological processes appear to be common across progressive kidney diseases of diverse etiologies, including progressive tubulointerstitial fibrosis, tubular atrophy, glomerulosclerosis, capillary rarefaction, and vascular sclerosis. Novel non-invasive biomarkers of CKD pathology may provide clarification of the underlying pathobiological processes responsible for CKD in an individual, enabling earlier diagnosis and more accurate prognosis. This proposal is an ancillary study to the CKD Biomarker Consortium (U01DK085660), a group of investigators with wide-ranging biomarker expertise who have access to samples from a number of cohorts of individuals with CKD. The proposal describes a prospective cohort study involving biological sample collection from patients undergoing native kidney biopsy, the gold standard determination for identifying the cause(s) of CKD. We will enroll participants from two hospitals in Boston, Massachusetts and measure urinary and tissue levels of novel biomarkers, including KIM-1, NAG, NGAL, MCP-1, L-FABP, IL-18, HGF, VEGF, IP-10, and CTGF. In Specific Aim 1, we will correlate urinary biomarker measurements with 13 histologic scores that capture inflammatory, fibrotic, and ischemic pathological processes in the mesangium, tubules, glomeruli, interstitium, and arterioles; further immunohistochemistry will be performed on the most promising biomarkers to identify their tissue distribution and correlation with urinary levels. In Specific Aim 2, we will test the prospective associations of urinary and tissue biomarkers with renal function decline, defined as a doubling of serum creatinine or the need for renal replacement therapy. We anticipate enrollment of over 900 participants with median follow-up of approximately 5 years. This ancillary proposal will be collaborative and integrated with U01 Consortium investigators and will facilitate multidirectional translation of findings across animal studies, large cohort studies, and the biopsy cohort. Urine, plasma, and DNA samples will be shared with the Consortium for future studies. PUBLIC HEALTH RELEVANCE: Chronic kidney disease is a major public health threat for which the current methods of diagnosis and assessing prognosis are inadequate. The development of new biomarkers of chronic kidney will be facilitated by studies such as the one proposed here, where we will compare urinary and tissue levels of novel biomarkers with actual pathological findings from individuals who have undergone kidney biopsy, which is the gold standard diagnostic tool. Better biomarkers are expected to translate into improved care for the growing number of individuals with chronic kidney disease.
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{
"pile_set_name": "NIH ExPorter"
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ADMINISTRATIVE CORE: ABSTRACT Karmanos Cancer Institute's (KCI) Administration is comprised of two integral parts: Cancer Center Administration and Senior Leadership. Cancer Center Administration provides a robust infrastructure that supports cancer research activities at KCI. The mission of Cancer Center Administration is to reliably and consistently provide cost-effective administrative services that further the conduct of efficient and effective research efforts. Overall responsibilities include fiscal accountability for the financial and operational activities of KCI, management of laboratory space, coordination of Shared Resources (Cores), management of information, assistance with faculty and personnel administration including coordination of faculty recruitment, coordination of institutional research projects, and the provision of necessary administrative support to the research related activities of KCI Leadership, Program Leaders, Shared Resources and the membership. CCSG funding of $147,682 is requested to support Administration. Notable accomplishments since the last grant submission include: ? Implementing significant upgrades to the Research Administration Application (RAA), the central repository of CCSG data, to ensure continuing compliance with latest NCI CCSG guidelines. ? Designing a Clinical Trials mobile app to display clinical trial information in real-time. ? Implementing iLab Solutions software for all CCSG Shared Resources. ? Expanding the marketing of CCSG Shared Resources through a Shared Resources handbook. ? Coordinating annual internal pilot project programs and the new Mock Study Section. ? Contributing to the integration of research and clinical operations with McLaren Health Care Corporation. Senior Leadership oversees the vision, mission and strategic plans for the KCI scientific community. The team ensures all KCI resources are optimally deployed to enhance the development of exceptional science. Senior Leadership is composed of scientific Associate Center Directors (ACDs) for Basic Sciences (Dr. Wei- Zen Wei), Clinical Sciences (Dr. Anthony Shields) and Population Sciences (Dr. Terrance Albrecht). They are supported by an ACD for Research Administration (Ms. Constance Claybaker) and overseen by the Deputy Center Director, Dr. Ann Schwartz. As President, CEO, and Center Director of the Karmanos Cancer Institute, Dr. Gerold Bepler bears ultimate responsibility for the administration and fiscal management of all KCI resources, including budgeting, personnel and faculty appointments, and space and equipment allocations. No CCSG funds are requested to support Senior Leadership activities.
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{
"pile_set_name": "NIH ExPorter"
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We seek the Institute on Aging Small Research Grant Program Award. The proposal, entitled Older Adult Safety in Surgery (OASIS) II, focuses on the role of self-reported physical functional status compared with Fried Frailty Criteria for the purpose of preoperative risk stratification in older adults undergoing major surgery. Current methods for identifying which patients will have an adverse postoperative course - i.e. the process of risk stratification - rely on inventories of comorbid disease that do ot account for disease severity or burden. In addition, these methods focus on discrete medical complications and not on major complications as a whole, discharge to nursing home, or readmission, which are the outcomes most important to patients and policymakers. Recently, multiple aging investigators have attempted to develop improved assessment methods for identifying older adults at risk for adverse postoperative course. They have shown an association between frailty and increased need for discharge to nursing home after surgery. Given the need to make point of care physical performance measurements and record time intensive inventories of exhaustion symptoms and activity items, frailty remains a suboptimal method for risk stratifying older adult surgical patients. Assessing self-reported functional stats offers the ability to efficiently capture a wide range of vulnerability to an adverse postoperative course. Investigators at Boston University with the support of the National Institute on Aging (NIA) have developed a tool for assessing physical function, the Late Life Function (LL-F) part of the Late Life Function and Disability Instrument which may be applied in the preoperative setting. From a large bank of potential items, a computer adaptive testing version of this instrument uses a patient's initial responses to guide subsequent inquiry (e.g. omitting questions of lower function for someone who's already reported high function). In this proposal, I will prospectively collect Fried Frailty Criteria and LL-F from patients visiting the Pre-procedure Clinic at Boston Medical Center. To track outcomes, I will leverage data already being collected by the Department of Surgery as part of a quality initiative sponsored by the American College of Surgeons National Surgical Quality Improvement Program. Comparing LL-F via computer adaptive testing with frailty head-to-head can clarify the path we should take regarding preoperative risk stratification. This proposal captures the unique experiences of the PI who has been leading medical consultation at the largest safety net institution in New England. It also includes the expertise of multiple leaders in aging research.
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{
"pile_set_name": "NIH ExPorter"
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Erythromycin has been shown to stimulate antral motility through motilin receptor stimulation, and serves as a beneficial therapy in gastroparetic patients. The effect of Octreotide on antral and small bowel motility is unclear. This study examines the effect of Erythromycin and Octreotide antroduodenal motility as measured by a hydrostatic perfusion device and electrogastrogram in patients with gastroparesis.
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{
"pile_set_name": "NIH ExPorter"
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Project Summary: Dr. Bart Scott is a Research Associate at the Fred Hutchinson Cancer Research Center (FHCRC) and an Acting Clinical Instructor at the University of Washington. His long-term career goal is to be an independent clinical investigator, and his research interests involve improving transplant outcomes in patients with myeloid malignancies. The Career Development Plan is a comprehensive program consisting of institutional meetings, a master's degree in epidemiology, continued training in responsible research, and appropriate evaluation and feedback from an Advisory Committee. Dr. Scott performed a retrospective analysis comparing results with myeloablative and nonmyeloablative regimens in patients with Myelodysplastic Syndrome (MDS) and Acute Myelogenous Leukemia (AML), which serves as the basis for this proposal. The data suggest there are distinct advantages and disadvantages with either approach; only a prospective trial can more definitively address the potential benefits in overall survival. With the mentorship of Drs. Joachim Deeg and Rainer Storb, Dr. Scott has designed and will implement a phase III trial comparing toxicity and efficacy of myeloablative and nonmyeloablative conditioning regimens in patients with MDS and AML. Dr. Scott will be the principal investigator of this trial and will be primarily responsible for all aspects, which include designing consent and assent forms, obtaining IRB approval, collaborating with outside centers, patient accrual, reviewing primary data for eligibility, reporting serious adverse events, and analyzing and publishing data. The specific aims are as follows: 1) determine if there is an overall survival difference in patients with MDS and AML conditioned with myeloablative (cyclophosphamide or fludarabine and busulfan) or nonmyeloablative (fludarabine and low dose total body irradiation) regimens, 2) determine the impact of chemotherapy used for pre-transplant "debulking" on mortality, relapse, and engraftment after myeloablative and nonmyeloablative conditioning, and 3) characterize pharmacokinetics (PK) of intravenous busulfan in the myeloablative regimens, and determine the impact of PK parameters on transplant outcomes. The FHCRC is a leading institution in the treatment of myeloid malignancies and provides an excellent environment to foster the development of Dr. Scott into an independent clinical investigator with expertise in developing and managing clinical trials designed to improve survival and quality of life in patients with myeloid malignancies. Relevance: The goal of this proposal is to improve survival and quality of life in patients with leukemia and related diseases by developing safer and more effective transplant options. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Mast cells are pluripotent effector cells that reside in tissue. They respond to diverse stimuli by releasing potent biological mediators into the surrounding tissue. These mediators include extracellular proteases, histamine and serotonin, as well numerous cytokines, chemokines, and growth factors. Immunologically, mast cells play a crucial role in the generation and maintenance of inflammation, in response to antigenic challenge. In addition to responses to immunological stimulation, mast cells also respond to polybasic secretagogues and physical stimuli. Through responsiveness to physical stimuli (thermal, osmotic and mechanical inputs), mast cells contribute to both tissue homeostasis, and the wounding process that follows noxious insults. Each mechanism for mast cell activation relies on calcium influx through specific plasma membrane cation channels. Here, we report that calcium-permeant cation channels of the TRPV family are expressed in mast cells. TRPV ion channels are a newly recognized family of sensors, which receive, and react to, physical environmental cues, including thermal, osmotic and mechanical stimuli. The central premise of the current proposal is that TRPV channels transduce physiological, and pathophysiological, signals that are functionally coupled to calcium signaling and mediator release in mast cells. We propose to study the regulation, and function, of TRPV cation channels in the mast cell context. Our first Specific Aim tests the hypothesis that expression of TRPV2 confers a specific, thermally-evoked, cation conductance upon mast cells. In the second Specific Aim, we will explore a novel regulatory mechanism for TRPV2. Our preliminary data show that TRPV2 is a target for phosphorylation by protein kinase A in mast cells, and that this interaction is mediated by a novel adapter protein, PAP7. PAP7 specifically bridges TRPV2 and PKA. We will explore the functional consequences and upstream regulatory mechanisms for phosphorylation of TRPV2 by PKA, and the contribution of PAP7. In Specific Aim 3, we will examine the representation and functionality of TRPV channels in dermal mast cells, exploring the hypothesis that TRPVs couple mast cell responses to environmental stimuli in the skin.
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{
"pile_set_name": "NIH ExPorter"
}
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Summary: A significant proportion of anti-parvovirus B19 is considered to be neutralizing antibody. We continued to assess the levels of anti-B19 in products by utilizing a commercial EIA kit that has wells coated with a recombinant parvovirus B19 capsid protein, VP2. A semi-quantitative assay utilizing an international standard for anti-B19 IgG was set up and the sensitivity of the assay was ca. 30 mIU of anti-B19/mL. We found that among Factor VIII concentrates (AHF), the lower the purity, the higher the levels of anti-B19. Of intravenous immune globulin (IGIV) products, significant difference in anti-B19 levels was observed among products made by different manufacturers. The three manufacturers' anti-D immunoglobulin products, used by some pregnant women to prevent the hemolytic disease of the newborn, also had appreciable levels of anti-B19. The apparent viral safety of immunoglobulins with respect to B19 can be attributed to the presence of appreciable anti-B19 antibodies in the products. However, further monitoring the products is needed to determine whether the varying levels of anti-B19 are due to different manufacturing procedures, B19 NAT screening implemented by manufacturers, or other factors. In collaboration with scientists in OVRR, we continued to monitor levels of anti-measles antibodies in IGIV products. Varying neutralizing antibody levels were found among IGIV products. Further studies are needed to investigate why the levels vary and to evaluate the neutralizing capabilities by the IgG fragments and subclasses. The CBER anti-D immunoglobulin standard, which was formulated many years ago and was recently found HCV RNA positive by us, is nearly depleted. The current International Reference Preparation of anti-D immunoglobulin was also formulated many years ago and has been found HCV RNA positive too. Hence we began recently a collaborative study with Dr. Susan Thorpe of NIBSC to formulate a standard not only as a new CBER standard but also as a replacement standard for the WHO International Standard. We acquired two candidate materials from two U.S. manufacturers. Purity and potency assays have been carried out. The pilot experiments by blending different proportions of the two materials to achieve appropriate anti-D potency and IgG concentration in a freeze-dried formulation are in progress. Once it is formulated, a worldwide collaborative study will be set up to determine the consensus levels of this anti-D standard. We evaluated our HPLC method so that it can sensitively monitor the molecular integrity for immunoglobulin and albumin products. In collaboration with Dr. B Golding's group, we evaluated the molecular integrity for some lots of Vaccinia Immune Globulin (Human)and their derived IgG subclasses. The purity of several other pending hyperimmune globulin products have also been subjected to this analysis and found satisfactory.
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{
"pile_set_name": "NIH ExPorter"
}
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Opiate drugs affect nervous system development by disrupting endogenous opioid systems (endogenous opioids and opioid receptors). Opiates can profoundly inhibit growth. Despite wide-spread licit and illicit use, the developmental mechanisms of opiate action are not understood. This proposal examines the developmental neurobiology of opiate drugs with abuse liability. Interrelated cell and molecular biological experiments will primarily use primary cultures of highly purified populations of cells from the mouse cerebellum and cerebral cortex to explore how opiates directly regulate brain growth. Growth, and opioid receptor and gene expression, will be studied in identified populations of dividing neuroblasts (cerebellar external granular layer cells) and glia (i.e., astrocytes, oligodendrocytes, and microglia) and their progenitors, in vitro, and to a limited extent in vivo. Alterations in growth will be demonstrated to be opioid receptor specific by showing that they are dose-dependent, antagonist-reversible, and stereospecific. Specific aim 1 will determine the effects of opiates on the generation (proliferation and survival) of identified populations of neural cells. Specific aim 2 will identify which opioid receptor types mediate growth. Growth will be manipulated using prototypic opioid receptor agonists and antagonists for both traditional mu (mu), delta-, and kappa- as well as non-traditional "immature', opioid receptor types and subtypes. Opioid receptors will be localized in developing neural cells using (i) a wipe-count method and radioligand autoradiography, (ii) fluorescent-labeled ligands and scanning laser confocal microscopy, and (iii) in situ hybridization for opioid receptor mRNA. Specific aim 3 will determine whether dividing cells intrinsically express opioids by examining opioid gene expression in identified cells in culture using in situ hybridization and immunocytochemistry. The production of opioids by dividing cells is proposed as an autocrine/paracrine mechanism of growth regulation, and is likely to be an important site for opiate drug action. Determining the direct mechanisms of action is crucial towards understanding the etiology of opiate-dependent abnormalities in neural and behavioral development. Our hypothesis is that opiates inhibit nervous system maturation by affecting the production of neurons and/or glia. The results will obtain a unified picture of how opiate regulate growth, correlating developmental effects with cellular sites of opioid gene and receptor expression. This study will have far- reaching significance towards understanding the basic mechanisms by which opiates regulate nervous system growth, and perhaps towards designing therapeutic interventions to prevent/correct developmental abnormalities in children resulting from maternal drug abuse.
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{
"pile_set_name": "NIH ExPorter"
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HIV and malaria are two of the most important infectious diseases in the world, and both inflict the greatest harm in sub-Saharan Africa. Interactions between these two illnesses are of tremendous public health importance. The greatest burden of malaria is in children, yet surprisingly, the effect of HIV on the incidence of malaria in this population is poorly understood. In both children and adults, the effect of HIV on malaria treatment outcomes is poorly characterized, and the introduction of trimethoprim-sulfamethoxazole (TMP/SMX) prophylaxis for HIV disease raises the possibility that cross-resistance may jeopardize response to first-line malaria treatments. Conversely, little information is available on the impact of malaria on the progression of HIV disease. We propose to investigate the epidemiology of HIV and malaria co-infection through a series of studies in Kampala, Uganda. These studies will be conducted at a time of rapid expansion of antiretroviral (ARV) therapy in Uganda, both allowing us to study the effects of ARV therapy on malaria/HIV co-infection, and ensuring that our findings will be relevant for future HIV care in sub-Saharan Africa. Our aims will be: 1) To determine if HIV-infected children are at increased risk for symptomatic malaria. We will test the hypothesis that HIV infection is associated with an increased incidence of symptomatic malaria among children in Kampala. Secondary aims will be to evaluate the effects of the level of immunosuppression, TMP/SMX prophylaxis, and the use of ARV therapy on malaria incidence. 2) To assess the effect of malaria on the progression of HIV disease. Using a cohort of 300 HIV-infected children, we will test the hypothesis that symptomatic malaria is associated with accelerated progression of HIV disease, as measured by change in CD4 cell count and progression to AIDS and death. 3) To compare the response to therapy for uncomplicated malaria in HIV-infected and uninfected children and adults. We will test the hypothesis that HIV infection is associated with a higher risk of treatment failure following therapy for uncomplicated malaria. 4) To assess the effect of TMP/SMX prophylaxis on the selection of drug-resistant malaria parasites. We will test the hypothesis that the use of TMP/SMX prophylaxis is associated with an increased risk of infection with malaria parasites containing mutations that confer resistance to antifolate anti-malarials. This proposal builds on an established collaboration between UCSF and Makerere University. A major focus of this project will be continued capacity building and technology transfer to Uganda. Specifically, we will establish a research infrastructure to implement comprehensive longitudinal studies in well defined cohorts, create an onsite data collection center, establish a molecular malaria laboratory, and foster training for independent research in Uganda. The work will be performed principally in Uganda, with Ugandans playing major roles. Our long-term goals are to develop effective and sustainable research capabilities in Uganda, to promote the establishment of improved clinical management strategies, and ultimately to improve local and international standards of care for HIV and malaria.
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{
"pile_set_name": "NIH ExPorter"
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The currently proposed project represents a phase I study in humans with metastatic kidney (renal) cancer to evaluate a novel form of gene modified tumor cell vaccine. This study is based on extensive preclinical experiments in animal tumor models for gene targeted immunotherapy. The animal tumor experiments have shown that introducing specific genes into tumor cells that encode stimulating growth factors (termed cytokines) for the immune system enhance the ability of these gene modified tumor cells to activate an immune response in the animal against the original tumor. Specifically, we have found that a particular cytokine gene, GM-CSF, when introduced into the tumor, results in the most effective generation of clinically relevant systemic antitumor immune responses. Using highly efficient and safety tested viruses as a vehicle to transfer genes into the tumor cells, we have been able to genetically modify human cancer cells to produce equivalent levels of the immunostimulatory GM-GSF as has been accomplished in the animal tumor cells. The current human protocol seeks to accomplish specific clinical and laboratory objectives. First, as a phase I protocol, it seeks to determine the maximum tolerated dose of irradiated autologous renal tumor cells, unmodified and after transfer of the human GM-CSF gene. Second, we seek to quantitate the toxicities, if any, of introducing increasing amounts of GM-CSF gene transduced tumor cells. Thirdly, we seek to quantitate the specific antitumor immune responses induced by these vaccination approaches. Fourth, we will seek preliminary evidence of therapeutic activity of the vaccine preparation. This trial is designed with two arms: one arm evaluates escalating doses of GM-CSF transduced tumor cells and the second parallel arm evaluates escalating doses of nontransduced tumor cells. While the primary objective of the two arm trial is to determine whether observed toxicity is due to cell administration of the GM-CSF gene transfer, we will also seek preliminary comparative evidence to determine whether the introduction of the GM-CSF gene into the tumor cells modifies their vaccination potential relative to untransduced tumor cells with regards to either immunologic or clinical antitumor responses.
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{
"pile_set_name": "NIH ExPorter"
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The goal of this project is to develop functional magnetic resonance imaging (fMRI) tools to assist the diagnosis and treatment of human patients with a brain tumor or other operable pathology. The specific focus of this proposal is to produce a practical, clinic-ready suite of MR imaging methods, analyses and display tools to solve the number one impediment to routine use of fMRI for guiding brain surgery and radiation treatment: risk of brain damage due to the treatment itself. Currently, the primary clinical use of fMRI is to identify healthy brain tissue that might be damaged by surgery or radiation treatment and thereby cause an unintended neurological deficit such as partial blindness or paralysis. Neurosurgeons who use fMRI for this purpose have reported that it allows them to be more aggressive in removing the tumor because they don't have to guess where the healthy brain tissue is located. However, the success of using fMRI for this purpose depends on its ability to reliably distinguish between healthy brain tissue and diseased tissue that can be removed without causing a deficit. Herein, lies a critical problem. fMRI signals are not generated by the brain cells themselves but, rather, by localized changes in blood flow and oxygenation that are triggered when the brain cells become active as the patient performs a sensory, motor or cognitive task. The cascade of cellular events that link changes in brain cell activity to changes in blood flow is complex and can be disrupted by a brain tumor or other disease process. Disrupting this cascade causes neurovascular uncoupling (NVU) and results in a localized loss of the fMRI signal even though nearby brain cells are still functional. If NVU is not detected, healthy brain tissue can be mistaken for diseased tissue and inadvertently resected or irradiated. This can result in treatment-induced deficits such as partial loss of vision or limb movement. Fortunately, there are two promising methods that can be used to detect NVU but they have not been fully tested with patients nor have they been developed into tools that are ready for routine clinical use and distribution to the health care community. Consequently, the specific goal of this project is to address this need through a collaborative effort between imaging scientists and physicians at the Medical College of Wisconsin, Johns Hopkins University and Prism Clinical Imaging, Inc. This Phase 1 STTR project will address the feasibility of combining the two most promising methods, testing the combined method with a small number of patients, and developing prototype software for acquisition, analysis and visualization of NVU-related data. Successful completion of this project will lead to a subsequent Phase 2 project that will focus on testing a larger range of patients and pathologies and creating a commercial product ready for release to hospitals and clinics. It is anticipated that the proposed technology will have a significant impact on the use of fMRI for guiding brain surgery and on the acceptance of fMRI as standard of care for this purpose.
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{
"pile_set_name": "NIH ExPorter"
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Several epidemiological studies indicate that cigarette smoking is associated with a significant anti-estrogenic effect, and this fact is related to the recent finding of decreased endometrial cancer among postmenopausal female smokers. Other estrogen-related physiological processes affected include earlier menopause, increased osteoporosis, and possibly, decreased breast cancer among female smokers. Elucidation of the underlying mechanism whereby smoking affects estrogen disposition and pathophysiology may prove useful in developing strategies toward the prevention of endometrial and breast cancer. We have demonstrated a powerful induction of estradiol 2-hydroxylation measured in vivo with the radiometric procedure in premenopausal females smoking at least 15 cigarettes per day, possibly accounting for the decreased estrogen bioavailability at hormone target tissues. Several constituents of cigarette smoke are known to be powerful inducers of the hepatic microsomal P-450 enzymes participating in estradiol metabolism. This laboratory has extensive experience using radiometric procedures for studying the three principal oxidative transformations of estradiol at the C-2, C-16yield, and C-17 position in humans. The C-2 hydroxylation of estradiol is unique among these metabolic steps in that the catecholestrogens formed in the reaction are virtually devoid of peripheral estrogenic activity. Thus, induction in this metabolic pathway would be expected to have a significant anti-estrogenic effect, and may explain the reduced risk of endometrial cancer in smokers. We will use the radiometric method to further explore the effect of smoking on estradiol 2-hydroxylation in the following groups: postmenopausal smokers versus controls; male smokers versus controls; and, female smokers with different levels of tobacco consumption. We will correlate changes in estradiol 2-hydroxylation with serum nicotine (or continine) levels, and attempt to induce changes in estrogen metabolism by administering nicotine transdermally. We will follow esstradiol 2-hydroxylation in female smokers who have stopped smoking for various periods of time. Finally, we will examine other metabolic pathways of estrogen metabolism, including the irreversible C-16yield hydroxylation step, and the aromatization of androstenedione to estrone, for further evidence of induction by cigarette smoking.
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{
"pile_set_name": "NIH ExPorter"
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Summary of Work: Exogenous DNA damaging agents such as cisplatinin, nitrogen mustard, and psoralen create interstrand DNA crosslinks. Such non-coding lesions must be repaired to ensure accurate replication of the genome and viability of the organism. The Drosophila mus308 mutation, which confers hypersensitivity to nitrogen mustard but not the monofunctional agent methyl-methane sulfonate, identified a DNA polymerase likely involved in processing DNA crosslinks. Based on homology to the Drosophila mus308 gene and another Family A DNA polymerase, DNA polymerase g, we cloned and expressed the cDNA for human DNA polymerase q. Amino acid sequence alignments predict DNA polymerase, ATP binding and/or hydrolysis, and 3' to 5' exonuclease functions for this enzyme. To verify these activities and to begin to characterize the role of this polymerase in DNA repair, we have generated a series of point mutations and deletions within the relevant motifs. Mutant and truncated proteins were purified from baculoviral infected insect cells. The substrate specificity and accuracy of DNA synthesis in vitro were determined. Unlike many of the other newly discovered DNA polymerases, polymerase theta synthesizes DNA with high fidelity.
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{
"pile_set_name": "NIH ExPorter"
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Our principal objective is to identify genetic variation that underlies liability to schizophrenia and other psychotic disorders (henceforth called Scz) in the Oceanic population of Palau. We will accomplish this goal by capitalizing on a wide range of scientific expertise and an incredible population sample obtained by enduring relationship between the investigators and the Government of Palau for more than a decade. We now have genealogical information on many thousand individuals, who form a compelling resource for genome-wide linkage and association mapping, and represents complete ascertainment from the Oceanic population of Palau. We propose a genome-wide study of linkage and association for risk to Scz by genotyping 500 individuals for 250,000 Single Nucleotide Polymorphisms (SNPs), using the Affymetrix chip technology. We use these genotypes to search for variation conforming to the conventional hypothesis - that risk to Scz is determined directly by genetic variation carried by affected individuals. We will also use these data to test a novel hypothesis - that initial liability to Scz is generated during fetal development by one or more environmental stressors, but it is facilitated by maternal genetic vulnerability to the stress, so that the placental environment cannot adequately buffer the developing fetus. To effectively search for variation affecting risk at either level (maternal or individual), we propose to genotype essentially all individuals affected by Scz, 250, 150 mothers of individuals diagnosed with Scz, and 100 control individuals. DMA from another 450 individual will be used for follow-up analyses. A rich set of risk maternal risk factors will be collected for this population and used in our analyses. Statistical analyses will target identification of extended haplotypes that are shared much more frequently than expected by chance (in either level of risk.) Identification of such haplotypes will be facilitated by extended linkage disequilibrium characteristic of this and other Oceanic populations. Molecular and fine-mapping studies will identify risk loci. We believe this study is unique among studies of Scz genetics for its population, sample, the research team and the novelty of hypotheses and approach. [unreadable] [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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Therapy for lymphoproliferative disease in immunocompromised patients is problematic. These patients often have a poor response to cytotoxic agents that also worsens their underlying immunosuppression. Nonetheless, there are distinct pathophysiologic features of these tumors that may be exploited as therapeutic targets. These lymphomas are often associated with gamma herpesviruses and dependent upon constitutive expression of NF-kappaB. We have identified a novel, pro-apoptotic therapy for Humanherpes Virus Type 8 (HHV-8), Primary Effusion Lymphoma (PEL) and Epstein Barr Virus (EBV) lymphomas. Azidothymidine (AZT) and Interferon alpha induce death receptor ligand mediated apoptosis in PEL. This occurs through a potent activation of the ligand TRAIL mediated by interferon alpha coupled with suppression of NF-kappaB by AZT. We hypothesize that death receptor signaling is potentiated upon suppression of NF-kappaB dependent anti-apoptotic factors. NF-kappaB blockade is effected by the monophosphate form of AZT which is preferentially generated in HHV-8 and EBV associated lymphomas. We have demonstrated the effectiveness of antiviral therapy for gamma herpesvirus associated lymphomas in both animal models and patients. We propose to investigate the cellular and viral factors that mediate this apoptosis by 1) defining the signal transduction pathways induced by antivirals in herpesvirus lymphomas; 2) determining the role of cellular and viral proteins (such as vFlip) in NF-kappaB mediated blockade of death receptor mediated apoptosis; 3) investigating the role of viral thymidine kinase in the phosphorylation of antiviral thymidine analogues and the initiation of apoptosis and; 4) studying the anti-lymphoma effects of antivirals in a recently developed SCID mouse model. Development of a therapeutic strategy based on antiviral therapy would represent a targeted, biological approach to lymphomas in resource poor settings. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
}
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Several paths of evidence converge in implicating a role for the cholinergic system in the pathophysiology of affective illness. In both unipolar depressed and euthymic bipolar subjects, cholinomimetic drugs (i.e., muscarinic agonists, acetylcholinesterase inhibitors) exacerbate depressive signs and symptoms such as dysphoria, psychomotor retardation, impairment of attention and memory, hypothalamic pituitary adrenal axis hyperactivity and sleep EEG abnormalities (Janowsky et al., 1974; Overstreet, 1993). In healthy subjects, the acetylcholinesterase inhibitor physostigmine elicits a range of depressive symptoms including dysphoria, anergia, psychomotor slowing, emotional lability, sleep disturbances, memory and concentration impairment, and with higher doses, tearfulness and depression. These effects have been shown to reflect stimulation of muscarinic receptors (Davis et al., 1976; Oppenheim et al., 1979; Risch et al., 1981a). Cholinomimetics also exacerbate behavioral despair in putative animal models of depression. Conversely, the anticholinergic agent biperidine improved symptoms of depression in a placebo controlled study (Fleischhacker et al., 1987). Moreover, muscarinic cholinomimetics and a choline rich nutrient, lecithin (phosphatidylcholine) exert antimanic effects in bipolar subjects. [unreadable] [unreadable] Potentially consistent with these observations, depressed subjects exhibit hypersensitivity to cholinomimetic agents. Administration of muscarinic cholinergic agonists, ACh releasing agents or acetylcholinesterase inhibitors induce exaggerated effects on REM density and latency in depressed subjects than in healthy controls (Berger et al., 1983; Gillin et al., 1991a; Nofzinger et al., 1997; Nurnberger et al., 1989). In addition, both manic and depressed bipolar subjects show increased pupillary sensitivity to the muscarinic cholinergic agonist pilocarpine relative to controls (Sokolski and Demet, 1996).[unreadable] [unreadable] Despite the data implicating the mAChR receptor system in mood disorders, no direct in vivo investigations of the central mAChR have been performed in depressed subjects. A novel PET radioligand, 18FFP-TZTP was recently developed by Eckelman (2001a; b) as a selective agonist of M2 receptors. Because the M2 receptor functions predominantly as a presynaptic release-controlling autoreceptor, decreased distribution volume (V) of this receptor could conceivably give rise to increased postsynaptic muscarinic receptor sensitivity. [unreadable] [unreadable] This project conducted a PET study of M2 receptor distribution volume in currently depressed subjects with major depressive disorder, currently depressed subjects with bipolar disorder, and psychiatrically healthy controls, about 25 subjects being studied in the past one year. The results confirmed the central hypothesis that M2 receptor V is decreased in regions where they are primarily located presynaptically in depressed subjects with bipolar disorder relative to healthy controls, namely in the cingulate cortex, insula, corpus callosum, and ventral striatum. These regions have been implicated in other studies as areas where impaired cholinergic regulation may result in abnormal emotional and attentional processing and altered emotional experiences including dysphoria, anxiety and euphoria. As such these data advance knowledge regarding the pathophysiology of depression. The magnitude of the cholinergic receptor abnormality correlated with the emotional salience which these subjects attributed to positively and negatively valenced words. Two scientific manuscripts have been prepared and submitted to describe these findings.[unreadable] [unreadable] During this past year, we demonstrated that this abnormality in bipolar disorder is specifically accounted for by subjects who are homozygous for a mutation of the M2 receptor. We are preparing a manuscript describing this seminal finding.[unreadable] [unreadable] We are now attempting to replicate this finding of the relationship between M2 receptor binding and M2 receptor polymorphism genotype. Moreover, we noted that the bipolar subjects who had this genotype and the associated reduction in M2 receptor binding manifested a more disabling and severe form of bipolar depression. We will thus be characterizing the bipolar disorder subgroup that exhibits this abnormality and genotype to identify clinical and biological correlates of this group. If these findings can be replicated in our expanded sample, we may be able to establish a particularly severe, genetically identified subtype of bipolar disorder. [unreadable] [unreadable] Finally, the genetic sampling from the subjects in this neuroimaging study were used to identify single nucleotide polymorphisms (SNPs) associated with unipolar depression (MDD) in a sample enriched for the likelihood of having genetic liability for MDD based upon having recurrent illness and early age-at illness-onset. The associations discovered in this very-well characterized, enriched MDD sample were then replicated in a larger sample selected more generally according to MDD criteria alone.[unreadable] [unreadable] One genetic marker showed a significant dominance genetic association effect with the MDD diagnosis which also remained significatnt (p<0.004) after correcting for multiple testing. Replication testing in a second, larger sample of healthy controls (n=739) and MDD cases (n=1,915) derived from the STAR*D cohort, confirmed the association with this marker(p<0.02). [unreadable] [unreadable] The marker so implicated mapped to the HTR3C gene, which encodes expression of the serotonin type 3C receptor subunit. The SNP variation (A/G) of this marker is located in the 3 untranslated region, which has a cis/trans control region known to subserve regulatory mechanisms such as translation and transport. Our results suggest that variation in HTR3C is a risk factor for MDD. A paper describing this finding is in the final stages of preparation.
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{
"pile_set_name": "NIH ExPorter"
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Details concerning the process of reverse transcription in mammalian cells newly infected with RNA tumor viruses are essentially unknown. The objectives of this research are to understand the process of RNA to DNA transcription as it occurs in RNA tumor virus-infected cells, with particular emphasis on determining the function and mode of action of RNase H in the process. These objectives will be achieved: 1) by determining the structural, catalytic, and antigenic properties and intravirion location of RNA-directed DNA polymerase-associated RNase H and free RNase H activities purified from mammalian RNA tumor virions, 2) by the use of model substrates synthesized in vitro to elucidate the mode of action of viral RNase H, and 3) by the isolation of a "reverse transcription complex" from newly infected mammalian cells and characterization of the enzymes and viral related nucleic acids associated with and synthesized by the complex.
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{
"pile_set_name": "NIH ExPorter"
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This research continues the investigation of the Navajo Ethno-Medical domain started by Werner. During the last three years this work has come to fruition through the substantive work of Martha A. Austin of the Navajo Health Authority (NEME project) and Oswald Werner's methodological and theoretical contributions. Volume 3 on Marriage, Conception, Pregnancy and Birth (first volume to be compiled) contains over 500 pages in Navajo. Volumes 7. Ts'i7i7hniido7o7, Disease and Sickness, and 8. Ti7di71ht'e7, Injuries are now being outlined. This project is based on the assumption that it is unrealistic to ask non-native anthropologists to make substantive contributions in Navajo. Only native speakers are in full control of the subtleties of their language as is necessary for a comprehensive encyclopaedic undertaking. (Obviously non-natives can learn a language well. The point is that to achieve subtlety, free from 'semantic interference', requires longer residence that is available to most ethnographers.) This research continues the following cooperative arrangement: Two parallel organizations are used. (1) A Navajo (Austin et al.) for substantive contributions in Navajo, and (2) An Anglo (Werner et al.) concerned with principles of theory and method of cultural knowledge systems and their practical presentation. This joint effort will result in two publications: (1) A ten volume 'Navajo Ethno-Medical Encyclopaedia', and (2) the 'Handbook of Ethnoscience: Ethnographies and Encyclopaedias'. The Encyclopaedia is being compiled by ethnoscience techniques, analyzed and organized according to ethnoscience theory, published according to a systematic adaptation of ethnoscience principles to practical presentation (i.e., readability), and will be used for the education of Native American Physicians and other medical personnel within the American Indian School of Medicine at Shiprock, NM. 'The Handbook of Ethnoscience' is codifying ethnoscience methodology for use in anthropological field work whereever a description of a social system or of a system of knowledge is necessary. This project will contribute to better medical communication (translation and interpretation) wherever folk-knowledge meets Western medicine.
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{
"pile_set_name": "NIH ExPorter"
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Muscle strain injuries are one of the most common conditions seen in sports medicine clinics. However, methods of treatment are variable and re-injury rates tend to be high which, in part, reflects a lack of fundamental understanding of the factors that influence injury risk. The prevailing theory is that injury occurs as a result of excessive strain of active muscle fibers. The goal of this study is to develop novel biocomputational tools to predict and analyze the strain distributions within skeletal muscles during movements associated with injury. Model predictions will be compared with strain measures obtained using state-of-the-art dynamic magnetic resonance imaging experiments. Once validated, we will use the biocomputational tools to investigate how morphology and coordination influence hamstring injury risk during running. Following are the specific aims. Aim 1 will use a dynamic magnetic resonance imaging technique to measure the strain distributions within the individual hamstring muscles during lengthening contractions, a loading condition commonly associated with injury. Comparisons between muscles will provide new insights into the propensity for hamstring injury to occur in the biceps femoris long head. Aim 2 will build a biocomputational framework to predict muscle strain distributions during movement. The framework will couple finite-element simulations of muscle tissue behavior with dynamic simulations of whole body movement. The methods will be validated by comparing strain predictions with those determined from the dynamic images in Aim 1. We will then use the framework to investigate the relationship between muscle excitations, hamstring tissue strains and skeletal movement during running. Aim 3 will evaluate whether computational models predict re-injury prevention strategies. We will build and validate models of subjects who exhibit residual changes in tissue structures as a result of a previous hamstring injury. We will then use the software framework to identify how movement coordination can be adapted to accommodate injury-induced changes in morphology. This research will establish a biocomputational framework that reveals the complex relationship between muscle morphology, coordination and injury risk, thus providing a new paradigm for identifying rehabilitation and injury prevention strategies. Muscle strain injuries are one of the most common conditions seen in sports medicine clinics. However, methods of treating muscle injuries are variable and re-injury rates tend to be high. This proposal couples novel biocomputational tools and imaging techniques to establish a scientific basis for preventing and rehabilitating hamstring muscle injuries.
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{
"pile_set_name": "NIH ExPorter"
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Muscle wasting is a serious complication of chronic renal failure (CRF) identified in virtually every survey of dialysis patients, and it is a strong predictor of morbidity and mortality. Although the mechanisms of CRF-induced muscle wasting have been intensively investigated, there are still many unknown aspects of the pathological processes. Our long-term goal is to identify cellular mechanisms of muscle wasting in chronic renal failure and to develop new therapeutic strategies. It has been known that the ubiquitin-proteasome proteolysis is one of important pathways in muscle protein degradation. The process of the proteolysis can be activated by uremia, a low extra-cellular pH, elevated glucocorticoid levels and inadequate insulin action. In our preliminary studies, we have observed that caspase-3, a protease activated by apoptotic stimuli, plays an initiatory role in muscle wasting; and inactivation of caspase-3 prevents the muscle wasting. X-chromosome linked inhibitor of apoptosis protein (XlAP) is an endogenous caspase inhibitor that regulates the function of the caspase signaling pathway. Over-expression of XlAP in L6 skeletal muscle cells can block muscle protein degradation induced by caspase-3 activation. Thus, we hypothesize that increasing of XlAP cellular function will block caspase-3 and CRF-induced muscle wasting, and that augmentation of XlAP cellular function will exert a therapeutic effect on CRF-induced muscle wasting. To examine our hypotheses, we propose: 1) to test if XlAP blocks muscle protein degradation induced by growth factor withdraw and staurosporine (activating caspase-3) in human skeletal muscle cells; 2) to examine if XlAP can inhibit the CRF-induced muscle protein degradation in vivo using a transgenic mouse with muscle specific XlAP expression; 3) to examine if the intramuscular injection of the recombinant XlAP lentivirus (Len-XlAP) prevents CRF-induced muscle protein degradation in mice. We believe that the experimental results will shed the insight on the mechanisms of muscle protein degradation, and provide a possible new approach to treat the pathological process of muscle wasting.
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{
"pile_set_name": "NIH ExPorter"
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Determine 1) the relationship between sleep stage and GH secretion in acromegaly; 2) whether spontaneous pulsatile and GHRP-stimulated GH secretion is normal for age when acromegalics are in remission; and 3) whether predictors for recurrence can be identified from these results.
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{
"pile_set_name": "NIH ExPorter"
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The long-range objective is to understand enough about the factors that control mammalian cell growth to be able to apply this knowledge to the control of malignant growth. As a result of recent work, a broad picture is now available of the growth control of mammalian cells. Application of this knowledge requires additional information and experimentation. For example, detailed information is needed of the specific factors that control the growth of the epithelial cells that are most important in the origin of malignant tumors, in order to design rational procedures to control these tumors. Among other things, it is important to know whether growth inhibitors such as a growth inhibitor produced by kidney epithelial cells, can be used to restrict tumor growth. The kidney epithelial cell growth inhibitor is active on certain lung and mammary cells in culture. With CCL64 mink lung cells, for example, 60% inhibition of [[unreadable]3[unreadable]H]thymidine incorporation is observed at a 0.1-nanogram/ml concentration of the growth inhibitor. Injection of the growth inhibitor in vivo into human mammary carcinomas growing in nude mice inhibits [[unreadable]3[unreadable]H]thymidine incorporation into the tumors. Recently, this kidney epithelial cell growth inhibitor has been found to be very similar to, or identical with, transforming growth factor beta. Each of these substances is active in the assays used for the other, and their general properties are identical. These results indicate that the same substance can be either an extremely active growth stimulator or an extremely active growth inhibitor, depending on the cells and assay conditions. The relevance of this in vivo and an understanding of the in vivo role of this growth inhibitor/transforming growth factor beta will be of major interest. (J)
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{
"pile_set_name": "NIH ExPorter"
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PROJECT I - SUMMARY/ABSTRACT Ischemia-reperfusion injury (IRI) remains the primary obstacle limiting the success of orthotopic liver transplantation (OLT) in patients with end-stage liver disease and those with tumors of hepatic origin. Our group has pioneered the concept that hepatic IRI requires activated CD4+ T cells to facilitate liver tissue damage. T cell immunoglobulin-3 (TIM-3; encoded by Havcr2 gene) is the central negative regulator of T cell activation. CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1; encoded by CC1 gene) has been identified as a new cellular ligand determining TIM-3 function. First, we found that compared with CEACAM1 proficient (WT) livers, CEACAM1 null-mutation (CC1-/-) exacerbated IRI in OLT. Second, we discovered that the benefit of recipient CD4+TIM-3+ signaling in IR-stressed OLT (WT?TIM-3Tg) was completely lost when CEACAM1 KO mice served as organ donors (CC1-/-?TIM-3Tg). These preliminary data have led us to central hypothesis that 1/ TIM-3 ? CEACAM1 negative regulation is essential to control IRI by imposing exhaustion-like dysfunction in OLT-infiltrating CD4+ T cells; and 2/ CEACAM1 in the donor liver promotes hepatoprotection. Project I will test this hypothesis through two interlocked specific aims: Aim 1: Define mechanisms of TIM-3 ? CEACAM1 negative T cell regulation in IR-stressed iso-OLT. A panel of mice available to us for Aim 1 experiments include CD4+ T cell mutants, which are: i/ CEACAM1Tg; ii/ double TIM-3Tg and CEACAM1-/-; as well as: iii/ TIM-3Tg and TIM-3-/- mice. Aim 1.1. Hypothesis: CEACAM1 - TIM-3 signaling on host circulating CD4+ T cells promotes exhaustion- type phenotype in IRI?OLT. Aim 1.2. Hypothesis: Under dominant CAECAM1 signaling, TIM-3+CD4+ exhausted T cells inhibit the development and progression of IRI in OLT. Aim 2: Define mechanisms by which hepatocellular CEACAM1 in donor liver regulates IRI in iso-OLT. Gene- targeted strains for Aim 2 studies include: i/ hepatic CEACAM1 inactivation (loss-of- function; L-CC1-/-); or ii/ forced hepatic CEACAM1 overexpression (gain-of-function; L-CC1Tg). Aim 2.1. Hypothesis: Enhancement of hepatocyte-specific CEACAM1 ? ?-catenin regenerative functions facilitates hepatoprotection. Aim 2.2. Hypothesis: TIM-3 ? CECACAM1 signaling enhances hepatocyte autophagy program. Integration with PPG: By providing novel insights into TIM-3 ? CEACAM1 checkpoint regulation at the innate ? adaptive immune interface in IR-stressed iso-OLT, Project I naturally informs/precedes studies assessing how host rejection regulates innate immune activation/IRI sequel in allo-OLT (Project II). Direct application of approaches blunting inflammation while promoting hepatoprotection in mouse OLT models will accelerate assessments of immune phenotypes in human liver transplants (Project III).
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{
"pile_set_name": "NIH ExPorter"
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DESCRIPTION (Applicant's Abstract):Cigarette smoking is an enormous public health problem. Although our understanding of the consequences of smoking is relatively advanced, our knowledge of the determinants of smoking cessation- and its frequent concomitant relapse-is far less complete. Enduring smoking cessation remains an elusive goal for most heavy smokers. The extraordinary addictiveness of nicotine is certainly a prominent cause of these difficulties, but cannot of itself explain the marked inter-individual differences in smoking cessation and relapse. The overarching goal of this proposal is to delineate etiological factors involved in smoking cessation and relapse. To accomplish this aim, we will: (I) obtain smoking history data on 40,000 individuals (20,000 twin pairs) from an existing twin-family registry based in Virginia, North Carolina, and South Carolina to (II) identify 16,000 individuals who are lifetime regular smokers (-62 percent current, -28 percent past) and obtain a comprehensive baseline set of predictors and correlates of smoking cessation and relapse; and (III) follow the smoking status of these lifetime regular smokers prospectively at yearly intervals for three years to identify smoking cessation and relapse events. These efforts will yield complimentary prospective and retrospective data sets. Analysis of these data sets will allow us: to describe changes in smoking behavior over time; to identify the predictors of progression in readiness to change, persistent reduction in amount smoked, serious attempts at smoking cessation, successful smoking cessation, and smoking relapse; to use multivariate classification techniques to derive a typology applicable to individual smokers that identifies more homogeneous subgroups; to model the smoking process (i.e., smoking initiation-regular smoking-smoking cessation OR smoking initiation-cessation-relapse) to determine the magnitude and overlap of the genetic and environmental sources of variation in the processes; and to investigate the co-variation of the etiological genetic and environmental sources of variation in these processes with measures of personality, self-esteem, self-efficacy, and psychopathology. Accomplishing the aims of this proposal will advance significantly our knowledge of the determinants of smoking cessation and relapse The use of a twin design, a large/powerful sample, multivariate assessment of the key phenotypes, and advanced analytic methods are key components of our design and approach.
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{
"pile_set_name": "NIH ExPorter"
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The ability to isolate and concentrate rare cells or pathogens is fundamental to numerous biological and medical applications, often forming the starting point for down stream bioassays. Current approaches for cell isolation include centrifugation, filtration, fluorescent- and magnetic-activated cell sorting, and dielectrophoretic (DEP) cell separation. Among them, the DEP separator provides the advantages of speed, flexibility, controllability, and ease of application to automation. However, conventional DEP cell separators, containing two-dimensional (2-D) planar electrode arrays, often cannot process large volumes of samples at a time that is essential for rare cell or pathogen separation. A new three-dimensional (3-D) DEP cell/pathogen separation system is proposed, which is capable of processing large volume of samples with significantly improved collection efficiency. As examples of the relevance of the new system to clinical oncology, infectious disease and biowarfare, human monocytic (U937) and cervic carcinoma (HeLa) cells will be separated from blood, Escherichia coli and Salmonella typhimudum bacteria will be separated from blood, and Bacillus globigii will be concentrated from water. There examples address the needs of lowering the threshold for detection of cancer cells in blood, with the goal of diagnosing infectious disease and biowarfare agents without the need to first culture the pathogens.
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{
"pile_set_name": "NIH ExPorter"
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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Center for Biomedical and Bioorganic Mass Spectrometry provides mass spectral service for academic customers around the United States. We can ionize samples by electron-impact (EI) ionization, chemical ionization (CI), electrospray ionization (ESI), fast atom bombardment (FAB [unreadable]retired this year) ionization, and matrix assisted laser desorption ionization (MALDI). We have analyzers based on magnetic sector, time-of-flight, ion trap, Q-TOF, FT-ICR, and FT-orbitrap technologies. We provide both nominal (LR), accurate mass (HR, 5ppm) and higher accurate mass (SHR, <1ppm) spectra to our customers as needed. We can also provide LC/MS and LC/MS/MS services in conjunction with ESI. In addition we have the computational tools needed to deconvolute the multiply charged ions in protein spectra. This provides the center with the capability to mass analyze samples with molecular weights ranging from small molecules all the way up to tens of kilodaltons.
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{
"pile_set_name": "NIH ExPorter"
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Some of the most fundamental concerns for health services research in the alcohol field revolve around understanding the long-term course of problem drinking, and how services, including alcohol treatment, other health and human services, and self-help impact that course. The proposed study identifies the determinants of 7-year time paths of consumption patterns and changing alcohol-related problems in large samples of treated and untreated problem drinkers drawn from the same community. Time paths, or "trajectories" of problem drinking are of both theoretical and applied relevance to the alcohol field, where drinking problems are increasingly viewed as chronic, cyclical and relapsing. The study addresses the underlying roles that a wide spectrum of health and human services play in the long-term course of alcohol problems -- in getting better, staying the same, or progressing to more serious problems over time. The study follows a large, representative sample of problem drinkers presenting to agencies throughout a single county's public and private treatment system (N=927) over 7 years. It similarly follows a large comparison group of untreated problem drinkers drawn from that same county's household population (N=672). Analyses draw on latent curve analysis techniques to compare the shape of trajectories of problem drinking, and their determinants , in treated and untreated samples. Analyses begin by establishing the roles played by individual characteristics (including markers of genetic and developmental predispositions towards alcoholism, demographic characteristics, maturation effects, and psychological vulnerabilities, including psychiatric severity, drug use and dependence) on trajectories of problem drinking in the two groups. Next, they examine the roles that social network responses and self-help play in altering the 7-year course of alcohol problems in treated and untreated problem drinkers, and the role of general health and welfare health and human services as well. A final set of analyses builds on upon these earlier stages by modeling the persistence and cumulation of specialty alcohol treatment interventions on trajectories of problem drinking. It then develops more integrated causal explanations of the roles that social networks, self-help, and interventions by general health and welfare service providers play in contributing to the long-term effects of alcohol treatment.
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{
"pile_set_name": "NIH ExPorter"
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The ?4 subunit of the ?6?4 integrin, which forms hemidesmosomes in quiescent normal cells, becomes phosphorylated in breast tumor cells that overexpress HER2 or EGFR. This phosphorylation converts the cytoplasmic domain of the integrin into a signaling scaffold that drives cell invasion, proliferation and survival. HER2 and EGFR are expressed in HER2+/ER- breast cancer, and EGFR is overexpressed in the triple- negative (HER2-,ER-,PR-) subtype. Both cancers are highly aggressive and resist treatments currently available in the clinic. Because these cancer types often overexpress the ?6?4 integrin as well, we have now examined their dependence on signaling from these receptor complexes. We have discovered that these signaling mechanisms are essential for the growth and survival on the cancer cells, and that their signaling requires the assembly of the integrin and HER2 or EGFR with syndecans, another family of matrix receptors. Indeed, syndecan-1 appears necessary for signaling by HER2??6?4, and syndecan-4 appears to be required by EGFR??6?4. Our goal is to define the molecular details of syndecan assembly with these signaling complexes, develop mutants and blocking peptides that disrupt the organizing function of these two syndecans, and test the mutants and peptides in tumor growth, angiogenesis and the activity of cancer stem cells in animal models of HER2+ and TN breast cancer. The outcome of this work will provide potential insight into the development of new therapeutics to target HER2+ and TN breast cancer.
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{
"pile_set_name": "NIH ExPorter"
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The consortium between the Massachusetts General Hospital and the Indira Gandhi Government Medical College Nagpur, India, affiliated with Lata Medical Research Foundation, has extensive experience in conducting international multicenter collaborative clinical trials/implementation science research dedicated to improving maternal and child health. Since 2008, the consortium has participated as a Research Unit (RU) in the Global Network (GN) for Women's and Children's Health Research and is applying to continue the productive and successful RU in Nagpur, India. The RU's goal is to continue to develop and implement GN sponsored common protocols that generate evidence and scientific knowledge to sustainably improve outcomes for women and children. The research that the MGH-Nagpur consortium proposes to develop is based on their current GN studies on the adverse effects of Household Air Pollution (HAP) on pregnancy and infant outcomes. Their GN research is focused in this area because 3 billion people (~ half of the world's population) and -80% of pregnant women in the GN still use traditional cook stoves and solid fuels that generate high levels of household pollutants, far exceeding World Health Organization (WHO) indoor air quality guidelines. WHO estimates that HAP causes 2 million deaths annually (mostly in women and children). They propose to develop a multicenter randomized clinical trial to evaluate whether pregnancy and infant outcomes can be improved by introduction of cook stoves/fuel that reduce HAP in households where pregnant women, in their first trimester, are using traditional cook stoves. The primary outcome is birth weight. Secondary outcomes are preterm birth, perinatal/neonatal mortality, childhood pneumonia/neonatal sepsis and adverse child growth/development. They also propose to measure HAP levels in women and children in a random subset of households to improve knowledge about the biological basis of the effects of HAP - including particulate matter < 2.5 micrometers, carbon monoxide and biomarkers 1-hydroxypyrene, methoxyphenol and cotinine (tobacco smoke). Successful introduction of cook stoves/fuel that reduce HAP has the potential to sustainably improve pregnancy outcomes and child health in resource limited settings.
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{
"pile_set_name": "NIH ExPorter"
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The list of neurological diseases is extensive: Alzheimer's, Parkinson's, ALS, and MS, to name just a few. The goal of this work is to understand how neurons work at the molecular level. Neurons communicate with each other by releasing neurotransmitters through the plasma membrane (PM) into the synapse in a process called exocytosis. The neurotransmitter is stored in small secretory vesicles (~50 nm diameter) docked at the PM. When a neuron depolarizes, an influx of Ca2+ to the cytosolic side of the PM causes Ca2+- triggered exocytosis in less than 1 ms. Release involves fusion of the lipid bilayer of the vesicle with that of the PM. The identity of many key components of this vesicle fusion machinery is established. However, little is known at the molecular level about how the components work together to carry out exocytosis. A central component is the trans-SNARE complex, comprising the v-SNARE protein synaptobrevin (Syb) anchored in the vesicle and the t-SNARE proteins syntaxin (Syx) and SNAP-25 anchored in the PM. Synaptotagmin (Syt), also anchored in the vesicle, is probably the Ca2+ sensor that evidently triggers fusion by altering binding relationships among proteins and lipids. [unreadable] [unreadable] The fusion machinery is so complex that mechanistic inferences drawn from in vivo studies are necessarily indirect. We and others have been working to develop a reconstituted model system that faithfully captures the Ca2+-triggered fusion observed in neurons. Such a system would enable very direct study of many key mechanistic questions by adding or subtracting components one by one. We now have a model that allows direct observation of single v-SNARE vesicle docking and fusion on a planar t-SNARE/lipid bilayer in real time by widefield fluorescence microscopy. Ours is the only in vitro system thus far that exhibits SNARE-dependent fusion on a 25-ms time scale, approaching that in nature. The goals of this work are: to determine the intrinsic SNARE-driven rate of fusion by optimizing protein and lipid components; to learn about the nature of the fusion pore itself by measuring the contents release and lipid mixing time scales with 1-3 ms resolution; and to recapture Ca2+ triggering by introducing Syt into the assay. Along the way, these experiments are poised to answer a wide variety of mechanistic questions in an unusually incisive manner. [unreadable] [unreadable]
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{
"pile_set_name": "NIH ExPorter"
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The Sylvester Cancer Center (Sylvester) is a membership-based, matrix organization which brings together 121 members and 133 affiliate faculty investigators from 18 different departments in two different schools of the University of Miami to conduct multidisciplinary cancer research. Interdisciplinary translational research is facilitated through the center's seven multidisciplinary research programs (MRPs). Total NCI funding is currently $19,297,201 million for 70 projects. Continuing development of Sylvester's Viral Oncology and Bio-behavioral Oncology MRPs are among the highest priorities of the Center. Each address unique community needs through cohesive research themes that take advantage of faculty strengths. Both are relatively small programs which have identified critical gaps that can be best filled through recruitment. We are proposing two candidates for recruitment - one into each of these Programs. The investigators named in this application were identified following strategic planning at the Center's membership retreat in the fall of last year, and had already made initial visits to the Center when recruiting was suspended due to financial challenges in the University. Fortunately (for Sylvester) they are still available and highly interested. Detailed plans for development of their research have been completed as has a plan for evaluation of overall success. The Sylvester research budget is highly dependent on fundraising, endowment, institutional support, and State funding. All four of these sources of support have been adversely impacted by the current recession. Funding of this P30 would provide critical support for strengthening two of the Center's seven MRPs. As a regional resource, the Center has a direct impact on a primary catchment area that includes 5.4 million people living along 125 miles of coastline in the three counties of southeast Florida. Because of its tourist oriented and growth based economy, this area has been particularly hard hit by the current recession. Sylvester is also highly interactive within the School of Medicine. All faculty recruitment is done jointly with the most appropriate academic department. Collaborations with other centers and institutes also abound, including the Interdisciplinary Stem Cell Institute, the Miami Institute for Human Genomics and the Miami CTSA initiative. It follows that enhancing these two MRPs will strengthen Sylvester and this Academic Health Science Center.
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{
"pile_set_name": "NIH ExPorter"
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The cornea, the outermost window of our visual system, is vulnerable to various types of infections and diseases. Corneal disease is one of the leading causes of visual deficiency and blindness.!There are nearly 5 million bilaterally corneal blind persons worldwide, and an estimated 23 million people affected by unilateral corneal blindness globally. In a conservative estimate, corneal diseases affect nearly 300,000 people in the United States, with Fuchs? dystrophy affecting 4% of people aged over 40, and 1 in 2,000 Americans affected by keratoconus. Since many corneal diseases can only be distinguished at the micrometer scale, there is a strong need for imaging tools that can noninvasively visualize the cellular changes in the cornea in vivo, to assist clinical diagnosis, evaluate progression of diseases, and treatment. We have developed a 3D high-definition imaging instrument based on Gabor-Domain Optical Coherence Microscopy (GD-OCM) for noninvasive visualization and quantitative characterization of the cornea over a large field of view. Our data suggest that GD-OCM has the following key advantages over existing in vivo corneal imaging techniques, which include optical coherence tomography (OCT) and confocal microscopy: 1) 6-10x increase in field of view compared to existing confocal microscopes used in clinics worldwide ? this will lead to more accurate qualification of the corneal tissue, since a larger area can be assessed; 2) non-contact operation, unlike confocal microscopes, greatly reducing patient discomfort and eliminating risks of corneal damage; 3) high-definition cross-sectional imaging at an order of magnitude better lateral resolution than OCT; 4) 3D imaging capability at the cellular level over the entire thickness of the cornea? this will enable advances in understanding the progression of the diseases. We hypothesize that GD-OCM can accurately conduct all the assessments currently performed with OCT and confocal microscopy, and additionally provide 3D visualization of the cornea structures over the full depth of the cornea for additional tissue characterization and diagnostic aid. In addition to aiding early diagnosis of corneal disease, the proposed GD-OCM instrument will aid in pre- and post-operative screening for refractive surgery, transplant, drug therapies, monitoring wound healing, corneal nerve assessment, and prevention of stromal rejection. We envision that in the future the GD-OCM instrument enabled by this Phase I SBIR proposal will provide the early foundation for an image-guidance method to assist clinicians in the assessment and treatment of corneal diseases and other diseases affecting the anterior segment of the eye, including diabetes and glaucoma.
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{
"pile_set_name": "NIH ExPorter"
}
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Cytomegalovirus (CMV) is the most common recognized cause of viral-induced cogenital malformations, accounting for neurologic injury in more than 5,000 infants each year in the United States alone. We have reported a CMV-specific lymphocyte profliferation defect in infants actively excreting CMV, and suggest that these infants provide a unique model for investigation of mechanisms of immune recognition and immune responsiveness to viral infaction. The immune response to CMV likely represents a complex process involving macrophages, effector T and B lymphocytes bearing specific receptors, and non-effector lymphocytes responsible for the regulation of T and B cell reactivity. Our initial investigations will therefore concentrate on the enumeration of mononuclear cell subsets in infants with active CMV infection and characterization of T and B effector functions in these infants using assays of lymphocyte proliferation, lymphokine production, cytotoxicity, and in vitro antibody synthesis. The specific role of macrophage-T cell interaction in the immune response to CMV will then be studied using purified cell populations from patients and HLA-D related and unrelated control donors. The role of the major histocompatibility complex in genetic restriction and/or regulation of the immune response will then be studied using established fibroblast cell lines from patients and HLA-A and/or B related and unrelated donors in a cytotoxicity assay. Knowledge of cellular immunity to CMV may eventually lead to prevention of primary infection of susceptible individuals and successful treatment of patients clinically affected by the virus.
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{
"pile_set_name": "NIH ExPorter"
}
|
PROJECT SUMMARY Chronic pain occurs frequently after burn injury and spinal cord injury (SCI). Currently available treatments are often ineffective or only partially effective. Relatively little is known about the mechanisms responsible for the onset and persistence of pain following burn injury, despite chronic pain being a frequent complaint of burn- injured and SCI patients. Our goal is to identify and characterize cellular and molecular mechanisms that contribute to pain following burn and spinal cord injuries, with the objective of delineating specific targets for more effective pain management. Our recent progress includes development of a burn injury model in rats that produces long-lasting mechanical allodynia associated with hyperexcitability of spinal cord dorsal horn (DH) neurons, and demonstration that pain and DH hyperexcitability occur concomitant with activation of DH microglia, with the spread of pain paralleled by spreading microglial activation in this model. In addition, we have shown that acute early inhibition of microglial activation attenuates burn-induced mechanical allodynia and DH neuronal hyperexcitability. We have also demonstrated that the maintenance of below-level pain following SCI is associated with activation of microglia, and that some microglial functions are regulated by Na channels and can be attenuated with Na channel blockade. Recently, we have also shown that activated polymorphonuclear neutrophils significantly increase excitability of DRG neurons, as manifested by lowered threshold and increased firing frequency. Our preliminary data indicate that macrophages infiltrate DRG following burn injury and SCI, at a time when pain behavior is evident. We have also shown hyperexcitability in DH neurons in vivo in conjunction with activation of DH microglia following burn injury and SCI. We now plan to build upon our progress, via the following specific aims. 1. Elucidate the molecular changes in DRG and DH neurons, and glia, following burn injury, and examine novel pharmacotherapeutic approaches to pain following burn injury. 2. Investigate the effect of Na channel blockade on the activity of microglia following SCI, and determine whether channel blockade can reduce microglial activation and attenuate pain behavior. 3. Examine if infiltration of macrophages into at- and/or below-level DRG is associated with the development and/or persistence of neuropathic pain following SCI. 4. Determine whether Na channel blockade attenuates macrophage infiltration into DRG following SCI and whether neutralizing TNF-a reduce macrophage infiltration into DRG following SCI. 5. Investigate the effects of macrophages and microglia on the excitability of DRG and DH neurons, and determine whether DRG neuron hyperexcitability following activation of these cells can be prevented via neutralization of cytokines which are expressed by immune cells. PUBLIC HEALTH RELEVANCE: There is a very substantial need, within the VA and within the U.S. population in general, for more effective treatments for pain associated with burn injury and spinal cord injuries. Relatively little is known of the mechanisms responsible for the onset and persistence of pain following burn injury despite chronic pain being the most frequent complaint of burn-injured patients. Chronic pain and dysethesiae that develop following burn injury, nerve and spinal cord injuries are often resistant to conventional therapeutic approaches. These forms of pain represent significant therapeutic challenges, often being unresponsive or only partially responsive, to existing therapies. Our goal is to identify and characterize cellular and molecular mechanisms that contribute to pain following burn and spinal cord injuries, with the objective of delineating specific targets for more effective pain management.
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{
"pile_set_name": "NIH ExPorter"
}
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The long-term goal of this project is to explore the mechanisms underlying the progression of transient to persistent B Lineage Clonal Expansions (BLCE) in middle-aged mice and the progression of persistent BLCE to late-life B cell neoplasms. We hypothesize that an ordered progression of genetic alterations underlies this age-associated progression. Our first aim is to isolate antigen-specific BLCE prior to the appearance of antigen-specific serum monoclonal immunoglobulin (mlg) and antigen-specific clonal BM plasma cells after the appearance of antigen-specific mlg or diffuse large cell lymphoma (DLCL) from DNP-human serum albumin (DNP-HSA)- or hen egg lysozyme (HEL)-immunized C57/BL/6 mice. The transient or stable antigen-specific BLCE will be isolated from cryopreserved, spleen cells obtained by partial splenectomy of 4-6- and 14-18-month-old mice. BM plasma cells secreting antigen-specific mIg will be isolated from bone marrow obtained from mice sacrificed at 19-months of age with antigen-specific mIg. The antigen-specific cells will be stained with fluorescent-labeled antigen. Results obtained in immunized mice will be confirmed by isolating spontaneous, stable BLCE, bone marrow plasma cell-secreting mIg, or DLCL. Spontaneous BLCE will be isolated from cryopreserved spleen cells obtained prior to the development of mIg or DLCL by their surface or intracellular binding of fluorescent-labeled anti-clonotypic antibody. The link between antigen-induced or spontaneous BLCE and neoplastic B cells will be established by identifying signature IgH/IgL CDR3 sequences of the neoplastic cells and their stable BLCE precursors by single-cell RT-PCR. Our second aim is to develop a genetic roadmap defining the progression of BLCE into late-life B-cell neoplasms. We shall define the genetic abnormalities that accumulate during the transformation of transient to stable BLCE and stable BLCE to mIg-secreting bone marrow plasma cells and/or to DLCL. BLCE isolated from mice prior to the development of B cell neoplasms will be analyzed for the presence somatic hypermutation of proto-oncogenes by single cell RT-PCR, for chromosomal translocations and aneuploidy by FISH and for proto-oncogene expression by quantitative RT-PCR.
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{
"pile_set_name": "NIH ExPorter"
}
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This study will evaluate the effectiveness of a unique comprehensive intervention program for families with in-utero cocaine-exposed infants. The Family Rehabilitation Program (FRP), newly funded by New York City, combines intensive social services with guaranteed drug abuse treatment slots for mothers at 28 community-based programs city-wide. Special efforts are made to involve extended family members and significant male figures in case management. FRP is designed to meet the special service needs of mainly minority-group, cocaine/crack-dependent women with drug- exposed infants and young children who have been largely excluded from the traditional drug abuse treatment system. The objectives of the study are: 1. To conduct an outcome evaluation of the effectiveness of FRP, focusing on: retention in the program; reduction of parental drug abuse; improvement of parents' social and psychological adjustment; prevention of foster care; improvement of infant's/children's physical and emotional care; reduction of re-reports for child abuse/neglect. 2. To conduct a process evaluation of FRP that will describe the case management and drug treatment process for FRP participants. 3. To determine predictors of differential outcomes, including both clients background and admission variables and during-treatment variables. 4. To examine how and to what extent FRP programs are involving significant male figures in case management and services, and to assess the impact of male involvement on outcomes for families. 5. To determine the impact of differences in program and treatment climates on outcomes for FRP families. A 15-month, prospective longitudinal cohort study of all families admitted to FRP during a one year period (N=777) will be conducted, using data on family admission characteristics, social service provision, drug abuse treatment, and case outcomes, available from existing reporting systems and instruments completed by service providers. A subsample of 250 mothers and 125 significant male figures will be studied in greater detail, through personal interviews conducted at admission and a 15 month follow-up. Instruments used will be: Addiction Severity Index, Parenting Stress Index, Family Risk Scales, Bates Scales of Infant Temperament, Beck Depression Inventory, and psychological scales of treatment readiness and self-esteem. The process study will be based on staff interviews, observation of program operations, and treatment climate scales (COPES).
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{
"pile_set_name": "NIH ExPorter"
}
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This proposal describes studies to specifically determine which blood elements in the coagulation cascade are activated by surface contact in extracorporeal perfusion systems. We will quantitate the initial changes in coagulation proteins of the intrinsic coagulation system caused by surface contact. We will study platelet membrane receptors and secreted proteins to determine the mechanism of platelet dysfunction caused by surface contact. We will define the activation and release of procoagulant substances by white cells. We will study specific inhibitors such as corn inhibitor, kininogen antibody, prostacyclin and lidocaine as well as ionically bonded herparin in an effort to selectively inhibit coagulation within extracorporeal perfusion systems without systemic herparin. Development of means selectively inhibit coagulation within extracorporeal perfusion systems will permit coagulation in wounds during perfusion, reduce morbidity due to released platelet and white cell granules and greatly extend clinical applications of extracorporeal perfusion systems for intermediate-term circulatory and respiratory support.
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{
"pile_set_name": "NIH ExPorter"
}
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PROPOSAL (Adapted from the applicant's abstract): Intractable childhood epilepsy remains difficult to treat and results in significant morbidity and cost to the individual, the family and society. With the KD, more than 50% of patients achieve clinically significant response. Despite the recent development of new antiepileptic drugs, the KD continues to play a prominent role in the treatment of intractable childhood epilepsy. The implementation and use of the KD is cumbersome and little is known about the effect of this highly restrictive diet on nutritional status. Moreover, there is limited information regarding early clinical predictors of KD response. This project will explore whether the KD implementation can be simplified without loss of efficacy, examine its impact on several key nutritional parameters, and investigate the usefulness of the electroencephalogram (EEG) as a predictor of the subjects response to the KD. In Phase 1, the current practices for initiation of the KD and its efficacy (defined as more than 50% reduction of seizure frequency at the three-month interval compared to baseline) will be investigated. The investigators propose that a gradual introduction of the KD, without a 24- to 48-hour preceding fast (Grad-KD), is as safe and efficacious as the currently recommended initiation of the KD (CR-KD) with fast. Safety of the two protocols will be evaluated during a six-day inpatient admission and the first three months of exposure. Efficacy will be determined at three months. Baseline and one-month changes in the routine EEG will be examined simultaneously. The predictive power of the changes between the baseline and one-month EEG for the three-month response (defined above) will be shown in those EEG records that have diminished background slowing or reduction of potentially epileptogenic discharges. In a pilot study, they will investigate the mechanistic hypothesis that the antiepileptic properties of the KD is due to an increase in CNS-gamma amino butyric acid (GABA), as measured by magnetic resonance spectroscopy (MRS) at baseline, 0.5-month and three-month in the CR- KD therapy. They will also examine the association between changes in CNS- GABA at 0.5 month and response to KD therapy (defined above). In Phase 2, subjects who have demonstrated a positive response at three months will be followed. The maintenance KD in responders on two caloric intakes will be evaluated. The currently recommended KD is hypocaloric, providing only 75% of subject's caloric needs, and does not allow any weight gain for the 12 months duration of treatment. The necessity of semi-starvation to maintain a positive clinical response has not been evaluated scientifically. They will examine if a eucaloric KD (Eu-KD), with adequate calories for the subject allowing for normal weight gain, is as effective as the currently recommended hypocaloric KD (Hypo-KD) with no weight gain or loss over 12 months. As part of this evaluation, comparisons of two methods used to estimate caloric needs, recommended daily allowance (RDA) tables and resting energy expenditure (by indirect calorimetry), will be made. In addition, the effect of the KD on several nutritional parameters including anthropometry, body composition and bone mineral status, will be evaluated, allowing the 12-month impact of the Eu-KD versus Hypo-KD diets on nutritional status to be compared.
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{
"pile_set_name": "NIH ExPorter"
}
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This project involves the design and field testing of questionnaires to be used as supplements to the NCHS's Health Interview Survey (HIS). The first such questionnaire was designed to collect information on the number of persons who had had a stroke, diagnosed or undiagnosed, and their hospitalizations. This questionnaire was included in the 1977 HIS. Questionnaires are being designed as supplemental modules for the HIS. One questionnaire will collect information on those persons with convulsive disorders. Another questionnaire will collect information on headache. The data collected will be used to provide national estimates of the prevalence of these disorders.
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{
"pile_set_name": "NIH ExPorter"
}
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