Datasets:

timaeus
/

pile-nih_exporter

Dataset card Data Studio Files Files and versions Community

Split (1)

train · 100k rows

text stringlengths 1 32.7k	meta dict
Project Summary?Abstract While total shoulder arthroplasty (TSA) is the preferred surgical treatment for advanced glenohumeral arthritis, a subset of patients does not experience improvement or sustains a complication. Mean TSA complication rates have ranged from 10-17%, with glenoid component loosening reported as the most common long-term complication of anatomic TSA and a common reason for revision surgery. While complications can lead to poor clinical outcomes, multiple studies have also shown associations between baseline demographic, disease- related and surgical factors, and clinical outcomes. Despite these prior studies, the factors associated with poor short- and long-term clinical outcome after anatomic TSA are still not well understood, in part due to the lack of large prospective cohort studies allowing for multivariable analysis. Our proposal?s objective is to identify the factors associated with short- and longer-term clinical and radiographic outcomes following anatomic TSA. Our approach will utilize two unique, prospective TSA cohorts ongoing at our institution to allow for simultaneous investigation of short and longer-term clinical outcomes, as well as the relationship between the two through the assessment of radiographic factors not possible with routine imaging: a larger cohort (over 1,200 projected cases) collecting baseline demographic, disease-related and surgical factors, together with 1 year clinical outcomes (Patient Cohort 1); and a smaller cohort (n=152) collecting CT imaging-based measures, as well as minimum 5 year clinical outcomes (Patient Cohort 2). Specific Aim 1 will use Patent Cohort 1 to identify the risk factors associated with short-term clinical outcomes at 1 year after primary anatomic TSA. Specific Aim 2 will use Patient Cohort 2 to conduct exploratory analyses of the incremental contribution of CT imaging-based radiologic factors to the prediction of longer-term clinical outcomes at minimum 5 years after primary anatomic TSA, beyond that provided by the perioperative risk factors identified in Specific Aim 1. We expect to show that baseline mental health status, pre-operative opioid use, pre-operative Penn Shoulder Score (PSS) or Single Assessment Numeric Evaluation (SANE) score, Walch classification, subscapularis management, and implant position will independently associate with 1-year patient-reported outcomes (PSS, SANE) after controlling for other demographic, disease-related, and surgical factors (Aim 1). We also anticipate that after adjusting for the risk factors investigated in Aim 1, glenoid component shift (translation and/or rotation) and central peg osteolysis at minimum 2 and/or 5 years post-operatively will associate with worse patient-reported outcomes (PSS, SANE) at minimum 5 years, and that the pre-operative Walch classification and joint line medialization and the presence of central peg osteolysis at minimum 2 years will associate with composite glenoid component shift at minimum 5 years (Aim 2). Identifying significant risk factors that are important to clinical outcomes following anatomic TSA would allow for the design of future prospective clinical trial(s) to investigate modification of these factors either directly or indirectly through pre-operative intervention or modification of surgical treatment and/or patient selection based on these factors, with the aim to improve outcomes and survivorship following anatomic TSA.	{ "pile_set_name": "NIH ExPorter" }
The Ly49 and KIR families are comprised of both inhibitory and activating receptors; the latter interacting and signaling through DAP12. We have continued our dissection of the proximal events associated with DAP12 signaling over the review period. The result of this work includes the definition of an apparent defect in DAP12 signaling in mice of the 129/Sv background that can profoundly effect the analysis of gene targeted mice, delineation of two levels of regulation of DAP12 signaling by CD45 (signal initiation and dephosphorylation of DAP-12 itself), and most recently, dissection of the overlapping use of the non-catalytic adaptor proteins Linker for Activation of T cells (LAT) and the Linker for Activation of B cells (LAB). TREM-1 regulates sepsis whereas TREM-2 can control the development of DC, microglia and osteoclasts. Our studies of the TREM cluster have led to publications regarding; 1) the identification of TREM-like transcript-1 (TLT-1), a protein that does not couple to DAP-12 but instead interacts with SHP-1; 2) characterization of TLT-1 as a platelet specific receptor sequestered in alpha granules; and 3) identification of soluble TLT-1 in serum, and solution of the crystal structure of the extracellular domain. We have also targeted TLT-1 in mice, shown its role in platelet aggregation in vitro, and defined some proteins that interact with it. Although platelets and thrombosis can regulate inflammation, cancer growth and metastasis, and adaptive immunity, we have chosen not to propose further TLT-1 studies under our base allocation here. If however, sufficient resources were available, we would continue biochemical study of TLT-1. We are currently, evaluating our TLT-1-/- mice in collaboration with a former fellow from our lab.	{ "pile_set_name": "NIH ExPorter" }
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Determine if Pioglitazone plus life-style intervention will be more effective than placebo plus lifestyle intervention in reducing triglyceride levels in antipsychotic treated schizophrenic patients with impaired (150 mg/dL) or elevated (200 mg/dL) triglyceride levels. Determine if Pioglitazone plus life style intervention will be more effective than placebo plus lifestyle intervention in increasing HDL and decreasing two atherosclerotic risk ratios (Total cholesterol/HDL and triglycerides/HDL). Determine if Pioglitazone plus life-style intervention will be more effective than placebo plus lifestyle intervention in reducing indices of impaired glucose metabolism (fasting glucose levels 100 mg/dL, Hb1Ac levels 6.4, 2 hr glucose values in glucose tolerance test 140 mg/dL). Determine if the degree of impaired glucose tolerance as assessed by fasting glucose levels and 2 hr glucose levels on glucose tolerance test will be significantly negatively correlated with immediate and delayed verbal memory performance (as measured by list learning and story recall on the RBANS.) Determine if the degree of improvement from impaired glucose tolerance (fasting glucose, and 2 hr glucose in glucose tolerance test)will be correlated with improved performance in verbal memory as measured by the tests noted in hypothesis. Determine if the degree of memory improvement (same day pre-glucose baseline vs. 2 hr) during GTT glucose load, on paired word and short story of the RANDT Memory Scale will be correlated with the degree of baseline cognitive deficit and the degree of impaired glucose tolerance.	{ "pile_set_name": "NIH ExPorter" }
This proposal is for the Fundus Photograph Reading Center for the Foveal Photocoagulation Study (FPS). This multicenter randomized controlled clinical trial is proposed to evaluate the efficacy of krypton red laser photocoagulation therapy in preventing severe visual acuity loss in patients with age-related macular degeneration (AMD) and subfoveal neovascular lesions. AMD is the major cause of uncorrectable loss of vision in the U.S. among individuals 60 years of age and older. Given the evidence that photocoagulation is useful for 75 to 90% of severe visual loss in AMD, it is reasonable to propose a prospective randomized trial of krypton laser photocoagulation treatment for subfoveal neovascular membranes. Two patient groups will be evaluated separately: first, eyes never before treated in which there is a discrete subfoveal noevascular membrane, and, second, eyes initially treated for extrafoveal or juxtafoveal membranes that present with recurrences under the center of the fovea. A registry will be maintained for patients otherwise eligible but whose visual acuity is too good to meet the eligibility requirements. The purpose of the registry is two fold: to serve as a source of future eligible patients and to document the natural history of visual acuity loss in AMD. It is anticipated that approximately 700 patients will be recruited over a two year period, half into each study group, and followed for a minimum of two years. Approximately 100 patients will be followed in the natural history registry. The Reading Center will review all fundus photographs and fluorescein angiograms of patients entered into the study at the twelve clinical centers for the following purposes: 1. To insure that patients selected for the FPS meet the eligibility criteria. 2. To insure that photocoagulation treatment is carried out according to the treatment protocol. 3. To determine complications associated with treatment. 4. To describe pathology present in eyes reaching defined points for both treated and not treated eyes.	{ "pile_set_name": "NIH ExPorter" }
The naturally occurring polyphenol, curcumin, has been shown to be a potent chemotherapeutic agent in a variety of tumor models; however, its effects on tumor-associated immunosuppression have not been investigated to any great extent. The accumulation of immature myeloid cells is a hallmark of cancer and our data indicate that exosomes produced by tumors can suppress the differentiation of immature myeloid cells (iMCs). We have now found that curcumin treatment inhibits the tumor exosome-mediated suppression of differentiation of IMCs in mice, preventing their accumulation in the spleen and delaying tumor growth. Analysis of the composition of the tumor exosomes using a proteomics approach identified CSN5 as a candidate regulatory molecule and siRNA knockdown of CSN5 resulted in attenuation of the tumor exosome- mediated induction of IMCs in the spleen, suggesting that exosomal CSN5 plays a key role in tumor exosome-mediated immune suppression. We have now found that curcumin treatment destabilizes exosomal CSN5. The exosomal CSN5 is stable when associated with phosphorylated STAM1. Curcumin treatment leads to the dephosphorylation of STAM1 permitting the recruitment of an E3 ligase BARD1/BRCA1 to the CSN5 complex, resulting in CSN5 degradation. Our in vitro kinase assay results indicate that the curcumin- mediated disassociation of STAM1 from CSN5 complex is most likely due to the inhibition of CSN5- associated kinase activity. The clinical relevance of these studies is supported by our recent data that indicate that CSN5 is packed in exosomes isolated from the peripheral blood of breast cancer patients but not healthy subjects. We propose: (1) To use siRNA technology to determine if tumor exosomal CSN5 is a primary target of curcumin in its prevention of breast tumor exosome-mediated immunosuppression; (2) To determine whether CSN5-associated kinase activity prevents BARD1/BRCA1 E3 complex degradation of CSN5 in TS/A tumor cells; and (3) To determine whether curcumin treatment of human tumor cells results in degradation of CSN5 and prevents blocking of differentiation of CD33 (equivalent to mouse GDI lb+) Grl+ cells. We will further determine if exosomes packed with CSN5 circulate in the peripheral blood of patients with breast cancer and if overproduction of exosomes is correlated with the accumulation of iMCs in patients with breast cancer. RELEVANCE: The data generated should identify a novel mechanism for the actions of curcumin and identify its molecular targets in preventing CSN5/tumor exosome-mediated blockade of myeloid cell differentiation.	{ "pile_set_name": "NIH ExPorter" }
A healthy moist ocular surface is essential for proper visual functioning. It requires cooperation of an integrated system called the Lacrimal Functional Unit, which consists of the ocular surface tissues, secretory glands, and the nervous system, that together produce and maintain a normal tear film. A disturbance in any group of these components leads to dry eye. The corneal sensory afferents are one of these components. Their activation is considered to be critical to basal tear production, and thus their dysfunction could cause a chronic lack of tears, resulting in dry eye. However, despite a great amount of work done to characterize the corneal afferents, their roles in the lacrimation reflex or dry eye disease still remain to be determined. Recently, we identified a special type of cold-sensitive corneal nerve that is activated by a variety of ocular stimulations thought to be critical for tear production: drying of the cornea, gentle cooling, tear evaporation, and hyperosmolar tears. Evidence indicates that dry eye disease is in part a dysfunction of the mechanisms that detect ocular surface conditions such as cooling of the cornea and the hyperosmolar tears. Thus, we hypothesize that activation of this special type of corneal afferent will induce tear production and their dysfunction contributes to dry eye disease. If so, a selective destruction of these cold-sensitive corneal afferents should lead to dry eye. Recent evidence indicates that members of the nerve membrane receptors (TRPM8, TRPV1, TRPA1) reside in this special type of corneal afferent involved in tear production; that TRPM8 receptors mediate the cooling response; and TRPV1/TRPA1 receptors mediate the hyperosmolar response. We hypothesize that activation of these receptors together constitutes the responses of these cold-sensitive corneal afferents to drying of the cornea and leads to tearing. Thus, genetically engineered mutant mice specifically lacking these receptors should produce very few tears and exhibit dry eye. We will also show that these mutant mice display other signs of dry eye diseases such as inflammation of the ocular surface, which would damage the integrity of the ocular surface tissues, producing the abnormal morphology of the corneal and the conjunctival epithelia. Finally, we will demonstrate immunohistochemically that some well-known models of dry eye disease lack these receptors, implying that this fact is one reason that they have dry eye. If the aims of these studies are achieved, the proposed research will lead to a better understanding of the cellular and molecular mechanisms underlying the activation of the lacrimation reflex and dry eye disease, and may provide new targets for pharmacological treatments for dry eye diseases.	{ "pile_set_name": "NIH ExPorter" }
As the number of minorities and older adults in the U.S. increase, it is increasingly important to reduce health care disparities among these groups. Recent reports have shown that ethnic disparities in health care utilization, such as successful patient-physician communication and engagement in preventative behaviors, are especially critical for older adults. Optimal health care depends on whether an individual chooses to follow their doctors' recommendations, adhere to prescribed treatments, and engage in health promotion programs. Understanding cultural and age-related differences in how these choices are made can help to reduce disparities in health care. The proposed program of research will test applications of Affect Valuation Theory (AVT) as a theoretical foundation for understanding cultural and age-related differences in how health care decisions are made. AVT suggests that variation in ideal affect (i.e., the affective states that people value) may contribute to cultural and age-related differences in health care decisions. The specific aims of the proposed research are to: (1) Examine the relationship between ideal affect and health care decisions; and (2) Determine whether cultural and age-related differences in ideal affect may in part account for differences in health care decisions. Using both survey and experimental methods, we will investigate the relationship between ideal affect and health decision making across the life span for two distinct cultural groups, European American and Chinese American. First, we will measure how ideal affect influences health care decision making among participants ranging in age from 21 to 80. Then, we will manipulate ideal affect and subsequently assess preferences for health care options. Findings from these studies will establish the role of ideal affect and health care decision making as a potential mechanism for explaining ethnic health disparities across the life span. PUBLIC HEALTH RELEVANCE: By understanding how ideal affect influences decision making across age and cultural groups, the proposed research will help to develop more culturally sensitive and appropriate treatments and interventions. Thus, integrating ideal affect into the improvement of health care programs may address the needs of diverse populations more adequately.	{ "pile_set_name": "NIH ExPorter" }
Abstract There is a growing consensus that protection against HIV infection will require BOTH antiviral antibody responses as well as polyfunctional CD4+ and CD8+ T cells with potent lytic activity. To stimulate the breadth, potency, and rate of response required, Profectus Biosciences intends to utilize its platform technologies based on DNA vaccines combined with genetic adjuvants. As a step in that direction, a phase 1 clinical trial, HVTN- 080, showed that electroporation in combination with our IL-12 adjuvant can double the CD4+ and CD8+ T cell response rate in vaccinees over electroporation alone1. Even with this success, we recognize that a truly effective HIV vaccine will require a regimen that can consistently provide high magnitude, long-lived responses at rates >95% with as few booster immunizations as possible. We believe that adjuvants are a key to developing such a vaccine. Under our phase I SBIRs, we identified a new class of genetic adjuvants that exploit the RIG-I signaling cascades and trigger potent anti-viral innate immune responses. These innate responses integrate with TLR evoked innate responses and DAMPs/alarmins to evoke potent anti-viral adaptive immune responses. Also under phase I, we found that the adjuvants attenuated vaccine antigen expression by negatively impacting the promoter. We have now developed an improved promoter to drive our vaccine antigen and adjuvant expression that is not attenuated by these adjuvants. Under this phase II SBIR, we intend to identify an optimal combination of DNA adjuvants using a macaque model. Upon completion of phase II, we expect to have identified a lead adjuvant combination worthy of testing with our HIV vaccine in phase I human trials. 2	{ "pile_set_name": "NIH ExPorter" }
The Mouse Genome Conferences have played a key role in organizing and focusing the interests and resources of the mouse genetics community. These meetings provided forum for disseminating recent mouse genomics research and a vehicle for addressing problems of common concern in the community. Tangible results of the previous conferences include: 1) a new journal, Mammalian Genome, that is devoted to mammalian genome analysis, is now in the fifth year of publication and has produced the chromosome committee reports; 2) The International Mammalian Genome Society has been established to better organize and coordinate the Mouse Genome Conferences and the chromosome committees; 3) the chromosome committees are functioning and they are integrating the efforts to develop consensus genetic and physical maps for each chromosome; and 4) the activities of other established committees have been integrated with the Mouse Genome Conferences including the Mouse Genome Committee of HUGO and the International Committee on Standardized Genetic Nomenclature for Mice. Both of these committees have independent standing but the membership of these committees actively participate in the Mouse Genome Conferences. The inclusion of the Nomenclature Committee into the conferences is particularly helpful given the rapid expansion of new technologies for identifying genetic variation and the substantial increase in mapped genes. Mouse Genome Conferences fulfill a unique mission by focusing specifically on l) presentation of significant new strategies and paradigms for genome analysis, 2) presentation of important new data, 3) chromosome committee meetings, 4) identification of problems that affect the mouse community and that require concerted efforts for resolution, and 5) an opportunity for chromosome committee chairs to meet collectively with the editors of the Special Issue of Mammalian Genome to review committee report format and content. These meetings provide an essential forum for direct interaction between both the producers and users of genome information and resources. More importantly, it provides direct interaction with the informatics resources such as the Jackson informatics program to verify and validate the database content and use by the community. These Conferences continue to play a key role in ensuring that mouse genome research provides comprehensive and rigorous genetic and physical maps. It is difficult to imagine how these maps could be developed in an integrated manner without these conferences.	{ "pile_set_name": "NIH ExPorter" }
Funds are requested for the acquisition of a liquid state 14.1 T NMR spectrometer to be used by a group of scientists from the Chemistry Department and the Biochemistry and Molecular Biology Department of The Pennsylvania State University. This instrument would update and expand the capabilities available within the Nuclear Magnetic Resonance Facility of the Chemistry Department. The primary investigators are all currently involved in projects that require the collection of high-resolution data on proteins in solution. A short description of their research interests follows. Stephen J. Benkovic - Investigation of the factors underlying catalysis and substrate binding in several classes of enzymes: catalytic antibodies, DNA polymerase I, T4 polymerase complex, dihydrofolate reductase, and trifunctional enzymes in the de novo pathway for purine biosynthesis. Donald A. Bryant - Molecular biology and biochemistry in photosynthetic apparatus from cyanobacteria and green bacteria; mechanism of electron transport of cyanobacteria; structure-function relationships in photosystem I; origins of chloroplasts. Christopher J. Falzone - Solution structure of proteins using multinuclear and multidimensional NMR methods; backbone dynamics; substrate dynamics and the effects of site-directed mutation on substrate specificity and protein structure. Juliette T. J. Lecomte - Structure and dynamics of partially folded proteins in solution; determinants of structure in small water-soluble proteins; modulation of structural and thermodynamic properties by specific and non-specific salt effects. C. Robert Matthews - Mechanism by which the primary structure of a protein directs the rapid and efficient folding to the native conformation; site- directed mutagenesis to target individual amino acids in the alpha subunit of tryptophan synthase, dihydrofolate reductase, and ras p2l. With the new instrument, these investigators and a group of several secondary investigators will be in a position to engage in and carry out competitive research related to structure and function in biological systems. The availability of a 14.1 T spectrometer will also allow other researchers to access the currently overbooked 11.7 T instrument.	{ "pile_set_name": "NIH ExPorter" }
The normal adult alveolar epithelium, comprised of type I (AT1) and type II (AT2) pneumocytes, forms a tight barrier to the passive leak of fluid from the interstitium and vascular compartments into the alveolar space. Maintenance of the relatively dry state of the alveolar spaces depends on the integrity of this barrier, as well as on active Na transport to create an osmotic gradient that drives fluid reabsorption across the alveolar epithelium. Despite the probable major role of AT1 cells (given the large internal surface of the lung that they cover) to alveolar epithelial function, the relative contributions of AT1 vs AT2 cells are not well delineated. The overall goals of this proposal are to evaluate the differential roles of AT1 and AT2 cells in alveolar epithelial function and biology, capitalizing on the recent development of new tools for studying alveolar epithelial (especially AT1) cells. Our central hypotheses are that 1) AT1 cells are important participants in active transepithelial ion transport (and therefore alveolar fluid clearance) across the alveolar epithelium, 2) AT1 cell contributions to alveolar function and biology are reflected in part by their spectrum of gene/protein expression and phenotypic plasticity, and 3) AT1 cells play major roles in the response to alveolar injury and recovery in adult lungs. We will exploit our recent success in obtaining highly purified AT1 cells, our ability to maintain them in primary culture, and availability of AT1 and AT2 cell-specific genes/promoters to conditionally delete ion transporters specifically in either cell type in vivo in order to address these hypotheses by exploring the following Specific Aims: 1) characterize functional properties of rodent AT1 and AT2 cells using isolated AT1 and AT2 cells and monolayers derived from these cells; 2) evaluate differential AT1 and AT2 cell contributions to alveolar function by conditional cell-specific knockout of transport genes in transgenic mice in vivo; and, 3) assess AT1 and AT2 cell-specific roles in lung injury and recovery in vitro and in vivo. Results of these studies will provide major new insights into the differential functional and biological roles of AT1 and AT2 cells in normal adult alveolar epithelium and during injury/recovery. Understanding how AT1 and AT2 cells in the adult lung maintain and differentiate their cellular functions, fates and lineages will constitute major advances in alveolar cell biology and could significantly impact the development of novel therapeutic approaches for modulating alveolar homeostasis in health and disease. [unreadable] [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Work in progress involves the development of chemical reactions which are applicable to the synthesis of antitumor natural products. These reactions include a new modification of the Ullmann reaction which permits cross coupling at room temperature, the coupled Cope Claisen rearrangement, and the alkylation of ketenethioacetals.	{ "pile_set_name": "NIH ExPorter" }
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This collaborative R01 involves parallel studies at the Universities of Wisconsin and Minnesota. The work explores the neurobiological bases of problems in attention/executive functions, sensory integration, and emotion/stress regulation exhibited by children who have experienced neglect/privation early in life. This project is motivated by renewed interest the effects of early deprivation/neglect on development, precipitated by the last decade's increase in international adoptions. In addition, it reflects concerns with brain development and neural plasticity coupled with continued controversies over the importance of early experiences and questions about sensitive periods. The marked change in living environments experienced with adoption allows a reasonable way to estimate both the duration and ages during which deprivation was experienced. To date, studies of these children have employed global measures that are difficult to map to specific neural systems. The General Aim of this project is to build upon extant studies and examine specific cognitive and affective systems. The specific aims identify (a) aspects of neural circuitry that are linked to the development of cognitive processes and self-regulatory behaviors and (b) later-developing neural systems that are likely to be affected by early deprivation/neglect. Specific Aim 1 examines frontally mediated systems believed to underlie aspects of attention, inhibitory control, and working memory. Specific Aim 2 examines sensory-motor processing including sensory integration, acquisition, and praxis. Specific Aim 3 explores systems thoughts to be compromised based upon the animal literature on early neglect: cortico-limbic-hypothalamic-brainstem systems that are involved in stress reactivity and regulation. Together, these studies will provide a relatively comprehensive and more specific picture of the potential neural compromises resulting from early deprivation in human children as well as highlighting those aspects of brain-behavior development that appear to be spared.	{ "pile_set_name": "NIH ExPorter" }
The dissemination e?orts of the Center have been tremendously successful over the last funding period. In the next funding period, the Center will continue its existing strategy, as laid out in Approach, while introducing innovative additions to enhance the e?ectiveness of its dissemination e?orts. The Center employs ten di?erent dissemination channels - the Center's website, software downloads, ?exible licensing, research publications, lectures and poster presentations, tutorials and case studies, image and movie downloads, brochures, media coverage, and streaming multimedia projects. Over the last funding period, the website received a huge volume of tra?c, serving over 1.4 million visitors, with 12.6 million page views, representing 28 terabytes of data transferred; few departments on our campus have busier websites. The website features rich content including, for example, 68 research highlights posted over the last funding period with links to movies, research pages and publications; another example is a series of 11 editorials of altogether 210 printed pages that cover key issues in molecular modeling and have had a deep impact on readers. A signi?cant portion of the Center's dissemination e?orts are dedicated to lowering barriers to learning of the Center's main software programs VMD and NAMD. To date, a total of 45 tutorials of altogether about 1,460 printed pages are available and have been viewed 228,000 times over the last funding period. User surveys of VMD and NAMD performed by the Center in 2016 indicated that 70% of respondents agree that the respective program is user-friendly. The Center has also been actively disseminating its research, producing 424 publications over the last funding period and giving 509 lectures and presentations at workshops and conferences. In the public sphere, the Center has continued to enjoy high publicity during the last funding period, through 111 instances of media coverage and science movies uploaded to its YouTube channel, which garnered 118,000 views. Images and movies on the Center's website have likewise been popular, attracting 25,000 downloads. Finally, new brochures have been printed to advertise the Center's revolutionary molecular dynamics program QwikMD. The Center has completed development of tools to create virtual reality (VR) visualizations of atomic level cellular structures made available already on YouTube by means of omnidirectional VR glasses such as Google Cardboard for smartphones, Oculus Rift and Samsung Gear VR. The Center is also participating, under the Cen- trality of Advanced Digitally Enabled Science (CADENS) supported by NSF, in the production of a documentary on computational and data-enabled discoveries in science. The Center's documentary, entitled ?Birth of Planet Earth?, will feature advanced digital visualizations of scienti?c data and will be screened to a wide public audience as ultra-high-resolution fulldome ?lms in museums, planetariums and science centers, as well as high-de?nition television programs on Hulu, Amazon, PBS, and NSF online television.	{ "pile_set_name": "NIH ExPorter" }
DESCRIPTION: This proposal outlines investigations of proteins and model systems with three innovation magnetic resonance techniques: high frequency dynamic nuclear polarization (DNP), 140 GHz electron paramagnetic resonance (EPR), and electron nuclear double resonance (ENDOR). (1) High frequency CW DNP/MAS spectra of a protein have recently been observed. The experiments will be employed to enhance signal strengths in MAS NMR spectra of large proteins -- T4 lysozyme, bacteriorhodopsin, and ribonucleotide reductase (RNR) -- containing exogenous and endogenous paramagnetic centers such as TEMPO and tyrosyl radicals, respectively. The experiments utilize a 140 GHz gryotron oscillator as a microwave source, and the principal investigator will develop a low temperature helium recirculating system for use with DNP/MAS probes. Signal enhancements of ca. 100 have been observed, resulting in a factors of 10,000 decrease in acquisition time. (2) Pulsed DNP experiments based on Hartman-Hahn cross polarization are planned for single crystals samples as well as powders. (3) 140 GHz CW EPR ENDOR investigations are planned for five different systems: (a) a fluorinated inhibitor of ribonucleotide reductase (RNR); (b) the X-intermediate formed on combining Fe and the apoRNR; (c) thiyl radicals associated with adenosylcobalamin; (d) glycyl radicals in pyruvate formate lyase; (e) tyrosyl radicals in photosystem-II (4) 140 GHz pulsed EPR experiments will be used to study the frequency, concentration and temperature dependence of the electron spin lattice relaxation (T1e) in model systems for DNP experiments and in proteins. The data are useful for optimizing the rate and magnitude of polarization enhancements in determining structures of proteins containing free radicals and paramagnetic centers, and are inherently interesting.	{ "pile_set_name": "NIH ExPorter" }
Melanoma incidence in the United States has increased 15-fold over the last 70 years. Despite prevention efforts targeting sun exposure, the major risk factor for melanoma, the mortality from this disease continues to rise. Therefore, other approaches such as chemoprevention have emerged as areas of investigation. One promising class of medications, non-steroidal anti-inflammatory drugs (NSAIDs), has demonstrated protective effects against melanoma in cell lines and mouse models. We have recently conducted a case-control study in humans that found that subjects taking non-aspirin NSAIDs had a 40% decreased risk of melanoma compared to those who did not take these medications. Based on these results, we propose a study dedicated to investigating our hypothesis that regular use of NSAIDs has a chemoprotective effect against melanoma. We will evaluate this hypothesis by pooling data from the Nurses Health Study and the Health Professionals Follow-up Study to address three specific aims: (1) To confirm our preliminary finding of an inverse relationship between the use of NSAIDs and risk of invasive melanoma; (2) To determine the effects of NSAIDs on risk of melanoma in situ, and (3) To assess the influence of NSAIDs on melanoma invasion, as measured by tumor Breslow depth and growth phase. By pooling these studies, effects can be determined in both men and women, and power is increased for more precise estimates of effect. Through our analysis of existing data from these studies, we hope to identify an agent for clinical chemoprevention trials. Due to the large numbers of individuals taking NSAIDs and the increasing incidence of melanoma, we believe that the detection of an effect will make a significant contribution to the public health. This study proposes to investigate whether regular use of non-steroidal anti-inflammatory drugs (NSAIDs) may protect individuals against melanoma. This question will be addressed using data from the Nurses Health Study and the Health Professionals Follow-up Study, two large groups of health professionals who have provided information about their health for up to 24 years. Due to the large numbers of individuals taking NSAIDs and the increasing incidence of melanoma, we believe that the detection of an effect will make a significant contribution to the public health. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Project Summary Abnormal or synchronous neuronal activity in the brain leads to epileptic seizures that, when repeated or prolonged, can cause neuronal damage resulting in delayed psychomotor development, intellectual disability and other neurological disorders. In neurons, action potentials are terminated by the inactivation of Na+ channels and by the repolarizing outward currents triggered by activation of K+ channels. One of the major potassium current in neurons is the muscarine-regulated M-current, a non-inactivating slow current that is activated at subthreshold voltages. The M-current, which is generated from the heteromerization of KCNQ2 and KCNQ3 channels (IKM), activates in the time frame of action potential initiation, providing a crucial role in controlling neuronal excitability. The slow kinetics of activation and deactivation of the IKM (KCNQ2/KCNQ3) channel regulates the membrane potential and impedes repetitive neuronal firing. A growing number of inherited mutations have been found in the IKM channel that cause a wide spectrum of early-onset epileptic disorders ranging from benign familial neonatal seizures to severe epileptic encephalopathies. I will determine the molecular mechanisms by which a set of epileptic-inducing mutations in KCNQ2 and KCNQ3 cause malfunction of the IKM channel. I will use a fluorescence assay, voltage clamp fluorometry (VCF), to simultaneously measure voltage sensor movement and gate opening during IKM channel activation in these mutations. Knowing the mechanisms that lead to defective channel function is essential to study how to modulate and ultimately restore function of these mutated channel. Furthermore, the IKM channel is an attractive pharmacological target to treat hyperexcitability-related diseases, such as epilepsy, because increasing the M-current stabilizes the resting and subthreshold membrane potential, thereby reducing membrane excitability. Lipophilic compounds, such as polyunsaturated fatty acids (PUFAs), have been shown to modulate neuronal function. In particular, PUFAs have been shown to improve the outcomes of epilepsy, therefore constituting very promising anti-epileptic agents. However, the molecular mechanism of action of PUFAs is unknown. For example, it is unknown whether PUFAs affect the voltage sensor movement, gate movement, or both. This is important because knowing the channel region where PUFAs act will allow designing PUFAs derivatives to more specifically tackle the IKM channel. Based on the molecular mechanism for each epileptic-inducing IKM channel mutation, I will assess, using VCF, which type of PUFAs variants would be most suitable to restore physiological channel activity in order to develop an antiepileptic drug. I will also use induced pluripotent stem cell (iPSC)-derived neurons, a simple but powerful model, to test the efficacy of PUFAs on the derived neurons expressing mutated KCNQ2 and KCNQ3 to test both the mechanistic implications of the proposed work and the therapeutic potential of the PUFAs. The anticipated results of these experiments will provide the basis to mechanistically understand how different mutations cause IKM channel defects, and should show proof-of-concept that PUFAs can act as antiepileptic drugs. This would be a milestone toward mutation-specific treatments of epilepsy and other neurological disorders caused by mutations in the IKM channel.	{ "pile_set_name": "NIH ExPorter" }
Alzheimer's disease (AD) is the most common form and devastating dementia. There are no effective approaches to specifically target and prevent AD pathogenesis. It is accepted that aggregated fibrillar forms of A? peptides are prominent hallmarks and the major cause of AD due to their toxicity to neurons. As such, A? aggregates are the potential targets for the intervention of AD, as targeting and removal of A? fibrils or plaques is expected to eliminate the toxicity of A? aggregates. Although the KLVFF peptide has been served as the major lead to develop potential probes to target A? aggregation, it only weakly recognizes and inhibits A? aggregation. Meanwhile, it is also susceptible to protease degradation. We recently developed a novel one-bead one-compound (OBOC) combinatorial library, from which we identified a potent peptidomimetic that is 100-fold more effective than KLVFF in disrupting A? aggregation in vitro. This peptide material also removes the cytotoxicity of A? aggregates towards N2a neuro-blastoma cells. Furthermore, this lead compound and its derivative can even significantly remove A? plague deposited in the brain of the AD transgenic mice ex vivo. As such, our long-term goal is to develop novel biomaterials that can prevent, halt and cure AD. The objective of this proposal, is to advance our preliminary work by developing new OBOC libraries with enhanced chemodiversity and novel structural classes, so as to identify and develop more potent and effective AApeptide biomaterials that can target and inhibit fibrillar formation of A? both in vitro and in vivo. We will first design and synthesize new OBOC gamma- AApeptide libraries with diverse functional groups and constraints by using a range of novel chemical approaches. Then we will use established screening assay to identify and optimize ligands that target and inhibit the aggregation of A? peptides. The compounds with activity equivalent or better than the lead compound identified from the preliminary study will be used to study their ability to inhibit A? aggregation both in vitro and in vivo on AD-transgenic mice. The proposed study is significant because there is no effective approaches for AD diagnosis and prevention. Our research will provide novel biomaterials to unravel AD pathogenies and to develop potential therapeutic agents for cure of AD. The proposed research is innovative because we not only provide a general approach for the development of novel class of biomaterials specially targeting A? aggregates, in addition, this combinatorial approach can be easily extended to identify new materials targeting other amyloid diseases such as Huntington's disease and diabetes diseases.	{ "pile_set_name": "NIH ExPorter" }
A detector for ions in a mass spectrometer is to be investigated in which ions are accelerated to high energy and then permitted to strike a metal surface where they produce secondary electrons. The electrons are then accelerated to a scintillator where they produce photons which actuate a closed photomultiplier tube. This type of detector is known as a Daly Doorknob detector when the photons are in the visible region. The present work addresses the possibility of using a scintillator that produced photons in the far UV rather than the visible spectrum. In this case, the photomultiplier need not have a photodynode with a low work function and its attendant thermal emission noise. Previous experiments on UV photon counters suggest certain materials as potential scintillator in the far UV. These will be tested and, if promising, be built into a Daly Doorknob configuration. Comparison of sensitivity and noise level of this detector with a conventional open channeltron detector would be made.	{ "pile_set_name": "NIH ExPorter" }
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Section A: Specific Aims The ultimate goal of research in my laboratory is to use various structural and biochemical techniques to understand the atomic basis of function of proteins of pathways like autophagy (a lysosomal targeting pathway for degradation of intracellular objects) and innate immune pathway;the cross-talk between these pathways;the role of these pathways/proteins in the defense against pathogens;and the inhibition of these proteins by different pathogens. One project focuses on investigating the structure, function and molecular mechanism of Beclin1, an essential autophagy effector. Our previous work, funded in part by an NIH R21 grant, investigated the molecular mechanism by which cellular and viral Bcl-2 proteins inhibit Beclin 1 and autophagy. We plan to continue our investigation of the Beclin 1 protein, including its interaction with other autophagy proteins and with pathogen-encoded proteins. Another project focuses on Rig-I, a key mediator of anti-viral innate immunity. We plan to investigate the atomic basis of the role of Rig-I in innate defenses by studying the intra-molecular, intra- pathway or cross-pathway interactions of the Rig-I CARD domains that transmit signals to trigger or repress various pathways. A prerequisite of all these studies is the availability of pure, homogeneous, recombinant protein. We are using COBRE funding to produce these reagents and obtain preliminary data to apply for other grants to further investigate these pathways. Specific aims of the COBRE funding are to overexpress and purify the recombinant proteins listed below, in quantities suitable for subsequent structural and biochemical studies: + Aim 1: Different Beclin 1 domains. + Aim 2: Rig-I CARD domains. + Aim 3: CARD domains of IPS-1, its downstream signaling partner. + Aim 4: Repressor domains of Rig-I and LGP2 (a Rig-I homolog). + Aim 5: The essential autophagy effector Atg5, which interacts with the Rig-I and IPS-1 CARD domains.	{ "pile_set_name": "NIH ExPorter" }
Blood pressure (BP) is a critical vital sign for health, including hypertension. Clinical practice, however, is largely limited to awkward, inconvenient arm cuffs that provide only point-in-time measurement. High- throughput screening as well as personal health monitoring would benefit from a simple BP monitor that obtains accurate measurements with no training-ideally in seconds rather than minutes-and includes electronic health record (EHR) integration to minimize patient reporting error and noncompliance. The overall objective of this project is to use ultrasound, and in particular, Acoustic Radiation Force Impulse (ARFI) techniques, to develop an easy-to-use, cuffless, noninvasive, ultrasonic BP monitor, consisting of a compact sensor pad and readout. Placement of the pad in contact with the skin over an artery will immediately provide self-calibrated, accurate BP. The operating principle is analogous to a conventional arm cuff, which relies on acoustically observing arterial wall deflection in response to the differential between internal arterial pressure and an externally applied cuff pressure; however, this project will instead use ultrasound energy to impose highly localized but safe ARFI pressure pulses on an arterial wall and measure the microscopic deflection that results from the pressure differential. Furthermore, the high speed of ultrasound will allow blood pressure to be conveniently assessed in just a few seconds. The first Aim will demonstrate the ultrasonic pressure measurement method, on a commercial ultrasound testbed, by: developing algorithms to noninvasively track the pulsatile waveform using measured displacements in response to ARFI pulses; calibrating the pressure strength of the ARFI pulses; and reporting the absolute systolic and diastolic equivalent pressures for fluid- and blood-filled arterial phantoms, finishing with human clinical evaluation o the method according to the IEEE 1708-2014 Standard for cuffless blood pressure devices. The second Aim will seek to replace the piezoelectric testbed transducer with a low-cost capacitive micromachined ultrasonic transducer (which is well-suited to affordable personal health monitoring applications), likewise ending with clinical evaluation according to the IEEE standard. The final Aim will translate the testbed into a prototype instrument with a specifically-designed readout unit, incorporate an EHR interface, and culminate with a clinical study to evaluate the complete health monitor system in a low-resource context. This project is particularly relevant to the Department of Health & Human Services' Million Hearts' Initiative, potentially touching a number of facets in the Million Hearts Logic Model by: improving measurement and reporting for large-scale epidemiological studies; increasing management effectiveness for ongoing hypertension medication; reducing hassle and barriers to use; and supporting further ambulatory research, both in the US and India.	{ "pile_set_name": "NIH ExPorter" }
Gyrate atrophy (GA) is a severe human recessive eye disease resulting in progressive loss of vision due to chorioretinal degeneration. The disorder is associated with a deficiency of the mitochondrial matrix enzyme, ornithine-delta-aminotransferase (OAT), which catalyzes the interconversion of ornithine and alpha-ketoglutarate to delta-pyrroline-5'-carboxylate and glutamate. GA patients with activity have 10- to 20-fold higher levels of plasma ornithine as compared with controls. Dietary and specific hormonal administration in rats and mice have been shown to modulate the regulation of this enzyme in different tissues, suggesting OAT's important physiological role in vivo. To test this hypothesis and to find out whether OAT could be a common mediator for retinal cell growth, we produced two transgenic lines, expressing the human OAT gene in strains of mice normally exhibiting a progressive retinal degeneration. Here we show that transgenic mice expressing human OAT exhibit less severe retinal degeneration than control mice littermates.	{ "pile_set_name": "NIH ExPorter" }
T helper cell (Th) function was assessed using peripheral blood leukocyte (PBL) from 47 patients newly diagnosed with Hodgkins disease (HD). The PBL were stimulated in vitro with recall antigens (REC), HLA alloantigens (ALLO) and phytohemagglutinin (PHA). Four distinct patterns of Th responsiveness were detected: patients who responded to all three stimuli; patients who did not respond to REC; patients who responded only to PHA; and patients whose PBL were refractory to all three stimuli. The more severely immune-compromised patients exhibited more severe hematologic parameters of HD.	{ "pile_set_name": "NIH ExPorter" }
Minorities in the United States experience higher prevalence of diabetes and obesity and have a higher mortality rate from these diseases than the rest of the population. For example, between 1980 and 2000, the age-adjusted prevalence of diagnosed diabetes was higher among blacks than whites and highest among black females. Between 1997 and 2000, age-adjusted prevalence of diagnosed diabetes for Hispanic males and females was similar to that of black males. A study published in the January 1,2003, issue of the Journal of the American Medical Association (JAMA), reported that the prevalence of obesity among US adults climbed from 19.8 percent to 20.9 percent between 2000 and 2001. This study also found that African Americans had higher prevalence rates of both obesity (31.1 percent) and diagnosed diabetes (11.2 percent) than did members of other racial and ethnic groups. These disparities are particularly great in minorities living in the south east region and require immediate attention. For example, in Alabama, age-adjusted diabetes prevalence was at least 50% higher in 2002 than in 1994 and it ranks number one among the other states. In this project, we propose to organize and conduct a regional conference targeting the African-American communities, who make up the largest cohesive minority in the Southeast region. This conference will be coordinated and implemented by the Minority Health and Research Center (MHRC) at the University of Alabama at Birmingham (UAB). The overall goal of this conference is to bring together academic scientists and community physicians to share their experiences, discuss and identify areas of basic, clinical and community-based research that may lead to the elimination of this disparity. The conference will feature both didactic lectures and interactive presentations on possible reasons for health disparities.	{ "pile_set_name": "NIH ExPorter" }
Our goal is to explore the frontiers for developing reconstruction strategies which overcome to the fullest extent possible the factors limiting detection and localization of lesions in emission imaging. The principle guiding our research is the use of task-based assessment of image quality. During the past funding period we have observed unexpected and intriguing results, the exploration of which we propose to further investigate. Our specific aims are to: 1) Probe what dictates when the use of attenuation correction (AC) in reconstruction will and will not improve the accuracy of lesion detection and localization in lung, soft tissue, and bone for chest and abdomen imaging; 2) Explore an innovative approach for formulating reconstruction strategies based on the enhancement of detection-task performance as opposed to exact restoration of the degradation; 3) Examine the hypothesis that priors based on anatomical information from dual-modality imaging will improve the accuracy of lesion detection in maximum a posteriori (MAP) reconstruction; 4) Investigate the hypothesis that respiratory motion correction methods can be developed which will promote AC, improve lesion detection, and facilitate the use of anatomical priors with neuroendocrine tumor imaging; 5) Explore extensions of multiclass (scanning) channelized human-model observers that will enable agreement with human observers for our proposed study conduction methodologies. The clinically significant procedures of single photon emission computed tomographic (SPECT) imaging for neuroendocrine tumors and for bone imaging of back-pain have been selected as the test-beds for our investigations. We will base our investigations on the hypothesis that the formulation of a reconstruction strategy is best performed when the slices and detection task closely approximate their clinical application. To closely emulate the clinic we emphasize free-response observer studies which employ representative populations of hybrid images and volumetric display. Hybrid images are actual clinical acquisitions to which Monte Carlo simulated lesions are added. The formulation and comparison of strategies for the quantification task will serve as a counter-point to our studies of detection. The result of these investigations will be the definition of ways to turn acquired pictures of molecular imaging tracers within a patient into clearer pictures of the three-dimensional location of these tracers from which physicians can make an accurate diagnosis and evaluate therapy options. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY/ABSTRACT The overall purpose of the CEC's Communication and Dissemination Core (CDC) is to create an infrastructure to develop and refine processes and products to convey information to internal audiences (within the DPC) and to external communities (broader educational institutions, diversity researchers, and policymakers) to share DPC evaluation outcomes and increase the visibility and uptake of products, lessons learned, best practices, and processes generated by the DPC. In addition, the CDC will enhance necessary internal communication processes to assure the optimal functioning of the DPC. It will also strategically support communication about and dissemination of products and processes to promote the best practices of consortium-wide programs including those for the CEC, BUILD and NRMN programs. The CDC will provide an important support function to the planned dissemination activities that will be identified by BUILD sites. The CDC will lead dissemination efforts for CEC-based evaluation outcomes, best practices and key processes in evaluation, measurement, and outcomes from its work in the DPC. The CDC will fulfill its purpose by executing these specific aims: 1) Refine DPC communication processes through collaborative planning with the DPC and NIH to develop and support dissemination of DPC site level and consortium-wide best practices; 2) Develop and support dissemination strategies to share CEC best practices, research outcomes, systems and tools among a variety of national stakeholders in diversity education and training, biomedical and health professional science research, and program evaluation; and 3) Support and amplify BUILD site-level plans for outcomes, products and process dissemination, to increase the visibility of best practices among a variety of stakeholders nationwide in higher education, biomedical and health professional science research dedicated to diversity education and training, and policy.	{ "pile_set_name": "NIH ExPorter" }
Dansylhydrazine reacted with aldehyde groups in collagen. The densyl fluorescence of aged collagen exhibited a weaker peak at 525nm, whereas that of young collagen had a stronger, broad peak at 500nm. Fibril formation in vitro was partially inhibited. During the turbidity lag phase, the dansyl fluorescence increased (30-50%). These changes reveal the telopeptide conformation changes occurring during this period. Upon UV(340nm)-irradiation, fibril formation was almost completely inhibited. With increasing temperature, the densyl fluorescence of young collagen decreased, whereas that of old collagen substantially increased. After denaturation, both fluorescences became similar in their intensity and position (490nm). The fluorescence (545nm) of turbium ions (Tb3ion) was enhanced by energy transfer from tyrosine to Tb3 ion bound to collagen and increased slightly during the turbidity lag phase. Tb3 ion also increased the rate of fibril formation, suggesting stronger binding sites on polymeric collagen. Hyaluronic acid (HA) from rooster comb, after chloroform treatment to remove proteins, exhibited no detectable UV absorption, but was degraded by UV (254nm) light, suggesting a highly efficient photo-degradation. The UV (255nm) peak persisted throughout deproteinization for HA from umbilical cord. This HA was also photo-degraded. UV-absorbing species are tissue-dependent and may not be correlated with proteins.	{ "pile_set_name": "NIH ExPorter" }
Cerebrospinal fluid concentrations of homovanillic acid, 5-hydroxindoleacetic acid, norepinephrine and 3-methoxy 4-hydroxphenylethylene glycol did not differ significantly between patients with Alzheimer's disease and age matched controls, and were not correlated with age in healthy men. Spinal fluid concentrations of choline increased with age in healthy men, and were higher in young adults with Down syndrome than in age matched controls. CSF biopterin, a coenzyme for tyrosine and tryptophan hydroxylase, was lower in patients with Alzheimer's disease than in controls, and was correlated with concentrations of 5-hydroxy-indoleacetic acid and homovanillic acid. Corticotropin releasing factor, a neuropeptide, was significantly reduced in the cerebrospinal fluid of patients with Alzheimer's disease as compared to controls, as was peptidyl-alpha-amidation activity, suggesting a loss of neurons which produce amidated neuropeptides. Ratios of albumin and immunoglobulin between cerebrospinal fluid and plasma were normal in Alzheimer's patients, suggesting that the blood brain barrier is intact. Neuron-specific enolase, a glycolytic enzyme enolase, was significantly reduced in cerebrospinal fluid of patients with Alzheimer's disease.	{ "pile_set_name": "NIH ExPorter" }
Spinal cord injury (SCI) results in a number of deficits of motor and sensory systems, including paralysis, anaesthesisa, hyper-reflexia and spasticity below the level of the lesion. The proposed research will address critical issues related to the treatment and mechanisms of hyper-reflexia and spasticity. Preliminary evidence suggests that the onset of hyper-reflexia is not immediate, but rather the animal transitions to a hyper-reflexive state between 7-21 days after injury, as measured by frequency-dependent depression of the H-reflex in terminal experiments at specific time points. We can now verify the exact time point using longitudinal surface electrode recording of the H-reflex measuring frequency-dependent depression changes in the same transected animal. We have developed the capacity to assess spasticity using windup and velocity protocols for the stretch reflex, providing an additional functional measure of symptoms after SCI. Research at the Center for Translational Neuroscience (CTN) determined that passive exercise treatment using motorized bicycle exercise training (MBET) can be used to normalize reflex changes after SCI in the human and the animal model. MBET, initiated 7 days post injury, for thirty days post-injury for 1 hour sessions 5 days/week was found to prevent the loss of frequency-dependent depression usually seen after injury in the animal SCI model. We now want to determine the efficacy of MBET in the chronic animal model of SCI and the amount of time required to show rescue of reflex changes using the longitudinal surface recording technique. Novel preliminary evidence also suggests that L-DOPA given orallly can prevent hyper-reflexia measured by normalization in frequency-dependent depression of the H-reflex. PUBLIC HEALTH RELEVANCE Currently, over 200,000 Americans suffer from spinal cord injury (SCI), many of who suffer from excessive reflexes and spasticity which prevent effective recovery and rehabilitation efforts. The proposed research will use an animal model of SCI to determine when these excessive reflexes and spasticity appear and whether they can be modified by Motorized Bicycle Exercise Training (MBET) or L-dopa, a drug commonly used for individuals with Parkinson's disease. [unreadable] [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Research has shown that institutions that recruit and graduate racial/ethnic minority students make an investment in the overall health of minority communities. Similarly, racial/ethnic minority researchers in the health sciences are likely to ask questions that impact their respective populations of origin. This will be a key long term outcome of the project. The specific aim of this comprehensive national project is to examine underrepresented student access to resources and forms of engagement that result in outcomes (skills, dispositions, and behaviors) necessary for a research career in the biomedical and behavioral sciences. The study draws on data from over 700 institutions that regularly participate in surveys at college entry, after the first year of college, and at the fourth year of college. This allows to monitor the experiences of a large number of African American, Hispanic/Latino, and Native American students at key transition points. Since institutions also receive their data on entering freshmen, and can participate in subsequent follow-ups, another aim is to encourage campuses to use the data and share findings on their own campuses with faculty in related disciplines. In an attempt to encourage broader conceptions of talent and its development in preparation for research careers, a study of introductory courses will also help determine whether students' grades are associated with the skills and dispositions needed for research career.	{ "pile_set_name": "NIH ExPorter" }
This is an R21 application to explore the development of an animal model to study the effects of parental and peripuberal exposure to nicotine. Nicotine remains an important drug of abuse worldwide. In the United States (U.S.), tobacco use is the single leading preventable cause of death. However, despite considerable negative publicity and health warnings, approximately 25 percent of the U.S. population still smoke. Aside from producing profound behavioral effects in the adult organism, nicotine also disrupts developmental processes in many species. Recent epidemiologic data suggest that fetal exposure to nicotine increases the risk for tobacco use during adolescence and adulthood, particularly in females. In addition, 75 percent of adult tobacco users report their first tobacco use occurred when they were "youngsters" (childhood or adolescence). In order to study this issue further, as well as to develop an animal model to elucidate potential underlying mechanisms, the effects of nicotine exposure during gestation on nicotine self-administration in adult male and female rat offspring will be studied. In addition, the effects of nicotine exposure during the periadolescent period on nicotine self-administration in adult offspring will be ascertained. It is hypothesized that nicotine exposure in utero will increase nicotine self-administration in adult offspring. Similarly, peripuberal exposure to nicotine also will increase nicotine self-administration during adulthood. The proposed studies will characterize the relationships between exposure to nicotine during critical periods of development and the acquisition, maintenance, extinction and re-initiation phases of nicotine self-administration. The results of these experiments should provide new and important insights on the relationships between prior nicotine exposure and nicotine-seeking behavior. In addition, since nicotine is considered as a "gateway" drug for the subsequent use of alcohol and other illicit drugs, the results of the proposed studies will lay the groundwork to further understand the factors which might increase vulnerability to drug addictions in general, and to nicotine abuse, in particular.	{ "pile_set_name": "NIH ExPorter" }
A technique is being developed for the visualization of specific mRNA sequences in living cells. This technique utilizes molecular beacons, which are oligonucleotide probes that become fluorescent upon hybridization. Use of molecular beacons allows the simultaneous mapping of the intracellular distribution of multiple mRNAs within living cells and imaging of gene expression on a cell-by-cell basis. Using a model system that employs syncytial embryos of Drosophila melanogaster, we will show that the dynamic changes that occur in the localization of different mRNAs can be observed in real-time in living cells. The distribution of multiple mRNAs will be mapped within the same cell by introducing a mixture of several different molecular beacons, where each molecular beacon is specific for a different mRNA target, and each molecular beacon is labeled with a differently colored fluorophore. The distribution of a group of mRNAs that localize at postsynaptic dendritic sites in response to the synaptic stimuli will be mapped by microinjecting molecular beacons into the perinuclear space of neurons maintained in primary cultures. The intracellular transport of mRNAs within neurons in the form of ribonucleoprotein particles will be monitored using confocal laser scanning microscopy, in order to shed light on the mechanism of mRNA localization. Techniques will be developed for an effective delivery of molecular beacons into cells in suspension and gene expression will be monitored in individual cells using fluorescence-activated cell sorting. The ability to detect mRNAs in living cells with molecular beacons will allow us to explore the molecular complexities of mRNA localization and transport within neurons and other cells with a sharper focus than has been possible before.	{ "pile_set_name": "NIH ExPorter" }
The Prader-Willi Syndrome (PWS) and Miller-Dieker Syndrome (MDS) are both associated with mental retardation and dysmorphic features. Recently, we have shown that both syndromes are caused by deletions of chromosomes 15 and 17, respectively. In both, the deletion is extremely small, requiring high-resolution techniques for detection. Even then, a percentage of patients appear to have normal karyotypes. Since submicroscopic deletions may be present, more sensitive and reliable diagnostic techniques are needed for these two disorders. Analysis at a molecular level would be beneficial, but few cloned genes or anonymous DNA segments have been mapped to chromosomes 15 or 17 and none falls within the critical region of these disorders. Recently, flow-sorted libraries for chromosomes 15 and 17 have become available from the Los Alamos and Lawrence Livermore National Laboratories. Additionally, a partial chromosome 15 library containing only the PWS critical region (q11-q13) is being constructed elsewhere. These materials make it possible to do detailed molecular analysis of these chromosomes, including dissection of the critical regions for PWS and MDS and the development of linkage maps for each chromosome. Using published cloned genes, arbitrary DNA segments, and flow-sorted libraries for chromosomes 15 and 17, the goals of the project are: 1) Molecular dissection of PWS and MDS. Identify DNA probes which fall into the critical region of deletion in PWS and MDS to be used as diagnostic tools in conjunction with high-resolution cytogenetic analysis. These probes will be tested against clinically affected patients with apparently normal karyotypes in order to determine whether submicroscopic deletions exist. For PWS, these probes will be useful in determining whether the breakpoints of all interstitial deletion patients are consistent, suggesting a "hot spot" for chromosomal rearrangement. These probes will help determine whether PWS patients with duplications and extra marker chromosomes have the same DNA segments affected. Finally, polymorphic probes within the PWS and MDS regions will be useful for determining the parental origin of de novo chromosome abnormalities. 2)\Development of linkage maps for chromosomes 15 and 17. Regionally map 20-30 DNA probes to specific regions of chromosomes 15 and 17 by somatic cell hybrid analysis using rearranged chromosomes 15 and 17, dosage analysis using unbalanced patient cell lines, and in situ hybridization. A search for restriction fragment length polymorphisms will be conducted to establish a genetic linkage map for both chromosomes which will be useful in the analysis of other human genetic diseases. Linkage analysis may be employed to determine the recombination distance between probes and their order on the chromosome.	{ "pile_set_name": "NIH ExPorter" }
Chronic myocardial ischemia leading to heart failure is a leading cause of morbidity and mortality in the United States. Experimental myocardial stem cell therapeutics have been performed largely in the acute ischemia model. The well established porcine hibernating myocardium model, which closely resembles the chronicity and stability of human coronary artery disease, will be used to develop and optimize therapeutic strategies based on combined stem cell and gene therapy. Bone marrow-derived porcine mesenchymal stem cells (MSCs) expanded in culture possess robust self-renewal and multilineage differentiation potentials, and are capable of producing many growth factors and cytokines. Although promising as regenerative medicine in aging and disease, MSCs await further analysis regarding the mechanisms governing their growth, differentiation, survival, tissue homing, and aging characteristics. Growth factor modulation of MSC multilineage potential, the influence of aging on the function of MSCs, and the use of allogeneic MSCs will be characterized. Central to these efforts is the use of recombinant adenovirus expressing genes involved in cytoprotection, angiogenesis, and MSC homing. The first part of the proposal relies on extensive cell culture characterizations of MSCs, building the foundation for the second part of the proposal that addresses the physiological effect of engineered MSCs. Aim 1 will determine the differential effects of multiple VEGF isoforms on the growth and multilineage potentials of porcine MSCs. Aim 2 will analyze the expression and regulation of MSC chemokine receptors involved in myocardial MSC homing. Aim 3 will characterize the influence of cellular and animal aging on MSC growth capability, cell survival capacity, multilineage potential, and chemotactic migratory potency. Aim 4 will optimize strategies for tracking and identifying the in vivo fate of implanted MSCs in the myocardium and evaluate the feasibility of using allogeneic and aged MSCs. Aim 5 will determine whether MSCs engineered for enhanced survival capacity, angiogenic potential, or homing potency can better improve flow and function in chronic hibernating myocardium. Long term, the translation between the MSC-based therapy in the porcine hibernating myocardium and regenerative medicine for humans with chronic coronary artery disease will lead to optimized MSC therapeutics that can be of clinical value in managing aging and curing disease.	{ "pile_set_name": "NIH ExPorter" }
As a fundamental component of MIT's new Computational and Systems Biology Initiative (CSBi;), a new interdisciplinary pre-doctoral graduate training program in computational and systems biology is proposed. Program faculty are concentrated in the three founding academic units - Biology, Biological Engineering (BE), and Electrical Engineering & Computer Science (EECS), with additional involvement of faculty from other departments, such as Chemical Engineering, Mechanical Engineering, and Brain and Cognitive Sciences. Faculty from these departments are actively involved in research programs spanning a broad set of topics in computational and systems biology, including gene and protein networks, cell and tissue engineering, imaging and image informatics, predictive toxicology and metabolic engineering, genomics and proteomics, nanobiology and microsystems, molecular biophysics, and cancer biology. Students will apply directly to the Ph.D. program from their undergraduate or Master's institution and receive interdisciplinary training in the emerging field of computational and systems biology. Unique aspects of the program include: (a) close association with the multi- and inter-disciplinary research agenda of CSBi, (b) a unique core formed from newly developed, interdisciplinary classroom subjects that combine biology, engineering, and computation, (c) a seminar program focusing on a broad range of research both within and outside of MIT and that includes student participation, (d) an annual retreat with participation of students and faculty focusing on research, leadership, and challenges to interdisciplinary research, (e) the annual CSBi symposium, which is an exceptional conference bringing leaders in the field to MIT each year, and (f) multi-disciplinary thesis committees.	{ "pile_set_name": "NIH ExPorter" }
The kinetochore is the specialized chromosomal region which forms the attachment point for the microtubules of the mitotic spindle. Little is known about either the composition or mode of action of this structure. This proposal presents a plan for the identification, structural mapping, and functional characterization of kinetochore proteins. These experiments will exploit autoimmune sera from patients with the CREST syndrome of scleroderma. These sera contain antibodies which bind specifically to the centromere region of human chromosomes. The possible significance of these antibodies in the pathogenesis of the CREST syndrome is currently unknown. In immunoblots they react with three minor chromosomal proteins CENP-A (Mr = 17kd), CENP-B (80kd) and CENP-C (140kd), as well as with several other antigens. We have used affinity-purified antibodies to show that the CENP species are found at the centromere. In an effort to obtain biochemical amounts of these extremely minor proteins, we will use the lambda GT11 expression vector system to clone cDNAs encoding the CENP antigens. This will enable us to purify sufficient amounts of CENP: B-galactosidase fusion proteins from bacterial lysogens for injection into rabbits. In this way we will obtain high titer antibodies of known specificity directed against individual CENP species. These antibodies will be used to probe isolated taxol-stabilized mitotic apparatus in experiments designed to show which, if any, of the CENP species interact closely with spindle microtubules. They will also be used in conjunction with an in vitro microtubule assembly assay in order to probe the role of individual CENP species in kinetochore function. The rabbit antibodies will also be used for immunoelectron microscopy to map the distribution of individual CENP proteins in the trilaminar kinetochore structure. In addition, the cDNA clones will be used to determine the pattern of synthesis of the CENP proteins throughout the cell cycle by Northern blotting, the organization of the CENP genes by Southern blotting, and finally, the sequence of the CENP proteins. These data will demonstrate whether the CENP species are the products of a multi-gene family, as suggested by previous immunoblotting experiments. The CENP species are not the only chromosomal proteins recognized by the CREST patient sera. The above experimental design will also be applied to the study of other antigens whose chromosomal location is currently unknown.	{ "pile_set_name": "NIH ExPorter" }
The Centralized Protocol and Data Management Unit (CPDMU) is a Cancer Center Shared Resource widely used by members performing clinical and translational trials. The CPDMU, which was formerly known as the Clinical Trials Office at the last CCSG review, plays a central role in clinical/translational research in the Cancer Center. The primary mission of the CPDMU is to provide the necessary components and support to conduct high-quality clinical cancer research at the Cancer Center at the University of Nebraska Medical Center, as well as UNMC affiliated hospitals (The Nebraska Medical Center, Children's Hospital Medical Center, Veterans Administration Hospital), and at Cancer Center Affiliate sites across the state. The CPDMU provides a centralized resource to facilitate the development, conduct, quality assurance monitoring, compliance with regulatory agency requirements, and evaluation of clinical research at the Cancer Center. All cancer related research proposals flow through the protocol development process established and administered by the CPDMU and are subsequently sent for review to the Protocol Review and Monitoring System (PRMS), the PRMS Scientific Review Committee (SRC) and to the IRB. The CPDMU database of clinical trials also provides support for the PRMS SRC and Audit Committee (AC) and for the Data and Safety Monitoring Committee (DSMC).	{ "pile_set_name": "NIH ExPorter" }
Malaria is one of the leading causes of mortality in the world. Current therapy for malaria is limited by drug resistance. Toxoplasma gondii is found worldwide and causes devastating eye disease and encephalitis. Current therapy for toxoplasmosis is poorly tolerated and does not eradicate infection from its host. Veterans are exposed to these infections during their military service and new treatments would greatly improve veterans' health. Dr. Doggett and his colleagues have developed new drugs, endochin like quinolones (ELQ), that are highly effective against malaria and T. gondii. He has chosen ELQ-316 as a lead drug because it is orally available, non-toxic and highly effective. Dr. Doggett's preliminary studies in T. gondii and Saccharomyces cerevisiae suggest that ELQs inhibit the mitochondrial cytochrome bc1 complex (cyt bc1). Dr. Doggett will test the hypothesis that ELQ-316 inhibits cyt bc1 and characterize how ELQ-316 interacts with the cyt bc1. He will also test if inhibiting both active sites of cyt bc1 reduces P. falciparum's ability to develop drug resistance. Dr. Doggett will test this hypothesis by measuring the rate of resistance against the combination of ELQ-316 with atovaquone and the rate of resistance of ELQ-316 against a P. falciparum clone that has atovaquone resistance. If mutations occur in both cyt bc1 active sites, he will measure the catalytic activity of the enzyme. Dr. Doggett will optimize the administration of ELQ-316 by finding synergistic combinations of ELQ-316 with clinically used drugs against toxoplasmosis and malaria, determining the pharmacokinetics of ELQ-316 orally and transdermally, and developing an ELQ-316 prodrug. Dr. Doggett is an infectious disease physician at Oregon Health & Sciences University and the Portland VA Medical Center. He is specifically interested in new treatments for neglected tropical diseases that effect veterans. He has carried out research in Dr. Mike Riscoe's lab investigating drug mechanism of action and drug resistance in protozoan pathogens. Dr. Doggett has developed proficiency in the biochemical and molecular methods that are needed for his current research plan. The VA CDA-2 award will provide Dr. Doggett further training in drug mechanism and new training in pharmacokinetics, drug design and computer based molecular modeling. As a part of a robust training plan, Dr. Doggett has selected a panel of senior scientists with expertise in molecular parasitology, T. gondii, Plasmodia, preclinical drug development and S. cerevesiae as a model system. This panel is experienced in mentoring early investigators and will meet with Dr. Doggett formally every 6 months. The panel will provide advice regarding experiments, publications and career development. In addition, Dr. Doggett will attend the Woods Hole molecular parasitology course, attend science courses at OHSU and present his research at conferences. This award will culminate in the establishment of Dr. Doggett's independently funded lab devoted to anti-protozoan drug discovery for veterans and his infectious disease career as a physician with parasitology expertise devoted to the care of veterans.	{ "pile_set_name": "NIH ExPorter" }
The present long term management of children with congenital adrenal hyperplasia (CAH) is non-physiological and suboptimal requiring administration of hydrocortisone (cortisol) or cortisone acetate three times per day and Florinef daily. The amount of cortisol administered must be carefully monitored and regulated depending on the child's size, growth rate, and bone age. Over suppression of the adrenal gland leads to growth failure , delay in bone age, and a Cushingoid appearance while under suppression leads to the over production of abnormal adrenal androgens which lead to accelerated growth , advancement of the bone age, premature fusion of the epiphysises and ultimately in virilization and very short stature. The purpose of this study is to determine if the physiologic replacement of cortisol by subcutaneous administration of cortisol will provide a new therapeutic modality for the treatment of CAH. Specifically, we want to determine if it will 1) reduce the amount of steroid required to achieve normal physiologic plasma concentrations of cortisol, 2)normalize the plasma concentrations of ACTH, and 3) normalize the circulating concentrations of abnormal hormones generated by the adrenal glands. This will be accomplished in three parts by: A. Refining the technique of administration and dosing of cortisol by a subcutaneous route utilizing a portable infusion pump in normal adult volunteers who are parents of children with CAH. B. Determining the pulsatile nature of ACTH, cortisol, delta-4- androstenedione, and 17-hydroxyprogesterone secretion in normal prepubertal children (who are siblings of children with CAH) and prepubertal children with CAH on present accepted therapeutic regimens. C. Determining whether subcutaneous boli of cortisol can mimic the pulsatile nature of cortisol secretion seen in normal children in the children with CAH. D. Determine whether subcutaneous boli of cortisol delivered in physiologic amounts can suppress the abnormal plasma concentrations of 17-hydroxyprogesterone, (4-androstenedione and ACTH in children with CAH.	{ "pile_set_name": "NIH ExPorter" }
The new Alzheimer's Disease Research Center (ADRC), at the University of California at San Francisco (UCSF) has made remarkable progress in just four years, developing new diagnostic approaches to dementia, supporting exciting translational and basic science research and providing training and education at a local and national level. We remain committed to four overarching aims: 1) to explore heterogeneous presentations of dementia in the early stages;2) to train new basic and translational science leaders in dementia;3) to educate regarding the non-AD dementias and atypical presentations of AD using conferences and novel web-based approaches;and 4) to bridge the gap between clinical and laboratory studies in dementia by supporting new treatment efforts for AD, FTLD and CJD. With renewal, we will develop better ways to diagnose patients with early FTLD, CBD, PSP, CJD and atypical AD. We will catalyze the development of rational therapies for FTLD with ubiquitin-TDP-43 inclusions, FTLD with tau inclusions, and FTLD associated with progranulin mutations. To generate successful new therapies, we will create effective animal models of disease (Farese project 3) and enhance the accuracy of early diagnosis using our Clinical Core and novel imaging approaches (Seeley project 1). We will involve investigators at the Gladstone Institute such as Lennart Mucke (project 2), Yadong Huang, Li Can, and Robert Mahley in the evaluation of ADRC tissue to bring laboratory findings to the clinic. We will also interact with the CJD drug development program in the Prusiner and DeArmond laboratories by providing well-characterized patients and biosamples. By developing an internet-based approach to education, we will reach tens of thousands of people every year, educating them about dementia, its symptoms, causes and available treatments. We will also host novel and exciting conferences and retreats that generate enthusiasm for our field. To spearhead dementia research, we will continue to attract, train and nurture future leaders in neurodegenerative disease, both in clinical and basic science research. To address the inequalities among aging minority populations, we will develop a unique cohort of Chinese-American patients and integrate this community into our treatment efforts. To understand why some people are able to stave off dementia, we will develop a large healthy aging cohort that will participate in new preventative research efforts. Finally, we will create ideal data management approaches for neurodegenerative conditions that will be shared with other ADRCs, beginning with UC-Davis.	{ "pile_set_name": "NIH ExPorter" }
This is a competing continuation proposal for the Florida Node of NIDA's CTN. The aim of the Node is to develop and implement protocols that test the effectiveness of treatments across a broad range of community-based treatment settings with diverse patient populations, and to disseminate knowledge in a manner that improves the practice of drug abuse treatment in the Node, Puerto Rico and the nation. The partnership funded by this grant is led by the University of Miami-Center for Family Studies, with its considerable clinical trials, multi-site, HIV+, race/ethnicity, and drug abuse treatment expertise, serving as the Regional Research and Training Center (RRTC). The Florida Node governance represents strong bi-directional collaboration between the RRTC and CTPs. Collaborators in this partnership are five Florida CTPs that are among the largest and most respected in the State, representing the north (Gateway in Jacksonville), west (PAR in the Tampa/St Petersburg), East (Center for Drug Free Living in Orlando) and south (The Village in Miami;Spectrum in Broward County) parts of the State. Accounting for 45% of Florida's drug abuse treatment dollars, and with the capacity to serve 20,000 patients, these agencies offer exceptional diversity of treatment modalities, patient racial and ethnic profiles, and drug abusing and addicted populations. The Node also includes strong collaborative relationships with the Florida Single State Agency Director, the Florida ATTC, and the Florida provider association. As our progress report shows, the past 4 years have yielded major accomplishments in our work with these partners. Additionally, this proposal presents a number of ancillary relationships to expand the diversity of treatment settings, including primary care and social service settings represented by two large Miami-Dade County based primary care networks, a methadone treatment network, a government-run drug abuse treatment network, a close collaboration with the Puerto Rico and Virgin Islands ATTC, and a network of providers in Puerto Rico, among others. The infrastructure we have established over the past 4 years have allowed us to meet the challenges we had originally confronted (e.g., communication and collaboration across geographic distance) and have allowed us to successfully implement six CTN protocols including leading a large trial of family therapy for substance abusing adolescents, (CTN-0014), and co-leading the first ever multi-site effectiveness trial of drug abuse treatment with a Spanish-speaking population (CTN-0021). A concept, ideally suited for the CTN, proposes to test two strategies in the dissemination of an efficacious HIV intervention.	{ "pile_set_name": "NIH ExPorter" }
Both prokaryotic and eukaryotic systems are represented in this study of transcription termination at the ends of genes, and of events associated with producing the mature 3' ends of transcripts in E. coli and S. cerevisiae. The molecular biology of such problems in these microorganisms has the advantage of both sophisticated genetics and biochemical accessibility. The focus will be guided by an additional desire to understand (1) what aspects of protein structure determine the strength and specificity of recognition and binding to nucleic acid, and (2) how RNA sequence and structure (more varied and less well understood than double-stranded DNA) relates to interaction with and recognition by protein. Proposed experiments will use current genetic, biochemical, and chemical approaches in vivo and in vitro. 1. Factors and signals involved in mRNA 3' end formation in E. coli and yeast: Recognition requirements of both wild-type and mutant E. coli rho proteins and RNA target sites for ability to catalyze transcription termination will be studied. Yeast mutat- ions identified in vivo as affecting transcriptional readthrough will be correlated with RNA cleavage and polyadenylation in vitro, and maturation of the RNA transcript. 2. Protein-nucleic acid interactions in transcription. Analysis of the functional interactions between RNA and proteins or macromolecular complexes that act upon it will be undertaken using chemical and enzymatic probes as well as crosslinking procedures. 3. Coupled interactions in transcription. The dynamic interactions that couple RNA binding and ATP hydrolysis to events producing mature 3' ends will be examined as will higher order overall coordination of polymerase termination and the processing events. Analysis of the interaction between regulatory sequences and the macromolecules or cofactors acting upon them is a prerequisite for understanding these processes at the molecular level. The basic principles involved in microbial systems will undoubtedly be applicable to the control of gene expression in higher organisms. Since many diseases are the result of regulatory mechanisms done awry, in the long run a detailed molecular understanding should contribute to the development of solutions to these health problems.	{ "pile_set_name": "NIH ExPorter" }
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. MHC class I-restricted cytotoxic T-lymphocytes (CTL) play an important role in controlling human immunodeficiency virus and simian immunodeficiency virus (SIV) infection. As a natural host of SIV, sooty mangabeys (Cercocebus atys;Ceat) are a valuable model for the study of AIDS pathogenesis. We have previously defined several immunodominant SIV-specific CTL epitopes in sooty mangabeys and shown that CD8+ T lymphocytes inhibit SIV replication in vivo in naturally SIV-infected sooty mangabeys. In order to identify the MHC class I restriction of immunodominant CTL epitopes, we have cloned and characterized MHC class I genes of sooty mangabeys. cDNA libraries generated from two naturally SIV-infected sooty mangabeys were screened for MHC Class I cDNA with an oligonucleotide probe which hybridizes to a conserved region in exon 4 of primate MHC Class I genes. Based on a minimum of two clones with identical full-length MHC class I sequence, to date we have identified five MHC-A locus alleles designated as Ceat-A0101 to Ceat-A0501, and 12 MHC-B locus alleles designated as Ceat-B0101 to Ceat-B1201 in sooty mangabeys. Sequence specific primers (SSP) have been developed for the defined MHC class I alleles to enable large-scale MHC Class I typing of sooty mangabey genomic DNA by PCR-SSP and are being used to MHC type the colony of SIV-infected and SIV-negative sooty mangabeys at YNPRC. To date, samples have been collected from 155 animals for processing.	{ "pile_set_name": "NIH ExPorter" }
We have developed novel approaches to create and evaluate murine immunodeficiency models and to identify genes potentially involved in such diseases. In particular, we have developed the RAG-2 deficient blastocyst complementation method to rapidly assess the role of almost any gene in lymphocyte differentiation of function. We will use these methods to generate models for certain human immunodeficiencies that affect lymphocyte differentiation and/or function. The human btk gene is implicated in X-linked agammaglobulinemia (XLA). We will determine the structure of the murine btk gene and then use gene targeted mutation techniques to generate mice that lack a functional btk gene in their lymphocytes (via RAG-2 deficient blastocyst complementation) or in their germline. These mice will serve as a murine model for XLA and as an experimental system to study the function of the btk protein. To elucidate the function of the btk protein, mutated forms of the btk gene will be used to replace the inactivated btk gene either by transfection of appropriated constructs into mutant ES cells and RAG-2 deficient blastocyst complementation or by transgenic complementation of btk germline mutations. Mice carrying the mutated forms of btk will be assayed for phenotype characteristics and used as a source of cells for biochemical studies of btk function. We also have employed a simple assay to rapidly identify mutations that impact on the VDJ recombination reaction. As a means to identify genes potentially involved in human SCID diseases, we will employ mutant CHO cells to identify, isolate, and characterize genes involved in VDJ recombination and DNA repair, such genes have been implicated in the mouse scid defect. We have already defined 3 such genes based on ability to complement CHO DNA repair/VDJ recombination defects and demonstrated that two of these map to human chromosomes 2 and 5, respectively. Once isolated, these genes will be disrupted in ES cells and assayed for affects on lymphocyte development by RAG-2 deficient blastocyst complementation. We will similarly assay cells from human severe combined immunodeficient patient (that are not X-linked of ADA-deficient) for defects in VDJ recombination with the ultimate goal of elucidating the involved gene.	{ "pile_set_name": "NIH ExPorter" }
Abstract The goal of this project is to build a state-of-the-science regional center of research excellence in Colombia, known as the Stanford-Colombia Collaboratory on Chronic Disease (S-C3), whose focus is on the prevention of major non-communicable diseases (NCDs) facing Colombia and the region, including cancer, diabetes, cardiovascular disease, and depression. Physical inactivity is one of the most important contributors to the global burden of NCDs, responsible for ~6-10% of major NCDs worldwide. The prevalence of physical inactivity and sedentary behavior is increasing in LMICs and approaching levels of higher income countries. In Colombia, 47% of adults and 74% of adolescents do not meet physical activity recommendations. Multi-level approaches that promote physical activity at the individual, family, community, and societal levels hold substantial promise for effectively preventing and controlling the major NCDs. We and others have demonstrated effective strategies for increasing physical activity across the socioeconomic gradient using approaches that leverage widely available information technologies such as smartphone applications (apps), SMS/text messaging, interactive voice response systems, and culturally tailored virtual advisors. This grant application builds upon an existing productive collaboration between scientists from Stanford University and the Universidad de los Andes in Bogot, Colombia. The S-C3 will enable scientists in the region to expand their expertise to include development and rigorous evaluation of behavioral interventions to promote physical activity that are effective and scalable. Our specific aims are the following: 1. Establish the necessary infrastructure to support a regional center of research excellence for NCD prevention in Colombia. The S-C3 will represent a partnership among key organizations in Colombia, including Universidad de los Andes, Universidad del Rosario, their partners from the sectors of Health, Sports and Recreation, as well as Transport and Urban Planning, and Stanford School of Medicine. 2. Build sustainable mechanisms for increasing research capacity aimed at developing and rigorously testing theory-based interventions that leverage culturally tailored, widely available technologies aimed at primary and secondary prevention of NCDs in Colombia. Capacity-building activities in Colombia will serve as a resource and a model for other Latin American countries. 3. Demonstrate the partnership's capability for engaging in cutting-edge behavioral intervention research to promote physical activity and prevent NCDs. We will conduct two initial pilot studies that target individual and community levels to increase physical activity in underserved populations. 4. Develop short-, mid-, and long-term trans-disciplinary research plans that focus on optimizing regional capacity for impactful change in the prevalence of NCD's in Colombia. We will leverage the partnership's expertise in creating effective physical activity interventions in diverse Latino populations.	{ "pile_set_name": "NIH ExPorter" }
The objectives are to study the inductive relationship between developing central and peripheral synapses, and the influence of these synapses on development changes in motoneuron properties. To ensure specific and long-term interaction between cells, differentiation of synapses is directed by gene expression as well as by influential signals passing in both directions across the synapse. Detailed information has accumulated describing the function of sensory-motor connection in adult vertebrates, but little is known about their development. Most knowledge comes from studies of developing central synapses in invertebrates, and non-mammalian vertebrates such as tadpole and chick. I will study the electrical and pharmacological properties of immature motoneurons in rat embryos, and investigate the roles of developing primary afferent-motoneruon contacts and peripheral nerve-muscle interactions on these properties. I shall study pattern and time course of development of spinal reflexes and the initial specificity of afferent-motoneuron contacts. Studies will be carried out in vitro, using isolated segments of thoracic spinal cord with or without their adjacent intercostal muscles. The small size of the embryonic spinal cord allows adequate penetration of oxygen and nutrients into the tissue. The advantages of using spinal cord-intercostal muscle explants are: 1) motoneurons are cultured in continuity with the muscles they normally innervate in vivo, and 2) we have previously described the chronology changes that occur during development of intercostal nerve-muscle contacts in vivo and in organ culture, and thus will be able to correlate these changes with the differentiation of motoneurons that innervate them. Maintaining these explants in the controlled environment of organ culture is valuable for isolating signals that influence motoneuron properties and the formation of central neuronal interactions. To broaden our knowledge about the way functional neuronal circuits develop, we must extend our information about the mechanisms involved in specificity of synapses during development. Such an understanding may give insight into mechanisms of long-term interactions between cells and reasons for their inability to re-establish these interaction following lesions in the central nervous system.	{ "pile_set_name": "NIH ExPorter" }
This project proposes to examine spatial and temporal linkages between human population and the environment in the Wolong Nature Reserve in China. Wolong is the largest reserve for conserving the world-famous endangered giant pandas. It also has more than 4000 local residents. The human population is organized around households (942 in 1998), which traditionally included several generations living together, but this tradition is being broken up. Since 1975, Wolong's human population has grown 66 percent, but the number of households has increased 115 percent. Each household garners resources needed to live, particularly fuelwood for cooking and heating, from the surrounding landscape. In this study, we view population-environment interactions as the interrelationships among five major components: human population, forests, giant panda habitats, socioeconomic and institutional factors, and government policies. Forests and giant panda habitats represent the environment, whereas socioeconomic and institutional contextual factors and government policies influence how human population and the environment interact with each other. Fuelwood consumption by local residents is now the single most important human factor affecting forests and subsequently giant panda habitats (forests are an important component of the panda habitats with trees as covers and bamboo as staple food). Thus, we treat fuelwood consumption as the main linkage between human population and the environment. We will take a systems approach to address five interrelated specific aims: (1) to understand human population processes and dynamics, (2) to examine the relationships between fuelwood consumption and household demography, (3) to identify spatial interactions between population and the environment, (4) to analyze reciprocal effects of population and the environment, and (5) to predict long-term spatial dynamics of population-environment interactions under different policy scenarios. To achieve these aims, we will use and integrate extensive household and socioeconomic surveys, interviews with local officials and residents, collection of historical data, field observations and measurements, data from previous and ongoing studies, statistical tools (e.g., event history analysis, multilevel modeling, logistic regression), graph theory and network analysis, spatial technologies (geographic information systems, remote sensing, and global positioning systems), and systems modeling and simulation. The completion of our proposed project will have significant implications for population- environment interaction theories, methods, and applications.	{ "pile_set_name": "NIH ExPorter" }
Over the last year Dr McBains lab has continued their investigation into the differential mechanisms of synaptic transmission and plasticity onto both principal neurons and inhibitory interneurons within the hippocampal formation of the mammalian cortex. In addition to this main goal we have also explored the role of intrinsic voltage-gated channels in regulating individual neuron as well as network excitability with the use of high-resolution whole-cell patch clamp recording techniques in brain slices of hippocampus. A new research endeavor explores the neurogenesis, migration and development of specific cohorts of local circuit GABAergic interneurons arising from the medial ganglionic eminence. Cells originating from the MGE give rise to distinct populations of interneurons that then populate the developing hippocampus. For all of these studies we use a combinaiton of high resolution electrophysiological tools, molecular and biochemical techniques as well as confocal and two-photon imaging. [unreadable] We have continued to explore the novel forms of long lasting plasticity (both long term depression and long lasting potentiation) observed at glutamatergic excitatory synaptic connections between dentate gyrus granule cells and interneurons of the CA3 hippocampus. In this cycle we have provided evidence demonstrating that the mossy fiber-CA3 system engages their interneuron targets via multiple parallel systems that differentially utilize glutamate receptors to endow distinct synaptic properties and computational outcomes for the postsynaptic target neurons. We are currently exploring the roles played by both feedforward and feedback inhibitory circuits in sculpting the overall CA3 network excitability and our data suggest that a fine balance between these two inhibitory systems maintains a narrow temporal window of excitation for CA3 principal cells. W have continued to explore the essential role for the presynaptically located metabotropic glutamate receptor, mGluR7, in controlling bidirectional synaptic plasticity at Ca-permeable AMPA receptors on hippocampal interneurons interneurons. Activation of mGluR7 by synaptically released glutamate triggers long term depression of synaptic transmission and subsequent internalization of the mGluR7 protein. This reduction of transmitter release is accompanied by a persistent depression of the voltage gated Ca transient. As a consequence, subsequent rounds of synaptic stimulation reverse or potentiate this synaptic depression providing a novel mechanism of bidirectional control at inhibitory neuron synapses. We have now elucidated the underlying mechanism of this long lasting potentiation, which results from a nascent cAMP dependent pathway that is sequestered when mGluR7 is present on the presynaptic surface membrane. Activity dependent internalization of this metabotropic glutamate receptor permits activation of a cAMP-dependent cascade which then acts to strengthen synaptic transmission on stratum lucidum interneurons by a mechanism involving an increase in transmitter release probability. During the course of this study we also identified a novel mechanism of long lasting plasticity at developing mossy fiber synapses onto CA3 pyramidal neurons. This previously unexplored form of plasticity mimics a naturally occurring developmental switch in the phenotype of glutamate receptor present at these synapses. Young, immature synapses typically signal via Ca-permeable, GluR2-lacking AMPA-preferring receptors, induction of this long lasting form of synaptic depression triggers a switch in the receptor type which is then replaced by GluR2-containing, Ca-impermeable AMPA receptors. [unreadable] In addition, we continue to explore the developmental profile of local circuit inhibitory interneurons both at the level of their GABAA receptor activity and their synaptic targeting within the hippocampal network. Using a combination of electrophysiology and neuronal modeling we have elucidated the role of muscarinic and kainate receptors in regulating synaptic transmission onto downstream partners as well as elucidating the roles played by these receptors in tuning interneuron firing preference with a particular attention to discreet interneuron subpopulations. Muscarinic receptors activate a number of intrinsic ion conductances; the interplay of these three conductances combine to tune the frequency response of the interneuron firing pattern toward specific frequency ranges.	{ "pile_set_name": "NIH ExPorter" }
This is a new Core developed to provide glass chip-based DNA microarrays to UMCCC investigators. The core will also maintain the Research Genetics human, mouse and rat cDNA libraries, and will assist UMCCC investigators with array hybridization, analysis and informatics. The Cancer Center has contributed of $90,000 for Microarray Core development. These funds were leveraged with contributions from the Medical Center for clone purchase and validation. The Core is also partially supported by the Prostate Cancer Spore grant.	{ "pile_set_name": "NIH ExPorter" }
Obesity among children is a major public health concern in the United States. A recent population-based survey estimated that 10% of children under age 2 are obese (at or above the 95th percentile of the weight-for-recumbent-length growth charts). Although imbalance between energy intake and energy expenditure is certainly a key determinant of weight gain, emerging evidence supports the hypothesis that environmental contaminant exposures, particularly those occurring in early-life, may interfere with homeostatic control and induce or exacerbate obesity. Organophosphate flame retardants (OPFRs) are commonly added to consumer products to reduce flammability and to meet fire safety standards. They have been detected with high frequency in recent studies of home, office, and automobile dust, suggesting that OPFRs leach from products and that widespread human exposure is likely. Although no comprehensive assessment of their distribution in the general population has been undertaken, our preliminary data and several small studies indicate that exposures are ubiquitous. Our proposed research will be the first to assess levels of OPFRs in biological samples from pregnant women and investigate individual characteristics associated with high exposure, identify subgroups at greater risk of high exposure, and assess the potential for adverse health outcomes among exposed offspring. Animal studies provide compelling evidence that OPFR exposures impact growth and development; however, the extent to which these animal studies extend to human populations is unclear. Our primary objective is to investigate the impacts of prenatal OPFR exposure on children's growth from birth through age 3, and in doing so, to fill a critical gap in the understanding of the impacts of OPFRs on human health. We will capitalize on previously collected data and biospecimens from the Pregnancy Infection and Nutrition cohort, a well- characterized cohort with carefully assessed longitudinal measures of growth from birth through age 3. Using stored second trimester urines, we will examine prenatal exposure during the developmental window that is thought to be critically relevant for environmental chemicals that disrupt growth patterning and the development of adiposity. Identifying a link between prenatal OPFR exposure and altered growth in early life could have a substantial public health impact given that even small changes in weight status have been associated with long-term adverse health. Such data is critically needed as use of OPFRs is predicted to increase as other flame retardant compounds are phased-out or banned.	{ "pile_set_name": "NIH ExPorter" }
The primary objective of this Mentored Patient-Oriented Research Career-Development Award is to allow Meredith R. Golomb (the candidate) to develop the skills necessary to become an independent investigator in perinatal stroke research. Dr. Golomb has completed fellowships in child neurology and pediatric stroke, as well as a Masters of Science degree in Clinical Epidemiology. She wrote her MSc thesis on walking after perinatal stroke. She is an Assistant Professor at Indiana University (IU) School of Medicine, which, with its affiliated 4 hospitals, is the largest provider of tertiary pediatric care in Indiana. Dr. Golomb will take advanced classes in biostatistics, study design, and research ethics though IU's Clinical Investigator Training Program, which is funded though an NIH K30 grant, and work with investigators at the Regenstrief Institute for Health Care, which houses one of the oldest and most complete electronic medical records systems in the country. Dr. Golomb plans to describe the clinical characteristics and the motor, cognitive, and epilepsy outcomes in a retrospective cohort of all children with perinatal arterial ischemic stroke or sinovenous thrombosis seen at the IU-affiliated hospitals over the past 10 years, and to initiate a prospective cohort study documenting patient-oriented outcomes in more detail. She plans to use logistic regression modeling and Cox proportional hazards models to assess clinical characteristics as predictors of cognitive, motor, and epilepsy outcomes. The long-term goal of this study is to lay the foundation for future clinical trials of interventions designed to improve the outcomes of these children. During the last year of the study, she plans to apply for an R01 grant to set up a pilot clinical trial. [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Transcortin will be treated with cyanogen bromide and the resulting peptide isolated by ion-exchange chromatography. The carbohydrate and amino acid composition of each will be determined. Another batch of transcortin will be affinity-labeled with 17-(5'-oxo-6'-diazohexanoyloxy)-4-pregnene-3,20-dione and then treated with cyanogen bromide. Isolation of the peptides will tell us which one is located at or near the binding site. Other experiments will identify the number of carbohydrate chains present and their role, if any, in binding of cortisol and progesterone.	{ "pile_set_name": "NIH ExPorter" }
The specific aims of this project are 1) to organize, implement and maintain a management program for reproduction in the three species of Galago at the Duke University Primate Center; 2) to obtain baseline data on selected parameters of reproduction in colony-housed Galago and 3) to test the effectiveness of gonadotropin therapy as a management tool to increase reproductive output in Galago. Techniques of management of reproduction derived from management programs used in the dairy and beef cattle industry will be modified and extended to apply to the breeding colony of Galago at the Duke University Primate Center. Experiments designed to determine the age of puberty in male Galago are described. Further experiments are detailed in which FSH and HCG are tested for their ability to induce estrus and ovulation in normal, but acyclic Galago. The long-term objectives of this project are to devise economically effective methods of breeding Galago in order to provide colony-bred animals at a reasonable cost to investigators using these animals in neuroanatomical, biomedical, behavioral and anthropological research.	{ "pile_set_name": "NIH ExPorter" }
Calponin (CaN) has been implicated in thin filament-based regulation of smooth muscle contraction by virtue of its similarities to skeletal muscle troponin I and troponin T. Little, however, is known about the structure and function of CaN. The objectives of this project are to gain information on the structural features of CaN and how it interacts with other smooth muscle proteins in the thin filament. A combination of site- directed mutagenesis, photocrosslinking and fluorescence spectroscopic techniques will be used. It is anticipated that such information will allow us to deduce the role that CaN plays in smooth muscle regulation and how it functions in this role. The specific aims of this project are as follows: (1) To determine the gross conformation of CaN. The secondary structure will be analyzed by circular dichroism (CD) spectroscopy, and its size and shape by electron microscopy and analytical ultracentrifugation. (2) To determine the proximity relationships between specific sites in CaN. Mutants containing pairs of Cys's at defined locations will be generated by site-directed mutagenesis, and the distances between them measured by resonance energy transfer (RET). (3) To characterize the properties of fragments corresponding to various regions of CaN. Fragments of CaN will be generated by genetic engineering techniques, and their properties such as secondary and tertiary structure, and modes of interaction with other thin filament proteins analyzed. (4) To identify and characterize sites of interaction with other smooth muscle thin filament proteins. Mutants of CaN containing single Cys's at putative interaction sites will be generated and labeled with photoactivable crosslinkers. Photocrosslinking with target proteins will be achieved by ultraviolet irradiation, and crosslinked products will be subjected to peptide and amino acid analyses to identify the amino acid residues that are involved. (5) To determine the parameters of interaction between CaN and its target proteins. Interaction parameters (stoichiometry and affinity constant) will be measured by fluorescence and fluorescence polarization techniques using single Cys mutants of CaN. (6) To determine the spatial arrangement of CaN in relationship to other proteins in the smooth muscle thin filament. Using single Cys mutants of CaN, the distances between various sites in CaN and various "landmark" sites in other thin filament proteins will be measured by RET. Such distance information will be used to deduce how CaN is "positioned" in the smooth muscle thin filament. Information from this project will contribute towards the understanding of normal smooth muscle function, and may eventually provide clues to the prevention and cure of smooth muscle related diseases.	{ "pile_set_name": "NIH ExPorter" }
I The overall goal of this research program is to understand the molecular events that bring about the transduction of photons into changes in membrane conductance in retinal cone photoreceptors and the mechanisms of receptor light adaptat on. Us ng so ated sing e photoreceptor cells, as wel as prote n molecules coned from them, we propose to continue our studies with the following specific aims. With the use of time- and space-resolved cell fluorescence we propose to determine the amplitude and time course of light-dependent changes in cytoplasmic Ca in intact, functional cone outer segments. We propose to test the relationship between these changes and the state of light-adaptation of the cells. The electrical response to light in cones arises from the activity of ion channels controlled by cytoplasmic cyclic nucleotides (CNG ion channels). The channel's sensitivity to nucleotide concentration changes depending on cytoplasmic Ca concentration. We propose to investigate the mechanisms of this Ca-dependent modulation by identifying the modulator molecule. This will be achieved through biochemical and molecular biology tools. Once the modulator molecule is identified, its functional properties will be characterized through functional studies both in cultured cell lines and isolated photoreceptors. This modulator is likely to play a central role in the ability of cone photoreceptors to operate over a wide range of light intensities. This thesis will be tested by exploring the functional consequences of overexpressing or underpexpressing the modulator protein in intact cones. Congenital achromatopsia is caused by specific genetic mutations in the cone CNG channels. Some of these mutations are in a structural motif known as $4. We have identified the reason these mutations cause functional channel failure and we propose to determine the cellular mechanisms of this failure. To this end, the consequence of ooint mutations in $4 will be studied with electrophysiological, biochemical and cell biology tools. Cone photoreceptors generate synaptic signals to inform other retinal neurons of the absorption of light. Extracellular, circulating paracrine molecules control these synaptic signals. We propose to test the thesis that synaptic regulation is achieved in part through control of a specific ion channels known as HCN. The possible control of the channels will be investigated in intact cells. The molecular identity of the channels will be determined through cDNA cloning techniques. The function of the cloned channels will then be characterized in cultured cell lines.	{ "pile_set_name": "NIH ExPorter" }
The main objective of this research is to establish the fact that prezygotic selection is one of the important selection mechanisms to maintain various genetic polymorphisms in nature. Special emphasis will be made for human populations, particularly for the ABO blood group system. This project requires long term efforts in time and is a long range goal. During the period of the present proposal, efforts will be made for the following specific points: 1) With the genetic and demographic informations already in my hands, together with some supplementary informations to be collected, the age-specific death-birth rates or reproductive values associated with each of the ABO types will be described, and based on this, with some other informations such as segregation frequencies, the overall fitness of each ABO type could be constructed. Special emphasis will be placed on the role of prezygotic selection in the total fitness. 2) Parental inbreeding data could be used to examine the average effect of prezygotic selection operating in a certain quantitative traits in man. With the existing data collected from Odate, Japan, together with some supplementary informations to be collected, the effect of parental inbreeding upon the height, and body weight, of their children will be examined. 3) As an experimental organism, Drosophila melanogaster will be studied for the Mr system in parallel to the human ABO study. The Mr system element exhibits several peculiar genetic phenomena, particularly distorted segregation. The frequency of this element in natural population is unbelievably high (50 percent or more), and this system seems to be really interesting and useful system in working with the components of fitness, and their interactions in the overall fitness.	{ "pile_set_name": "NIH ExPorter" }
Recent observations made in our laboratory and by others suggest that defects in development of the fine structure of cerebral cortex, especially of terminal axons, synapses and dendrites, form the anatomic basis in some cases of mental retardation. The present study is designed to further explore this finding. Dendritic development in brains from the retarded will be studied by use of the Golgi-Cox method. Whenever possible, cases in whom the etiology of the mental defect is known will be selected for study. Frontal cortex, hippocampus, and neocerebellar cortex will be examined. Extent of dendritic branching will be assessed by use of the quantitative Golgi method. In addition, the adaptability of the Bloom-Aghajanian electronmicroscopic method for synapses to human postmortem brain tissue will be explored. Synapse counts will be obtained from areas adjacent to those studied by the Golgi-Cox technique. Ganglioside sialic acid and activity of Na ion - K ion ATPase - two possible biochemical markers for neuronal and synaptic membranes - will be measured in the same brain areas. It is hoped that from these studies will emerge patterns of developmental defect which will provide a neuropathologic basis for cases of mental retardation that fail to show significant lesions by gross examination or by standard light microscopy. The data also should provide further information regarding the special vulnerability of developing cerebral cortex.	{ "pile_set_name": "NIH ExPorter" }
The localization of chitin in yeast cell walls was studied by fluorescence and electron microscopy, with the use of specific markers attached to fluorescein or colloidal gold, respectively. Chitin was also labeled by incorporation of (14C)glucosamine in growing cells. The results confirmed that most of the chitin is in primary septa and that its distribution on the septum is uniform. Some chitin (7-8% of the total) is dispersed over the cell wall. Further studies on the regulation of yeast Beta(1 yields 3) glucan synthetase have shown that both ATP and GTP activate the enzyme, but by different mechanisms. GTP seems to act by simply binding to the enzyme or to a regulatory subunit. ATP, on the other hand, would modify, in an enzymatically catalyzed reaction, a small molecular weight compound attached to enzyme, thereby converting it into an activator. The effect appears to be reversible, inasmuch as incubation with Mg ions of highly active enzyme, prepared in the presence of 5-10 mM EDTA, leads to an inactivation, which can be reversed either by ATP or GTP. It is suggested that this reversible activation process may be of great importance for the regulation of cell wall growth in vivo.	{ "pile_set_name": "NIH ExPorter" }
Cellular signaling by double-stranded (ds) RNA will be investigated in this project. DsRNA is an important regulator of gene transcription, protein synthesis and cell growth. The multiple cellular functions of dsRNA have been discovered in the contexts of the interferon system, different aspects of which will be studied in the other projects of this program. Transcriptional signaling by dsRNA will be analyzed using defective cell mutants. One such mutant has been already isolated and more will be generated during the course of the proposed investigation. The mutant cells will be examined for defects in candidate proteins which might be involved in dsRNA signaling. If no such defect is found, the cells will be complemented with cDNA expression libraries made from wild-type cells and the mutated protein will be cloned. In two other aims, cellular functions of two dsRNA-binding proteins, which were cloned during the last funding period, will be investigated. One protein, PACT interacts with PKR and activates it. The mechanism of the activation process and its consequence on transcriptional signaling, protein synthesis and cell growth regulation will be studied. P9O, the other dsRNA-binding protein, is a nuclear phosphoprotein. Its possible involvement in cell cycle regulation and nuclear RNA metabolism will be investigated. The proposed studies will provide important mechanistic information about the cellular functions of dsRNA, a major regulator in virus-infected cells.	{ "pile_set_name": "NIH ExPorter" }
Allogeneic bone marrow transplantation (allo-BMT) is an effective therapy for hematologic malignancies through T cell-mediated graft-versus-leukemia (GVL) effects, but allogeneic T cells often lead to severe graft-versus-host disease (GVHD) as well. Because GVHD and tumor relapse are two major concerns when allo-BMT is used as a therapy for hematologic malignancies, the broad and long-term goal of our research is to prevent GVHD and tumor relapse, which would greatly enhance the therapeutic potential of allo-BMT. Glycolysis and oxidative phosphorylation (OXPHOS) are two basic cellular metabolic pathways to generate of adenosine-5'triphosphate (ATP) as a source of energy. Because OXPHOS generates ATP with high efficiency, normal cells rely on OXPHOS for ATP under normoxia conditions, and only switch to glycolysis under hypoxic conditions. However, malignant cells primarily utilize glycolysis for energy production even under normoxia conditions, known as aerobic glycolysis. Similar to malignant cells, activated T cells also switch to glycolysi to acquire sufficient energy. Cell metabolism plays a key role in T-cell activation, differentiatio and function, which is essential for the induction of GVHD. As a consequence, targeting T-cell metabolic pathways to control GVHD has recently become an interesting strategy. However, very little is known about T-cell metabolic pathways under allogeneic BMT. The objective of this project is to understand the metabolic pathways of T cells after being transplanted into allogeneic recipients, and to identify/validate potential targets on T-cell metabolic pathways in controlling GVHD as well as tumor relapse. The central hypothesis is that a certain metabolic pathway likely within glycolysis is shared by pathogenic T cells and malignant cells such as leukemia, and thus targeting this specific pathway will control T-cell mediated GVHD and prevent leukemia relapse. To determine T-cell metabolic pathways (Aim 1), we will use and metabolomics technology to define the complex biochemical processes of T cells under allogeneic responses in vivo, and to identify metabolic processes or products as potential targets to specifically inhibit T-cell allogeneic responses. Using Seahorse bioenergetics approach, our preliminary study has shown that T cells after allogeneic BMT dramatically increased aerobic glycolysis, and blocking glycolysis by genetically ablating mammalian target of rapamycin (mTOR) essentially prevented GVHD. Thus, we will further validate glycolic pathway as potential target in controlling GVHD and leukemia relapse (Aim 2). Using preclinical murine models of allogeneic BMT and leukemia, we will determine whether targeting glycolic pathway is effective in controlling GVHD and leukemia relapse.	{ "pile_set_name": "NIH ExPorter" }
The purpose of this application is to investigate, at the molecular and integrative levels, the biology of the newly cloned mu and delta opioid receptors, attempting to understand how these molecules function as units, and how they interact with other brain molecules to modulate critical functions such as pain control and drug reward. A major emphasis is to understand the function of these receptors in the context of the multiplicity of ligands in the endogenous opioid family. Given that mu and delta receptors interact, to varying extents, with the products of the three opioid precursor gene family, and given that the endogenous ligands share a common "message" sequence, the question arises as to how the receptors achieve their affinity and selectivity for the various ligands. Thus, the first specific aim of this application is to carry out structure-function studies of the mu and delta receptors, investigating the nature of the binding pocket for the peptides and alkaloids, and the mechanisms of discrimination or selectivity among the various type of agonist ligands, and between agonists and antagonists. These studies will be conducted using molecular modeling and site-directed mutagenesis, as well as relying on an empirical approach with construction of chimeras to indicate critical domains, followed by mutagenesis of unique residues within these domains. The second specific aim is to place these opioid receptors in their anatomical context and to describe their expression relative to the expression of the 3 opioid precursors. Using dual in situ hybridization, receptor immunohistochemistry, or a combination of in situ and immunohistochemistry, we will study the expression of the mu and delta receptors and their ligands in order to determine the existence of autoreceptors, and to describe the interface between the opioid transmitters and the two receptors. We will survey the brain and spinal cord, and then focus on the anatomy of the opioid system in areas implicated in nociception and analgesia. This information will form the foundation of regulatory studies whereby stimuli known to alter pain responsiveness will be used to modulate the expression of the opioids, the receptors, or both. The third specific aim will move our knowledge of the mu and delta receptors at the molecular and anatomical level to a more complete description of the anatomy of drug reward, the role of opioid receptors in the reward circuits, and the modulation of these molecules by treatment with drugs such as morphine and cocaine. Using a combination of anatomical tools, we shall describe the co-expression of opioid molecules with other molecules relevant to reward mechanisms, particularly dopamine receptors. We shall define, at a finer level, subpopulations of cells in the nucleus accumbens with unique combinations of these molecules, and will study their projections. Then we shall relate these anatomical/biochemical entities to drug responsiveness, and determine changes in expression under conditions which promote drug tolerance versus drug sensitization. Taken together, these studies will permit us to use the cloning of the opioid receptors to achieve a better understanding of the biology of opioids and their role in drug use and abuse.	{ "pile_set_name": "NIH ExPorter" }
Dr. Dodds will be entering the Dentist Scientist program on July 1, 1994. Her first year will consist of laboratory rotations, didactic courses, literature review, and journal club. She will also attend the DSA Ethics and Oral Biology seminar offerings, and meet with the program director, Dr. Holt, monthly to discuss progress in the program. A mentor will be chosen following selection of a research project.	{ "pile_set_name": "NIH ExPorter" }
Title: REGULATION OF CERAMIDE SYNTHASE BY PROTEIN-PROTEIN INTERACTION Abstract: Ceramides form the backbone of all sphingolipids, and ceramide synthases (CerS) are critical enzymes for de novo production of ceramides. In spite of the key role of CerS in ceramide generation, there is a serious deficiency in understanding how these enzymes are regulated. The long-term goal of this project is to uncover and understand the fundamental molecular mechanisms of how CerS enzymes are regulated. Using a proteomics approach, we discovered that the small heat shock protein Hsp27 interacts specifically with CerS1. Based on our preliminary data we generated the novel hypothesis that Hsp27 is a negative regulator of CerS1 activity via direct interaction that can be modulated by p38-MK2 MAPK mediated phosphorylation of Hsp27, and that down-regulation of Hsp27 induces CerS1/C18:0-ceramide mediated cellular responses. To test this hypothesis, we propose the following Specific Aims: Aim 1: Define the biochemical significance of Hsp27 mediated CerS1 regulation in cells with respect to sphingolipid metabolism and signal transduction. Aim 2: Define the biological significance of Hsp27 mediated CerS1 regulation. Aim 3: Determine the mechanism of Hsp27- CerS1 protein-protein interaction. Overall, these studies will establish Hsp27 as an endogenous modulator of CerS1 and uncover a novel mechanism of how Hsp27 regulates CerS1 and CerS1/C18:0-ceramide governed mitophagy and cancer cell death. The knowledge generated from this study will help design mechanism-based novel therapies against cancer and other pathologies in which C18:0-ceramide is the key mediator by identifying new methods to modulate CerS1 activity.	{ "pile_set_name": "NIH ExPorter" }
Polypeptides synthesized in the hypothalamus have been implicated in the control of hypophysiotrophic hormone release by the anterior pituitary. Recent evidence, however, has shown unequivocally that these polypeptides, referred to as releasing hormones or factors, have significant extrahypophyseal activity that is independent of their effect on the adenohypophysis. These observations have prompted speculation that the polypeptides, and particularly pyroglutamyl-histidyl-prolineamide (thyrotropin-releasing factor, or TRF), in fact may be brain neuroregulatory agents. Preliminary data obtained in our laboratory are in accord with this hypothesis, and specifically suggest that TRF may regulate brain catecholamine metabolism. We have demonstrated, for instance, that TRF directly inhibits the enregulatory role of TRF in brain and the specific regulatory role of TRF on catecholamine metabolism. Evaluation of the effects on catecholamine function will begin by investigating potential interactions of TRF on isolated portions of the metabolic pathways, including synthetic and degradative enzymes, uptake systems, and release mechanisms. These studies will serve as an essential preface to analysis of interactions in the living animal. The second portion of this proposal, while intimately related to the studies on catecholamine metabolism, will proceed independently. We reason that if TRF is a neuroregulatory agent, then TRF should possess metabolic characteristics similar to those of known neuroregulatory compounds. Thus, we will define the neuroanatomic distribution of TRF-containing cells by immunofluorescence, the subcellular distribution of TRF in various brain regions, the nature and localization of enzymes controlling its formation and destruction, and possible mechanisms responsible for its transport and storage in cells. While the specific effects of TRF are of immediate interest in this proposal, the overall significance of potential neuroregulatory roles of other central nervous system polypeptides may be of far greater import. BIBLIOGRAPHIC REFERENCES: Stolk, Jon M. and Peter C. Whybrow: Clinical and Experimental interrelationships between Sympathetic Nervous System Activity and Pituitary-thyroid function. Neuroendocrinology, 1976, in press.	{ "pile_set_name": "NIH ExPorter" }
Sublines of transplantable rat adenocarcinoma MTW9 have been developed which differ in content of estradiol and prolactin receptors, ovariectomy response, and growth rate. These will be studied in serial transplant and in culture for genetic stability, and growth control by hormones. The regulation of receptor content and nuclear transport of these sublines will be studied in terms of hormonal environment in an effort to explain why some tumors that possess estradiol receptor do not regress after ovariectomy. We have developed techniques which may distinguish receptor sites which are apparently homogeneous by Scatchard analysis. These techniques will be utilized in measuring receptor sites for estradiol and prolactin in tumors growing under different hormonal conditions. MTW9 grows in only 20% of hosts with a normal serum prolactin but increased environmental temperature increases the number of tumors which grow out. This phenomenon is under further study. MTW9-MtT (MTW9 growing in a rat with co-implanted mammosomatotropic MtT does not regress after ovariectomy). The factor inhibiting ovariectomy regression is not prolactin or growth hormone. We will attempt to identify it.	{ "pile_set_name": "NIH ExPorter" }
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. To examine the effects of smoking cessation and smoking resumption on insulin resistance and its markers, cardiovascular risk factos, changes in body composition and fat distribution, and changes in the metabolism of body fuel substrates in humans.	{ "pile_set_name": "NIH ExPorter" }
The ability of hepatic stem/progenitor cells to respond to severe liver injury is dependent upon a finely tuned balance of progenitor cell renewal, expansion and differentiation. In the liver, developmentally activated signaling pathways including the hedgehog, wnt and notch systems have been implicated in the regulation of hepatic progenitor cell renewal, tissue repair and the development of hepatocellular cancer. Until now, lack of specific genetic tools to modulate these pathways in adult hepatic progenitors has precluded the analysis of the specific contribution of these pathways to the progenitor cell response to liver injury. We have discovered that the winged helix transcription factor Foxl1 uniquely marks bipotential hepatic progenitor cells and have derived both Foxl1-Cre and tamoxifen inducible Foxl1-CreERT2 mouse strains to enable gain- and loss-of-function studies through the conditional deletion of individual components of the hedgehog, wnt and notch signaling pathways in hepatic progenitor cells. Using this approach, we will dissect the complex autocrine and paracrine signaling events that contribute to progenitor cell recruitment, differentiation and proliferation as well as paracrine effects on fibrogenesis during tissue repair in response to liver injury. We propose the existence of a signaling hierarchy in which the winged helix factor Foxl1 mediates the hedgehog and wnt/b-catenin signaling pathways necessary for progenitor cell renewal and proliferation. We predict that Notch acts downstream of wnt/b-catenin and promotes progenitor cell differentiation towards the biliary lineage and may also be required for biliary tubule morphogenesis. Understanding the biology of hepatic progenitor cells will have wide-ranging implications for the development of therapeutics for acute liver failure, chronic cirrhotic liver disease and hepatocellular carcinoma. PUBLIC HEALTH RELEVANCE: The studies proposed in this grant application will elucidate the signals that regulate hepatic progenitor cells and their response to acute or chronic liver injury. Understanding the biology of hepatic progenitor cells will have wide-ranging implications for the development of therapeutics for acute liver failure, chronic cirrhotic liver disease and hepatocellular carcinoma.	{ "pile_set_name": "NIH ExPorter" }
Antitermination increases the transcription of genes that are located downstream of terminators. Phage HK022 antiterminates transcription of its early genes by converting host RNA polymerase to a terminator- resistant form. This requires cis-acting phage sequences called nut sites. The isolation and characterization of nut region mutants argue that the sites are about 70 bases long and act as RNA transcripts consisting of two adjacent stem-loops. These transcripts presumably interact with a recognition site on RNA polymerase. The properties of polymerase mutants that are unable to antiterminate HK022 early transcription but are normal in all other respects tested suggest that the recognition site is located in a zinc-binding domain of the beta' subunit. The modified polymerase is capable of reading through at least three successive strong transcription terminators with much increased efficiency. The efficiency is greater than 90% per terminator in vivo and about 50% in vitro, consistent with the hypothesis that the requirements for persistent antitermination in the HK022 system are extraordinarily simple. The integrase proteins of phages lambda and HK022 are closely related site-specific recombinases that recognize different nucleotide sequences in the core regions of their substrates, the attachment sites of the two phages. The two proteins differ by 92 amino acid substitutions. We have found that no more than 5 of these substitutions are critical for the different DNA sequence specificities of these enzymes. Two of these substitutions act principally by reducing specificity, while the other three act principally by specifically decreasing activity on one of the substrates.	{ "pile_set_name": "NIH ExPorter" }
The generation and maintenance of a B cell repertoire that is tolerant to self rely on multiple mechanisms that regulate the differentiation of autoreactive B cells. These mechansims are controlled by the nature and duration of interaction between B cell receptor (BCR) and its antigen. Multivalent membrane-bound self antigens or double-stranded DNA causes immature self-reactive B cells to undergo deletion or receptor editing. Lower valency antigens, such as a soluble antigen or single-stranded DNA, induce anergy. Anergy is characterized by a profound block in proximal antigen receptor signaling resulting in an antigen-unresponsive state. The induction and maintenance of anergy require chronic binding of antigen and signal transduction. Therefore, the strength of BCR signals dictates the fate of developing B cells. However, the molecular events that translate BCR signal strength to fate choice in tolerance induction and underlying signaling pathways are not well understood. We propose to map these signaling pathways genetically using a genome-wide shRNA screen. The rationale of our approach is that induction of a specific form of B cell tolerance will be defective when a component of the underlying signaling pathway is reduced by shRNA knockdown. As in a genetic screen, cells having desired phenotypic changes are first selected after they are transduced with a retroviral shRNA library. Then the shRNAs carried by these cells are amplified and sequenced resulting in the identification of underlying genes. Since it is technically challenging to carry out shRNA screen in vivo, we will first carry out the screen on WEHI-231 B cell lymphoma cell line. Crosslinking of BCR on WEHI-231 cells results in their apoptosis and it has been used as a model of tolerance induction. Our specific aims are i) to identify novel molecules of BCR signaling pathways in WEHI-231 cells; and ii) to establish the roles of identified genes in different in vivo models of B cell tolerance including clonal deletion, receptor editing and anergy. The goal of this project is to identify new components of differential BCR signaling pathways in tolerance induction. Mutations that affect BCR signaling can alter B cell fate decisions and result in autoimmune diseases. Defective tolerance induction has been demonstrated in both animal models and patients of autoimmune diseases. These studies should provide a refined signaling map which will be essential in dissecting the defects of B cell tolerance in autoimmune diseases. Such knowledge should offer new opportunities to develop novel immunomodulatory drugs. PUBLIC HEALTH RELEVANCE: Using a genetic approach, we propose to screen and identify new genes in signaling pathways that are important for establishing or maintaining tolerance in B cells. This refined signaling map will be essential to dissect tolerance defects in autoimmune diseases and to develop novel immunomodulatory drugs.	{ "pile_set_name": "NIH ExPorter" }
Numerous human disorders arise from microdeletions and microduplications of relatively large genomic regions. These rearrangements can result in copy number alterations (CNAs) of one or more genes. Conditions arising from microdeletions and microduplications can manifest as multiple congenital anomalies (MCA) in patients. The most common phenotypic features observed in these patients include global developmental delay, mental retardation, cardiac defects and cranio-facial differences. Thus, diseases mediated by CNAs are referred to as 'genomic disorders' as large regions of the genome are altered leading to disorders in multiple organ systems. Many of the recurrent genomic disorders are mediated by aberrant homologous recombination between highly identical blocks of DNA sequences referred to as low copy repeats or segmental duplications (SDs) which may comprise up to 5% of the human genome. Despite this known correlation between SDs and genomic disorders, a significant proportion of SDs have not yet been associated with disease- causing genomic rearrangements. This observation suggests that other SD-mediated genomic disorders may exist but are currently undetectable mainly due to the low resolution of diagnostic techniques standardly used in molecular cytogenetics. We hypothesize that a significant proportion of children with MCA have a submicroscopic CNA that is not evident on standard genetic testing. We believe that a substantial number of these CNAs may be mediated by SDs or other unstable architecture within the human genome. We will test this hypothesis by analyzing patients with MCA using high resolution microarrays in order to detect disease-causing CNAs. Further, analysis of the genomic sequences at the rearrangement breakpoints will help determine what proportion of CNAs are mediated by predisposing genome architecture like SDs. The goal of this proposal is to identify previously undetectable, disease-causing CNAs in patients with MCA. The identification of genomic regions altered in MCA patients will allow a better understanding of the mechanisms underlying this group of disorders. We can then begin to assess the role of gene(s) in these regions that may be critical to early human developmental pathways. Relevance: Many children have genetic diseases that are undiagnosed due to limitations in current methods used for testing. These children may have very small deletions and duplications which can only be detected by techniques that allow a genomewide analysis. The identification of genomic regions that are deleted or duplicated in patients with birth defects will allow the identification of genes that are important in normal development. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Abstract ? The Admin Core will support all grant management functions and prepare regulatory submissions for the proposed clinical studies and tissue collection. The Admin Core director will work closely with the BETRNet Steering Committee on network programs and with the PIs of Cores A and B on Research Center issues. This Core will support all grant management functions of the Research Center including budgeting, sub-contracts, and financial accounting, and to communicate with the Internal and External Advisory Committees, NCI scientific staff, BETRNet Steering Committee, and BETRNet Coordinating Center. This Core will also support all regulatory submissions for clinical studies, including human subjects protocols, Investigational New Drug (IND) Applications, analytical validation, pharmacology & toxicology studies, and standard operating procedures (SOP). This Core will also facilitate participation in all network activities, including Patient Registry-Virtual Biorepository, Pilot Projects, and Cross-BETRNet projects and communications with the Coordinating Center, Steering Committee, and NCI scientific staff.	{ "pile_set_name": "NIH ExPorter" }
A study of subcellular distribution of cadmium within animal lung cells after toxic inhalation exposure will be undertaken. Special emphasis will be placed on characterizing the molecular structure(s) surrounding the metal. The relationship between cadium and the interacting macromolecule(s) will be investigated in order to understand the mechanism of toxicity (mechanism of action) of the metal. Electron paramagnetic resonance (EPR), nuclear magnetic resonance (NMR), and other spectroscopic techniques will be used to probe the immediate environment around cadmium. The subcellular distribution pattern of cadmium in normal lungs and those subjected to chronic inhalation exposures of cadmium oxide has been established in these laboratories. The cumulative data demonstrates a shift of cadmium, normally associated with the connective tissue and cell membranes of the lung, into the cell cytoplasm. Further effort will be expended to separate and to categorize the molecular components to which cadmium is bound. Established purification procedures such as gel filtration chromatography and electrophoresis will be employed to obtain electrophoretically and ultracentrifugally homogenous components, anticipated as being primarily low molecular weight molecules. The specific absorption attributes to metallothionein(s) at 250 millimicrons, characteristic of cadmium mercaptide bonds, will be used as monitor for isolation of that component when present. A comparison between the binding of cadmium to molecular constituents of the lung and kidney will be made.	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY The successes of proteasome inhibitors bortezomib (Velcade(R)) and carfilzomib (Kyprolis(R)) as FDA-approved cancer drugs have invigorated efforts to devise new cancer therapeutics targeting upstream pathways involving enzymes of ubiquitin and ubiquitin-like protein modification systems. Our long-term goal is to provide therapies that inhibit the NEDD8 system and the major NEDD8 targets cullin-RING ubiquitin ligases to directly benefit cancer patients. The NEDD8-activating enzyme (NAE) inhibitor MLN4924 is currently the most advanced of these potential new classes of drugs. MLN4924 has extensive published pre-clinical data demonstrating strong anti-cancer efficacy and several Phase 1 clinical trials are in progress for hematologic and advanced non-hematologic malignancies. Given the clear benefit MLN4924 provides by selectively inducing cancer cell death with no noted toxicities or side-effects on experimental cancers, it remains highly significant to understand how cells respond to NEDD8 system inhibition. As an important step towards this goal and in response to PAR-12-145 NCI Exploratory/Developmental Research Grant Program, the overall objective of this proposed study is to use insight derived from experiments examining on-target MLN4924 resistance to identify new NAE inhibitors that function through mechanisms of action distinct from MLN4924. Through our strong preliminary data, we have devised a highly innovative strategy that uniquely positions us to describe the types of on-target mutations possible for MLN4924 resistance. We have also generated a new screening assay based on differential scanning fluorimetry. This uses a ligand competition strategy to evaluate the mechanism of action of molecules that bind NAE. Our central hypothesis is that the diversity of biochemical changes in NAE sufficient for cancer cell resistance to MLN4924 can be overcome by small molecules that inhibit NAE through mechanisms not involving the enzyme's catalytic pocket. To test this hypothesis and achieve the overall objective of the research, two Specific Aims are proposed: 1) to characterize the biochemical diversity of MLN4924-resistant forms of NAE and 2) to identify small molecule inhibitors of these drug-resistant NAE complexes. IMPACT: This work is highly innovative with direct translational relevance due to the extraordinary opportunity it provides to describe MLN4924 resistance mechanisms in detail before they are reported in relapsed patients and to use this information to discover new NAE inhibitors that function through distinct mechanisms of action. Collectively, these efforts will significantly contribute to providing much needed new treatment options for cancer patients.	{ "pile_set_name": "NIH ExPorter" }
We propose to configure a DNA-based assay to detect mutations in the gene related to cystic fibrosis (CF), encoding the cystic fibrosis transmembrane conductance regulator protein (CFTR). The assay will have an extended mutation detection for two domestic ethnic groups, non-Caucasian Hispanics and African Americans compared with the standard assay based on the 25-mutation panel recommended by American College of Medical Genetics. The aim of the current project is to improve the detection rates above 80% for both the Hispanic and African American groups. The goal is also to improve the coverage for Caucasians. The assay will detect 52 CF mutations and will be implemented on small electronic chips, microtransponders. Each microtransponder is composed of photocells, antenna and memory to store information that identifies the sequence of the DNA probes attached to the microtransponder surface. In the assay, fluorescently labeled target DNA binds to DNA probes. The fluorescence intensity of the microtransponder surface is quantified in a flow-based instrument, which also reads the ID of the chip. The chips, as well as the instrumentation to read the chips, have been built by PharmaSeq. The benefits of the program include improved detection of disease-related mutations leading to better health care for cystic fibrosis-affected individuals and extending clinical understanding of the genetic basis of cystic fibrosis.	{ "pile_set_name": "NIH ExPorter" }
The present application is to enhance research efforts at the University of Chicago that aim to advance the understanding and treatment of sleep disorders by obtaining support for the hiring of a new tenure-track faculty at the Assistant Professor level. The focus of the world renowned Sleep Research Program at the University of Chicago is the relationship between sleep and health, particularly cardio-metabolic risk. The program is directed by Eve Van Cauter, Ph.D., Professor of Medicine. This multidisciplinary program currently includes 13 tenured or tenure-track faculty members from the Sections of Pulmonary and Critical Care, Endocrinology, Diabetes and Metabolism and Emergency Medicine of the Department of Medicine, from the Department of Pediatrics and from the Department of Health Studies who are actively involved in research efforts related to the health impact of sleep and its disorders. The program includes basic scientists using animal models of hypoxia, clinical investigators conducting laboratory studies in small groups of normal adults and children as well as In patient populations, neuroscientists developing novel quantitative analyses of the sleep and wake EEG, physician scientists using genetic approaches and epidemiologists conducting primary and secondary analyses of existing data bases. The individual to be hired under this award would bring to the existing program crucial missing expertise in prospective cohort and intervention field studies in larger subject populations with rigorous assessments of sleep and cardio-metabolic measures. Such studies would bridge the findings from basic science and short-term clinical research center studies with the body of epidemiologic evidence linking steep disturbances and adverse health consequences. Consistent with the funding priority of the National Heart, Lung and Blood Institute for this RFA, the present application has identified a highly qualified and exceptionally promising new independent investigator. Our commitment is to appoint Kristen L. Knutson, Ph.D. (Biomedical Anthropology) as Assistant Professor (tenure track) in the Section of Pulmonary and Critical Care Medicine of the Department of Medicine at the University of Chicago for a 4-year period starting on September 1, 2009. The present application seeks to obtain 2 years of funding to cover 80% of her salary and to provide a start-up package with the resources needed to develop a pilot research project and enable her to secure independent funding. Dr. Knutson has a superb track record of research activities, including field studies of sleep in diverse populations, and career development in sleep disorders and cardio-metabolic risk, a research area relevant to the primary mission of the NHLBI. This application is to request funds to enhance the University of Chicago Sleep Research Program by recruiting tenure-track faculty at the Assistant Professor level. The new recruit will be mentored to ensure success in academic medicine with a focus on sleep disorders and cardio-metabolic risk.	{ "pile_set_name": "NIH ExPorter" }
This proposal was designed to examine the effects of sex and estrogen on the self-administration of intravenously (i.v.) delivered drugs in rats. Four animal models will be used to investigate acquisition, maintenance, regulation/dysregulation, and reinstatement of drug self-administration. The effects of sex and estrogen on the acquisition of heroin, methamphetamine and d, 1-3, 4-methylenedioxymethamphetamine (MDMA) self-administration will be examined using an autoshaping procedure. A progressive ratio (PR) schedule of reinforcement and a behavioral economic analysis of demand will be used to evaluate the effects of sex and estrogen on the maintenance of cocaine and heroin self-administration. Breakpoint (BP) measures obtained on a PR schedule, along with slopes and P-max values obtained from demand curves will be used to evaluate the reinforcing efficacy of several doses of cocaine and heroin. Differential access to cocaine self-administration will be used to investigate the effect of sex on the regulation/dysregulation of cocaine intake. Dose-response curves for cocaine will be obtained to examine group differences in shifts in the dose-response curves. The effect of estrogen on the reinstatement of cocaine self-administration will be examined using a priming model of drug reinstatement in which lever pressing for drug is extinguished by replacing cocaine infusions with saline infusions. Reinstatement of responding is then tested by administering a priming injection of cocaine.	{ "pile_set_name": "NIH ExPorter" }
This is a proposal dealing with medical consequences (pulmonary arterial hypertension) associated with HIV-1 and drugs of abuse. The advent of antiretroviral therapy has clearly led to improved survival among HIV-1 infected individuals yet this advancement has resulted in unexpected increase in the prevalence of vascular complications including pulmonary arterial hypertension. Development of HIV-associated PAH (HPAH) results in early mortality and serves as an independent predictor of death in patients infected with HIV-1. While intravenous drug use accounts for one-third of all new cases of AIDS in the United States, it has been identified as the most common risk factor in the individuals diagnosed with HPAH. Furthermore, our recent study showing enhanced pulmonary vascular remodeling in HIV-infected lung tissues from IV heroin and/or cocaine abusers indicate that IVDU and HIV-1 potentially act in concert to cause pulmonary arteriopathy. Abnormal smooth muscle cell proliferation/migration are considered to play a key role in vascular remodeling leading to increased pulmonary vascular resistance associated with PAH but the mechanisms and pathogenesis involved remain elusive. Our recent study supports an additive effect of cocaine on the HIV-Tat mediated increase in proliferation of human pulmonary arterial smooth muscle cells (pSMCs). Based on our recent strong preliminary findings we hypothesize that this Tat and cocaine mediated increase in proliferation of pSMCs involves down-modulation of anti-proliferative bone morphogenetic protein receptor (BMPR) protein expression through post-transcriptional regulation by micro-RNAs (mi-RNAs). This hypothesis will be tested by pursuing three specific aims. In the first aim we will evaluate the effect of HIV-Tat and cocaine on the BMPRs expression and its down-stream signaling pathways. In the second aim, we propose to delineate the post-transcriptional mechanism(s) involved in Tat and cocaine mediated regulation of BMPR expression, through modulation of specific miRNAs. In order to further confirm the interactions of HIV-1 and cocaine on BMPR axis, the third aim will be focused on ex-vivo and in-vivo investigation of miRNA mediated regulation of BMP/BMPR axis in pSMCs. These studies are innovative because this will be a first attempt to understand the miRNA mediated effect on anti-proliferative signaling pathways involved in the interaction of cocaine and viral protein that results in smooth muscle hyperplasia and linking these changes to the pulmonary vascular and right heart dysfunction associated with HPAH. The proposed research is significant because it will provide a more complete understanding of pathogenic mechanisms involved in the development of HPAH in the presence and absence of cocaine abuse. Thus, important advances in the development of targeted therapies and understanding of complications associated with HIV and drugs of abuse associated PAH are expected in the future which is relevant to the NIH's mission of developing fundamental knowledge that will potentially help reduce the burdens of human disability.	{ "pile_set_name": "NIH ExPorter" }
Abstract Neurodevelopmental disorders (NDDs), such as Intellectual Disability (ID) and Autism Spectrum Disorder (ASD) are more prevalent in males than females. Despite the fact the NDDs are diagnosed more frequently in males, the molecular mechanisms underlying sex bias in these disorders are still unknown. Until recently, sex had not been routinely considered in data analysis, and females were often excluded from behavioral experiments altogether. In parallel, for decades most genes identified for NDDs were on the X chromosome because of the relative ease to identify linkage regions in X-linked pedigrees, leading to mouse models that were studied only in one sex. Thus, testing of sex differences in preclinical animal models of NDDs has been limited. Our work introduces mice deficient for Coiled-coil and C2 domain containing 1a (Cc2d1a) as a novel mouse model for molecular and behavioral sex-specific deficits in NDDs. We found that CC2D1A loss of function (LOF) mutations cause ASD, ID and seizures in humans, and that Cc2d1a knock-out (KO) male mice display behavioral features of the human phenotype, including cognitive and social deficits, hyperactivity and anxiety. In contrast, our preliminary data show that female KO mice only present with milder cognitive deficits. Cc2d1a KO males also display a sex-specific reduction in CREB activation due a disregulation of the upstream PKA pathway. These findings raise the possibility that males and females may use different molecular strategies for encoding information and building neuronal circuits for the same behaviors, and in particular lead to our hypothesis that CC2D1A regulates CREB signaling in a sex-specific manner leading to sex-specific behavioral deficits. The overall goal of this proposal is to prove our hypothesis. In Specific Aim 1 we will explore morphological, physiological and behavioral differences between male and female Cc2d1a KO mice which correlate with reduction in CREB signaling. In Specific Aim 2 we will modulate the PKA/CREB pathway to demonstrate that all these phenotypes are connected by a sex-specific CREB signaling defect. Finally, in Specific Aim 3 we will explore the molecular mechanisms underlying sex-specific CC2D1A signaling. If successful our studies will identify a novel sexually dimorphic mechanisms for CREB signaling regulation and link molecular and cellular deficits to behavior. Our findings will be highly significant as they will establish the ID/ASD gene CC2D1A as a critical regulator of sex-specific signaling in the brain.	{ "pile_set_name": "NIH ExPorter" }
The Biolmaging Resource Core (BIRC) provides leadership, scientific expertise and instrumental support in quantitative imaging techniques, including MRI, CT, DXA and PET, critical to the WFU OAlC's research theme: Integrating pathways affecting physical function for new approaches to disability prevention. The BIRC is well integrated with all WFU OAIC cores and leads the development and implementation of imaging techniques and protocols to quantify and characterize body composition, particularly skeletal muscle and total, regional and organ-specific adipose tissue and to obtain non-invasive measures of vascular and neuromuscular function. The Core also supports a large image repository that it developed during the present cycle in order to facilitate cross-study comparisons and exploratory analyses of established imaging outcomes as well as rapid development and validation of new imaging phenotypes. Methods developed during the current cycle have been demonstrated to be robust and of high precision in both human research and in translational models using rodents and nonhuman primates. During the present cycle, the BIRC has strongly contributed to the strong success of the OAIC by supporting 3,431 image acquisitions (DXA, CT, MRI and PET) in 10 externally-supported studies, 8 pilot studies and 3 research development projects. The specific aims of the Biolmaging Resource Core are to: 1) determine the associations of age and disease-related changes in skeletal muscle and adipose tissue depots with biological, genetic, co-morbid, psychosocial and behavioral factors, and other outcomes relevant to physical function; 2) measure the effects of interventions (dietary/nutritional, exercise, pharmacologic, and combinations) on adipose and skeletal muscle tissue depots; 3) use in vivo bio-imaging to enable bi-directional translation of key findings between humans and animal models (including nonhuman primates) in support of mechanistic research; 4) develop and validate novel non-invasive imaging metrics which allow improved characterization of skeletal muscle, adipose tissue and vascular function in older adults to be integrated with a standardized battery of physical function and laboratory/molecular assessments. The BIRC leadership team are highly experienced and successful investigators who have strategically complementary skill sets and strong records of collaborative scientific and administrative achievements. In the initial year, the BIRC will support 7 externally-funded projects, 2 pilot projects and 1 developmental project. The BIRC will also provide training and mentoring to RCDC-supported faculty to develop expertise in imaging techniques and to advance their development as independent translational research scientists.	{ "pile_set_name": "NIH ExPorter" }
The objective of this project is to understand the role of hyaluronan- cell interactions in limb vasculogenesis. Previous results suggest that such interactions modulate blood vessel formation, in that hyaluronan- endothelial cell interaction appears to be essential but high concentrations of hyaluronan, e.g. in the peripheral mesoderm of the embryonic limb, are inhibitory. A key factor in these events is a newly discovered hyaluronan-binding protein (HABP) that possesses the peptide motif, arg-gly-asp, that is recognized by several integrins. It is hypothesized that hyaluronan is bound to the surface of endothelial cells via a complex of this HABP and an integrin; it is further proposed that this interaction influences endothelial cell behavior, especially migration, which is an essential component of new vessel formation. The specific aims of the project for this grant period are: 1. to produce full-length cDNAs for the HABP variants expressed by endothelial cells, and to use these cDNAs to obtain purified recombinant HABPs and to study their properties; 2. to isolate and characterize the integrin with which the HABP interacts, and to examine the role of interactions between this integrin, the HABP, and hyaluronan in endothelial cell attachment and migration; 3. to begin several approaches to further elucidation of the role of hyaluronan and HABPs in limb vasculogenesis. The results of these studies will lead to a better understanding of angiogenesis and vasculogenesis in limb development, and in other normal and disease processes. In addition this work will give new insights into the molecular mechanisms underlying cell migration and invasion, which are fundamental to tissue remodeling in development, healing and several diseases.	{ "pile_set_name": "NIH ExPorter" }
T cell lines and T cell hybridomas for ovalbumin and beef insulin have been produced and tested for specificity by proliferation, radiobinding and helper/suppressor assays. Characterization of the factor(s) released by these cell lines is currently underway. For the insulin system, B cell hybridoma antibodies are being used to prepared idiotypic reagents which will be tested for T cell reactivity. In other studies being initiated, patients with recurrent herpes will be characterized in terms of their cellular immune status to HSV-2.	{ "pile_set_name": "NIH ExPorter" }
This project is designed to explore the feasibility of using somatic cells to screen for chemically induced chromosomal aberrations. Included in this project are comparisons with respect to the efficiency of detection of chromosome aberrations in germ cells (testicular preparations and sperm morphology), cultured lymphocytes and bone marrow preparations.	{ "pile_set_name": "NIH ExPorter" }
Understanding cellular mechanisms that preserve totipotency or pluripotency in cells are central goals of both stem cell and developmental biology. Primordial germ cells (PGCs), precursors to the gametes, are the ultimate stem cell as they retain full developmental potential while other embryonic cells become progressively restricted in their fates. Our long term goal is to establish the key regulatory pathways that operate during the specification and differentiation of PGCs in the vertebrate model organism Xenopus. Genetic studies in Drosophila and C. elegans and molecular embryological studies in Xenopus have identified gene products in PGCs that appear to protect them from inappropriate somatic differentiation. How the identified gene products are "networked" and importantly, the real mechanism through which these preserve totipotency, remain largely unknown. The present application will address this gap in our knowledge. Xenopus is the system of choice for these studies because it offers a unique combination of total accessibility at both the biochemical and embryological level. Based on our previous data, we hypothesize that during the time that germ layers are established in the embryo, totipotency is preserved in PGCs by a combination of targeted translational repression of specific maternal mRNAs and global repression of mRNA transcription controlled by Xcat2. To test our central hypothesis we will complete the following specific aims: 1) Determine how maternal VegT function is restricted in PGCs by assessing VegT stability, translation, and nuclear localization in isolated PGCs using a combination of Real Time RT-PCR, injected tagged VegT transcripts and immunolocalization. 2) Determine the role Xcat2 plays as a translational repressor in preserving PGC totipotency by identifying the RNAs that co-purify with Xcat2 using a pull-down, RT-PCR, and cloning approach. Mis-expression of these RNAs in PGCs will be tested for their effects on PGC fate. 3) Determine what mechanism is responsible for the transient repression of transcription in primordial germ cells by selecting for Xcat2 interacting factors that repress transcription in vivo functional assays. Relevance of this research to public health: Stem cells are of high therapeutic value because of their ability to develop into a wide variety of cell types that could be used in the treatment of degenerative diseases including diabetes, Parkinson's and cardiovascular disease. Our studies on PGCs, prime exemplars of totipotency, will explore how different mechanisms cooperate to preserve genetically naive or ground states in cells. Our findings will be relevant to how adult somatic stem cells maintain a pluripotent condition, important issues in stem cell biology. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The objective of this research program is to define the molecular mechanisms that regulate the inflammatory process. One of the key mediators of the inflammatory process is TGF-beta1 and our recent studies have focused on TGF-beta-1 homozygous null mice in which the gene for TGF- beta-1 has been inactivated as a model of autoimmune-like pathology similar to Sjogren's Syndrome. Moreover, beyond its immunoregulatory role, TGF-beta-1 is considered essential to healing and tissue repair because of its growth and synthetic influences on mesenchymal cells. In this regard, we have taken advantage of TGF-beta-1-gene targeted mutant mice which over-express TGF-beta-1 (TGF-beta-1 transgenics) or in which TGF-beta-1 has been inactivated (TGF-beta-1-/-; null mutation) or partially inactivated (TGF-beta-1+/-; null mutation heterozygotes), to explore the role of this peptide in a model of fibropathology. Schistosomiasis mansoni, an extracellular parasite, which is a major cause of hepatic fibrosis in many developing countries, triggers a granulomatous inflammatory reaction in response to its eggs that lodge in the liver. The persistent granulomatous response leads to prolonged matrix synthesis and hepatic fibrosis during which TGF-beta-1 is produced. Matrix production and fibrosis in response to natural infection with S.mansoni worms exhibited distinct profiles in TGF-beta-1 gene targeted mice, consonant with their levels of TGF-beta-1. Although fibrotic manifestations appear dependent on TGF-beta-1 levels, fibrosis is not eliminated in the absence of TGF-beta-1, nor overwhelming in mice overexpressing this fibrogenic peptide. These observations emphasize the multifactorial nature of the signals involved in the complex pathophysiology of this and other fibrotic diseases. Nonetheless, as a link in the cytokine cascade, TGF-beta-1 represents a potential target for novel therapies in fibrotic disorders. Another effector molecule in inflammation and repair is nitric oxide (NO), the synthesis of which is regulated by TGF-beta.	{ "pile_set_name": "NIH ExPorter" }
To advance the development of therapeutic strategies for rare and/or emerging viral diseases of medical importance in targeted patient populations by 1) characterizing contemporary course of the clinical disease, and/or 2) directly evaluating a therapy.	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY This application focuses on investigating signals in the subventricular zone (SVZ) that regulate oligodendrocyte (OL) development in the healthy and injured brain. OLs arise from neural stem cells in the SVZ and produce the myelin sheaths that insulate neuronal axons, thus allowing proper synaptic transmission. Damage to or loss of OLs results in many neurological problems, as can be seen in multiple sclerosis and other neurodegenerative diseases. Therefore, identifying endogenous mechanisms that regulate OL development and repair in vivo may lead to new therapeutic targets for treating such conditions. Our previous studies found that the signaling peptide, Endothelin-1, delays OL maturation after demyelination of white matter in the rodent brain. This suggests that Endothelin-1 is a novel regulator of OL development in vivo. To test this hypothesis, the function of Endothelin-1 in the SVZ during normal development and after white matter injury will be determined. First, the mechanism of Endothelin-1 in the postnatal SVZ will be genetically dissected using conditional knockout mice and RNA sequencing. Then, using the lysolecithin model of demyelination and viral tracing, the role of SVZ-derived Endothelin-1 in OL regeneration will be defined.	{ "pile_set_name": "NIH ExPorter" }
This is a continuation of ongoing work to understand the genetics and pathogenesis of pectus excavatum in Lemur variegatrus, the black and white ruffed lemur. The condition appears to be inherited as an autosomal dominant and is, at times, expressed as flat chest. It is not manifest at birth but develops in some individuals towards the end of their first year of life. A few lethal skeletal anomalies have also occurred in this pedigree but do not appear to have a direct relationship to pectus. In addition, severe hemosiderosis, occasional hemochromatosis, and hepatomas have been observed in this colony. These are studied through selective breeding.	{ "pile_set_name": "NIH ExPorter" }
This publication project will involve the development of a monograph encompassing a critical review of existing validation studies concerning individual and group estimates of empirical data. A review and critique of this literature in terms of relevance to health care, scientific validity, and accessibility is vital in expanding our capabilities for meeting both local and national needs involving quality assurance, health policy development, and health care delivery. The project will encompass a reformulation of this validity problem, an extensive review and critique of available published and unpublished materials on the subject, and the preparation of a monograph (150 to 200 pages) to be submitted for publication.	{ "pile_set_name": "NIH ExPorter" }
The principle objective of this study is to determine the effects on neurological improvement of patients dosed with 2000 mg/day versus placebo over a 6 week treatment period and a 6 week follow-up (post-treatment) period in patients with acute ischemic stroke.	{ "pile_set_name": "NIH ExPorter" }
HIF-1 controls the transcription of many genes that are involved in key aspects of cancer biology. Overexpression of HIF-11, an inducible subunit of HIF-1 in response to hypoxia, is a prognostic factor in many cancers. The major signaling axis mediated by PI3K, PDK1, and Akt (also referred as protein kinase B) plays a significant role in the regulation of HIF-11 expression. Both GSK32 and MAPKs are also known to directly phosphorylate HIF-11, thereby affecting its stability and/or nuclear localization. To understand the regulatory network that suppresses tumor development and tumor angiogenesis, we have recently identified HIF-11 as a new in vitro substrate of Polo-like kinase 3 (Plk3). Plk3 strongly phosphorylates HIF-11 on serines 576 and 657 in vitro, two residues that lie in the oxygen-dependent degradation domain and near the nuclear export signal, respectively. By studying primary isogenic murine embryonic fibroblasts (MEFs), we have shown that PLK3-/- MEFs are hyper-sensitive to the induction of HIF-11 under hypoxia or treated with nickel, a hypoxia mimetic. Compared with that of wild- type MEFs, PLK3-/- MEFs contain a high level of Akt1/PKB activities, which is tightly associated with the inhibitory phosphorylation of GSK32. Consistent with the potential role of Plk3 in regulating the hypoxia signaling network, PLK3-/- mice develop tumors in various organs at an advanced age and PLK3-/- tumors are large in size and highly vascularized, suggesting active tumor angiogenesis. On the basis of these observations regarding physical and functional interactions between Plk3 and HIF-11, we hypothesize that Plk3 negatively regulates the hypoxia regulatory network and HIF-11-dependent tumor angiogenesis. To test the validity of this hypothesis, we will (i) study functional interaction between Plk3 and known signaling molecules, including GSK32 and MAPKs that phosphorylate HIF-11, and identify additional Plk3 target(s) upstream of Akt1, (ii) determine whether PLK3-/- mice are prone to tumorigenesis under hypoxia, and (iii) investigate whether mice harboring Plk3 phosphorylation-resistant mutant alleles of HIF-11 are more susceptible to tumorigenesis after nickel exposure. The combined in vitro and in vivo studies will greatly facilitate the elucidation of a new mechanism by which HIF-11 is regulated by Plk3 during hypoxic responses or after exposure to environmental carcinogens such as nickel compounds. A detailed understanding of the molecular regulation of HIF-11 will add significantly to the existing knowledge of tumor angiogenesis and tumor cell resistance to anti-cancer therapies.	{ "pile_set_name": "NIH ExPorter" }
`DESCRIPTION (provided by Project Leader): Brain tumors are the most common group of solid malignancies in children, causing devastating mortality and morbidity in a very understudied patient population. The goal of this program project is to improve understanding and treatment of pediatric brain tumors. During the first funding period, Project leaders worked together to develop novel mouse models that provided key biological insights into cerebellar growth regulation and medulloblastoma development, used these models for relevant preclinical testing of new therapeutic agents, and translated research results into clinical trials. The current application expands the focus of the program to encompass additional pediatric brain tumors. Five interactive projects plan directed studies of growth regulation in the brain using human tumors and mouse models to study signal transduction, gene expression and function in the context of pediatric brain tumors and brain development. An Administrative Core, a Bioinformatics and Biotechnology Core and a Neuropathology Core provide essential support to all projects. In Project 1, S. Baker investigates the molecular pathogenesis of pediatric high-grade glioma taking advantage of a large collection of primary tumors and novel mouse models for glioma that she developed. In Project 2, T. Curran investigates the mechanism of action of a small molecule inhibitor of Hedgehog signaling that he showed ablated medulloblastoma in his model systems. In Project 3, R. Gilbertson will define molecular subgroups of medulloblastoma through analysis of human tumors and development of mouse models, and will characterize cancer stem cells and their niches in subclasses of medulloblastoma. In Project 4, P. McKinnon will further his analysis of defective DNA damage response underlying brain tumor development and will determine the effects of DNA repair inhibitors as a therapeutic approach for brain tumors. In Project 5, M. Roussel and C. Sherr will explore mechanisms of oncogenic transformation in medulloblastoma, and will define microRNAs that modulate gene expression in cerebellar development and medulloblastoma. Integrated analyses within the group will identify common and unique signal transduction pathways in pediatric brain tumorigenesis.	{ "pile_set_name": "NIH ExPorter" }
Protection from infectious agents known to be major causes of death worldwide, such as influenza, tuberculosis, and malaria, as well as potential release of bioweaponized agents which cause plague, tularemia, and meilloidosis require vaccines that generate humoral and T-cell responses. Effective component vaccines require the addition of adjuvants to increase their immunogenic capacities. Until recently alum salts, which require repeated applications and tend to be skewed towards T helper TH2-based immunity (humoral) rather than TH1, (cellular) were the only adjuvants approved for use in human vaccines. Recently, the lipid A mimetic (monophosphoryl lipid A, MPL) adjuvant has been combined with alum (AS04) in two FDA-approved vaccines (Cervarix (Human Papilloma Virus), and Fendrix (Hepatitis B Virus)). Additionally, synthetic lipid A mimetics aminoalkyl glucosaminide phosphates (AGPs) that also signal through Toll-Like Receptor 4 (TLR4) are being studied as both adjuvants or stand-alone immunogenic compounds. Thus, TLR4 agonists show great promise for use as adjuvants in component vaccines. However, the approved TLR4 agonist, MPL has distinct deficiencies both in potency and structural consistency and AGPs are labor intensive and costly to synthesize. Therefore, the goal of this revised R21 application is to undertake a novel approach of using Bacterial Enzymatic Combinatorial Chemistry (BECC) to make rationally-designed lipid A structures by modifying the lipid A structure of the lipooligosaccharide (LOS) from an attenuated (BSL-2 approved) Yersinia pestis (Yp) strain. This approach will use the non-stimulatory, hypoacylated, and bisphosphorylated lipid A structure present in LOS synthesized by this Yp strain as a scaffold to be modified by heterologous in trans expression of lipid A biosynthesis enzymes obtained from a wide variety of bacterial backgrounds with specificities for the removal or addition of fatty acid chain, phosphates moieties, and carbohydrates to the lipid A backbone, This process allow for the safe, cost effective, and efficient design of molecules with immunostimulatory potential. We will test the immunotherapeutic potential of these new molecules in vitro and in vivo to identify novel molecules representing adjuvants and/or immunomodulating reagents. We will also include well-characterized immunostimulants, such as MPL and known LPS structures as comparisons to the molecules synthesized by BECC. The successful demonstration of protective innate/adaptive immune responses by this novel approach of creating new adjuvants, in highly relevant in vivo animal model(s) could have important implications in the field of antigen recognition, formulation, and vaccine design.	{ "pile_set_name": "NIH ExPorter" }
The work in this proposal addresses two issues of fundamental importance to our understanding of neuronal function and development. The first is to determine how membrane glycoproteins synthesized in the cell body of a neuron are selectively routed to the pre- and postsynaptic terminal. These processes are central to the proper assembly of surface membranes and to how distinct regions of the neuronal plasma membrane are maintained in a functionally and structurally differentiated state. The second concerns the possible role of glycoproteins as mediators of interneuronal recognition and adhesion. My colleagues and I intend to explore these points using a single cell - R2 the giant neuron of Aplysia californica. R2's cell body, presynaptic, and postsynaptic regions are anatomically accessible. Moreover, R2 synthesizes only ten membrane glycoproteins, a remarkable simplicity relative to the complexity of glycoprotein fractions from vertebrate nervous tissue. Two of R2's glycoproteins have distinctly different destinations in the cell: glycoprotein-I is the major glycoprotein on the cell surface of the cell soma whereas glycoprotein-V is preferentially exported into the axon where it is rapidly transported towards R2's synapses. The site at which the two glycoproteins are sorted will be determined by immunocytochemistry at the ultrastructural level using monoclonal antibodies raised to each glycoprotein. In addition, the oligosaccharide chains of both glycoproteins will be characterized with the idea that a modification of a glycosyl moiety is the signal responsible for the sorting. R2, GCN, and R15 are three large identified neurons that differ in function, location, and neurotransmitter type. We will examine the glycoproteins present on the surface of the cell body and terminals of these neurons in order to test directly the hypothesis that neurons can be distinguished by their glycoprotein constituents. Many features of the proposed experiments are unique and present a unified, multidisciplinary approach to issues of major importance to our knowledge of the neuron.	{ "pile_set_name": "NIH ExPorter" }
Inactivity and excessive and inappropriate caloric intake stimulate development of visceral obesity that is associated with increased risks of type 2 (non-insulin-dependent) diabetes, hypertension, and coronary heart disease (CHD). Both exercise and dietary restriction have been successfully used as a means of body fat loss. In addition, exercise, with or without fat loss, can reduce risks of diabetes, hypertension, and coronary heart disease but its effects have not been consistent. In particular, there is little consensus regarding the effects of exercise intensity on either body fat loss, insulin sensitivity, or risks of hypertension and CHD. Our premise is that if exercise is to be used effectively to prevent or reduce different health risks or to be used as a tool in mechanistic analyses of hormone action, a better understanding of dose-dependent effects of exercise intensity is needed. Our pilot study, initiated two years ago, focused on the question of whether exercise training at high walking intensities could increase basal pulsatile growth hormone (GH) secretion. This hypothesis was supported by our results (8,9), but in addition, we discovered that exercise intensity bears an inverse relationship to body fat loss (assessed by indirect, anthropometric methods), has an apparent diabetogenic effect (assessed with an indirect measure of insulin sensitivity), and is associated with increased androgenic action (ratio of plasma testosterone to sex-hormone-binding globulin, SHBG). Since all three observed changes, the accumulation of visceral body fat (6,44), GH oversecretion, and increased androgenic index (47), either antagonize insulin action (GH oversecretion) or are accompanied by increased risk of diabetes, hypertension, and CHD (high androgenic index and increased postprandial insulin response), our results suggested that exercise intensity during 15 weeks of training had a deleterious effect on at least two risk factors: it was inhibitory to body fat loss and to peripheral sensitivity to insulin. These serendipitous observations redirected our interest from the role of exercise in alleviating somatopause, the exponential decline in spontaneous GH secretion with age, to the mechanism by which exercise intensity affects insulin resistance, hypertension and CHD in postmenopausal women.	{ "pile_set_name": "NIH ExPorter" }
We propose to continue a multidisciplinary project to understand the mechanisms by which aberrant RNA and DNA structures trigger gene silencing. Two general approaches are intertwined in this work. First, we examine molecular responses following introduction of novel RNA or DNA into the organism. We propose to use multiple high resolution analyses (molecular, genetic, and biochemical) and to apply these where possible on a genomewide scale. The capability and potential impact of such analysis relies on tools and protocols developed during the previous granting period (and the subject of many of our publications during this period), including whole genome approaches to chromatin structure, transcription, steady state mRNA and small RNA patterns, and precise profiling of translation and other transient RNA dynamics. Second, we examine the natural roles of the above informational surveillance pathways in developmental and pathological contexts. Approaches parallel those above, including biochemical, molecular, and phenotypic analysis of silencing-treated molecular machinery in development and response to pathological conditions. The specific aims of the proposed research are: 1. Investigate the roles, mechanisms, and triggers of gene silencing in response to foreign/noncannonical RNA. 2. Investigate recognition and specific gene silencing in response to unusual DNA structures and sequence. 3. Investigate the interplay between RNA-triggers of gene silencing and chromatin associated nuclear events. 4. Continue to develop and refine tools and assays for studies of genetic activity and silencing in C. elegans. Studies of gene silencing have considerable potential for long term impact. First, as we understand mechanisms of gene silencing, we acquire the ability to specifically and effectively silence genes within cells or in an organism, generating a significant toolkit for functional genomic research, and aiding in the development of tools for gene-based therapeutics. Second, an understanding of gene silencing mechanisms allows improved design of systems for deliberately expressing specific genes in vivo. Such expression can provide significant advantages for investigations of biological function, for experimental elucidation of disease pathways, and for eventual intervention in biological systems (gene therapy). Third, because gene silencing mechanisms are indicative of a variety of cellular gene regulation mechanisms, work on gene silencing has provided valuable insights into normal gene regulation. Fourth, many gene silencing mechanisms reflect the response of the cell/organism to DNA or RNA that is viewed as foreign. PUBLIC HEALTH RELEVANCE: A greater understanding of signals that activate and silence regions of the genome will illuminate the fundamental mechanisms that our cells use (i) to properly control the activity of each of their genes and (ii) to protect themselves from unwanted genetic activity in the form of viruses and other genomic parasites. This research program applies a variety of information-based and experimental approaches directed toward that understanding.	{ "pile_set_name": "NIH ExPorter" }
DESCRIPTION: Chronic fatigue syndrome (CFS) is associated with orthostatic intolerance which often takes the form of postural orthostatic tachycardia syndrome (POTS) in adolescents. Preliminary data suggest the novel concept that defective vasoconstriction produces POTS in CFS with cardiac autonomic changes as a secondary response. CFS patients will be compared to healthy controls and to controls with simple faints to test 3 hypotheses: 1) Blood is redistributed peripherally and redistribution is enhanced during orthostasis producing increased microvascular filtration and dependent edema. Central hypovolemia causes decreased cardiac output, reflex tachycardia and reduced cerebral blood flow. This is enhanced during orthostasis producing increased microvascular filtration, dependent edema, and peripheral pooling. These changes alter the interstitium, and cause reflex tachycardia, reduced cerebral blood flow and often hypotension. Blood volume and cardiac output using the indocyanine green dye dilution technique will be measured supine, during conventional 700 head-up tilt, and during low angle head-up tilt. Cerebral blood flow velocity (CBFv) will be estimated by transcranial Doppler ultrasonography. Thoracic, splanchnic, and pelvic vascular volumes will be measured by impedance plethysmography, and limb blood flow, arterial flow, venous volume-pressure relation, and venous pressure will be measured by venous occlusion strain gauge plethysmography. These will show increased blood flow to lower extremities when upright. Central hypovolemia will occur and will reduce CBF and produce symptoms of CFS. Cardiac autonomic status including baroreflex will be assessed by heart rate and blood pressure variability and transfer function. Baroreflex and heart rate variability will be decreased and blood pressure variability will be increased related to circulatory deficit 2) The defect in vasoconstriction is heterogeneous comprising abnormal arterial baroreflex mediated sympathetic vasoconstriction in one subgroup of CFS patients and abnormal local vasoconstriction in a second subgroup with defective veno-arteriolar reflex (arterial baroreflex insensitive dysfunction). Low angle tilt will be used to activate baroreflex mediated and local reflexes. Local reflexes including myogenic, metabolic and veno-arteriolar will be sorted out through use of supine testing designed to specifically stimulate a specific reflex (limb hang, large pressure step and reactive hyperemia) and measuring peripheral resistance. 3) Cardiac autonomic findings are secondary to circulatory changes. Thus, tachycardia relates to vagal withdrawal because of circulatory insufficiency. CFS patients will be treated with midodrine or placebo in a cross-over study. Using supine and low angle tilt experiments, circulatory measurements and psychological instruments will be combined to demonstrate that circulatory abnormalities, autonomic abnormalities and symptoms correct in a subgroup of CFS patients with low resting peripheral resistance.	{ "pile_set_name": "NIH ExPorter" }
The regulation of tissue growth is of paramount importance for the understanding of vertebrate, including human, embryogenesis. It has also a tremendous medical significance. Deviations from normal tissue growth lead to severe patterning defects during embryogenesis, and to cancer in the mature organism. Abnormal patterns of cell proliferation produce a number of developmental eye disorders in humans. One of them is microphthalmia, which in the most extreme cases leads to the complete absence of eyes. To identify the genetic causes of human microphthalmia and anophthalmia, we chose to use an animal model, the zebrafish. Genetic screens in zebrafish identified many mutations that result in a decreased eye size. One of the most severe small eye phenotypes is caused by defects of the out of sight (out) locus. The goal of this project is to identify the molecular nature of the out gene, and to develop an understanding of its function on the cellular level. Once the molecular identity of this gene is known, we will investigate whether its defects are responsible for human microophthalmia by screeing DNA samples from diseased individuals. Similar animal model-based approaches have been very productive, leading to the identification of microphthalmia-causing mutations in several genes, including rx, six6, and sox2. This project is the first step on the way to use zebrafish mutants on a broader scale to identify the genetic causes of eye growth disorders. In the long run, the zebrafish is likely to become a major source of candidate genes to study this class of abnormalities. This strategy is likely to make a major contribution to the understanding of the genetic causes of microphthalmia and its more severe form, anophthalmia, in humans. We will use animal model, the zebrafish, to identify genes responsible for microphthalmia, a human eye disorder characterized by a reduction of eye size, and in the most extreme the absence of eyes. These studies will provide insight into the genetic basis of this debilitating disorder that affects 1 in 5,000 children. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The goal of this project is to characterize the acute toxicity of trimethylsilyldiazomethane (TMSD). TMSD is a methylating agent used in synthetic chemistry. It is marketed as a safe replacement for diazomethane, primarily due to a lower explosive potential. However, two chemists died of acute pulmonary edema while working with TMSD in the laboratory. NTP is performing toxicity studies in order to further examine the potential for TMSD to induce pulmonary toxicity. These studies are in progress. Keywords: toxicity, inhalation, trimethylsilyldiazomethane	{ "pile_set_name": "NIH ExPorter" }
The following results were reported: (1) Establishment and characterization of origin-defective SV4O genome immortalized adult human prostatic epithelial cells; (2) neoplastic transformation of SV4O T- antigen immortalized human keratinocyte (RHEK-1) line by two human herpesvirus (HHV)-6 DNA clones; (3) demonstration of altered epidermal growth factor signal transduction in activated Ha-a transformed human keratinocytes; (4) demonstration of the use of RHEK-1 immortalized human keratinocytes for detection of induced mutation at the hypoxanthine- guanine phosphoribosyltransferase locus; (5) demonstration of the role of chromosome 5 in immortalization and tumorigenesis of human keratinocytes; (6) development of an in vitro model of human corneal epithelium for ocular studies; and (7) immortalization of mouse bone marrow-derived mast cells with Ad12-SV4O virus.	{ "pile_set_name": "NIH ExPorter" }
The long term objective of our research is the elucidation of the molecular mechanism of the acute and chronic effects of opiate narcotic analgesics. Our approach to this problem continues to be the study of the endogenous opioid system. Knowledge of the workings and functions of this system will have wide-ranging implications and, we feel, will lead towards an understanding of the actions of exogenous opiates. The focus of our research is an interdisciplinary exploration of the properties and functions of the major types or opioid receptors. Based on our success in purifying mu binding sites to homogeneity, we plan to apply our experience to the purification of other opioid receptor types, such as kappa and delta. The cloning and total sequencing of the mu receptor is in progress. We also plan to study the structure of the non-protein portion of receptor molecules, i.e., the carbohydrate portions of these glycoproteins and the nature of lipids essential for binding and function. The ultimate goal of all these studies is the knowledge of the complete structure of the binding portion of the opioid receptors. The interaction of the binding sires with G-proteins and second messenger systems as well as the possible role of receptor phosphorylation in receptor regulation will also be examined. The production of antibodies to purified receptors and to peptide fragments derived from them will be of enormous utility in many of our proposed studies and will be actively pursued. When the total amino acid sequence of one or more opioid receptor types is known, we plan to study receptor mRNA distribution and regulation using appropriate cDNA probes. Once we have established that the "wild type" receptor cDNA or gene can be expressed in a suitable mammalian cell line, we plan to use site-directed mutations and deletions to study the functional domains of the receptors. Purified receptors will be reconstituted into artificial and natural membranes and into cells normally devoid of opioid receptors. This has proved to be a powerful technique for the study of interactions of receptors with second messenger systems and ion channels. On the behavioral level we shall try determine the types of receptors and opioid peptides involved in analgesia, feeding and reward mechanisms. Behavioral as well as binding assays will be used to test new compounds synthesized by organic chemists collaborating with us.	{ "pile_set_name": "NIH ExPorter" }

Subsets and Splits