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The Comparison of AMD Treatments Trial (CATT) is a multi-center, randomized clinical trial investigating the treatment of neovascular AMD with Lucentis and Avastin. The primary objective of the activities proposed in this fundus Photograph Reading Center (FPRC) application is to provide support to the CATT. This support includes A) the evaluation of follow-up visit fundus photographs to assess the changes in the choroidal neovascularization and the changes in retinal vessel diameters following both treatments; and B) assessment of the correlation of these findings with the findings of the CATT OCT Reading Center. This continuation grant proposal has two Specific Aims: Aim 1, to complete in all participants the two-year follow up of the original CATT grant application. Because enrolment of patients in the CATT trial was delayed, the period needed to complete the two year follow up must be extended beyond the time period proposed in the original plan. Aim 2, to perform an observational study for an additional 3-year follow up to investigate the long term effects of the study medications. Throughout the proposed studies the FPRC will provide reliable, timely, reproducible, and systematic grading of submitted color photographs and angiographic image materials for CATT. These activities will be similar to those provided in the first years of the CATT study. The FPRC will also continue to implement study specific materials and methods for reader, photographer, clinic coordinator, and ophthalmologist certification and training. FPRC activities will also support study goals as an image resource for recruitment, Study Group meetings, presentations and publications. RELEVANCE (See instructions): Age-related macular degeneration (AMD) is the leading cause of blindness in the United States and most western countries. These studies of the relative effectiveness and safety of the two drugs under study will determine how patients with neovascular (wet) AMD are treated in the future.	{ "pile_set_name": "NIH ExPorter" }
The overall objective of this application is to design and develop enantioselective phosphine-catalyzed annulations and to apply them in the chemical syntheses of natural products, their analogs, and unnatural small organic molecules of medicinal significance. Building on our successful studies of phosphine catalysis reactions and their applications in the total syntheses of medicinally useful natural products [(+)-ibophyllidine (?)-alstonerine, (?)-macroline, (?)-hirsutine, 3-deoxyisoochracinic acid, isoochracinic acid, isoochracinol], here we propose the development of new phosphine catalysis reactions and novel chiral phosphines. Specifically, we will prepare three families of new [2.2.1] bicyclic chira phosphines from a naturally occurring amino acid (trans-4-hydroxy-L-proline) and a terpenoid (carvone). These new phosphines have already displayed tremendous potential in facilitating enantioselective [3+2] and [4+2] annulations between allenes and imines. To build on these exciting preliminary observations, we propose to synthesize (-)-actinophyllic acid and (+) - ajmaline through enantioselective allene-imine [3+2] and [4+2] annulations, respectively, and another indole alkaloid, (+)-hirsutine, through the catalytic asymmetric allene-imine [4+2] annulation. In addition, we wish to develop new phosphine-catalyzed and -mediated reactions. We will examine and expand upon two hypotheses in the area of new reaction development: (i) tandem umpolung addition/Wittig olefination and (ii) sequential phosphine/transition metal catalysis. The proposed research could significantly expand the principles of organic reactions and provide new methods and reagents for the synthetic organic chemist's toolbox. Many innovations will be necessary to successfully implement our three specific aims. Our preliminary results have been strong in all three specific areas, supporting the likelihood of further successes. The small molecules described in this proposal are medicinally important, possessing activities related to diseases such as thrombosis [(-)-actinophyllic acid], hypertension and arrhythmia [(+)-ajmaline], influenzas [(+)-hirsutine], neurological disorders (glutamate receptor antagonist), and cardiac diseases and cancer (shihunidine). Successful completion of this proposed study would significantly impact the synthetic organic chemistry community by introducing a series of new chiral phosphines readily accessible from inexpensive natural products; in fact, we are collaborating with Sigma-Aldrich to make these chiral phosphines available to the scientific community. In addition to using these chiral phosphines in the proposed organocatalysis reactions, they should also serve as ligands for transition metal-catalyzed processes. Therefore, the proposed research could significantly increase the efficiencies of various synthetic organic processes.	{ "pile_set_name": "NIH ExPorter" }
A growing number of studies indicate that religion exerts a beneficial effect on health and well-being in late life. Moreover, this literature reveals that the salubrious effects of religion may be especially evident in racial minority groups. However, virtually all of this work has been conducted with older African Americans. Much less is known about the relationship between religion and health among older Mexican Americans. Part of the reason for this oversight arises from the fact that no one has developed a comprehensive set of religion indicators that are designed specifically for use with older Mexican Americans. The purpose of this study is to address this gap in the literature. In particular, the proposed research has the following objectives: 1.To conduct a series of qualitative studies (e.g., in-depth interviews and cognitive interviews) in order to identify the content domain of religion as it is lived by older Mexican Americans; 2. To use the data obtained from these qualitative methods to craft closed-ended survey items on religion; 3. To conduct a nationally-representative survey of older Mexican Americans in order to quantitatively evaluate the factor structure and psychometric properties of the newly devised religion measures; 4. To merge the quantitative data with a comparable nationwide survey that has already been conducted by the Principal Investigator to assess the relationship between religion and health in older Whites and older African Americans; and 5. To compare and contrast race differences in religiousness as well as race differences in the relationship between religion and health among older Whites, older Blacks, and older Mexican Americans. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The CIFASD Informatics Core provides a research data resource to be used by investigators to explore the relationships of face, brain, and behavior to improve the diagnosis of FAS and work towards therapies for alcohol exposed children. The Informatics Core has the following specific aims for the next five years: Aim 1: Continued cyber infrastructure support. The Informatics Core will consult and collaborate with CIFASD clinical projects, cores, and developmental projects to support the existing data submission to the CIFASD Central Repository, manage those data, and make those data available for use to accomplish CIFASD goals. Aim 2: Collection of additional data sets. The Informatics Core will securely bring new data sets into the CIFASD Central Repository for cross-study and cross-modality data analysis amongst CIFASD projects. Aim 3: Support of affiliated projects. The Informatics Core will develop technical solutions that will allow the comparison of data between CIFASD and affiliated projects. The Informatics core will use the following methods to support the goals of this project and CIFASD: The ongoing management of standardized data in the CIFASD Central Repository will provide core consortium resources including; data input and bulk upload tools, a data dictionary to standardize terms across studies, a cross query tool so that data from multiple studies can be integrated for synthetic studies, and data browsing tools that enable investigators to ensure data quality. Expert consultation will support the input, export, and integration of data in the Central Repository to lower the barrier of use for investigators and improve data quality of synthetic studies. This consultation will also regularly report on data in the Central Repository to track consortium progress goals. Expert software engineering will provide new input tools and modify existing tools to support the changing needs of CIFASD projects as well as new research projects that are added as part of the third Phase of the CIFASD program. PUBLIC HEALTH RELEVANCE: The goal of the CIFASD Informatics Core is to support research on the relationships efface, brain, and behavior in order to improve the diagnosis of Fetal Alcohol Spectrum Disorders and work towards therapies for alcohol exposed children. By providing a central, standardized data resource for CIFASD, the Informatics Core will provide a means by which comprehensive diagnostics and therapies can be developed.	{ "pile_set_name": "NIH ExPorter" }
nHuman mothers interact emotionally with their newborns through exaggerated facial expressions and mutual gaze, a capacity that has long been considered uniquely human. A few years ago we initiated a research program on early face-to-face interactions in rhesus monkeys after we had serendipitously discovered that very young rhesus monkey infants did, in fact, engage in extensive face-to-face interactions with their mothers, but only during their first month or so of life. This past year we characterized face-to-face interactions between mothers and their newborn infants in a naturalistic setting. We found, similar to our previous results from mother-infant pairs housed in social pens, large individual variability in rates of maternal/infant face-to-face interactions. Further analysis revealed that newer mothers who had only one or two infants engaged in mutual gazing/lipsmacking in the first 30 days of life significantly more than more experienced mothers with three or more infants. Conversely, the more experienced mothers let their infants out of their arms reach significantly more in the first 30 days of life than newer mothers. We also discovered that during their first week some (but not all) infants could accurately match certain facial gestures produced by a human experimenter, even after a delay. For those infants who could imitate in this fashion, this capability was evident on their first postnatal day. We have since been investigating brain activity during periods of imitation vs. exposure to a dynamic but nonsocial stimulus (a rotating disk) and during non-stimulus baseline periods, using scalp electrodes to record EEG activity, and found a distinctive EEG signature involving significant suppression of mu rhythm activity at low frequencies in frontal and parietal brain regions exclusively during periods of imitation. This pattern of EEG activity intensified through that first week, and it was significantly stronger in mother-reared than in nursery-reared neonates. We also documented the emergence of EEG rhythms in one-week-old infant rhesus macaques under both light and dark conditions, showing that the 57 Hz frequency band responds reliably to changes in illumination and that EEG in higher frequencies (1220 Hz)significantly increase between dark and light conditions, similar to the increase in the beta band of humans during cognitive tasks. These findings demonstrate similarities between infant human and infant monkey EEG. More recently, we have demonstrated, using eye-tracking technology, that week-old infants readily respond to a computer-generated dynamic monkey avatar, and that those infants who imitate tend to focus on different aspects of the aviatars face (eyes and mouth) than those don't (mouth only). We also compared neonatal imitation abilities in mother-reared and nursery-reared monkeys, focusing on D3 performance only. Even though NR infants show an imitation effect when tested over the first week, they do not exhibit imitation specifically on D3. In contrast, MR monkeys responded to facial gestures with more gestures themselves, consistent with our previous EEG findings that MR infants show larger mu suppression than NR infants when viewing facial gestures. Given the potential impact of neonatal imitation on infants' social, cognitive, and emotional development, we devised an intervention whereby NR infants either received additional facial gesturing from a human caretaker, received additional handling (but did not see facial gestures), or remained in standard nursery rearing. We found that only the group that had received facial gesturing showed improved performance on the standard neonatal imitation task on D7 and showed greater sensitivity to facial identity of others in a standardized stranger task. Infants from the facial gesturing group also appear to be less inhibited in their latency to touch a toy in a novel object task and had better memory for social stimuli compared to infants in the handling and standard nursery rearing groups. A second attempt to increase infants social perception and social sensitivity looked at the effects of oxytocin on infants social interactions. Nursery-reared infants were nebulized with oxytocin or saline, and were then tested in an imitation recognition task. Salivary assays confirmed increased levels of oxytocin, and infants showed increased affiliative gesturing towards a human experimenter following oxytocin administration. We further explored infants facial processing strategies by presenting them with various faces and facial configurations on a remote eye tracker. Rhesus macaque infants generally prefer faces with normally arranged features over faces with linearly arranged featured, suggesting a special sensitivity to faces and face-like stimuli. These preferences are particularly strong for rhesus rather than human faces, which furthermore indicates a genetic predisposition towards rhesus faces in particular. We also found particular sensitivities towards the eye region in neonatal imitators but not in non-imitators, which may indicate that neonatal imitation and differential social sensitivity are intricately linked. A new project begun this year has involved the analysis of mothers milk in rhesus monkeys with respect to parity and early life history (i.e., rearing condition). In collaboration with Dr. Katie Hinde at Harvard University, we have begun collecting milk samples from mothers within the first 30 days of her infants life, and we are currently analyzing these samples for cortisol content and nutrient composition. Dr. Hinde has examined mothers milk for infants aged 3-4 months and has found that mothers yield milk of differing quality and quantity based on the sex of the her offspring. At this age, cortisol in mothers milk is also predictive of infant temperament. At the LCE, we are in the unique position to study, for the first time, similar relationships in neonatal milk (the first 30 days of life). Similar to Dr. Hindes studies of mothers milk in older infants, we have found that parity predicts milk yield volume (MYE) in the first month of life. Our preliminary findings also indicate that mothers with higher hair cortisol during pregnancy have a higher MYE in the first 30 days of life. We are currently examining this relationship with respect to both the mothers and infants hair cortisol reflective of long-term HPA axis activity in the neonatal period. We are further examining the extent to which constituents in mothers milk (cortisol and nutrient content) during the neonatal period influence infant social behavior, cognitive development, and temperament. Another focus is on the incidence of alopecia and related physiological processes. We have previously observed that many females undergo severe hair loss during pregnancy, only to regain full hair growth in the two months postpartum. In collaboration with Drs. Novak and Meyer at U-Mass, we are examining the role of chronic HPA axis activity as assessed by hair cortisol concentrations in alopecia. Our early results indicate that overall concentrations change across pregnacy and that monkeys that exhibit hair loss have higher hair cortisol concentrations than those that do not. We stared a research program on personality and facial characteristics with our Cebus monkeys, ide 5 personality dimensions (Assertiveness, Openness, Neuroticism, Sociability, and Attentiveness), and found that Cebus monkeys facial width-to-height ratio as well as their face w), and found that Cebus monkeys facial width-to-height ratio as well as their face width/lower face height are positively and significantly associated with Assertiveness. Lower face/face height ratio was also associated with neuroticism and attentiveness, suggesting that facial morphology reflects personality domains.	{ "pile_set_name": "NIH ExPorter" }
In our initial studies, we found that the human neuroblastoma (SH-SY5Y) cell line consistently expresses a high density of mAChRs (about 220 fmol/mg protein or 25,000 receptors/cell). Most of the mAChRs in the SH-SY5Y cells showed high affinity for [3H]PZ, suggesting an M1 nature of these muscarinic receptors. To our knowledge, the SH-SY5Y cell line is the only cell line with high affinity [3H]PZ binding to_the mAChRs on the intact cells. The SH-SY5Y cells have a functional PI system and an adenylate cyclase system. The existence of these effector systems offers a unique opportunity to study the effector coupling mechanisms of the M1 receptors in a homogeneous neuronal cell line. Our initial data suggest that these M1 receptors are coupled to the PI system. In this proposal, we will further examine the pharmacological properties of the mAChRs in SY-SY5Y cells using several new selective muscarinic ligands and compare these data with a transfected B82 system which is thought to contain only one type of mAChr. i.e., M1. Furthermore, we will also examine the second messenger system coupled to the receptors in order to obtain a better understanding of the molecular basis for the function of the mAChRs in both cell lines. Although our initial pharmacological studies indicate that the predominant mAChRs on the SH-SY5Y cells are of the M1 type, since there could be more than one mAChR showing high affinity for PZ (40), an unambiguous definition of this receptor requires a knowledge of its primary structure by gene cloning and sequencing. However, this will only be done if the mAChRs in the SH-SY5Y cell line is unique. This study will yield clone(s) which express a constant population of M1 receptors with pharmacological and biological properties of the native M1 receptors. This study will provide in depth information on the functional mechanism of neural mAChRs and their genetic regulation. This knowledge will have importance in the drug treatment of CNS disorders with cholinergic deficits such as senile dementia of the Alzheimer's type (SDAT), Huntington's disease (HD) and Parkinson's disease(PD). Furthermore, these cell lines will be used as models for the development of selective drugs which act at a single type of mAChR thereby improving therapeutic effects while reducing side effects.	{ "pile_set_name": "NIH ExPorter" }
Retroviruses induce a variety of tumors in vertebrates. A host origin for retroviral oncogenes is implied by the discovery that vertebrate DNA contains highly conserved genes (denoted "c-oncs") that are homologous to retroviral oncogenes. Recent experiments suggest that induction of increased transcription from one of th c-oncs, c-myc, directly causes lymphomas in chickens. The similarity between viral oncogenes and c-onc genes implies a normal role for c-onc proteins in growth cotnrol and/or differentiation. The long-term aim of my research is to elucidate the presumptive roles of c-oncs as cancer genes and as regulators of normal cell growth. Specifically, I will 1) analyze cloned chicken c-myc DNA to determine which regions of c-myc give rise to mRNA and characterize the maturation of c-myc mRNA from larger precursors to determine whether c-myc expression may be controlled at the level of splicing. 2) Determine the sequence of about 2000 bases in c-myc and thus deduce the amino acid sequene of the hitherto unidentified c-myc protein. 3) Investigate a possible role for a c-onc gene in development. Chicken thymus apparently contains a cell fraction enriched for the expression of c-myb, one of the c-onc genes. I will determine whether the expression of c-myb, a putative regulatory gene, fluctuates in parallel with stages in development and/or differentiation in the thymus; such findings would imply a role for c-myb during thymus development. Using cloned c-myb DNAs from mouse and chickens, first I will measure c-myb RNA from organs of various vertebrate species to determine whether elevated levels of c-myb RNA are a general feature of vertebrate thymus. If so, this observation would indicate that c-myb expression is indeed important in thymus physiology. Next, I will fractionate mouse (rather than chicken) thymus into cellular components to determine whether c-myb RNA is expressed primarily within epithelial cells, immature t cells, or immunocompetent T cells. I will follow variations in the expression of c-myb in different fractions of thymus cells during various stages in thymus development. I will monitor specific fluctuations in c-myo expression as a means of assessing the response of cultured thymus cells to the addition of putative mediators of T cell differentiation and thymic involution. If these studies do not prove to be feasible using mice, instead I will investigate c-myo expression in chicken thymus.	{ "pile_set_name": "NIH ExPorter" }
Many biological processes are regulated at the level of initiation of transcription of genes. The goal of the experiments in this proposal is to increase our understanding of the mechanisms by which transcription is regulated in eukaryoptic cells. To this end, an in-depth study of the cis-acting DNA sequences and the trans-acting components required for transcription of mammalian Beta-globin genes will be performed. Cloned globin genes with mutant regulatory sequences will be introduced into tissue culture cells to delineate both promoter elements and DNA sequences responsible for the tissue-specific transcription of Beta-globin genes in erythroid cells. Cell-free transcription experiments will be used to confirm the in vivo analysis, and both in vitro transcription and DNA binding assays will be used to identify the trans-acting promoter and erythroid cell-specific factors responsible for the regulated transcription of the gene. These assays will be used to purify the transcription components and to study in detail their interactions with wild type and mutant DNA templates. The identification of the DNA sequences and factors required for transcription of the Beta-globin gene and the establishment of a reconstituted regulated transcription system in vitro will provide a useful model for analyzing the detailed molecular mechanisms of eukaryotic gene regulation.	{ "pile_set_name": "NIH ExPorter" }
The acquired immune deficiency syndrome (AIDS) is reaching epidemic proportions in the United States. The disease is caused by a retrovirus, commonly called HTLV-III/LAV, that is tropic and cytopathic for T lymphocytes. Infection with the AIDS virus is dependent upon the action of an RNA dependent DNA polymerase (reverse transcriptase). In contrast, the replication and repair of DNA in the target T lymphocyte is mediated primarily, if not exclusively, by DNA polymerase-alpha. This major enzymatic difference between virus and host is a logical candidate for the selective development of anti-viral agents. Indeed, several deoxynucleoside analogs have recently been described that apparently inhibit viral replication by this mechanism. Unfortunately, many deoxynucleosides are not phosphorylated efficiently by the resting T lymphocytes that are potential targets for HTLV-III/LAV infection. An ideal agent for the treatment of AIDS should accumulate in both non-dividing and proliferating T lymphocytes, as well as inhibit reverse transcriptase. Previous experiments in this laboratory have shown that deoxyadenosine, and related compounds, are selectively phosphorylated and "trapped" by human T lymphocytes, compared to other cell types. The purpose of the present studies is to design and evaluate specific analogs of deoxyadenosine that are similarly taken up by T lymphocytes, that resist degradation by cellular enzymes, and that render the T cells resistant to infection by HTLV-III/LAV. Intially, the uptake and metabolism of each deoxynucleoside will be examined in cultured human CEM T lymphoblasts, in mutants deficient in individual purine metabolic enzymes, and in normal T cells. Subsequently, the capability of selected agents to inhibit the in vitro infection of wild type and enzyme deficient CEM T cells by HTLV-III/LAV will be assessed. Finally, the ability of the drugs to protect normal peripheral blood T cells from HTLV-III/LAV infection will be determined. The anti-viral studies will be performed in association with Dr. Douglas Richman at the San Diego Veterans Administration Medical Center, with whom the principal investigator has an established collaboration.	{ "pile_set_name": "NIH ExPorter" }
Recent research in the laboratory has led us to study the lysosomal disposal of glycogen. Our work has suggested that the glycogen-binding protein Stbd1 may participate in mediating an autophagy- like mechanism for the transfer of glycogen to lysosomes and furthermore may allow for the preferential trafficking, hence removal, of glycogen that accumulates abnormal structure, as characterized by increased covalent phosphorylation and decreased branching. Certain cardiac glycogenoses, as observed in Pompe disease, Danon disease, and Wolff-Parkinson-White syndrome, have in common the hyper-accumulation of glycogen in vesicular compartments within cardiomyocytes.. The current proposal is aimed at understanding how Stbd1 is involved in this process. Aim (i) will utilize cell-based experiments to examine the mechanism of glycogen trafficking and to test whether manipulations of cells to disturb glycogen structure influences glycogen trafficking. The goal of Aim (ii) is to test whether genetic manipulation of vesicular glycogen trafficking can affect the phenotypes of two established mouse models of cardiac glycogenoses. The first, the Gaa-/- mouse, duplicates most features of Pompe disease. In the second, transgenic over- expression in heart of mutant Prkag2, provides a rodent version of Wolff-Parkinson-White syndrome. Both mouse lines are associated with massive cardiac overaccumulation of glycogen, in lysosomes or late endosomic vesicles, which is believed to be important to the pathology of the corresponding human disease. We are working on two other genetic mouse models that could impact intracellular glycogen trafficking. The first is a conditional knockout of the Stbd1 gene;based on our proposal for the function of Stbd1, we would anticipate that the absence of Stbd1 would severely limit the transport of glycogen to the lysosome. The second mouse line is malin-/- mice, generated primarily as a model of another disorder, Lafora disease. The function of malin in vivo is still under active investigation but one hypothesis for its function is that it promotes lysosomal disposal of abnormally structured glycogen. Thus, both of these mouse models could suppress vesicular glycogen trafficking. By crossing these mice with either Gaa-/- or Prkag2 mutant mice, we hypothesize that limitation of glycogen trafficking to the lysosome may alleviate symptoms of Pompe or WPW. Instead, one would expect a build-up of cytosolic glycogen, more similar to what is found in McArdle disease, which has a much less severe phenotype. In crude terms, we would hope to convert one glycogenosis to a less serious one. The most important outcome though would be to indicate whether therapies that specifically target glycogen trafficking might be a fruitful approach for the treatment of Pompe disease or Wolff-Parkinson-White syndrome. PUBLIC HEALTH RELEVANCE: Glycogen is a storage form of sugar that normally accumulates in cells. However, in certain disease states abnormal glycogen accumulates and impairs cell function. This proposal examines whether manipulation of a particular mechanism for glycogen disposal (the lysosomal pathway) can alleviate the damage caused by abnormal glycogen in certain heart diseases.	{ "pile_set_name": "NIH ExPorter" }
Human suffering and health care costs resulting from acute and chronic microbial diseases could be reduced by the use of molecular tools for microbial detection. AndCare scientists propose to design, fabricate, and test a electrochemical gene probe system for the detection of microbes in water that can affect human health via the infections and toxins they cause. Pfiesteria and Cryptoperidiniopsis are representative microbes that are difficult to detect by traditional means and whose detection by the electrochemical system will demonstrate feasibility of the proposed approach. The overall goal is to prove the feasibility of direct (non-PCR amplified) detection and identification of specific microbes in water. This cannot be done with currently available technology. Phase I work will be led by AndCare scientists, in collaboration with Duke University and Boston Probes.. Initial work will be done with a manual version of an eight electrode system that will be automated with novel fluid-handling methods for increased ease of use and system reproducibility. A successful Phase I effort will lead to Phase II integration and optimization of the prototype automated system and electrochemical platform for direct detection of potentially hazardous microbes in water. PROPOSED COMMERCIAL APPLICATIONS: According to Anthony S. Fauci, M.D., Director of NIAID, "The problem posed by emerging and reemerging infections is one of unparalleled complexity." New products designed to detect the emerging and reemerging pathogens of many infectious diseases are essential. Products under development now via an AndCare/Boston Probes venture and those whose development will be assisted by the current project will address a rapidly emerging market tied to such basic necessities as safe food and water that is growing at 100% per year. When this type of testing to counteract these new threats to human health becomes routine, the market could easily reach $10 million/yr for each major disease-causing microbe.	{ "pile_set_name": "NIH ExPorter" }
The focus of the first year Dentist Scientist Awardee is to complete the didactic phase of the basic science curriculum. The Curriculum in Neurobiology requires that students take courses and complete three laboratory rotations related to areas of interest. To this end, an interest in nociceptive transmission have engaged me in relevant course work and laboratory rotations. The first rotation was completed under the guidance of William Maixner, DDS, PhD, associate professor in Pharmacology. The laboratory sought to understand the mechanisms underlying Temporomandibular Joint Disorders. The human psychophysiological studies examined the processing and perceptions of thermal and ischemic pain in chronic pain subjects. The current second rotation is being directed by Alan Light, PhD, associate professor in Physiology. The laboratory focuses on understanding the mechanisms underlying pain pathways. The animal behavioral studies examine the effects of opiods and spinal cord lesions on nociceptive transmission during electrocutaneous stimulation. Before beginning clinical training in Pediatric Dentistry later in the summer, it is hoped that a third rotation involving the nervous system cytoarchitecture will be completed.	{ "pile_set_name": "NIH ExPorter" }
People in recovery from serious mental illness are often excluded from making decisions about their own mental health care, frequently resulting in poor alliances with their care providers. While the Patient Protection and Affordable Care Act addresses this issue by requiring programs to be person-centered and recovery-oriented, providers often struggle to identify concrete ways to change their practices. Person-Centered Recovery Planning (PCRP) is an evidence-based approach to collaborative service planning among an individual, the treatment team, and supports. PCRP maximizes client choice and ownership in the treatment process in order to achieve recovery goals. Dissemination of PCRP in the mental health field has been gradual, and training opportunities are limited. As a result, many people with mental illness continue to experience low self-efficacy in participating in their treatment planning. Recovery Roadmap: A Collaborative Multimedia Tool for Person-Centered Recovery Planning is a highly interactive web-based tool that provides guidance for providers and people in recovery, and promotes widespread implementation of PCRP. The Recovery Roadmap prototype was developed and tested by the Center for Social Innovation (C4), in partnership with Yale University's Program for Recovery and Community Health (PRCH), under a Phase I Small Business Innovative Research (SBIR) grant funded by the National Institute of Mental Health (NIMH) (1R43MH100712). The interactive web-based tool consists of two modules: one for service providers and one for clients. The provider module presents didactic information and exercises to help providers gain the necessary knowledge and skills to implement PCRP. The client module helps people in recovery engage in their care by identifying priorities and life goals, as well as gaining skills in advocating for themselves and driving their own care. Response to the Roadmap was overwhelmingly positive; providers and clients noted concrete changes in the treatment planning process. Providers and clients reported statistically significant improvements on the Person-Centered Care Questionnaire (PCCQ), a measure of PCRP implementation. Phase II will refine the prototype Roadmap to streamline content, provide additional handouts and exercises for providers and clients to complete together, expand the audio/video vignettes and case studies, and add interactive online coaching and support for providers. Phase II also involves a robust evaluation of the Roadmap, using a quasi-experimental design in a fully powered trial. Approximately 25 community-based mental health programs will be randomized into Recovery Roadmap (RR) or Treatment as Usual (TAU) conditions. A total of 230 providers will be recruited (RR n=115; TAU n=115). Each provider will recruit two clients to participate (RR n=230; TAU n=230). Data will include one pre-test and three post-tests with providers and clients, interviews with providers and administrators, network analytics, and administrative client data. This phase will culminate with the dissemination of findings and preparation for Phase III commercialization.	{ "pile_set_name": "NIH ExPorter" }
Kinases function as molecular routers in the transmission of signaling events from the cell surface (e.g. the binding of a hormone or cytokine) to the cellular response in the nucleus (e.g. transcription). One kinase, Akt1, serves as a node in multiple downstream signaling pathways and has been shown to be overexpressed and/or hyperactivated in a wide variety of cancers. Ligands which bind to this kinase and its alternate activation states would enable the development of diagnostic platforms for the study and early detection of cancer. [unreadable] [unreadable] We propose to design a series of peptide-based bi-ligands that recognize the phosphorylation states of Akt with high affinity. By combining chemical one-bead, one-compound peptide libraries and in situ click chemistry with ATP analogs, we can rapidly generate a series of bi-ligands that recognize each of the common phosphorylation states of Akt1 and its isoforms. These ligands will be integrated into microarrays and used to quantitate the concentration and activation states of Akt in complex biological samples. [unreadable] [unreadable] To begin, small libraries of azide-modified Akt1 inhibitor peptides will be screened against fully active Akt1 in the presence of a second library of alkyne-modified ATP-mimics. The Akt1 target will direct the conjugation of the optimal peptide and small molecule to form a high-affinity triazole-linked bi-ligand. Following characterization of the "hit" molecules, the phosphorylation state specificity and affinity of the lead compounds will be increased by re-screening with larger and more diverse peptide libraries. The final round of ligand maturation will employ in situ click chemistry in conjunction with a second library of alkyne-bearing small molecules to select multi-functionalized ligands with improved affinity and biostability. Bi-ligands against each of the phosphorylation states of Akt1, Akt2, and Akt3 will be generated in a similar manner. Following optimization, the affinity capture agents will be immobilized in a microarray and used to measure the concentrations of each phosphorylation state of Akt1, Akt2, and Akt3 in mammalian cell extracts. [unreadable] [unreadable] Increased Akt expression and/or activity has been observed in ovarian, breast, thyroid, prostate, lung, and colon cancers. The ligands generated in this study will be combined into an inexpensive, high-throughput microarray device to map the changes in the activation state of Akt1 in normal, pre-cancer, and cancer tissues, The ability to rapidly determine the concentration and activation state of Akt isoforms in tissue samples would be a major step toward the early diagnosis of many cancers and the determination of optimal therapeutic regimes for cancer patients. [unreadable] [unreadable] [unreadable]	{ "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. A new form of data collection has been developed that exploits the high precision and reproducibility of the MD2's x,y,z sample centering stages to make better use of crystals larger than the beam size, and to distribute x-ray damage over the entire volume of large crystals. This is accomplished by defining a 3-space scanning vector that spans the sample and scanning along this vector as part of the data collection process. Complete or partial data sets may be acquired at a number of discrete points along the scanning vector (discrete vector scanning) or continuously along the scanning vector, with small steps along the scanning vector asserted after acquisition of each frame in a data set (continuous vector scanning). We show that for both discrete and continuous vector scanning data collection, radiation damage effects on data quality are largely mitigated by equipartitioning damage across the volume of samples larger than the beam. Modifications of vector scanning methods are used to locate optimally diffracting regions of a heterogeneous crystal or to align samples embedded in opaque frozen, cryosolution.	{ "pile_set_name": "NIH ExPorter" }
Duchenne Muscular Dystrophy (DMD) is an inherited disorder caused by mutations in the dystrophin gene. For many years gene therapy has been considered a potential way to cure inherited diseases such as DMD. Despite the effort to make gene therapy safer and more applicable, the host immune response to the vector and the therapeutic gene product is still considered a major barrier to successful gene transfer. We hypothesize that manipulating the immune system of dystrophic mice to recognize dystrophin as a self- component before gene transfer will lead to successful delivery and long-term expression of dystrophin in muscles of these mice. We propose to expand Tregs in the presence of dystrophin protein and to adoptively transfer them into vector-recipient mice to induce dystrophin-specific tolerance in these mice. Results will be obtained from parallel studies to investigate the difference between treating dystrophic mice with high- capacity adenoviral (HC-Ad) and adeno-associated viral (AAV) vectors. These studies will enhance our understanding of effects of inducing antigen-specific tolerance on gene delivery and will ultimately facilitate successful delivery of the dystrophin gene to DMD patients. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The Methodology, Data Management and Analysis Core (MDMAC) will provide technical support and training of investigators developing or performing intervention and other geriatric research projects examining the aging phenotype and outcomes research. It will also develop new instruments, methodologies, and data archives to enable future studies. Thus the MDMAC will both address techniques for appropriate design and execution of current experiments and build the foundation for future research studies. Building on our experience with the UM Pepper Center, the MDMAC will address the needs of OAIC investigators, and especially junior investigators, for assistance in the design of intervention experiments, and maintenance, analysis, and interpretation of their data. We propose under this Core Development Project to create and maintain over a two year period a unique data set detailing long-term care in the State of Michigan, as part of the OAIC Methodology, Data Management and Analysis Core (MDMAC). The Michigan Master Long-Term Care Data Archive (MMDA), to be developed in a phased plan over several years, will be available to the UM research community and particularly to junior researchers in aging. It will link multiple disparate but critical data sources describing long-term care (LTC), including longitudinal descriptions of individuals in institutional and community-based care programs, with program eligibility, vital statistics, community characteristics, cost information, census information, etc. The proposed MMDA will expand a successful four year partnership between the State of Michigan and the UM Institute of Gerontology (IoG) which has demonstrated the value of the data sets proposed as the core of the new archive. Thus, the funding of a core capability, including both data and expertise, has the potential to encourage and improve research into clinical- and policy-relevant research in the delivery of effective and efficient long-term care. As such, it can create a visible focal point within UM for researchers involved in study of Michigan's publicly-funded LTC services and a model of how other states can develop such archives and these data can be used nationally and internationally.	{ "pile_set_name": "NIH ExPorter" }
Project Summary Down Syndrome (DS) model mice have three abnormalities in synaptic functions that can explain the cognitive deficits in DS: Decreased long-term potentiation (LTP) and enhanced long- term depression (LTD) of glutamatergic synapses, as well as enhanced inhibitory transmission at GABAergic synapses. Our recent published work indicates that all three observed synaptic dysfunction should be caused by the hyper-phosphorylation of the Ca2+/calmodulin-dependent protein kinase II (CaMKII) seen in DS mice. Our recent unpublished work indicates an intriguing possible mechanism for the cause of CaMKII hyper-phosphorylation in the DS mice. Importantly, this mechanism could also be targeted by pharmacological intervention. As a proof of principle, this project will focus on two aspects of the overall question: (Aim 1) Testing the underlying cause for CaMKII hyper-phosphorylation in DS mice (using a new mutant mouse line that has been generated in the lab and that is incompetent for the proposed underlying mechanism); (Aim 2) Restoration of normal LTP by genetic reversal of T305/306 hyper-phosphorylation in DS mice (using a CaMKII T305/306AV mutant mouse line currently available to the lab). Future studies will test restoration of the other synaptic dysfunctions in DS, restoration of cognitive behavioral tasks, and restoration also by pharmacological means. Notably, while it may seem preposterous to propose a reversal of cognitive impairments in a very complex syndrome by a rather simple intervention, there is actually prior precedent for success: In a model of a different genetic condition that causes cognitive impairments, Angelman Syndrome (AS), CaMKII is also hyper-phosphorylated, and even heterozygous T305/306AV mutation was sufficient to restore both normal LTP and spatial learning. In contrast to AS, the hyper-phosphorylation of CaMKII in DS could be targeted also pharmacologically (if our hypothesized underlying mechanism is correct). Thus, this project will not only provide significant further insight into the mechanism underlying normal synaptic functions and their impairments in DS, but also has tremendous potential for directly opening a new therapeutic avenue for restoring cognitive functions in patients with DS.	{ "pile_set_name": "NIH ExPorter" }
Our proposal tests the hypotheses that polybrominated diphenyl ethers (PBDEs) and their metabolites (OH- PBDEs) accumulate differently in the human fetal liver and placenta than in maternal serum, and that fetal exposure to PBDEs affects fetal metabolic capacity. Virtually all pregnant women in the United States are exposed to at least one and often multiple PBDEs, a class of persistent organic chemicals widely used as flame retardants in consumer products since the 1970s. PBDEs are an important public health concern, as in vitro and in vivo studies show that in utero exposure can adversely impact fetal development. Human exposure and epidemiologic studies have largely characterized in utero exposure by measuring chemicals in proxy biological specimens, such as maternal blood and umbilical cord blood collected at delivery. These samples are not necessarily representative of fetal exposures earlier in pregnancy, particularly to target organs of concern such as the liver and placenta. We propose to measure levels of PBDEs and OH-PBDEs in human maternal and fetal biological specimens from women undergoing voluntary, second trimester pregnancy terminations. Because data on chemical-induced changes in human fetal metabolic capacity are critical to understanding mechanisms of in utero toxicity but are currently lacking, we will also generate original human data on whether fetal exposures to PBDEs alter gene expression of cytochrome P450 (CYP) enzymes in the second-trimester human fetal liver and placenta. At the end of this study, we will produce unique information about human fetal exposure to PBDEs and OH-PBDEs during the second trimester, including empirical relationships between maternal and fetal exposures which can be used to estimate fetal exposures when only maternal levels are available. Further, this will be the first ever in vio data on the relationship between PBDE exposure and human fetal CYP activity. Collectively, this information will help bridge the gap between experimental toxicology and human observation studies and will improve our understanding of population risks to PBDEs as well as other structurally similar environmental chemicals that interact with xenobiotic-sensing nuclear receptors.	{ "pile_set_name": "NIH ExPorter" }
Hypoplastic left heart syndrome (HLHS) is a severe and devastating heart defect that affects ~ 1 in 5-10,000 children born each year and accounts for 25% of all neonatal deaths from congenital heart disease. The significant impact of HLHS stems from a compromise of left- sided cardiac structures (mitral and aortic valves and the left ventricle or LV). Etiologic mechanisms leading to HLHS are unknown making advances in prevention challenging. Genetic studies show that HLHS does not follow simple Mendelian genetics but exhibits "complex inheritance" with contributions from both genetic and environmental factors. Our long- term goal is to help eradicate HLHS through better understanding of the mechanism(s) involved in its pathogenesis. We propose a novel hypothesis for the pathogenesis of HLHS. Our hypothesis is that HLHS is an expression of a form of rheumatic heart disease (RHD) in the fetus. RHD is a serious sequelae of pharyngeal ss-hemolytic group A streptococcal (strep) infection, most commonly manifest in the form of "strep throat". In RHD anti-strep antibodies are generated in response to the strep infection "cross-react" with human valvular and myocardial antigens, through a mechanism known as molecular mimicry. This initiates an inflammatory and immunologic cascade that ultimately damages the aortic/mitral valves and the adjacent LV. We propose a similar mechanism for the pathogenesis of HLHS in which maternal antibodies produced in response to antecedent (and recurrent) strep infection, cross the placenta and damage the fetal heart in the susceptible host. The injury to the fetal valves and LV causes alterations in flow that then leads to LV hypoplasia. Lack of animal models of HLHS has hindered research that could translate to humans. The goal of this proposal is to develop a animal model of HLHS. In a novel rat model we will show that transplacental passage of antibodies produced in response to prior immunization with strep antigens can lead to offspring with HLHS-like pathology. Preliminary studies in a small number of animals have confirmed the feasibility of our proposal. Development of an animal model that clinically recapitulates this disease is essential to furthering the efforts to improve understanding of HLHS and define its mechanisms of pathogenesis as well as potentially identify therapeutic targets. A good animal model for HLHS will have a sustained impact on this field of research. PUBLIC HEALTH RELEVANCE: Heart defects are the #1 cause of birth defects. Hypoplastic Left Heart Syndrome or HLHS accounts for 25% of all neonatal deaths from heart disease. The proposed studies to develop an animal model of HLHS, currently unavailable, will significantly impact research into causes of HLHS and management of this devastating disease.	{ "pile_set_name": "NIH ExPorter" }
Venous thromboembolism (VTE), including both pulmonary embolism and deep vein thrombosis, is an age related condition with potentially catastrophic clinical consequences. Although VTE occurs as frequently as stroke in some populations, its epidemiology is not well understood. The broad goal of this application is to clarify the determinants of VTE, with a particular focus on those influenced by age, both to identify modifiable risk factors as well as those people who would benefit most from preventive strategies. Studies will utilize data from five populations: 39,876 women who participated in the Women's Health Study for over 10 years;29,071 men who participated in the Physicians'Health Studies I and II, of whom 22,071 have over 20 years of follow-up since the initiation of the first trial;8,171 women at high risk of cardiovascular disease who participated in the Women's Antioxidant Cardiovascular Disease Study for an average of 8 years, 508 people with prevalent, idiopathic VTE who participated in the PREVENT trial;and 17,802 people from 26 countries currently participating in the JUPITER trial of statins and cardiovascular disease. Parallel, centralized methods were used to review records and confirm reported cases of VTE. Archived blood samples are available for over 75% of participants. Main relationships to be evaluated include: whether intake of whole grains, fruit, and vegetables influence risk of VTE;the influence of comorbid conditions, especially cancer, cardiovascular disease, stroke and diabetes, on risk of VTE;interactions of genetic determinants of VTE, including factor V Leiden, the prothrombin mutation and polymorphisms in the MTHFR gene with age;whether genetic variants in adhesion molecules influence risk of VTE;and the impact of statins and the metabolic syndrome on risk of VTE. The combined populations will include over 1,500 incident cases of VTE. Studies will consider both a first occurrence of VTE as well as the risk of recurrent VTE. Methods of analysis include Kaplan-Meier estimates of cumulative risk and estimates of the relative hazard of VTE based on proportional hazards models. Strengths of the studies include their prospective design, high follow-up rates, detailed and reliable, long-term exposure and outcome information, and availability of blood samples on a high percentage of the population. Results will help identify high-risk older people, and potentially modifiable behaviors that influence risk of	{ "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. Tris(dithiolene) transition metal complexes have piqued considerable interest in inorganic chemistry due to their fascinating physical properties and electronic structures. Moreover, tris(dithiolene) complexes were the first discrete molecules to possess trigonal prismatic geometry, a feature observed in the oxidized active sites of several molybdenum and tungsten enzymes that catalyze a wide variety of fundamental biochemical reactions. In material sciences, certain dithiolene complexes are candidate materials for nonlinear optical devices and laser applications due to their enormous absorption bands in the near-infrared region. Their reversible redox chemistry has been utilized in the production of semi- and superconducting materials and sensors. The understanding of enzymatic reaction mechanisms or the design of new functional materials should be based on a thorough understanding of the electronic structures of such compounds. Dithiolene ligands can exist as radicals and diradicals and are described as ?non-innocent?, which makes the oxidation and spin states of the central metal ambiguous and difficult to determine experimentally. The existence of oxidized diselenolene ligands is still unknown. Thus, x-ray absorption spectroscopy at the metal K-, and L-edges as well as the sulfur and selenium K-edges is ideally suited to provide fundamental electronic structure insights into the function of these systems. Additionally, metal and ligand EXAFS will assist in defining metric parameters in the solid and solution state. Sulfur and selenium EXAFS are perfectly poised to gauge the geometric change from trigonal prismatic to octahedral as the complex is sequentially reduced by measuring interligand distances. The experiments will be combined with quasi-relativistic density functional as well as multiplet calculations in order to obtain more insight into the experimental data.	{ "pile_set_name": "NIH ExPorter" }
This is a resubmission of a competitive application for funding for the General Clinical Research Center at University Hospital, a part of the Medical Center of the State University of New York at Stony Brook. The primary objective of the Unit is to provide an environment that fosters clinical investigation and education. It will offer resources such as space, hospitalization costs, laboratories, equipment and supplies for clinical research by any qualified member of the faculty in the School of Medicine. In this resubmission application we have a diversity of Departments who will be utilizing the GCRC including Medicine, Surgery, Pediatrics, Psychiatry, Neurology, Dermatology and Orthopedics. The criteria for use of the Center include quality and significance of the research, the special need for Center resources, as well as the collective justification for personnel or facilities. The Center will serve as a resource for teaching students, stimulating interest in academic medicine and developing new methodology in patient diagnostics and therapeutics.	{ "pile_set_name": "NIH ExPorter" }
The purpose of this proposal is to continue support of a two-year postdoctoral fellowship that will attract individuals with a strong commitment to transdisciplinary research from a wide variety of backgrounds from medical, biological, social, behavioral, and policy sciences to help build the next generation of scientific leaders in tobacco control. The need for tobacco control experts continues to grow with Congress' recent legislation granting the US Food and Drug Administration authority to regulate tobacco products, passage of health care reform, with its emphasis on disease prevention and the implementation of the WHO Framework Convention on Tobacco Control, the world's first public health treaty. Our program is emblematic of the innovative cancer prevention training needed to give young scientists the tools to conduct research in collaborative transdisciplinary settings and to be future leaders in catalyzing the integration of multiple disciplines and translation of science to policy and clinical practice. We have one specific aim: Train postdoctoral fellows who will be qualified and well-positioned to become leaders in the future development and implementation of research-based tobacco control policy and interventions. This aim is met through a combination of transdisciplinary coursework related to health policy, biostatistics, tobacco control policy and prevention and treatment of tobacco-related addiction and disease combined with research directed by a primary and secondary mentor from different disciplines. The coursework forms the foundation for an intensive research program directed by two mentors from different disciplines. Fellows also learn how to prepare, submit, and review grant proposals and submit proposals to external funding agencies for their third year of fellowship funding. Research mentors include 30 faculty members with active research programs from all four schools at UCSF (Medicine, Nursing, Pharmacy and Dentistry). Mentors' research interests include tobacco control policy development and evaluation, the tobacco industry as a vector for cancer and other diseases, tobacco industry marketing strategies, tobacco addiction and its prevention and treatment, health effects of smoking and secondhand smoke exposure, smokeless tobacco, and individual- and community-based smoking cessation. The great variety in backgrounds and interests of fellows housed in the UCSF Center for Tobacco Control Research and Education provides a strong transdisciplinary environment that creates continuous opportunities to learn from and appreciate the work done by colleagues working in other disciplines. Each fellow's program is individually developed by the fellow, his or her mentors, and the Fellowship Advisory Committee that monitors his or her progress. The Committee also selects fellows from a very competitive pool: In 2010 there were 40 applications for 4 slots. During the first 4 years of this R25T our fellows have produced 77 papers in 48 different peer reviewed journals. The training program actively supports the career development and placement of our fellows; 8 assistant professors, 3 with nongovernmental health organizations, 1 with a health research consulting firm and 1 full time mother of a small child with an unpaid academic affiliation. We use the R25T mechanism rather than a discipline-based T32 because our program emphasizes transdisciplinary training and communication through unique individually tailored programs of didactics and mentored research.	{ "pile_set_name": "NIH ExPorter" }
Project Summary Atherosclerosis is an inflammatory disease that underlies heart attacks and stroke, a leading cause of death in the world. Disturbed flow (d-flow or OS) promotes, while stable flow (s-flow or LS) inhibits atherosclerosis by differentially regulating endothelial genes, which in turn regulate endothelial function by the mechanisms that are still not fully understood. We previously reported that kallikrein-related peptidase-10 (KLK10) is the most flow- sensitive gene based on a microarray study using endothelial RNA isolated from mouse arteries. However, it is unknown whether it plays any role in EC biology and atherosclerosis. Our preliminary studies indicate that KLK10 produced by s-flow in ECs appears to regulate several anti-atherogenic responses including EC inflammation, and barrier function, potentially in a protease activated receptors (PAR1 and PAR2)-dependent manner. Here, we propose to test the overall hypothesis that s-flow stimulates endothelial KLK10 production, which provides anti-inflammatory and barrier protective function, via PAR1- and PAR2-mediated pathways, leading to atheroprotection. In contrast, Klk10 expression in EC is reduced by d-flow, resulting in induction of EC inflammation, barrier disruption and atherogenesis. We will test this hypothesis in three Aims. Aim 1 will determine the role of KLK10 in flow-dependent EC function. First, flow-dependent KLK10 expression in cultured human and mouse ECs (HAEC, HUVEC, MAEC) under LS vs. OS and in mice will be determined. Effect of KLK10 on flow-dependent EC function (EC inflammation and permeability) will be determined using rKLK10, KLK10 expression vectors, or siRNA. Next, the role of PAR1/2 in these KLK10- dependent EC functions will be determined using specific PAR1 and PAR2 agonists or antagonists, siRNAs and overexpression vectors. Aim 2 will determine the role of PARs in mediating the role of KLK10 in ECs. We will test whether PAR1/2 mediate the anti-inflammatory and barrier protection function of KLK10 in ECs via PAR1/2-biased agonisms by using pharmacological inhibitors and gene-manipulation approaches. BRET and TANGO assays will determine interaction between PAR1/2 and ?-arrestin vs G-proteins. Aim 3 will determine the role of Klk10 in atherosclerosis by the PAR1/2-dependent mechanisms in mouse. ApoE-/- mice will be treated with rKLK10 protein or AAV-KLK10, or KLK10 siRNA in 7C1 EC-targeting nanoparticles. The partial carotid ligation model of atherosclerosis (2 weeks) will be used first, and confirmed in a standard high-fat diet model (3 months). Both pharmacological agents, PAR1-/- and PAR2-/- mice will also be used. Successful completion of these studies would identify KLK10 as a flow-sensitive protein produced and secreted into the circulation, whereby it serves as an autocrine and systemic anti-atherogenic mediator and therapeutic target of atherosclerosis.	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY/ABSTRACT This is an NIMH R01 proposal entitled, ?Neuroprogression across the Psychosis Spectrum in the Early Course of Illness.? Neuroprogressive changes that occur through the early years of illness have been described using neurocognitive testing, PET, CT, fMRI, and post-mortem brain studies; however, these studies rely mainly on cross-sectional data, and longitudinal studies involving frequent measurements are rare, limiting our understanding of the actual timing and trajectories of these measures within this critical time period. The development and implementation of targeted and effective treatments is critically dependent on clear understanding of the timing and nature of disease progression in order to target processes amenable to intervention. Thus, there is an urgent need to carefully characterize neuroprogression in the early course of psychosis if we are to develop effective interventions to target areas of preserved functioning, potentially preventing further decline and chronic loss of functioning. This knowledge gap severely limits our ability to develop targeted treatments when they may be most effective, and to tailor treatment to patients' needs. The aim of the present proposal is the systematic, multimodal characterization of neuroprogression throughout the early course of illness in a cross-diagnostic sample of patients with psychosis using an accelerated longitudinal design. First, we will measure neurocognitive and neurobiological change over the first eight years of illness in order to characterize variability of timing and magnitude of neuroprogression across key measures. Second, we will assess the predictive utility of neuroprogressive trajectories on clinical and functional outcomes. We will also leverage the heterogeneity in baseline cognitive and brain measures to characterize patients by neuroprogressive profile and test whether baseline profiles offer improved prediction of clinical and functional course. The richness of these data will also position us to explore heterogeneity of neuroprogressive trajectories and their associations with clinical and functional outcomes. It has been argued that combining data from clinical, structural and functional imaging, and cognitive measures is superior to monomodal data in the prediction of course and outcome (6). Findings from this project will hasten identification of actionable treatment targets that are closely associated with clinical outcomes, and provide guidance for individualized treatment implementation during a critical period where early intervention strategies may be most effective. Notably, this proposal aims to build on the Human Connectome Project for Early Psychosis (U01MH109977, PI: Shenton) by utilizing the same high-quality methodology and adding longitudinal assessments to baseline data collection already underway, maximizing both the power of the present study and the utility of the HCP-EP data. The PI is an early stage investigator and K23 Awardee with a career focus on characterization of the nature and course of multidimensional symptom domains (e.g. cognition, reward processing), and targeted treatment approaches in psychosis.	{ "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. MR microscopy - shared MR mouse brain data with the investigator. Develop methodology for automated segmentation of the mouse brain into neuroanatomical structures. In specifically, we are planning to explore the applicability of Support Vector Machine (SVM) and Markov Random Field (MRF) for mouse brain segmentation. SVMs have been applied to numerous fields including classification of brain PET images, optical character recognition, object detection, face verification, text categorization and so on. To our knowledge, applying SVM to classify mouse brain structures has been less touched. Recently literature indicates Ali et al (2005) explored the use of Markov Random Field (MRF) for mouse brain segmentation. Given smaller sample size of subjects, SVM tends to have superior results than techniques such as MRF. Therefore, we would like to first conduct comparison study. We will compare the segmentation results of SVM with that of MRF.	{ "pile_set_name": "NIH ExPorter" }
Autoimmune diseases such as Type 1 diabetes (T1D) manifest themselves clinically when insulin producing cells are already completely lost. At this point, the only cure for diabetes is the replacement of these insulin-producing cells. However, patients receiving such grafts suffer from lifetime immunosuppressive treatments, leading to increased propensity to infection. If functional resistance.- to allograft rejection could be conferred on these grafts the requirement for immunosuppression would be avoided aid the quality of life for those receiving allografts would be greatly enhanced. In this exploratory R21 application we propose a new hypothesis for the induction of resistance to islet allograft rejection. Since these grafts are destroyed by cytokines produced by infiltrating cells, we proposed to engineer islets that are unresponsive to these pathogenic cytokines. Two different experimental designs are proposed in this application. In the first set of studies we will determine whether targeted expression of SOCS-1, a protein that inhibits interferon signaling, can ameliorate graft responses. We present compelling preliminary data stating that this aim is feasible and shows promising results. In the second set of studies we plan to down-modulate SOCS-3 gene expression to determine if modulation of IL-6 responses, with increased responses to IL-10, could play a pro-survival role in graft survival. Extensive mechanistic studies, to understand both systemic and localized allograft responses, are proposed for each paradigm. These studies should lead to new methods for inducing resistance to islet allograft rejection.	{ "pile_set_name": "NIH ExPorter" }
See Instructions): This application requests continuing support for Predoctoral Training in Systems and Integrative Biology with specific emphasis on Graduate Training in Neuroscience. The training program proposed encompasses Neuroscience from the study of such complex systems as the brain's mechanisms for regulating social interactions to the cellular circuitry that underlie the brain's processing of visual information to the molecular biology of synaptic transmission. Trainees receive intensive research experiences including formal classes and individual mentoring over the full range of contemporary Neuroscience including access to state of the art facilities for interactive human functional brain imaging, high resolution optical imaging of cells and molecular complexes, genetic analysis of brain development, electrophysiology of ion channels and computational analysis of neurogenetics, neurophysiology and behavior. Trainees leam these approaches for normal brain function and for brain disorders. The program's goal is to prepare students for research careers in Neuroscience with an emphasis on multi-dimensional approaches. With this goal in mind we have assembled a faculty able to direct research projects in behavior, development, biochemistry, electrophysiology, biophysics, molecular biology, photonics, psychophysics, large scale neuronal network analysis, computational neuroscience and brain disease. We have implemented a new core curriculum that encompasses molecular genetics and cell biology, nervous system development, sensorimotor integration and higher brain function and the neurobiology of disease. Our program offers a unique opportunity for students to train in the study of brain function across the phylogenetic spectrum, including in humans in health and disease. We have prepared a series of exams to test our progress in this endeavor and we have mechanisms in place to promote collaborative interactions between the students and the faculty in many departments spanning basic and clinical science. We believe that Predoctoral Training in Neuroscience meets an important national need in modern biomedical and behavioral research, the training of a cadre of students with an integrated understanding across the full spectrum of contemporary molecular, cellular, systems and behavioral neuroscience. Ph.D.students also need to gain a clear appreciation of the many disease-related opportunities in Neuroscience research for which we have added a neurobiology of disease course track along with opportunities for students to interact with patients, learn about animal models of human disease and carry out collaborative and jointly mentored training that spans basic to translational research. RELEVANCE (See instructions): This proposed continuation of Predoctoral Training Program in Neuroscience address a major national health need to train biomedical scientists to carry out cutting edge research on the function, development and diseases of the brain. Trainees learn contemporary molecular, genetic, physiological, behavioral and computational research approaches to brain function in human and animal health and disease.	{ "pile_set_name": "NIH ExPorter" }
This proposal is concerned with: 1) how actin is maintained in an insoluble, yet unpolymerized form (profilactin) in echinoderm sperm, 2) what controls and what is the packing of actin filaments in Limulus sperm, in stereocilia of the ear, in gut microvilli, and in the cnidocil, 3) how are the length and polarity of actin filaments controlled, and 4) how do fluxes of ions into the cell control the actin antics in eggs and sperm. The five proteins making up the profilactin will be isolated and characterized, and their role in inhibiting actin polymerization and/or in making the actin insoluble will be determined using NBD chloride actin. The role that nucleotides play in polymerization in vitro and in vivo will be assayed. I will also assay the profilactin for bound lipid. I propose to compare the chemical composition of the filament bundle in Limulus sperm in its three states, the coil, the false discharge, and the true discharge so that we can see what changes occur in the bound proteins when there is a change in pitch of the actin helix. We will also try to locate the position of the 95,000 molecular weight protein. A detailed study in collaboration with David DeRosier using optical diffraction and 3d reconstruction techniques on the actin filament bundles in the stereocilia of the ear, on the filament bundles in microvilli, and on filament bundles in the cnidocil will be carried out. We propose to use fast freezing, freeze fracturing, and etching techniques, and to study embryonic ear stereocilia. I will also try to determine what ions must enter the cell locally to allow polymerization of actin in eggs and sperm. I propose to use microinjection techniques. I will attempt to see how the filament bundles become ordered in fertilization cones and cytokinesis.	{ "pile_set_name": "NIH ExPorter" }
PROJECT ABSTRACT Non-invasive monitoring of deep-tissue developmental, metabolic, and pathogenic processes will advance modern biology. Imaging of live mammals using fluorescent probes is more feasible within the near-infrared (NIR) transparency window (NIRW: 650-900 nm) where hemoglobin and melanin absorbance significantly decreases and water absorbance is still low. Chromophores in genetically-encoded probes can be formed either autocatalytically from amino acids, as in a case of green fluorescent protein (GFP)-like proteins, or be bound to apoproteins. The most red-shifted fluorescent proteins (FPs) of the GFP-like family have excitation and emission spectra completely or partially outside of the NIRW and suffer from low brightness and modest photostability. Natural bacterial phytochrome photoreceptors (BphPs) utilize low molecular weight biliverdin as a chromophore. BphPs binding biliverdin provide many advantages over other chromophore containing proteins. First, unlike the chromophores of non-bacterial phytochromes, biliverdin is ubiquitous in mammals. This makes BphP applications in mammalian cells, tissues, and whole mammals as easy as conventional GFP-like FPs, without supplying chromophore through an external solution. Second, BphPs exhibit NIR absorbance and fluorescence, which are red-shifted relative to that of any other phytochromes, and lie within the NIRW. This makes BphPs spectrally complementary to other existing biophotonic tools such as all GFP- like FPs and available optogenetic tools. Third, independent domain architecture and pronounced conformational changes upon biliverdin photoisomerization make BphPs attractive templates to design various photoactivatable NIRFPs. Based on our analysis of the photochemistry and structural changes of BphPs we plan to develop three new types of the BphP-based NIRFPs. These include three bright and spectrally resolvable NIRFPs, putatively called short-, medium-, and long-NIRFPs (Aim 1); photoactivatable with non- phototoxic NIR light PA-NIRFPs that are initially dark but become fluorescent either in short-, medium-, or long- NIR spectral regions, and photoswitchable either irreversibly (PS-NIRFPs) or repeatedly (RS-NIRFPs) between these NIR regions (Aim 2); and NIR reporters for protein interactions and phosphorylation based on a reversible bimolecular fluorescence complementation approach utilizing monomerized versions of NIRFPs (Aim 3). We will apply directed molecular evolution approaches based on rational structure-based design and random mutagenesis of candidate proteins, followed by flow cytometry bacterial cell sorting, screening colonies on Petri dishes, and multiwell plate protein characterization. These conventional techniques will allow screening for standard FP properties such as excitation and emission wavelengths, overall brightness, photostability, pH-stability, and folding at physiological temperatures. New high-throughput screening methods will be developed to specifically optimize BphP-based NIRFPs. Selection of NIRFPs with high quantum yield, a crucial parameter for BphP-derived FPs, will be performed using time-resolved fluorescence lifetime measurements of thousands of colonies simultaneously. To screen for a high affinity to biliverdin, which does not penetrate through the inner bacterial membrane, a pulse-chase production of biliverdin using heme oxygenase co-expression and targeting of BphP NIRFPs to bacterial periplasmic space accessible for exogenous biliverdin will be employed. Promising NIRFP candidates will be directly screened in mammalian cells using shuttle vectors to optimize protein folding and stability in mammalian cells, affinity to endogenous biliverdin and low cytotoxicity. Optimized NIR probes will be tested in mouse tumor models and applied to studies in living mammals. The resulting NIR probes will extend fluorescence imaging methods to deep-tissue in vivo macroscopy including multicolor cell and tissue labeling, cell photoactivation and tracking, detection of enzymatic activities and protein interactions in mammalian tissues and whole animals.	{ "pile_set_name": "NIH ExPorter" }
Previous research has shown that an illusory correlation--the perception of an association between two variables that does not exist in the information on which observers' judgments are based--can be based on the co-occurrence of distinctive stimuli, and that this information processing bias can result in the differential perception of social groups. This research program traces the development and consequences of stereotypic beliefs established through this illusory correlation bias. The research investigates the effect of this bias on how people process and interpret subsequent information about group members, evaluate the products of group members, and behave toward group members. Mechanisms for preventing and undermining the stereotypic beliefs formed from illusory correlations are also investigated. The research will be informative about a process by which stereotypic conceptions of groups can develop and the consequences of holding such beliefs on information processing and interpersonal behavior.	{ "pile_set_name": "NIH ExPorter" }
The Clinical Psychopharmacology Section conducts preclinical and clinical research into the mechanisms of action of cocaine. A major component of this project, conducted in collaboration with investigators at NIDDK AND NIMH, is the synthesis and evaluation of analogs of GBR12909 as putative cocaine antagonists. The initial phase of this project has identified several promising novel agents, including the most selective dopamine uptake inhibitor reported. Another component involves investigation of the possible heterogeneity of DA transporter binding sites. This project has identified multiple binding sites for [3H]GBR12935 and [3H]BTCP, and a single binding site for [3H]mazindol. Another component of this project addresses the role of classical conditioning in cocaine-induced behavioral sensitization. These studies have demonstrated that associative learning mechanisms are involved in the acquisition of context-specific behavioral sensitization to cocaine. Another component of this project tested the leading theory of cocaine addiction, which supposes that chronic cocaine use depletes brain dopamine ' Chronic cocaine did not significantly alter the basal levels of DA or 5-HT, or the rate of L-DOPA or 5-HTP accumulation in any region. However, in chronic cocaine-treated rats, the inhibitory effect of cocaine on DA and 5-HT synthesis was greatly reduced. These data suggest that the compensatory feedback mechanisms regulating biogenic amine synthesis are desensitized by repeated cocaine injections. Clinical protocols, currently underway, 1) test the DA hypothesis of cocaine addiction by acute administration of cocaine to subjects who are on various DA receptor antagonists (ARC-170); and 2) attempt to develop a human model of context-specific behavioral sensitization to cocaine (ARC-174). The significance of these findings to drug abuse research is that increased understanding of how cocaine works will lead to the development of improved treatments.	{ "pile_set_name": "NIH ExPorter" }
[unreadable] Basic and clinical research indicate a close association between ingestive behavior and drug-seeking. This laboratory has shown that sustained negative energy balance augments behavioral and cellular effects of psychostimulants. A key endocrine response to negative energy balance is hypoinsulinemia, evident in both plasma and cerebrospinal fluid (CSF). The objective of this proposal is to determine whether central insulin replacement, as delivered by osmotic minipumps, prevents the augmentation of amphetamine effects in chronically food-restricted rats. The first aim is to determine if the output of the minipump remains stable throughout a 14-day period, and whether addition of an anti-aggregant or protease inhibitor helps to maintain delivery of the intended dose. The second aim is to determine the dose of insulin that restores CSF insulin level of food-restricted rats to the control level, as measured by CSF sampling and radioimmunoassay. The third and fourth aims are to determine if intracerebroventricular insulin replacement during food restriction reverses the augmentation of amphetamine's stimulant and rewarding effects. Stimulant effects are measured using photobeam interruption to count horizontal and vertical movements. Rewarding effects are measured by the degree of threshold-lowering for electrical brain stimulation reward. Results of this research may shed light on the high comorbidity of drug abuse and eating disorders. [unreadable] [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
A broad goal of the proposed research is to develop personal exposure monitors comprising wearable, wireless sensor arrays to detect harmful air pollutants within the breathing zone, and improve the assessment of causative exposure-dose-response relationships in epidemiological studies. Heterogeneous orthogonal detectors comprising functionalized carbon nanotube (CNT) based resonators will be designed and integrated with a smart phone platform, targeting the detection of ambient ozone (O3) in Phase 1, particulate matter, NOx and VOCs in Phase 2. Ozone not only influences climatic changes globally through greenhouse gas emissions, but also produces adverse health effects in humans as a secondary pollutant in urban smog through precursors generated in traffic and industrial pollution. As a powerful oxidant gas, O3 can elicit a range of physiological responses once inhaled, including reduced lung function and inflamed airways, exacerbating respiratory diseases such as asthma. Numerous research efforts address the relationship between ozone exposure and health outcomes including mortality and morbidity (hospitalizations, decrements in lung function, and asthma status), but due to lack of personal exposure data, considerable error may be introduced in assessing dose-response relationships. Currently, there are no existing sensors in the market suitable for real- time O3 exposure measurement within the breathing zone. Phase 1 research develops an ozone exposure detector array, fabricated using two classes of nanosensors: (a) polybutadiene polymer-functionalized CNT thin-films, (b) CNT thin-films decorated with Pt or Pd metallic nanoparticles. Both offer maximum sensitivity to ozone while reducing cross-sensitivity to interferents such as NOx. These detector films are integrated with compact radio-frequency (RF) resonators, which respond with unique resonance shift caused by molecular level gas adsorption on the film interface. In contrast to a chemiresistor, both amplitude and frequency shifts in this RF nanosensor may be used to minimize false positives for robust discrimination. Differential signaling between the primary sensor and a reference sensor (pure CNTs) compensates for environmental factors such as humidity, further improving selectivity. The differential signal is wirelessly interfaced through a microcontroller t a smart phone for data display of concentration levels and data communication to public health professionals or regulatory bodies via participatory and ubiquitous sensing. Additionally, sensors embedded in the mobile phone yield information on motion, physical activity, time stamp and GPS location of the subject that can be correlated to the exposure. A prototype O3 sensor will be developed and its performance characterized under controlled laboratory and ambient conditions. Field tests will be conducted on human volunteers to detect ambient ozone as a function of time over several days, and results compared directly with measurements made by standard badge samplers. Causative relationships will be explored by measuring eNO concentrations following the exposure, in collaboration with an environmental epidemiologist.	{ "pile_set_name": "NIH ExPorter" }
The goal of the research projects in this program application is to improve the therapeutic results for patients with multiple myeloma. Important to the success of this project is the biostatistical collaboration with individuals in this Core who provide computerized entry and quality assurance of the data. The purpose of the Biostatistics Core (Core B) is to provide the following services that will be utilized by all of the research projects included in the Program Project. 1. To provide biostatistical collaboration for clinical research protocols. This includes all aspects of design, conduct, analysis and reporting of the clinical protocols, and direction of data processing and quality control assurance. 2. To provide biostatistical collaboration for animal and laboratory research studies. This includes all aspects of design, conduct, analysis and reporting of such studies, including the coordination of laboratory results with parameters and outcomes from clinical studies. 3. To supervise the data processing and data quality assurance, and to coordinate the efforts of the data specialist and those of the data managers in maintaining an accurate computerized database. 4. To provide a data specialist to maintain a computerized database, with assured quality control and standardized reporting. 5. To provide computing resources for data processing, forms design, and statistical analysis. 6. To assure computerized recording of laboratory results on research specimens.	{ "pile_set_name": "NIH ExPorter" }
Following on last years work--UPATSU (Understanding Protein Architecture Through Simulated Unfolding)--we have been computationally studying protein folding. There are two approaches. One is to computationally simulate the refolding of partially unfolded structures. The other is to grow a polypeptide amino acid by amino acid in the same way as it occurs in ribosomal translation. Molecular dynamics is done for 100 picoseconds after the addition of each amino acid. These computations for several known crystallographic proteins will have taken several monts of array processor time. The goal is to mesh the results from the two approaches and to thus obtain the ability to build a model of any new protein.	{ "pile_set_name": "NIH ExPorter" }
Our goal is to investigate the miRNA profiles of prostate tumors and the normal epithelium to evaluate if there are differences in specific miRNA expression patterns of tumors with different grades of differentiation and different biological beavior. Prostate cancers will be manually microdissected to obtain pure populations of tumor cells as well as available normal epithelium distant to the tumor area. Histological features such as Gleason score, presence of perineural, lymphatic invasion, extracapsular or seminal vesicle invasion were also evaluated and recorded for each case. Differentially expressed miRNAs will be defined as those with a two-fold change (up or down) and a p value <0.05. Different miRNAs expression profiles will include comparison between the low grade tumors (Gleason score 6 and 7) vs. high grade (Gleason score 8 and 9) lesions. Then, each tumor group will be compared with their respective group of normal epithelium.	{ "pile_set_name": "NIH ExPorter" }
In the spring and summer of 1987, 1,150 youth aged 17 to 22 and a parent of the youth are being interviewed in the third wave of the National Survey of Children. The child and his or her parent were initially interviewed in 1976 and re-interviewed in 1981. Consequently, a wealth of information exists about the early lives of these young people, including demographic data; measures of personality; a test of intellectual ability; information about family relationships, closeness to parents, religion, and school progress; attitudes about school; behavior problems; and relationships with siblings and peers. In 1987, information was again obtained on a variety of topics, including if and when the youth initiated use of alcohol, of marijuana, and of other illicit drugs; on drug use during the past year; and on the occurrence of varied problems associated with drug use. It is proposed to use these data to examine the etiology of adolescent drug abuse. The antecedents of age at initial use of each drug would be assessed using proportional hazards life table models. Multivariate logit models would be used to investigate the antecedents of current use, frequency of current use, and problems associated with current use. Hypotheses to be tested are based on a synthesis of previous research with elements drawn from personality theory, alienation theory, social learning theory, and developmental theory. It is hypothesized that youth will be more likely to be drug users if they manifested personality deficits as children, if they were alienated from family, school, and religious institutions as children, and if they were exposed to models of substance use in the home and among friends. Based on developmental theory, we anticipate experimentation with substance use as a part of the identity formation process; however if youth experienced personality problems or alienation as children or if they were exposed to models of substance use, it is further hypothesized that youth will have initiated use at younger ages and that they are more likely to be heavy users experiencing problems related to drug use.	{ "pile_set_name": "NIH ExPorter" }
PROJECT SUMMARY/ABSTRACT ? ADMINISTRATIVE CORE The goal of the Center for Advancing Point of Care Technologies (CAPCaT) in Heart, Lung, Blood, and Sleep Diseases is to develop and optimize novel point of care technologies (POCT) to improve the diagnosis and management of heart, lung, blood and sleep (HLBS) diseases. A strong Administrative Core provides a foundation for the proposed center, which will allow program resources to achieve maximum impact. This structure is necessary to: provide the appropriate scientific and fiscal oversight of CAPCaT; promote interactions within and between the Center and the Point of Care Technology Research Network (POCTRN); and communicate and manage stakeholders including users, clinical systems and funders. The Admin Core will have the primary responsibility for interactions with NIH staff, including working with NIH to advance national efforts to improving the diagnosis and management of HLBS diseases.	{ "pile_set_name": "NIH ExPorter" }
Parkinson Disease (PD) causes progressive motor and non-motor deficits with 75% of patients developing dementia after 10 years. A growing body of evidence suggests the involvement of amyloid beta (A?) in the cortical pathology of PD that has been linked to accumulation of ?-synuclein and neurodegeneration. Yet, how regional A? accumulation relates to decline in specific cognitive domains and to deficits in regional neuromodulatory transmitter systems remains unknown. This study will test the hypotheses that 1) A? measurements made by Pittsburgh Compound B (PiB) correlate to postmortem proteinopathy. 2) deficits in selected cognitive domains (executive function, attention, visuospatial ability, and memory scores) will correlate regionally to PiB and postmortem measures of tau, A?, and ?-syn proteinopathy; 2) PiB imaging and cognitive deficits will correspond to regional neurochemical changes in postmortem tissue; and 3) accumulated A? predicts deterioration in specific cognitive domains and the time to dementia. To test these questions, I will use a large, previously-recruited cohort of 287 participants with PD and controls that have undergone baseline PET imaging scans of PiB and ongoing longitudinal (baseline and 3-year follow-up) comprehensive cognitive testing. Cross-sectional analyses of PiB uptake and selected cognitive deficits will be assessed using appropriate statistical techniques with a data-drive, whole-brain, parameterized voxel-based approach to identify regions associated with PD-driven cognitive impairment. Neurochemical and pathophysiological analyses will be conducted on postmortem tissue from the 47 deceased participants with pathologically confirmed PD who had PET PiB scans in life to determine the relationship between PiB uptake and post- mortem deposition of A? and ?-synuclein. Correlations between PiB uptake in life and changes in regional distribution of pathological protein aggregates and behaviorally-relevant neuromodulators (dopamine, serotonin, norepinephrine, and acetylcholine) will then be compared in regions associated with cognitive impairment. Whether PiB uptake can predict decline in specific cognitive domains or time to onset of dementia will be determined using baseline PiB imaging and longitudinal cognitive data. This study and my mentorship team will provide training relating to neuroimaging, neurochemistry, and behavioral manifestations of PD and analytical approaches to integrating questions of transmitter activity and cognitive output. This training will provide the basis for moving forward toward a career as an independent investigator through formal and informal professional development and laboratory management training. This study will help elucidate the relationship between cognitive dysfunction in PD and regional pathology, identifying new targets for treatment in PD. Findings from this study will inform the development of PET tracers and help identify deficits related to specific behavioral symptoms including depression, apathy, executive dysfunction, or visuospatial dysfunction.	{ "pile_set_name": "NIH ExPorter" }
In keeping with the original grant title "Bedside to Bench", we emphasize the translational aspect of the conferences, especially the use of clinical issues to frame, define and drive basic inquiry, and to promote interactions between basic and clinical investigators. This proposal continues many of the successful features of the original program;topics developed with input from the geriatrics community and the NIA, multidisciplinary conferences with ample time for interaction, participation by young investigators, minority investigators and established leaders from within and outside the traditional aging research community, dissemination through Symposia at the Annual Meeting of AGS and publication through the Journal of the American Geriatrics Society (JAGS), as well as other media. Based on priorities developed by AGS and NIA over the last year, we propose three new topics to be addressed in 2007-2009;1) "Thinking, moving and feeling: do cognitive, mood and movement disorders of aging have overlapping causal mechanisms?", 2) "Idiopathic Fatigue of Aging," and 3) "Inflammation and nutrient metabolism: interaction as a key to effective intervention." This is an application for a three year U13 cooperative conference grant on behalf of the American Geriatrics Society (AGS) in cooperation with the Geriatrics and Clinical Gerontology branch of the National Institute on Aging (NIA). The mission of this conference grant program remains unchanged from the initial proposal;"to heighten research attention on clinical geriatrics issues that are of pressing concern clinically or have the potential to greatly improve clinical care or prevention for older adults". Our long term goal remains to "translate research results into clinical care that will improve the health outcomes of older adults."	{ "pile_set_name": "NIH ExPorter" }
The contractor will accomplish the following for hemodialysis and peritoneal dialysis: The specific target is to evaluate the in vivo intercompartmental weight. Study mass transfer-molecular weight characteristics of peritoneal dialysis and study the effect of solute-protein activities in the dialysate on solute transfer. Optimize peritoneal dialysis dosage cycles and schedules as a result of the above findings.	{ "pile_set_name": "NIH ExPorter" }
Project Summary/Abstract Single-cell technologies have contributed to the understanding of underlying order in heterogenous samples, providing a platform to identify rare cell types with implications to human disease. These platforms are largely based on analyses of single-cell suspensions and do not provide a spatial context to the findings. It is becoming increasingly evident that the tissue architecture is a critical parameter and contributes significantly to the overall biological state of a sample. High-parameter imaging technologies enable the identification and localization of rare cell types in heterogenous tissue samples. Currently available high-parameter imaging technologies are time-consuming, limited in the number of parameters that can be reproducibly imaged and/or are cost-prohibitive for broad use in the scientific community. Akoya Biosciences Inc. has exclusively licensed a new high-parameter imaging technology termed CODEX (CO-Detection by indEXing) that is robust, inexpensive and can quickly acquire fluorescence data across tens of parameters. The CODEX platform consists of a set of specialized biologic reagents, including tagged antibody and oligo products, as well as a fluidics robot that can be attached to any inverted microscope. The broad applicability of this technology will enable any lab, ranging from small academic labs to large pharmaceutical research entities, to acquire high-parameter imaging data. These analyses will enable a deep understanding of disease mechanisms, therapeutic targets and overall tissue architecture. The goal of Akoya Biosciences Inc. is to commercialize this technology through the production of the fluidics robot as well as reagent kits for antibody staining and signal rendering. The current version of the technology was developed for the analysis of fresh-frozen (FF) tissue samples; however, most tissue samples are stored as formalin-fixed paraffin embedded (FFPE). The conditions for analyzing FF tissues using the CODEX platform do not result in effective antibody staining for FFPE tissues. The goal of the research outlined in this grant proposal is to develop conditions and methodologies that enable the analysis of FFPE tissues with the CODEX platform. This will increase the overall applicability of the technology and drastically increase the customer base for Akoya Biosciences Inc. We plan to use the information learned in this Phase SBIR I grant proposal to develop antibody panels validated for FFPE tissues that will be commercialized. This greater development effort will be the focus of a follow-on Phase II SBIR grant application.	{ "pile_set_name": "NIH ExPorter" }
The purpose of this project is to investigate the molecular structure of fibrous proteins and proteoglycans, and to study intracellular gelation of Hemoglobin S. The structural information obtained will be correlated with function. Areas of present interest are 1) Molecular structure and dynamics of collagen. 13C and 2H magnetic resonance techniques are being used to study the structure and interactions in collagen fibers. 2) Proteoglycan structure. 13C magnetic resonance is also being used to study the molecular mobility of the polysaccharide and protein chains in cartilage proteoglycans. 3) 13C magnetic resonance is being used to study the extent and mechanism of hemoglobin S gelation in erythrocytes. For these studies, magnetic resonance spectrometers have been assembled which give 2H, 13C and 31P spectra of solids. High power decoupling, cross-polarization, magic angle spinning, and solid echo experiments are all performed.	{ "pile_set_name": "NIH ExPorter" }
Our laboratory has shown that amphetamines trigger the internalization of the dopamine transporter (DAT) by a series of intracellular events that are distinct from the generally established actions of amphetamines to inhibit dopamine (DA) uptake or to increase DA efflux. We have found that when applied to cell lines, cultured DA neurons or midbrain slices, amphetamine activates the small GTPases, RhoA and Rac-1 and triggers internalization of the dopamine transporter (DAT) by a specialized internalization pathway that requires the activation of the small GTPase, RhoA. Intriguingly, amphetamine must be transported into the cell to have these effects and its actions are actually blocked by cocaine, a drug that inhibits DAT and prevents amphetamine entry. We have also found that elevation of cAMP, via DA receptors or by amphetamine-induced adenylate cyclase activation, inactivates RhoA and serves as a break on carrier internalization, thus demonstrating an important interaction between PKA- and RhoA-dependent signaling in mediating the actions of amphetamines. These observations also imply the existence of a novel intracellular target for amphetamines and suggest new cellular pathways to target in order to disrupt amphetamine action. In recent studies we have established that a G-protein coupled trace amine receptor (TAAR1) serves as a direct intracellular target for amphetamines in dopamine neurons. Using transgenic mouse lines lacking the TAAR1 receptor we have shown that the intracellular effects of amphetamine, including both the elevation in cAMP and the increased RhoA activity, depend absolutely upon TAAR1 activation. We are currently investigating where within the cell amphetamines activate TAAR1 and which G-proteins serve as downstream effectors. In other experiments we have shown that G-protein beta-gamma subunits released when G-protein-coupled receptors are activated bind directly to the DAT and enhance dopamine efflux. Using cell permeable peptide fragments and mutagenesis of the DAT we have been able to define the transporter domains required for this interaction and to develop structural models for how this interaction may facilitate dopamine efflux by the transporter. We have also observed that the same amphetamine-activated RhoA-dependent mechanism downregulates a glutamate transporter, EAAT3, present on the surface of dopamine neurons. We have identified the EAAT3 peptide sequence responsible for this regulation, generated a cell-permeant fusion protein that blocks internalization and have used it to explore the effects of amphetamine on excitatory neurotransmission in brain slices. These studies have provided new tools to distinguish the effects of amphetamine on dopaminergic and glutamatergic signaling. We have also compared the effects of various amphetamine compounds on the activation of cellular signaling pathways. Comparison of the effects of methamphetamine on glutamate transport activity to those of amphetamine indicate that while both treatments lead to a loss of cell-surface EAAT3, the effects of methamphetamine are much broader and do not depend on the expression of the DAT. These findings suggest an explanation for the broader, more devastating effects of methamphetamine: unlike amphetamine, methamphetamine has the capacity to alter EAAT3 surface expression and regulate excitatory neurotransmission, not only in dopamine neurons, but also in many other neuronal cell types within the brain. Glutamate transporters (also known as excitatory amino acid transporters or EAATs) present at the surface of neurons and supporting glial cells regulate the extracellular concentration of glutamate, the major excitatory neurotransmitter in the brain. By transporting glutamate back into the cell, these carrier proteins prevent glutamate from reaching toxic levels and also limit the extent and duration of transmitter signaling during glutamatergic neurotransmission. These carriers have an additional function in that they possess an anion channel activity that can regulate cellular excitability, which enables them to serve as sensors of glutamate levels outside the cell. Our laboratory has used site-directed mutagenesis, sulfhydryl modification, and chemical cross-linking approaches together with biochemical, and electrophysiological analyses of the mammalian carriers to examine the structural domains required for substrate transport and ion permeation. Although there has been progress towards understanding the mechanism of glutamate transport, little is known about the mechanism and structural basis of anion channel activation. We recently identified a conserved positively-charged arginine residue in transmembrane domain 7 as an essential element in the anion channel gating mechanism. Substitution of this residue with a negatively-charged amino acid, eliminates sodium- and substrate-dependent anion channel gating, and drives the channel into a substrate-independent constitutively open state. Our data suggest that anion channel gating occurs through a transition from intermediate conformations that are closely linked to transport. We hypothesize that interactions of this crucial cationic residue with nearby anionic groups are required to stabilize the closed state of channel and thus are intrinsic elements of the channel gating mechanism. We created a homology model of the transporter and have identified potential interacting residues that also appear to be required for anion channel gating in EAATs. The neuronal glutamate transporter isoform, EAAT3 has also been shown to transport the amino acid cysteine and has been proposed to be a primary mechanism used by neurons to obtain cysteine for the synthesis of glutathione, a molecule critical for preventing oxidative stress and neuronal toxicity. This year the laboratory also completed work examining the mechanism of cysteine transport by EAAT3. Our results support a model whereby cysteine transport through EAAT3 is facilitated through cysteine de-protonation and that once inside, the thiolate is rapidly re-protonated. Moreover, these findings suggest that cysteine transport is predominantly unidirectional and that reverse transport does not contribute to depletion of intracellular cysteine pools.	{ "pile_set_name": "NIH ExPorter" }
The research capabilities of NIH investigators here at Brown are compromised by limited confocal microscope access and technology. Over the past several years, researchers have had to postpone experiments, travel to other institutions outside of the region to perform experiments, minimize the experimental protocol to accommodate the existing limitations, and farm out the confocal imaging of projects. This deficiency has created greatly inefficient use of NIH research support of investigators here. The applications of microcopy have also expanded for many investigators and the available, limited instrumentation is incapable of meeting these demands. We see no other way of acquiring such instrumentation without receiving this support from the NCRR. Twelve-faculty members, identified as Major Users, have collaborated to create this request for funds to support the acquisition of an integrated confocal microscope system. The Major User Group consists of twelve different laboratories with 86 researchers. The proposed instrument system contains the following: 1. Zeiss LSM 510 META confocal microscope, 2. Imaged through a Zeiss Axiovert 200 MOT inverted microscope, 3. Equipped with Narishige micromanipulators and an Eppendorf microinjector, 4. Microscope stage temperature control system. The requested instrument would significantly enhance the research productivity of the broad, interdisciplinary community of researchers described here, and greatly improve the biological image capabilities in the region. [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Hepatitis C virus (HCV) is an important cause of liver disease. Current therapies are inadequate. A cocktail of multiple drugs, each targeting an independent viral function offers the best chance for effective pharmacologic control. Our long-term objectives are to better understand the molecular virology of HCV, increase the repertoire of new targets that can be translated into novel drug classes for inclusion in future anti-HCV cocktails. We have identified and genetically validated two new targets within NS4B: an arginine-rich-like motif mediated RNA binding activity that is specific for the 3' terminus of the viral negative RNA strand (3'term(-)); and an amphipathic helix, termed 4BAH2, with a dramatic ability to promote lipid vesicle aggregation, an ideal candidate biochemical activity for creating the membranous web, the HCV replication platform. We then identified small molecule inhibitors of these two targets with significant anti-HCV activity: a first generation H1 antihistamine-clemizole-potently inhibits NS4B RNA binding and exhibits dramatic in vitro synergy with the HCV NS3 protease inhibitor SCH503034. Two distinct compounds-A2 and C4-were each found to be potent specific inhibitors of 4BAH2-mediated lipid vesicle aggregation. Finally, we recently developed a new method for rapidly mapping RNA secondary structure, affording new ways to study RNA elements such as the target of NS4B binding. Our overall hypothesis is that two different newly described functional activities within NS4B are each essential for mediating NS4B's role in HCV replication. Thus approaches designed to disrupt the function of these domains may be potentially used to inhibit HCV replication in patients. More specifically, we hypothesize: 1) clemizole's antiviral activity is distinct from its antihistamine activity; 2) the mechanism of action of A2 and C4 can be further validated by analysis of resistant mutants and derivatives of these compounds; 3) while A2 and C4 both target the same 4BAH2, they do so via distinct mechanisms, where one targets 4BAH2's interaction with membranes, and one targets 4BAH2's interaction with itself; 4) NS4B RNA binding inhibitors are broadly synergistic with NS3 protease inhibitors; 5) both NS4B RNA binding inhibitors and 4BAH2 inhibitors represent attractive potential new drug classes for future anti-HCV therapeutic cocktails designed to maximize efficacy and minimize resistance; 6) 4BAH2's lipid vesicle aggregating activity reflects a biochemical activity important for membranous web formation that is amenable to both genetic and pharmacologic disruption, and the specific amino acids critical for this activity can be identified by mutational analysis; 7) the secondary structure of the HCV 3'term(-) is altered upon interaction with NS4B or clemizole, or upon mutation to escape clemizole inhibition; 8) expression of wild-type and drug-resistant forms of NS4B can enable critically important 3D structure determinations. To test these hypotheses, we propose to further study these compounds' mechanism of action, determine their potential as critical components of future anti-HCV cocktails, and better understand the structure and function of these compounds' targets within NS4B and the HCV life cycle.	{ "pile_set_name": "NIH ExPorter" }
On the basis of gene expression profiling, the laboratory proposed that the most common form of lymphoma, diffuse large B cell lymphoma (DLBCL), is a composite of three molecularly distinct diseases that are indistinguishable by standard diagnostic methods. These diseases, termed germinal center B cell-like (GCB) DLBCL, activated B cell-like (ABC) DLBCL, and primary mediastinal B cell lymphoma (PMBL), arise from B lymphocytes at different stages of differentiation by distinct oncogenic pathways. The curative response of patients with DLBCL to chemotherapy is highly variable, and the DLBCL subtype distinction accounts, in part, for this heterogeneity. With CHOP multi-agent chemotherapy, the 5-year survival rates of ABC DLBCL and GCB DLBCL are 60% and 30%, respectively. This clinical disparity likely reflects the host of genetic differences between these DLBCL subtypes. A recurring theme that emerges from our molecular profiling efforts in lymphoma is that the curative response to treatment and the length of survival following diagnosis are dictated by molecular features of the tumors at diagnosis. In DLBCL, we developed a multivariate model of therapeutic outcome based on gene expression signatures, which quantitatively reflected distinct aspects of tumor biology. This study was performed in the era of CHOP chemotherapy for DLBCL, which has subsequently been supplanted by regimens that add the anti-CD20 monoclonal antibody Rituximab (R-CHOP). Since R-CHOP improves survival in DLBCL by 10-15% compared with CHOP, it was important to investigate whether our previously identified prognostic gene expression signatures were still applicable. We therefore profiled gene expression in 233 pre-treatment biopsy samples from patients with DLBCL treated with R-CHOP as well as in 181 biopsies from CHOP-treated patients. We initially demonstrated that the distinction between the ABC and GCB DLBCL subtypes remained prognostically significant in R-CHOP-treated cases, with each subtype benefiting equivalently from the addition of Rituximab. In fact, 3 of the 4 gene expression signatures that were associated with survival in the CHOP cohort retained their prognostic significances in the R-CHOP cohort. This suggested that several aspects of tumor biology that influence the curative response to therapy remain important after the addition of Rituximab. We therefore constructed an optimal survival model based on gene expression signatures identified using the CHOP-treated cohort and then tested the model in the R-CHOP-treated cohort. The final model consisted of three signatures, termed germinal center B cell, stromal-1, and stromal-2. The model was strongly associated with both overall and progression-free survival in the CHOP and R-CHOP cohorts as well as in a third CHOP cohort for which gene expression profiling data were available. The gene expression-based survival model could divide the patients in the R-CHOP cohort into quartile groups that had 3-year progression-free survival rates of 84%, 69%, 61% and 34%, respectively, indicating that the model captures much of the heterogeneity in therapeutic response. Remaining heterogeneity could be ascribed to clinical factors in the International Prognostic Index, which were statistically independent from the prognostic gene expression signatures in predicting survival. An important feature of this study was our ability to associate the prognostic gene expression signatures with particular aspects of tumor biology. The germinal center B cell signature mirrored the distinction between GCB and ABC DLBCL and therefore reflects the myriad genetic and epigenetic differences that exist between these two subtypes24. On the other hand, the stromal-1 and stromal-2 signatures reflected different aspects of the tumor microenvironment. The stromal-1 signature, which was associated with favorable outcome, identified tumors that were fibrotic and rich in histiocytic cells of the myeloid lineage. The stromal-2 signature, which was associated with poor outcome, included a host of genes that are characteristically expressed in endothelial cells and was correlated with increased tumor blood vessel density, revealing an unanticipated role for angiogenesis in DLBCL. Based on current understanding of tumor angiogenesis81, a hypothetical angiogenic switch in DLBCL can be envisioned. In this model, high expression of the stromal-1 signature is presumed to be the initial state. Such tumors express thrombospondin-2, a potent inhibitor of angiogenesis. However, the stromal-1 signature also includes KIT ligand, which can mobilize bone marrow-derived endothelial precursor cells, and the matrix metalloproteinase MMP-9, which can liberate KIT ligand from the extracellular matrix, possibly initiating angiogenesis. Tumors with high stromal-2 signature expression characteristically express the chemokine SDF-1, a chemoattractant for mobilized CXCR4+ endothelial precursor cells. These findings have several implications for the diagnosis and therapy of DLBCL. First, it is evident that clinical trials in DLBCL need to assess these signatures to allow the patient cohorts in different trials to be assessed. Second, several therapeutic strategies are suggested by the survival associated signatures and their component genes. It is certainly conceivable that anti-angiogenic therapy may be selectively active in cases with high expression of the stromal-2 signature. Given the expression of SDF-1 in these tumors, agents that block its receptor CXCR4 should also be considered. For tumors with high stromal-1 signature expression, it is conceivable that therapies that purge the myeloid-derived cells from these tumors may help to eradicate the malignant cells by eliminating trophic signals derived from these immune cells. An intriguing component of the stromal-1 signature is connective tissue growth factor, (CTGF), a key mediator of fibrotic responses. Antibodies to CTGF are being developed for the therapy of fibrotic disorders and should also be investigated in the intensely fibrotic DLBCL tumors with high stromal-1 signature expression. Array-based comparative genomic hybridization was used to identify genomic changes in copy number that influenced survival. Two genomic alterations that occurred exclusively in ABC DLBCL were deletion of the INK4a/ARF tumor suppressor locus and trisomy 3. These genetic aberrations, considered separately and together, identified a subset of patients with ABC DLBCL with inferior prognosis relative to other patients with this DLBCL subtype. This ABC DLBCL subset was also characterized by oncogenic mutations in the CARD11 gene, which encodes a scaffold molecule required for NF-kB signaling downstream of the B cell receptor. These mutations are responsible for constitutive NF-kB activity in ABC DLBCLs. The co-occurrence of CARD11 mutations, INK4a/ARF locus deletions and trisomy 3 in a subset of ABC DLBCLs suggests that this subset may arise by a distinct pathogenetic pathway. We are currently investigating several platforms to deliver the molecular diagnostic and prognostic distinction to patients with lymphoma. The goal is to utilize formalin-fixed and paraffin-embedded tissue for these analyses since most lymphoma biopsies are routinely stored in this fashion.	{ "pile_set_name": "NIH ExPorter" }
This is a competing renewal application whose overall objective is to define the mechanisms of interferon (IFN)-signaling in malignant cells. IFNs exhibit important antineoplastic properties in vitro and in vivo and are key elements in the immune surveillance against cancer, but the mechanisms by which they generate such effects remain to be defined. We have provided the first evidence that the mTOR signaling cascade is engaged by IFN receptors (IFNRs) and regulates cap-dependent mRNA translation via control of the eukaryotic initiation factor 4E (eIF4E) and the eIF4F complex. Remarkably, our studies have provided evidence for signaling specificity and differential use of mTORC2 complexes by IFNs, as compared to oncogenic signals. Our studies suggest dual regulatory roles for mTORC2 complexes in IFN-signaling, controlling downstream pathways that regulate both transcription and mRNA translation of ISGs. The current proposal is a systematic approach to dissect the functions and roles of these complexes in IFN-signaling and to define their relevance in the generation of the antineoplastic effects of IFNs. Specific aim 1 will identify upstream IFNR-generated signals that lead to mTORC2 activation and will determine the mechanisms of specificity of mTORC2 engagement by the Type I IFNR. It includes studies to dissect the role of IFNR-associated kinases in the process; studies to define whether differential engagement of distinct Sin1 isoforms accounts for specificity in the IFN-system; and screening efforts to identif novel Rictor- and Sin1-interacting IFN-signaling elements. Specif ic aim 2 will define the mechanisms by which mTORC2 complexes control ISG expression and their roles in the generation of IFN-inhibitory responses in malignant cells. It includes experiments to define the roles of mTORC2 complexes in IFNR-activated signaling cascades that regulate transcriptional activation and mRNA translation of ISGs; experiments on the effects of IFN?-activated mTORC2 complexes on AGC kinases; and studies to dissect the requirement of distinct downstream effector elements of mTOR pathways in the generation of IFN-antiproliferative responses. Specif ic aim 3 will examine the roles of mTOR-dependent signals in the antineoplastic effects of IFNs in Ph (-) myeloproliferative neoplasms (MPNs). JAK2V617F mouse models will be established in AKT -/-, S6K -/- and Pdcd4 -/- KO mice, and the ability of IFN? to induce antileukemic responses in vivo, in the presence or absence of distinct effectors of the pathway, will be assessed. The activation of IFN-dependent mTOR pathways in primary hematopoietic precursors from patients with MPNs in vitro and in vivo will be assessed and correlated with IFN-sensitivity. Altogether, these studies will advance our understanding of the signaling mechanisms controlling generation of IFN-antitumor responses and will provide important information on the events that lead to malignant cell resistance to IFNs. Ultimately, they may form the basis for new approaches to overcome IFN-resistance.	{ "pile_set_name": "NIH ExPorter" }
The goal of this investigation is to increase the success of clinical islet transplantation. Two interrelated hypotheses are put forth to achieve this goal. First, Nitric Oxide (NO) is postulated to be a prime mediator of islet dysfunction. Second, a previously unrecognized ability of insulin to decrease NO production is proposed to explain its islet-protective activity. Both of these hypotheses will be tested directly by focusing the experiments in this investigation on the use of inducible Nitric Oxide Synthase "knockout" mice (iNOS-/-). Specific Aim I will determine the role of macrophage NO in islet graft rejection. Based on preliminary and published results from this laboratory, NO is proposed to inhibit islets in 2 "waves": an early wave that occurs 1-2 days posttransplant of syngeneic or allogeneic islets is responsible for early islet dysfunction. A second wave NO-mediated islet dysfunction is proposed to occur during classical allograft rejection. The early wave will be investigated by comparing syngeneic islet function in diabetic iNOS-/- and iNOS-/+ mice; the second wave will be investigated using allogeneic islets in these hosts. Inhibitors of NO will then be used to determine how to best increase islet function. Preliminary and published evidence from this laboratory suggests that inhibiting NO in a clinically applicable way does increase the success of islet transplantation. Islets will be implanted in the intraperitoneal cavity in this investigation because intragraft NO production and allograft rejection responses can readily be followed at this site. Mice will be the primary species used because, like humans, they are "low" producers of NO; rats will be used in selected experiments because of their larger size. Specific Aim II will determine the mechanism by which insulin promotes islet function. A dominant paradigm has been that insulin allows transplanted beta-cells to "rest". However, preliminary evidence indicates that insulin administration to diabetic mice or rats decreases macrophage iNOS mRNA expression and NO production. Therefore, specific experiments will again compare iNOS- /- and iNOS-/+ mice to directly test the hypothesis that the islet- protective activity of insulin results from decreasing NO production. Finally, hyperglycemic clamps will be employed to maintain hyperglycemia in order to determine if the islet-protective activity of insulin depends on its ability to lower blood glucose. The results of this investigation should provide important new information that will increase the success of clinical islet transplantation as well as providing basic information on what role NO plays in rejection of cell transplants, and how NO production is regulated by insulin.	{ "pile_set_name": "NIH ExPorter" }
Childhood leukemia (CL) rates among children with Hispanic ethnicity are among the highest in the world, although little is known about the mechanisms and determinants underlying this observation. CL risk is widely thought to be modulated by the interplay between individual genetic susceptibility and exposure to environmental and endogenous substances. The long-term goal of the proposed research is to identify combinations of specific genetic factors and environmental exposures that cause children with Hispanic ethnicity to have elevated rates of CL. With its diverse Hispanic population comprised primarily of pure indigenous and mixed (mestizo) indigenous-European peoples, as well as the widespread use of biomass fuels for heating and cooking, Guatemala presents an ideal model system to study exposure to suspected carcinogenic combustion products in an ethnic population most at risk for this disease. Before such a study could be undertaken in Guatemala, however, a pilot study is required to establish the feasibility of conducting an epidemiologic assessment in this largely understudied population. Such a pilot would also provide the opportunity to identify unforeseen barriers and subsequently modify the protocols to overcome such barriers. Therefore, the aims of the proposed pilot study include identifying and enrolling 50 children with incident CL from a tertiary pediatric oncology hospital in Guatemala City and 50 age, sex, and urban vs. rural status matched healthy controls (identified as healthy siblings of cardiovascular pediatric patients at an adjacent tertiary cardiovascular referral center). Additional aims include the development, validation, and administration of a valid survey instrument to record exposure to relevant epidemiologic and environmental exposures for CL, and to collect diagnostic bone marrow and remission peripheral blood on cases, and saliva on cases, controls, and their parents. Success for this pilot study will be measured by response rates for subject recruitment and biospecimen collection, results of a survey instrument validity sub-study, and comparability of cases and controls. If successful, this proposed study will motivate a full case-control study of the genetic and environmental epidemiology of childhood leukemia in Guatemala. Such a study would fill in gaps in what is known about the tumor biology of CL, of inherited genetic susceptibility associated with Hispanic ethnicity, and of putative causal relationships with products of combustion and risk. PUBLIC HEALTH RELEVANCE: A case-control study of childhood leukemia in Guatemala can fill important gaps in our knowledge about its epidemiology, including the observed elevation in rates among children of Hispanic ethnicity and the putative causal relationship of exposure to environmental products of combustion. To establish the feasibility of conducting a full epidemiologic study of the genetic and environmental epidemiology in Guatemala, a largely understudied population, we propose to conduct an initial pilot study. In this pilot, we plan to enroll 50 cases of childhood leukemia and 50 matched healthy controls, to obtain detailed information on relevant environmental and epidemiologic exposures, and to collect biospecimens for future molecular and genetic studies. The success of the pilot project will provide preliminary data to motivate a future full case-control study.	{ "pile_set_name": "NIH ExPorter" }
The physical and biological processes governing the flow of blood are intimately responsible for safety and efficacy of all blood-wetted medical devices. The quest to design improved cardiovascular devices is however stifled by the inadequacies of current understanding of blood trauma and thrombosis. In spite of decades of experience with device design, hemotrauma research, and computational fluid dynamics modeling, it is virtually impossible to avoid deleterious hematological effects without experimental trail-and-error. Contemporary design relies upon venerable mathematical formula for blood that is more descriptive than predictive. Furthermore, we now understand that that these three phenomena are more closely coupled that previously appreciated. Indeed, previous models virtually neglected the existence of any coupling between these three processes. The objective of proposed project is to advance the accuracy and utility of a predictive model for thrombosis in blood-wetted cardiovascular devices. The research is built upon a combination of a previous model developed by the PI and colleagues for shear- mediated thrombosis and recent progress in modeling cellular-scale hemodynamics. Further incorporation of a model for synergy of platelet agonists is intended to yield a comprehensive design tool that is practical for design optimization of cardiovascular devices. Computer simulations will predict the dynamic interaction of red blood cells (RBCs) with platelets (Plts) in blood flow, and will rely upon a sophisticated theory of interacting continua that can predict the distribution of cells in any arbitrary flow path. The model will be validated and calibrated by both micro-scale computer simulations and microscopic visualization of blood cells in micro-channels. The predictive capacity of model will be demonstrated in three benchmark applications motivated by the development of cardiovascular devices for children: (1) parallel plate study incorporating various microscopic steps and crevices, (2) flow within blade tip of rotary blood pump, and (3) hydrodynamic bearing for pediatric rotary blood pump. Successful completion of these aims will produce a comprehensive computational model for blood trauma in cardiovascular devices, which we believe will contribute to the paradigm shift in the way these devices are developed: replacing trial-and-error with prescriptive bioengineering methods. Combined with computer optimization, the use of this model will greatly accelerate development of innovative new devices, and will reduce the occurrence of adverse complications. We also envision that the models will also be informative for diagnosing various clinical events, and help guide management of anticoagulation therapy. PUBLIC HEALTH RELEVANCE: Cardiovascular devices that are implanted today carry a risk of un-intended blood clotting, which may cause serious injury including stroke. The purpose of this project is to create a computer simulation program that will predict when this might occur, and thereby guide developers of these devices to produce more safe and effective devices.	{ "pile_set_name": "NIH ExPorter" }
available to the Administrative Core includes 599 sq.ft, off-campus space for Core C (Dr. Albers, Speer, McGowan and Kennedy) located at the Cochran Building. This space is leased by the University of Washington for the use of the investigator and is located with 1 block of the Lake Union Building (Project 1 location) and within 1 mile of the University campus. Adequate space is available for Ms. Aspen for the Program purchasing/secretarial needs for on-campus projects (Project 2, 4 and parts of Project 1 and 3, and Core B) to include manuscript word processing and on-line purchasing. Computers available to the Program Project for administrative purposes include PC's with printers for Albers, Speer, McGowan, Aspen and Kennedy. All computers have both DOS and Windows operating systems. Along with the computer hardware available to the program is a large variety of software for word processing, program communications, and financial recordkeeping, graphics, and data analysis programs for statistical analysis and production of tables and graphs for publication purposes. All computers are linked via an Ethernet connection to University of Washington administrative systems and the internet. Conference room facilities in the Cochran Building are large enough for most monthly Program meetings. Other University meeting rooms are available at a small cost when meeting attendee numbers necessitate a larger meeting room. Location of parts of the Program off-campus actually makes attendance at the meetings significantly less complicated and costly since ample free parking is readily available. Project researchers and staff have been located at the Cochran Building since 1990 and often consider this a positive factor when arranging regular meetings with on-campus investigators. The Cochran and Lake Union Place Buildings are both serviced daily by both US Postal Service and internal University services for daily pick-up and delivery of written materials meeting the needs of the program. PHS 398/2590 (Rev. 05/01) Page 375 Continuation Format Page	{ "pile_set_name": "NIH ExPorter" }
The Department of Chemistry of the University of South Carolina is requesting matching funds for the purchase of a high-mass, high resolution Gas Chromatograph/Mass Spectrometer with Fast Atom Bombardment (FAB) and Field Desorption capabilities (FD) and a supporting Data System. A total of ten primary and five secondary users for the GC/MS instrument are identified in this proposal which documents (1) a strong need for a second instrument in the Chemistry Department's GC/MS Center to meet the increasing demands on our present GC/MS capability, and (2) a compelling need for more sophisticated GC/MS instrumentation to meet the needs of existing and developing research programs in the areas of Analytical, Biological, Organic and Organometallic Chemistry. Specific research programs in which the requested instrument will play an essential role include: Synthetic Organic Chemistry Characterization of biologically active natural products, synthetic antibiotic, anti-tumor and antimetabolic compounds, and synthetic vitamin D analogs. Organometallic Chemistry Characterization of high nuclearity metal carbonyl cluster compounds, organoboranes, transition metal (Ta, W) catalytic intermediates, organo-main group (B, Si, Ge, Se, Sn) compounds, and iron and other alkylmetal complexes. Biological Chemistry Characterization of fluorescent, protein crosslinking compounds formed during the Maillard reaction; studies on the active site and mechanism of action of mono-oxygenase enzymes; characterization of selenium analogs of sulfur-containing biomolecules, glyconjugate labels used in studies of protein catabolism, leukotriene metabolites and peptide regulators of connective tissue metabolism. Analytical Chemistry Analysis of organochlorine and petroleum compounds in the environment, and of bacterial cell wall components.	{ "pile_set_name": "NIH ExPorter" }
Our laboratory is interested in immune activation and, in particular, the molecular mechanisms underlying the activation of T cells, a key step during an immune response. Our initial approach to gain fundamental insights into the early phase of activation of human primary T lymphocytes involved the isolation by subtractive technologies of novel mitogen- and antigen-inducible genes. Characterization of several of the more than 60 genes isolated in this way has led to the discovery of several cytokines, transmembrane proteins, a phosphatase, a ras-related protein and six different transcription _factors. Two of these originally cloned factors are part of the NF-kappaB transcription factor complex called p50 and p50B. NF-kappaB is an important component of the immune response; it is essential to the regulated expression of many immunomodulatory proteins, acute phase proteins and viruses, including the human immunodeficiency virus (HIV). Upon antigen- or cytokine- or stress-mediated signalling NF-kappaB is released from a cytoplasmic inhibitor called I-kappa-B to translocate to the nucleus. We have determined that NF-kappaB activity is created from different homo-and heterodimeric complexes which all differ in their functions. The homodimeric p50 complex actually acts as a repressor in the nucleus opposing the transactivating heterodimeric complexes. Both types of complexes are independently regulated by members of the I-kappa-B family: I-kappa-B inhibits transactivating complexes and the oncoprotein Bcl-3 inhibits the repressing complexes; these inhibitory molecules therefore have opposite effects on gene expression. The complexity of the NF-kappaB transcription factor suggests an equally complex regulatory role during immune activation.	{ "pile_set_name": "NIH ExPorter" }
1. Lectin-like receptors on murine pulmonary alveolar macrophages which bind to Aspergillus fumigatus conidia in vitro were found to be inhibited by attaching macrophages to glass surfaces coated with saccharides. The most inhibitory saccharides were those which are also known to inhibit the mannose-fucose receptor and chitotriose, a component of chitin. In studies using fluid-phase inhibitors, the monoclonal antibody against a macrophage surface antigen, designated F4/80, was found to be an effective inhibitor of the conidia receptor on macrophages. 2. In studies of the new allylamine antifungal drug, terbinafine (SF 86-327), absorption after oral administration to mice was found to be greatly enhanced by dissolving the drug in 1% Tween and 5% DMSO. One patient was studied after 20 months of receiving this drug. Half-life of plasma clearance was strikingly slow, being 50 days. 3. Purification of the Aspergillus fumigatus complement inhibitor was facilitated by the discovery of anionic properties on ion exchange chromatography. In other studies on fungal opsonins, deposition of the third component of complement on Cryptococcus neoformans in normal human serum was much less in the variety gattii. This may help explain the poorer host defenses, as found in the slower response to chemotherapy among patients infected by this variety. Complement fragment C1q was found to enhance phagocytosis and killing of opsonized C. neoformans blastospores by human monocytes. It is feasible that this interaction may contribute to host defense against cryptococcosis. 4. Improvement in the rapid chemical assay of flucytosine in serum has been achieved using a deaminating enzyme. Ammonia release can be measured by automated techniques. Application for patenting this device has been made.	{ "pile_set_name": "NIH ExPorter" }
The CCR established a Microarray Facility in 1999 with support from a supplement to this CCSG in recognition of the growing importance of being able to study gene-expression changes on a genome-wide level. Since that time, this microarray technology has expanded to include the use of arrays to study DNA-protein and protein-protein interactions in a global manner. To remain on the cutting edge of this technology, the MIT CCR Faculty has pooled resources with other groups at MIT to generate a single Microarray Technologies Core Facility. Its mission is to provide researchers with integrated facilities for microarray fabrication, scanning, processing and image analysis. The predominant focus of the Microarray Technologies Core Facility is on mRNA analysis. Most CCR Investigators choose to use commercial arrays for gene profiling experiments and the facility provides outstanding technical and intellectual support for both Affymetrix GeneChips and Agilent microarrays. In addition, the facility fabricates high-density oligo, cDNA and genomic arrays for a variety of CCR Investigators. In most cases, these arrays are custom designed to address a specific research problem that is not well served by commercial reagents. Finally, the facility helps users with the pre-processing and normalization of their expression array data and up-loads it onto an Iobion LabShare database. CCR researchers can then access their data in a standardized format through specialized workstations within the CCR building, creating a smooth transition from the wet-bench to the data analysis phase. CCR researchers can then conduct their own statistical analysis and data mining using software packages provided through the Microarray Technologies Core Facility or can seek expert advice from the a Bioinformaticists Specialist in the Bioinformatics and Computing Core Facility.	{ "pile_set_name": "NIH ExPorter" }
The speed of advancement in data-rich biomedical sciences is limited in part by the lack of awareness and expertise in big data sets and tools, the absence of approaches that productively intersect disparate domains, and the underrepresentation of diverse groups in the workforce. To address these disparities, we propose CREEDS: Community Research Education and Engagement for Data Science. This title represents our commitment to our overall goal of fostering practical skills for a national, diverse and interdisciplinary community of early career researchers. Our three specific aims are: (1) to provide biomedical researchers with the practical skills and insight needed to harness the power and advance the promise of big data science to accelerate scientific discovery, (2) to develop an online social environment to facilitate the exchange of big data ideas, approaches and techniques between novices and experts, and (3) to enhance the diversity of the biomedical big data workforce through targeted recruitment and retention of disadvantaged and underrepresented student populations. We will personally engage 150 graduate students through an intensive, self-tailored, two-week summer school in NYC that will showcase interesting, and current, collaborative case studies in activities at schools throughout NYC. It wil also include training in the Responsible Conduct of Research. Participants will employ active learning techniques to develop their skills of specific new methods and tools through both individual and group tasks on real-life large data sets. The training will raise the skills of students of varied backgrounds to a sufficient level for additional graduate research and will not require any prior computing experience. Students will also receive experience and materials to help them teach others when they return to their home institutions. Additionally, we will mentor another 30 NYC-based graduate students for team participation in four month long DREAM challenges. The DREAM challenge teams will help develop computational skills on solving difficult, real-life biomedical problems. An online social environment, Kipin Hall, will enable participants to discover job opportunities, find data sources and tools and communicate with peers as well as big data experts. We will leverage our extant relationships with organizations serving underrepresented groups to boost participation and enhance this exciting field. To continually improve our program we will actively seek and respond to feedback from our participants, program faculty and our advisory panel. As a result of this project, we will train ovr 180 people to develop team skills to understand, select and use genomics data tools and approaches. We will reach more individuals by placing the summer school on Coursera. Armed with this knowledge, we plan that the next generation of genomics scientists will be better placed to design, analyze and interpret high-throughput genomics datasets. This is the important need for research education that we aim to address with CREEDS.	{ "pile_set_name": "NIH ExPorter" }
This multicenter, randomized clinical trial will examine the effect of iron depletion (phlebotomy) on serum aminotransferases (biochemical parameters), viremia levels (virologic parameters), and hepatic histology in hepatitis C patients. The possible genotypic-determined differences in response to both iron depletion (phlebotomy) and alpha interferon in patients with chronic hepatitis C will also be examined.	{ "pile_set_name": "NIH ExPorter" }
Rates of opioid use during pregnancy are at epidemic proportions. Neuroimaging suggests potential adverse effects of opioids on the developing brain. Children exposed to opioids in utero can have behavioral problems and lower school achievement and are also often exposed to toxic stress, maternal depression, and adverse home environments. It is important to determine the relative impacts of multiple adverse exposures and identify potentially modifiable factors. Our consortium, comprised of 4 highly performing centers of the NICHD Neonatal Research Network (NRN), proposes the Outcomes of Babies with Opioid Exposure (OBOE) study to quantify effects of prenatal opioid exposure on the trajectory of brain development, determine associations of neuroimaging parameters with developmental and behavioral outcomes, and examine how specific factors (differing substance exposures, severity of neonatal opioid withdrawal, maternal stress/ depression/ parenting) modify these effects. We hypothesize that neural connectivity and neuroanatomical volumes are altered by prenatal opioid exposure and that the magnitude of these alterations correlate with developmental and behavioral outcomes. To test this hypothesis, the consortium will enroll 200 opioid exposed infants and 75 healthy appropriately matched controls in a longitudinal cohort study. We anticipate enrolling approximately 75 infants (55 opioid exposed and 20 controls) in our clinical site at the University of Alabama at Birmingham. The Specific Aims are: Specific Aim 1: To determine the impact of prenatal opioid exposure on brain structure and connectivity over the first two years of life. Specific Aim 2: To define medical, developmental, and behavioral trajectories over the first 2 years of life in infants exposed to opioids. Specific Aim 3: To determine how the home environment, maternal mental health, and parenting modify trajectories of brain connectivity and neurodevelopment over the first two years of life. Our clinical site at the University of Alabama at Birmingham (UAB) is a Regional Perinatal Center that can enroll a large cohort of newborn infants both exposed and non-exposed to opioids in utero. A particular strength of UAB is the comprehensive addiction in pregnancy program (CAPP) that facilitates enrollment. As a participating center in the NICHD Neonatal Research Network, UAB has long-standing established and highly productive collaborations both with the other clinical centers in this application, and with the Administrative and Research Support Core at RTI International. Our center has an excellent track record of enrollment of high-risk infants in clinical studies with successful participant retention and follow-up to 2 years of age and beyond, with trained certified examiners for neurodevelopment as well as pediatric neuroradiologists and established infrastructure for neonatal and infant cranial magnetic resonance imaging.	{ "pile_set_name": "NIH ExPorter" }
DESCRIPTION: (applicant's abstract) Human focal cortical dysplasia (FCD) is a developmental brain malformation characterized histologically by disorganized cerebral cortical cytoarchitecture and lamination. FCD is associated with several mental disorders including mental retardation (MR) and autism. FCD likely results from abnormal neuronal migration during corticoneogenesis although the molecular events that culminate in aberrant cortical lamination are unknown. Previous work has suggested that FCD neurons may have failed to terminally differentiate prior to migration since these cells express proteins, such as the intermediate filament nestin, that are typically identified in immature neuronal precursor cells. These, dysplastic neurons may not express other developmentally appropriate genes necessary to complete migration and lamination. This proposal will describe three approaches to study the developmental and molecular pathogenesis of FCD. First, expression of transcription factors, neurotrophic factors/receptors and cell adhesion molecules mRNAs will be assayed in single nestin- or MAP1B-immunolabeled cells in human FCD specimens and compared with normal cortical neurons, and neuronal precursors in the ventricular zone. Because expression of these candidate genes varies during early development and because they have been implicated in corticogenesis, coordinate change in their relative abundance will provide insights into molecular pathways altered in FCD. Second, developmentally inappropriate genes such as s nestin or MAP1B will be overexpressed in cortical slice cultures and effects on migration will be assayed. Finally, dynamic changes in gene expression will be assayed in actively migrating neurons in animal models of FCD including administration of exogenous neurotrophins or antisense oligonucleotides. It is anticipated that these studies will provide for the first time a view of altered gene expression in FCD, which will shed light on the pathogenesis of these lesions. Furthermore, in identifying alterations in specific genes, the relationship between FCD and mental disorders can be rendered. These analyses may point toward new avenues for therapy.	{ "pile_set_name": "NIH ExPorter" }
Stem cells and tissue regeneration have great therapeutic promise for treatment of a wide variety of medical conditions. For example and relevant to the current application, extremity injuries compromise 50-60% of all combat casualties observed in Iraqi/Afghanistan War Veterans resulting in large soft tissue defects (muscle and skin) and high amputation rates. Skeletal muscle, while being the tissue most vulnerable to ischemic damage in the extremities, also has an amazing potential to regenerate due to satellite cells. Satellite cells are stem cells that reside in skeletal muscle and with muscle injury, can proliferate and fuse together or with damaged muscle fibers to regenerate muscle; macrophages are critical to this process. Multiple types of macrophages participate in the regenerative process; classically activated (M1), wound healing (M2a) and regulatory (M2c) macrophage subsets are present in skin wounds and regenerating skeletal muscle; promoting inflammation, tissue repair and resolution of inflammation, respectively. However, the regulation of macrophage specialization (polarization) by microRNAs (miRNAs), and the consequential effects of macrophage subsets on muscle regeneration have not been elucidated. MiRNAs are small, noncoding RNAs that inhibit gene expression, thereby regulating many processes, including immune cell differentiation. A better understanding of the mechanisms of skeletal muscle regeneration and skin wound healing, and the influence of macrophage polarization on these events, could lead to new and adjunct therapies for limb salvage. Our long-term goal is to define the influence of inflammation, including the chemokine system, in angiogenesis, wound healing and skeletal muscle regeneration. We have extensively studied the importance of the CC Chemokine Receptor 2 (CCR2) in muscle regeneration. CCR2 is crucial for monocyte/macrophage recruitment and differentiation. Following muscle injury, CCR2-/- mice have impairments in macrophage recruitment, angiogenesis and muscle regeneration compared to wild type (WT) mice. Importantly, macrophage recruitment and muscle regeneration defects can be reversed in CCR2-/- mice by providing WT bone marrow, thus, CCR2 expression on bone marrow-derived cells is critical for normal muscle regeneration and macrophages are the likely BM-derived cell that mediates this outcome. The following 3 specific aims will be tested: 1) Define the in vitro regulation of macrophage polarization by selected miRNAs (miR-21 and -147). 2) Establish the mRNA targets of selected miRNAs (miR-21 and -147) during macrophage polarization and 3) Determine the biological effects of individual miRNAs on in vivo macrophage subsets and coordinated biological events during wound healing. By modulating macrophage polarization via increased or decreased miRNA expression, we seek to improve wound healing and decrease the adverse affects of acute and chronic inflammation present in many diseases that affect the Veteran population. The proposed studies are innovative because they will help define the contribution of miRNAs to macrophage polarization and subsequent muscle regeneration and wound healing. The combination of in vitro and in vivo studies will collectively identify miRNAs important for macrophage polarization, determine the mRNA targets, and assess any effects on muscle regeneration and wound healing. Given the availability of locked-nucleic-acid-modified oligonucleotide (LNA-antimiR), RNA oligonucleotides complementary to specific miRNAs that can be used in animals to decrease miRNA expression, a new therapeutic class of agents could become available for humans. The significance of this research is that a better understanding of the mechanisms of skeletal muscle regeneration and wound healing could lead to the design of novel primary or adjuvant treatments for improved limb salvage using miRNA- altering compounds.	{ "pile_set_name": "NIH ExPorter" }
We have measured dopamine-beta-hydroxylase (DBH) in cerebrospinal fluid (CSF) from psychiatric patients. DBH is the enzyme which catalyzes the conversion of dopamine to norepinephrine; its level in CSF may be an index of central noradrenergic activity. No significant differences have been found among the the mean CSF DBH values for 4 diagnostic groups--primary affective disorder, schizophrenia/schizoaffective disorder, alcoholism, and personality disorders. Preliminary evidence indicates that DBH activity in CSF may be related to mood state and to treatment with psychoactive drugs.	{ "pile_set_name": "NIH ExPorter" }
For millions of workers participating in the U.S. economy, their regular or secondary work shifts largely take place outside of traditional hours, including evening, night, rotating, irregular, and/or weekend shifts. Nonstandard and unpredictable schedules may have important implications for maternal well-being, work-family balance, and child care arrangements. However, knowledge on the association between nonstandard work schedules and indicators of maternal well-being is not consistent or comprehensive. More knowledge is needed on how nonstandard work schedules complicate the child care decision making processes of low-income mothers, and little is known about the supply of child care at nontraditional times. The research approach of the three papers in this dissertation aims to address these gaps in the literature and is described below. Nonstandard Work Schedules, Maternal Parenting Stress, and the Moderating Role of Child Care This paper explores the association between nonstandard work schedules and maternal parenting stress, with particular attention to variation by: type of work schedule, mother?s relationship status, and family income. This paper also explores the moderating role of child care arrangements between nonstandard work schedules and maternal parenting stress. Child Care Preferences and Choices for Mothers Working Nonstandard Schedules This paper seeks to better understand the role and importance of social, economic, and structural factors driving child care decisions for low-income mothers working nonstandard schedules. This paper will focus on two subpopulations of low-income mothers working nonstandard schedules: (1) mothers with irregular schedules and (2) mothers not living with a romantic partner. In addition, this paper explores what factors would make center-based child care during nontraditional hours more accessible and appealing to mothers working nonstandard schedules. Availability of Center-Based Child Care at Nontraditional Times in Texas: Supply Over Time This paper will explore the availability of licensed, center-based child care at nontraditional times for children preschool age and younger in Texas over time, describing trends on the timing of nontraditional hours of care, as well as availability by age group, accreditation/Texas Rising Star status of the provider, and the provider?s acceptance of child care subsidies. To begin to explore the mismatch between supply and demand, this paper also compares trends in the supply of care at nontraditional hours to several relevant labor force trends. To address these questions, this dissertation takes a mixed-methods approach. Quantitative data from the Fragile Families and Child Wellbeing Study, Child Care Market Rate Survey in Texas, and Bureau of Labor Statistics will be integrated with qualitative data from focus groups with mothers in Austin, Texas. Analytical techniques will include ordinary least squares, difference-in-difference, and multinomial logistic regression models. Qualitative data will be analyzed using themes from existing literature as well as identifying new emerging themes.	{ "pile_set_name": "NIH ExPorter" }
[unreadable] [unreadable] This application is to support the development of Dr. Dayna Groskreutz into an independent investigator and an accomplished physician-scientist. Dr. Gary Hunninghake will assume the responsibility as mentor to ensure the success of the career development plan. Dr. Groskreutz has already taken courses in general research principles and graduate immunology, and career development during this K-08 award will include further coursework in immunology, virology, cell and molecular biology, study design, and statistical methods. The proposed research investigates the effect of RSV infection on double-stranded RNA (dsRNA) receptors in airway epithelial cells and examines how these dsRNA receptors modulate p53 to delay apoptosis and allow for enhanced viral replication. In Aim 1, Dr. Groskreutz will investigate the effect of RSV on the expression and location of dsRNA receptors. In addition, their effect on p53 will be determined by blocking the expression of the dsRNA receptors and evaluating for a change in p53 protein. In Aim 2, Dr. Groskreutz will examine the biological effect of RSV-induced changes in p53 protein. To this end, p53 will be over-expressed and the biological effect of preserved p53 on cell viability, RSV replication, and inflammatory pathways will be determined. Finally, Dr. Groskreutz will evaluate whether specific RSV genes play a role in modulating dsRNA receptors and p53 protein. She will develop and add RSV strains void of the expression of single genes to airway epithelial cells and evaluate their effect on dsRNA receptors and p53. Throughout Dr. Grosrkeutz's career development, she will consult with experts in immunology, virology, cell biology, pharmacology, pediatrics, and pulmonary medicine. This environment will facilitate her development into an independent investigator, and her colleagues will serve as important resources as well as provide critical review of her work. Both her mentor and her institution are committed to her success and career development. As a sign of this commitment, she joined the faculty of the Division of Pulmonary and Critical Care Medicine in July 2006. Dr. Groskreutz's research proposal has great clinical significance. RSV continues to be a perplexing and challenging clinical problem in young children, in patients with chronic lung disease, and in immunosuppressed patients. This research could provide insight into to the cell signaling pathways important to the pathogenesis of RSV infection and identify key therapeutic targets for future clinical trials. [unreadable] (End of Abstract) [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Acute effects of putative cocarcinogens and tumor promoters were investigated on cultured human esophageal epithelial cells in serum-free LHC-8 and 199 media with 10% fetal calf serum. The effects were evaluated on clonal growth rate (CGR), cross-linked envelope (CLE) formation, and the enzymatic activities of ornithine decarboxylase (ODC) and plasminogen activator (PA). 12-0-Tetradecanoylphorbol-13-acetate (TPA) and the other compounds were inhibitory to clonal growth. None of the compounds induced CLE formation under the conditions used. TPA significantly induced ODC in media 199 but inhibited it in LHC-8 media. Capsaicin induced ODC in both media. TPA and capsaicin induced PA in both media. The other compounds did not change or rather decreased ODC and PA activities.	{ "pile_set_name": "NIH ExPorter" }
DESCRIPTION (adapted from the application): The proposed work will make use of newly gained information concerning the gene responsible for the defect in cystic fibrosis (CF) to develop approaches to use the cystic fibrosis transmembrane conductance regulator (CFTR) as a direct protein replacement therapy for treatment of the disease. The eventual goal is the development of an aerosol containing a fusogenic, targeted preparation of purified CFTR in a lipid-containing formulation capable of directly delivering protein to defective cells of the airway epithelia in CF patients. The cDNA for CFTR has been isolated and cloned into a number of mammalian cell expression vectors including those with inducible promoters. DNA will be transfected into C127, Chinese hamster ovary (CHO) cells, and cells capable of apocrine secretion. Cells expressing reasonable levels of protein will be isolated. Polyclonal and monoclonal antibodies have been raised to fusion proteins containing sequences from CFTR. Plasma membranes containing CFTR from non-recombinant and recombinant cells and membrane vesicles or blebs resulting from apocrine secretion from recombinant cells will be prepared. These will be fused chemically to nasal epithelial cells from CF patients and changes in ion efflux measured. The ability to correct the CF phenotype in tissue culture cells will indicate production, isolation, and delivery of active CFTR. This will be followed by experiments to purify the CFTR protein and to reconstitute it into lipid containing-vesicles. In parallel, attempts will be made to increase the efficiency and specificity of fusing the CFTR protein into cells. The approach will be to use the hemagglutinin and fusion glycoproteins of viruses with tropism for airway epithelial cells. Plasma membrane preparations, blebs, virosomes, and liposomes all containing viral glycoproteins and CFTR will be used to target to CF airway cells and ion efflux measured. Following successful transfer of CFTR into cells, the incorporated protein will be characterized immunologically and electrophysiologically in addition to being tested in the efflux assay. Conditions for growth of recombinant cells in large scale production will be established and the purification process developed to at least the 10 mg scale. Successful completion of the experiments will allow initiation of animal experiments and refinement of the CFTR protein delivery system.	{ "pile_set_name": "NIH ExPorter" }
Relatively little is known about reducing HIV risk behavior among HIV+ persons, despite the fact that high levels of such behavior have been documented within the HIV+ population, and despite the fact that development of effective risk reduction interventions for HIV+ persons has been identified as a top NIH priority. The need to design effective interventions to reduce high risk behaviors among HIV+ individuals is pressing, since HIV+ persons who practice risky behaviors are certain sources of HIV risk to uninfected sexual and needle-sharing partners, and may transmit treatment resistant strains of HIV and other pathogens to HIV+ partners, and are themselves vulnerable to coinfection with other pathogens (e.g., STDs) through high risk behavior. Moreover, reducing high risk behavior among HIV+ persons is especially critical in the current era of highly active antiretroviral therapy (HAART). Because HAART may result in greatly increased health and lifespan for HIV+ persons, it also affords the possibility of transmitting HIV to others over greatly extended periods of time and may therefore enhance transmission of strains resistant to antiretroviral therapies (e.g., AZT, 3TC, and possibly HAART). The proposed research applies the Information--Motivation--Behavioral Skills (IMB) model of HIV risk behavior change to design a physician- delivered intervention to reduce HIV risk behavior among HIV+ adult men and women receiving care in HIV treatment settings. Standard treatment for HIV necessitates regular and frequent visits to clinical care settings, often resulting in close patient-clinician relationships. Thus, this setting is an exceedingly powerful context for prevention interventions for HIV+ individuals. The study will be conducted at two very comparable HIV clinics which together treat over 1800 HIV+ adults. On clinic will serve as an intervention site and one as a control site. The intervention will be designed and conducted by an experienced group of behavioral scientists and HIV clinical researchers. The overall goal of this program of research is to design, to implement, and to rigorously evaluate a conceptually based, easily disseminated, clinician- delivered intervention to promote HIV risk reduction among HIV+ adults receiving care in HIV treatment settings. This population is at enormous risk for transmitting HIV to others who are HIV-, and for acquiring other pathogens themselves and it is a population for whom there are few if any effective HIV risk reduction interventions.	{ "pile_set_name": "NIH ExPorter" }
(Revised Abstract) DESCRIPTION (provided by applicant): Nearly all synthetics - including nano-scale liposomes, particles, or our recently developed polymer vesicles - are limited in in vivo performance due largely to phagocytes (eg. RES macrophages) which recognize all as foreign. To delay such recognition, the last decade has seen widespread efforts to effectively mimic the cell's glycocalyx with hydrophilic polymers such as PEG. However, at least 50% of PEG-modified STEALTH liposomes - as a prime example - still end up being phagocytosed in the liver and spleen (even when a tumor is present elsewhere to take up vesicles through enhanced permeability. While circulation times of our pure polymer vesicles with their denser and more stable PEG brushes are longer in rats than STEALTH liposomes, the vesicles still circulate much less than the organism's own blood cells. Inhibiting clearance of the body's own cells are specific mechanisms of protein-protein interaction that involve the ubiquitous membrane protein CD47: knockout mouse studies together with our own work on yeast-displayed human CD47 suggest 'marker of self' CD47 transiently binds to phagocyte receptors and turns off motile processes. We propose to engineer particles - primarily our polymer vesicles and more recent worm micelles - to display recombinant mouse or rat CD47 and demonstrate truly long-circulating, phagocyte-inhibiting particles in mice. Beyond this R21 proposal, our long-term objectives are to exploit such hybrid systems for controlled degradation/release carriers of drugs and imaging agents. One therapeutic direction envisioned is the delivery of various antioxidants targeted to the endothelium of oxidant stressed lung. This is a difficult-to-target tissue that is often inflamed (i.e. rich in activated leukocytes). In addition to providing a means of avoiding phagocytosis, CD47 could prove therapeutically useful in any such inflamed tissue by passivating accumulated leukocytes. Our newest biocompatible and hydrolysable PEO-PLA copolymers were synthesized crudely but already demonstrate kinetic control of degradation/release; these and related copolymers will be re-fined, made in larger quantities, and modified for protein attachment. Separate studies with a recombinant -100-aa extracellular immunoglobulin domain of human CD47 already demonstrate phagocyte inhibition in vitro, but the mouse protein must be expressed, purified, attached to particles, and also tested in vitro. Finally, we propose assessing the pharmacokinetics of CD47-displaying particles - primarily our own PEG-based copolymer vesicles and micelles -by IV injection into mouse and following circulation time, biodistribution, among other key responses.	{ "pile_set_name": "NIH ExPorter" }
There is compelling evidence for a role for two types of chemical, tobacco specific nitrosamines and heterocyclic amines, in the induction of pancreatic ductal adenocarcinoma in humans. There are animal models for this form of cancer involving tobacco specific nitrosamines and other experimental evidence implicating heterocyclic amines. We propose to examine the actions of a representative amine (2-amino-1-methyl-6- phenylimidazo [4,5-b] pyridine (PhlP) and nitrosamine (4- (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL)) and the carcinogenic nitrosamine MIP (N-nitrosomethyl (2-oxopropyl)amine) in pancreatic duct epithelial cells (DEC) from humans and from the appropriate rodent species and strains. We will concentrate on the metabolic activation of these chemicals focussing on the enzyme(s) that are involved in their metabolism. Our aim here is to determine those enzymes that may predispose humans to the pancreatrophic actions of environmental chemicals and to attempt to define groups who are at risk for pancreas cancer induction. We will determine the profile of metabolizing enzymes in both human and rodent DEC, focussing on those enzymes that are known to be involved in the activation of carcinogens. Having established this profile we will examine the ability of specific enzymes, mostly P450 isoforms, to metabolize MIO, NNAL and PhlP using the profile of metabolites and the ability to induce mutations. The ability of these carcinogens, activated by enzymes that are identified in the earlier aims, to neoplastically transform DEC will be measured. Lastly we will examine the capacity of DEC to repair different forms of DNA damage. A crucial factor in determining susceptibility might be the ability of DEC to repair DNA damage caused by the putative carcinogens. Our understanding of chemically induced pathological conditions is seriously hindered by a lack of knowledge of the ability of these cells to metabolize chemicals and to repair the genetic consequences of this activation. This proposal seeks to address some of these concerns and issues.	{ "pile_set_name": "NIH ExPorter" }
Engraftment has been established in nonsensitized major histocompatibility complex (MHC)-matched recipients with nonmyeloablative conditioning regimens. Recipients previously transfused with blood products become sensitized to donor minor histocompatibility antigens, increasing the risk of graft rejection. Patients with inherited red blood cell diseases require blood transfusions and have a higher probability of graft rejection. In this proposal, nonmyeloablative conditioning regimens will be developed for sensitized recipients which 1) lack the toxicities characteristic of myeloablative regimens, and 2) could be safely administered in an outpatient setting. The development of the nonmyeloablative regimen for sensitized patients will be based on two hypotheses: 1) host-versus-graft (HVG) reactions of sensitized recipients can be suppressed with immunosuppressive agents other than high-dose chemoradiotherapy; 2) T cells from the graft can suppress the host immune system including sensitized immune effector cells. These hypotheses will be tested and nonmyeloablative regimens will be developed in a preclinical canine model of transfusion-induced sensitization. Sensitizing recipients with blood transfusions prior to transplant results in uniform graft rejection with conventional high-dose conditioning. In Aim 1, it will be determined if further stepwise intensification of pretransplant immunosuppression in addition to CSP will successfully promote engraftment in a dose de-escalation study of TBI. Posttransplant immunosuppression (MMF/CSP) will be assessed separately to determine if it prevents graft rejection in sensitized recipients at TBI 920 cGy. If effective, intensification of posttransplant immunosuppression will be done at the maximal TBI dose at which graft rejection occurs at the completion of Aim 1A. In Aim 2, it will be determined if promoting GVH will achieve engraftment by suppressing sensitized host T cells and "creating space" in the marrow. If these studies are successful, then donor T cells will be ex vivo expanded and transduced with a "suicide (HSVtk) gene" for prevention of severe GVHD. In Aim 3, the optimal immunosuppressive regimen will be combined with a GVH enhancing regimen. The ultimate goal of this proposal is the elimination of cytotoxic agents from the conditioning regimen. By lessening the morbidity and mortality associated with conventional conditioning, these studies could significantly change the management of selected inherited red blood cell diseases.	{ "pile_set_name": "NIH ExPorter" }
This proposal seeks renewed funding for an NRSA Institutional Predoctoral Training Grant from the Molecular Biophysics Program of the National Institute for General Medical Sciences. This grant will support predoctoral training in Structural and Computational Biophysics (SCB) leading to a Ph.D. degree at Wake Forest University. The SCB Training Program is a multi-disciplinary, multi-departmental program that has as its goal the training of physicists, mathematicians, and chemists in biological science and the training of biological scientists in structural biology and biophysics. Furthermore, the graduate programs at Wake Forest University and the SCB Training Program in particular occupy a unique training niche in this country, drawing students from all over the nation and the world, but with a specific focus on students from the Piedmont and Appalachian regions of the United States. These regions are underserved by our major research universities, but are rich in highly motivated students from high quality smaller institutions for whom Wake Forest University provides a gateway to the greater research enterprise of our country. The SCB training faculty is highly collaborative, and includes faculty members from Biochemistry, Biomedical Engineering, Chemistry, Molecular Medicine, and Physics. Students are trained in structural biology, computational biology, proteomics, and biophysics. The major scientific themes of the SCB Training Program include the overlapping areas of nucleic acid metabolism, redox biology, and molecular signaling. The Program has a flexible curriculum that is designed to meet the individual needs of the students in the program, and includes a well-developed system of interdisciplinary seminars and tutorials to promote interaction among members of the program. We offer specific training in skills for a wide variety of potential careers for our Ph.D. graduate, including academic careers, careers in the biotechnology and pharmaceutical industries, as well as careers in the intellectual property arena. We have developed a pipeline in which Master's degree students in Biomedical Science, Chemistry, Physics, Mathematics and Computer Science can transition into Ph.D. training in our program. We also have a certificate program in Structural and Computational Biophysics that provides a pipeline for students to enter the SCB Program. Funding is requested for 4 predoctoral positions out of the steady state level of 25- 30 students in the laboratories of the training faculty. These positions will be used to support students in the second year or beyond, at which time it will be apparent that their dissertation research is consistent with the goals of the Program. Support will be for a maximum of two years, after which the students will be supported by the research funding of their advisors or their own individual fellowships.	{ "pile_set_name": "NIH ExPorter" }
The objectives of this research are to further purify and characterize exonuclease VII of E. coli and to examine its role in DNA replication, recombination and repair. The work will exploit mutants which we have isolated having reduced levels of exonuclease VII activity to study 1) the joining of Okazaki fragments; 2) excision of thymine dimers and repair synthesis and 3) whether or not a single protein catalyzes the hydrolysis of single-stranded DNA from both 5' and 3' termini. The previous work that we have done on this enzyme has resulted in a 1700 fold purification. We will now purify it to homogeneity and study its subunit structure. In order to facilitate the purification of the enzyme we will attempt to isolate an E. coli strain which produces increased amounts of exonuclease VII. The homogeneous enzyme will be used in in vitro studies employing model compounds that may be structurally similar to intermediates in DNA replication and recombination to test mechanisms by which exonuclease VII could function in these processes according to current models. Studies will also be carried out related to the practical application of exonuclease VII as a general enzymatic reagent for certain types of modifications of polynucleotide structures.	{ "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. A proportion of patients continue to experience symptoms of Lyme disease after antibiotic treatment and in chronic infection, yet the organism can rarely be detected from blood or skin biopsy. The effectiveness of antibiotic treatment for borreliosis has been explored in murine and canine animal models. Spirochetal persistence post-treatment is evident, but the recovery of intact organisms has been infrequent. Further, the infectious capacity of persistent organisms remains indeterminate. This study examines the question of antibiotic efficacy in two controlled experiments with nonhuman primates. Macaques were infected by needle inoculation of B. burgdorferi. At four to 6 months post-inoculation, a portion of the animals received aggressive antibiotic therapy regimens. Multiple methods were employed for the detection of residual organisms, including the placement and feeding of lab-reared ticks on monkeys for xenodiagnosis. The antibody responses to the Borrelia-specific C6 diagnostic peptides were measured longitudinally to include before, during, and after antibiotic treatment. Antibody responses to C6 declined in all antibiotic-treated animals but remained elevated in controls. Intact spirochetes were recovered by xenodiagnosis from 2 of 3 treated monkeys. Active spirochetal transcription was also frequently detected in the tissues of treated animals. These results demonstrate an ability of B. burgdorferi spirochetes to withstand antibiotic treatment, administered post-dissemination, in an incidental host[unreadable]the nonhuman primate. Importantly, we have not discerned whether these organisms remain infectious and continue to cause objective signs of disease.	{ "pile_set_name": "NIH ExPorter" }
This training grant in developmental endocrinology and metabolism at Children's Hospital Boston intends to provide funding for postdoctoral fellows to engage in research training leading to independent careers in academic pediatric endocrinology. Trainees enter the program with either the MD, MD/PhD or occasionally the PhD degree, and are selected from highly qualified pediatric endocrinology fellows at Children's Hospital. The 28 trainers include 13 pediatric endocrine physician-scientists (including 5 MD/PhDs), 13 physician-scientists of other disciplines, and 2 PhD scientists. Children's Hospital is the major training site, along with laboratories at Massachusetts General Hospital, Beth Israel Deaconess Medical Center, Brigham and Women's Hospital, and Joslin Diabetes Center. The major areas of research emphasis in this training grant include clinical, translational and basic laboratory research in diabetes mellitus, obesity, neuroendocrinology, thyroid disease, bone health, stem cell biology, bioinformatics, ion transport, and human outcomes research. Training consists of didactic courses, including in quantitative methods and the ethical conduct of research, and an intensive period of individually mentored research. The intended average duration of training is two years. Of the 15 trainees during the past 10 years who have completed this training program, all but 1 are faculty in academic pediatric institutions (1 Professor, 2 Assistant Professors, 9 Instructors) and 2 are in industry. Of these, 3 have had independent NIH K awards, and 1 has an R01. Their record of publication is strong. This program takes advantage of the breadth of scientific expertise of the training faculty within the Division of Endocrinology, as well as in the larger Children's Hospital Boston community and surrounding Harvard environment.	{ "pile_set_name": "NIH ExPorter" }
Genetic Studies of Loci Associated with Atrial Fibrillation Atrial fibrillation (AF), which is characterized as the quivering of the atria instead of coordinated contraction, is the most common cardiac arrhythmia, and it is associated with a 2- fold increased risk of mortality and morbidity and a 4- to 5- fold increased risk for stroke. Many risk factors have been identified for AF, however the discovery of heritable components suggests that genetic variation may play a role in AF development. In published genome-wide association studies (GWAS), an AF susceptibility locus has been identified in an intergenic region of chromosome 4q25. We and others have replicated this finding. Additionally, we are part of a consortium that has performed a meta-analysis that has identified five SNPs in this 4q25 region that are independently associated with AF. One of these SNPs is just ~27 Kb downstream from PITX2, the closet gene to this region. PITX2 appears to be an excellent candidate for an AF-causing gene as it is the closest gene to the culprit 4q25 region, and it's known to be important in left/right asymmetry of the heart during development. In addition, Pitx2 +/- mice have been described which are susceptible to arrhythmias when subjected to cardiac electrical pacing. A GWAS identified SNP associated with coronary artery disease on chromosome 9p21 is also located in an intergenic region;and, this region has been shown to have enhancer activity affecting the expression of the nearest genes that are more than 60 Kb away. Thus, we hypothesize that there may be long range enhancers and/or silencers in the 4q25 region and that these may directly affect gene expression of PITX2 or other neighboring genes. To investigate this hypothesis, in vitro and in vivo experiments will be performed to identify and test functional transcriptional regulatory elements in the 4q25 region determine their effects on gene expression of Pitx2 and neighboring genes. PUBLIC HEALTH RELEVANCE: The goal of this project is to identify functional genetic variants associated with atrial fibrillation and thus expand the existing knowledge regarding the genetic changes and their mechanisms that increase ones risk for developing atrial fibrillation. The information gained in these studies may be useful the development of novel and effective treatments, diagnostic tests, and preventative measures for atrial fibrillation.	{ "pile_set_name": "NIH ExPorter" }
In each division cycle of a normal somatic animal cell, a crucial block to further proliferation has to be passed at the point where the decision is made to replicate the genetic material, DNA. In contrasts, in cancer cells, proliferation control is deranged. It is therefore of great importance to identify the molecules which trigger this decision point and to understand the signals upon which their action depends. Drosophila is an excellent organism in which to investigate these molecular checkpoints using genetics, cell biology and molecular probes. The phenotypes of mutations at the pan gu (png) locus of Drosophila indicate that this gene is of key importance in te regulation of embryonic DNA replication. png is one of three, interacting, maternal genes whose products are essential to prevent DNA replication in the unfertilized egg. In fertilized eggs from png mutant mothers DNA synthesis is uncoupled from mitosis, producing giant, polyploid nuclei. The aim of this proposal is to determine the molecular mechanism whereby DNA synthesis is repressed, and how this repression is overcome by fertilization. To do this it will be necessary to clone the png gene and to analyze its sequence and spatial and temporal expression. The role of the png+ product in coupling DNA synthesis to mitosis will be tested by determining whether DNA synthesis in png mutant embryos occurs in discrete cycles. For this, nucleotide pulse-labelling will be used in vivo and in extracts. The dependence of one gene product upon another will be inferred from the effect of duplications and mutations of other genes upon both the png mutant phenotype and the distribution of the png+ product. These experiments will locate the wild type png+ gene product within the hierarchy of genes coordinating DNA replication with meiosis, fertilization and mitosis in the early embryo.	{ "pile_set_name": "NIH ExPorter" }
This is a 5-year renewal application to support 5 postdoctoral trainees for a broad program in Academic Endocrinology. This program has been in place at the Brigham and Women's Hospital/Harvard Medical School since 1985 and has been highly successful in training physicians and basic scientists, many of whom remain in academic endocrinology. In the last 10 year period, 67% of the 18 trainees completing their training remain in a research career. The faculty combines both the physician-scientists of the Endocrinology Division with those basic scientists on the Harvard Medical School faculty whose research is significantly related to endocrinology. Over the last ten years, these 16 faculty individuals have trained over 300 postdoctoral fellows, over 95% of whom remain in research-related careers, either in academic positions or in industry. The faculty are well-funded, with estimated modified direct costs per annum of over $50 million. The 5 trainees appointed each year will be individuals with MD, PhD, or MD/PhD degrees and they will undertake 3-4 years of an intensive research experience in either basic or clinical investigation. The goal of the program is to provide the trainees with sufficient expertise to apply successfully for mentored research awards, such as K23, K08, or K01 awards, or R01 grants, when indicated. The areas of research interest of the faculty include regulation of energy expenditure, insulin sensitivity, mechanisms of calcium signaling, the role of the iodothyronine deiodinases in controlling differentiation and cell division via local activation and inactivation of thyroid hormone, the process of bone development and its clinical implications, the study of G-protein-coupled receptors, particularly in the hypothalamus, the control of skeletal muscle repair and mechanisms of muscle proteolysis in cachexia and muscle atrophy, the mechanisms of steroid hormone action and the epidemiology of diabetes, obesity, and vitamin D-deficiency disorders. Individuals in this program will also be trained to translate the insights they gain to develop strategies for the treatment and prevention of common endocrinology-based public health problems, including diabetes, obesity and osteoporosis. PUBLIC HEALTH RELEVANCE: This Endocrinology Training Program assembles a group of outstanding clinical and basic scientists to oversee the training of fellows with an interest in understanding the regulation and mechanism of hormone effects and how they are disrupted to produce clinical disorders. Fellows will be trained in either basic or clinical research techniques in order to contribute to the understanding and treatment of major endocrine-related disorders, such as obesity, diabetes, and osteoporosis.	{ "pile_set_name": "NIH ExPorter" }
Over the past two decades, forensic DNA profiling has become an important tool in the investigation of human rights abuse and genocide. There is, however, little understanding of the ethical, historical, political, psychosocial, or policy dimensions of this application of genetic technology. The lack of a well-developed body of relevant research, and few regulations to guide the implementation of humanitarian DNA identification projects, means that organizations and individuals must develop their own ad hoc rules and procedures for the identification process. The same problems have arisen in numerous situations, often leading to continued hostility and a lack of closure. This project will contribute to empirical literature on humanitarian DNA identification, and inform the development of concrete policy recommendations. Using theoretical approaches from science and technology studies, anthropology, history, subaltern studies, and studies of trauma and reconciliation, the principal investigator will systematically analyze two major cases: the use of DNA profiling by the International Commission for Missing Persons in the former Yugoslavia to identify the 40,000-plus missing people from the Balkan Wars of the 1990s; and the South African National Prosecuting Authority's efforts to locate and identify the 2000-plus missing people from the apartheid era. The specific aims are to: 1) describe the historical development of these DNA identification programs; 2) examine the ethical and political dimensions of individually identifying victims of human rights abuse; 3) investigate the psychosocial dimensions of DNA identification; and 4) analyze the impacts of DNA identification projects at the individual, community, inter-group, and political levels. This project will also explore several major ethical issues that are inextricably linked with these aims, including, for example: the role of private biotechnology companies (and the profit motive) in humanitarian projects; how to determine whether or not DNA testing is in the best interest of a particular community; balancing the evidentiary needs of the legal system with the therapeutic needs of families and communities; and what should be done about the discovery of bodies from previous conflicts that are still symbolically and politically relevant. This research will combine documentary and archival research with oral history interviews and ethnography. Care will be taken to ensure that the voices of ordinary people affected by DNA identification are heard and that their perspectives are incorporated into all final work products. In addition to being published as individual case studies, academic articles, and presentations, this research will become part of the principal investigator's larger project on science and human rights, entitled Accounting for the Dead: Science, Politics, and Identity in the Aftermath of Genocide and Mass Violence. [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
Nonalcoholic Fatty Liver Disease (NAFLD) is a rapidly emerging disease estimated to afflict 30 to 70 million people in the U.S. in general. NAFLD, if allowed to progress, can result in premature death. Heritability is strong in NAFLD, and those with genetic predisposition are at higher risk for disease progression. Little is known about symptoms and health-related quality of life (HRQOL) in persons with NAFLD. This pilot study will examine the symptom experience and HRQOL in persons at risk of NAFLD progression as defined by the presence of one or two copies of the PNPLA3 gene (rs738409-G) allele vs. persons at lower risk of NAFLD progression as defined by the absence of PNPLA3 gene (rs738409-G) allele. The specific aims of this study are to: 1) Identify the presence of symptoms in persons with NAFLD at lower risk versus higher risk of disease progression based on PNPLA3 (rs738409) genotype;2) Compare the extent to which the frequency, intensity, and distress of symptoms in persons with NAFLD differ between persons at lower risk versus higher risk of NAFLD progression based on PNPLA3 (rs738409) genotype;3) Determine the difference in health-related quality of life in persons at lower risk versus higher risk of NAFLD progression based on PNPLA3 (rs738409) genotype;and 4) Describe the relationship between symptom distress and health-related quality of life in persons at lower risk versus higher risk of NAFLD progression based on PNPLA3 genotype. The design is a cross-sectional descriptive pilot study of 59 persons ages 21 years and older with previously diagnosed NAFLD via imaging or liver biopsy using convenience sampling. The methods used will be the completion of patient questionnaires and saliva sample collection;genotyping for the presence of PNPLA3 (rs738409-G) allele via saliva sample will be conducted to stratify the population. The Memorial Symptom Assessment Scale, the Charlson Comorbidity Index, and the CDC's Healthy Days Measure will be used to measure symptoms, comorbid conditions, and HRQOL. Chi-square, odds ratios, and general linear modeling techniques will be used to analyze the data using PASW 18 software. This study will provide foundational work upon which the applicant will base her research trajectory examining symptoms, genetics, and HRQOL in persons with NAFLD. The significance of this study is that there is a dearth of knowledge regarding disease symptom burden and HRQOL in persons with NAFLD. This study will provide a foundation of knowledge regarding symptoms and HRQOL in persons with NAFLD to build a symptom profile that may be used to measure the impact of interventions for those at risk of progression, early detection of persons at risk, and management of symptoms in the later stages of NAFLD using HRQOL as a measurement of interventions. PUBLIC HEALTH RELEVANCE: This study will describe symptoms and quality of life in persons with nonalcoholic fatty liver disease who are genetically predisposed versus those who are not genetically predisposed for worsening of the disease to liver cirrhosis or liver cancer. We will compare symptoms in persons with nonalcoholic fatty liver disease who have a particular version of the PNPLA3 gene to persons with nonalcoholic fatty liver disease who do not have that version. Identifying a difference in symptoms and quality of life in those with the PNPLA3 (rs738409-G) may enable the use of these findings to screen and intervene earlier to prevent premature death from nonalcoholic fatty liver disease.	{ "pile_set_name": "NIH ExPorter" }
Large coronary artery vasospasm can induce angina pectoris and even myocardial infarction. The mechanisms responsible for coronary vasospasm are not known, but potentially involve an increase in alpha adrenergic tone or reduction in beta adrenergic tone. Most prior studies in this field examined the coronary circulation as a whole by measuring coronary blood flow and calculating coronary vascular resistance, and have not examined the role of large coronary arteries, which is the major locus of coronary vasospasm. Furthermore, the results of the few prior physiological studies examining large coronary arteries have been conflicting and inconclusive. The goal of this research proposal is to study large coronary arteries for the first time with receptor ligand binding techniques and elucidate the affinity (KD) and the number of receptors per mg protein of the Alpha1, Alpha2, Beta1, and Beta2-adrenergic receptors in the normal large coronary artery. By using bovine coronary arteries, which are relatively large and the high affinity ligands 125I-pindolol for Beta-adrenergic studies, 125I-CP63,155 (analog of prazosin) for Alpha1-adrenergic studies, and 3H-rauwolscine for Alpha2-adrenergic studies, the study of coronary arteries will be feasible, despite the relatively small amounts of protein and small numbers of receptors in those vessels. An additional unique feature of this research proposal is to correlate the findings from the biochemical studies with physiological studies in conscious, chronically instrumented calves. Specifically, Alpha and Beta-adrenergic agonists and antagonists will be delivered intracoronary (to avoid systemic effects) to conscious calves instrumented for direct and continuous measurement of left circumflex coronary artery diameter and blood flow. This research proposal represents the first identification and quantitation of adrenergic receptors in coronary arteries utilizing receptor ligand binding techniques and the first correlation of these measurements with physiological measurements of responses of large coronary arterial dimensions to adrenergic agonists and antagonists. The results of these studies will provide basic information relating to potential adrenergic mechanisms involved in coronary artery spasm.	{ "pile_set_name": "NIH ExPorter" }
Development of a Novel Aerosol Mass Spectrometric Technology for Rapid and Sensitive Biomonitoring of Environmental Toxicants Name, address, and telephone number of the Principal Investigator: Dr. Manjula Canagaratna, Aerodyne Research, Inc., 45 Manning Rd., Billerica, MA 01821-3976, 1-978-663-9500 ext. 285 Names of other key personnel: Prof. Qi Zhang, Prof. Fumio Matsumura Participating institutions: Aerodyne Research Inc.; University of California, Davis This SBIR project will develop a novel technology for biomonitoring of environmental toxicants that cause adverse health effects in humans. The new technology will couple micro-nebulization with high resolution aerosol mass spectrometry to allow sensitive detection and chemical characterization of specific toxicants as well as toxicant classes in microvolumes of blood or biofluid samples. A droplet-on- demand (DOD) interface will convert microvolumes of liquids into aerosol particles of tightly controlled sizes and quantitatively deliver them into an aerosol mass spectrometer (AMS) for detection and chemical characterization of toxicants. DOD is a proven means of generating particles and aerosol mass spectrometry has been used for over a decade for sensitive and quantitative analysis of particle phase air pollutants with very fast time resolution (seconds to minutes). The coupling of DOD with AMS will allow characterization of liquid-phase species in real time, either directly in biological samples or after liquid chromatographic (LC) separation. The combined DOD-AMS instrument will be capable of simultaneously detecting and quantifying a wide variety of relevant toxic chemicals including inorganic, metal, and organic species. A prototype DOD interface will be developed and characterized in the laboratory with test aerosols containing known toxicants, and with archived blood samples from epidemiological studies. Data mining techniques will be developed to identify organizing patterns within the mass spectral distributions, understand correlations between detected species and health outcomes, and examine the association of multiple analytes and analyte classes with systemic effects. For the purpose of demonstrating the usefulness of this technology we will conduct an epidemiological pilot study on previously collected serum samples from asthmatic and non-asthmatic groups of children. We will first, simultaneously detect many toxicants or their metabolic products in the blood samples, second, use data mining techniques to characterize the toxicants, third identify the types and/or classes of toxicants in the serum that are most significantly associated with asthma using epidemiological assessment techniques. PUBLIC HEALTH RELEVANCE: This project addresses the critical need for new biomonitoring technology for rapid, sensitive, and simultaneous measurements of multiple known and suspected toxicants that can be ultimately linked with systematic effects in epidemiological and exposure studies. The proposed technology will allow sensitive detection and characterization of chemical species in small volumes of blood or other biofluids.	{ "pile_set_name": "NIH ExPorter" }
Cellular markers for prognosis in human bladder cancer will be identified and validated. The goal is to predict accurately the probability of recurrence or progression for individual tumors, thereby contributing to patient management and understanding of the pathobiology of bladder tumors. Nuclear DNA content, cytokinetics, chromatin distribution patterns, oncogene expression, and in situ cytogenetics will be quantified by image and flow cytometry. Additional markers, developed by colleagues, the marker network, or elsewhere will be evaluated when appropriate. Retrospective studies will access tissue blocks from 200 cases with known outcomes. DNA content, chromatin pattern, and oncogene expression will be quantified. The diagnostic and predictive power of these measurements will be established. Prospective studies will access 100 new patients per year. Specimens include initial surgical biopsies, urines, and bladder irrigations, and four-monthly follow-up urines and irrigations. Adequately cellular specimens will be incubated with 5-BrdUrd for cytokinetic studies. Analysis includes: cellular DNA content, proliferation index, chromatin pattern, oncogene expression, in situ cytogenetics by hybridocytochemistry, and immunocytochemical markers. Cells in urine and irrigation specimens will be pelleted and embedded: sections from these samples and from the tissue blocks will be available for subsequent analysis and for distribution to other network laboratories. In all cases, marker results will be evaluated against histologic and cytologic findings and against patient outcome in order to identify markers that accurately predict tumor recurrence, progression, and therapeutic response, and patient morbidity and mortality.	{ "pile_set_name": "NIH ExPorter" }
NLM has implemented a Local Area Network (LAN) for distributing data and video communications throughout Buildings 38 and 38A. The subject contract will provide technical support for the Data Communications Services of the NLM LAN, and general data communications analysis for various requirements at NLM. Areas of support to be provided for the LAN include: network management, installation, operation, maintenance, documentation, user assistance, and trouble shooting. In the area of general data communications analysis, tasks will be issued as required to evaluate existing and envisioned communications requirements, and prepare alternative recommendations for support. Requirements or problems relative to any aspect of NLM data communications may require analysis. Possible area of work include: 1) addition of new types of data services to the LAN; 2) extension of LAN data services to other buildings; 3) study of alternatives for implementing communications services for the IBM 30XX host computers and related systems including IBM 3270 communications; 4) evaluation of new communications product capabilities and features, which may include setting up tasks and initial configurations of hardware and/or software products; and 5) provide training sessions or seminars or various aspects of communications at NLM.	{ "pile_set_name": "NIH ExPorter" }
Cysteamine has been shown to suppress kindled seizures at doses from 90 mg/kg to 300 mg/kg when given to rats kindled to stage V. This project will involve the careful evaluation of the alterations in brain chemistry and the onset of the suppression of seizures in order to better understand the mechanism of action of cysteamine. The aims of he present project are to better understand the mechanisms by which cysteamine eliminates seizures. Rats which are kindled and sham operated will receive a single intraperitoneal injection of cysteamine (200 mg/kg). Following the administration of the drug, the animals will be observed for behavioral changes as well as seizure suppression. The rats will be killed at known time intervals following cysteamine administration, the brain removed, and the cortex, cerebellum, midbrain, pons-medulla, and hippocampus will be dissected. These tissues will be extracted and evaluated for peptide, amino acid, receptor, and catecholamine levels. The correlation of the chemical changes to the seizure suppression may allow the identification of one chemical alteration with a decrease in seizure activity. The next step in the study will be to determine if antagonists of the compound will also suppress seizures. Studies at present from our laboratory as well as others shown that both somatostatin and norepinephrine are decreased as a result of cysteamine administration. The results of the cysteamine experiments have shown that following administration of the drug, the suppression of seizures occurs not at the point where somatostatin is the lowest, but at a point where the levels of somatostatin are the closest to the control levels. These observations suggest that the suppression of the seizures following administration of the drug may be due to a receptor resensitization rather than the decrease of the somatostatin itself. Currently an investigation of the receptor activity and its interactions with other receptors is underway to better understand the mechanism of action of cysteamine.	{ "pile_set_name": "NIH ExPorter" }
Pathogenic species of mycobacteria and corynebacteria possess a thick and impermeable cell wall that helps these organisms evade detection and treatment by denying drug and probe molecules access to their cytoplasm. A distinctive feature of this barrier is an additional hydrophobic outer membrane composed of mycolic acids, which are covalently bound to an arabinogalactan (AG) polysaccharide that provides a link to the cell surface peptidoglycan. Efforts to identify weaknesses in these outer defenses would benefit greatly from a more complete understanding of how its components are assembled and maintained throughout the life cycle of the organisms. This project aims to develop chemical tools that specifically target and exploit the extracellular mycolyltransferase enzymes Ag85A-C, which mediate the covalent attachment of mycolic acids to the cell wall AG. First, we will detail a strategy to covalently label the cell wall mycolyl-arabinogalactan (mAG) complex with fluorophores to enable the visualization of mAG biosynthesis in real time. Our approach involves the design and synthesis of mimics of the endogenous mycolyl donor trehalose monomycolate (TMM), which carry fluorophore-conjugated mycolic acids. These substrates will be added to living bacteria, where we will evaluate their recognition by Ag85 and the incorporation of their fluorescent cargo into the growing cell wall. We anticipate that this robust labeling strategy will allow the localization and dynamics of cell wall assembly to be studied in unprecedented detail. Finally, we describe an approach to develop an Ag85-triggered turn-on fluorescent probe designed to fluoresce specifically in the presence of low concentrations of mycobacteria. We envisage that such a chemical tool would facilitate the early detection of pathogenic mycobacteria.	{ "pile_set_name": "NIH ExPorter" }
This study will be conducted among a large multiethnic cohort (N = 11,432) from the nationwide Women's Health Initiative (WHI), which includes an observational study and three clinic trials. The age range of this cohort is between 50-79 years at the baseline, and it has multiple minority groups: 1583 black, 739 Hispanic and 149 Native American women. The maximal follow-up time of this cohort will be 9 years by 2005. Dual-energy x-ray absorptiometry (DXA) is used to measure bone mineral density (BMD) and body composition. The randomized clinical trials and longitudinal nature of the WHI study provide a unique opportunity to investigate: 1) treatment effects of hormone replacement therapy (HRT) and calcium and vitamin D supplementation on hip structural geometry; 2) longitudinal changes in skeletal muscle mass as a factor in hip fragility; and 3) ethnic differences of mean and rates of changes in hip geometry and muscle mass. Special computer software will be used for analyzing hip scans by dual-energy x-ray absorptiometry (DXA). Cross-sectional area, subperiosteal width, estimated endocortical diameter, estimated mean cortical thickness, buckling ratio and section modulus at the femoral neck, at the intertrochanteric and the femoral shaft regions will be assessed. MRI scans will be used as references to calibrate total, appendicular and leg skeletal muscle measurements from DXA subregion analyses. Prevalence rates of sarcopenia (low muscle mass) among each age and ethnic group will be studied. Random Effects Models will be used to analyze the longitudinal data. This proposed study is not funded by the WHI program. Recourses that the WHI will provide include DXA scans, fall and fracture data, and information on covariates. Since the majority of data collection work has been or will be done by the WHI, we will be able to cost effectively test multiple important scientific hypotheses in this study. The novel approaches in this ancillary study will enhance scientific contributions of the WHI program. The significance of the proposed study is that it may demonstrate the utility of bone structural analysis in addition to bone mass measurements for understanding ethnic differences in fracture risk and/or for assessing the effect of pharmacologic therapy (i.e. HRT) on bone health. Furthermore, if the muscle variables are found to be a strong determinant of bone structure in the proximal femur and the risk of fall, then it may be important to develop interventions to increase muscle mass in this region to prevent hip fracture.	{ "pile_set_name": "NIH ExPorter" }
ABSTRACT Epidemiologic studies have linked obesity to poor breast cancer outcomes, and it has been suggested that obese women may experience poorer outcomes, in part, due to inadequate dosing of cytotoxic agents among obese women. Specifically, most cytotoxic agents are dosed according to body surface area, and therefore, the larger the woman, the higher the absolute dose. However, evidence shows that clinicians are more likely to depart from recommended dosing among heavier women for fear of inducing chemotherapy-associated toxicity. In 2012, the American Society of Clinical Oncology (ASCO) released guidelines stating that obese women should be dosed according to their full body surface area, largely based on evidence that suggested that fully- dosed obese women do not appear to experience more toxicity than fully-dosed normal-weight women. However, the guidelines acknowledge that data are extremely limited with regard to more severe obesity and in the real-world context of comorbidities. Furthermore, these guidelines cite that this practice of `dose reducing' obese women may be one reason contributing to the poorer outcomes observed in this group. However, to date, no empirical investigations have sought to determine if, and to what extent, dose-reduced chemotherapy may explain differences in breast cancer survival. These guidelines were met with some criticism, citing the need for further evidence, and data suggest continuing uncertainty about proper dosing of obese cancer patients. Understanding the drivers of dose reductions may help better inform our understanding of this practice and efforts to disseminate guidelines; however, we know little about factors driving dose intensity, and how these factors may vary by body size. We therefore propose to address these gaps using data on nearly 34,000 Stage I-IIIA breast cancer patients diagnosed and treated at Kaiser Permanente Northern California and at Group Health. Specifically, we will use the rich data from these integrated healthcare delivery systems to examine the relationship between body size and dose intensity, and will further examine how the factors contributing to dose reductions vary by body size (Aim 1). We will also evaluate if, and to what extent, dose reductions mediate the association between obesity and breast cancer recurrence and survival (Aim 2). Lastly, we will evaluate the association between body size and toxicity among women identified as receiving the full BSA-determined dose of chemotherapy (Aim 3). Our findings will provide critical and timely information to support or to warrant modification of current recommendations for chemotherapy dosing for obese breast cancer patients. Given the high and increasing prevalence of obesity in the United States, it is critical that we improve our understanding of chemotherapy dosing. The knowledge gained from this study can be used to better inform optimal treatment for the estimated 102,000 obese women diagnosed with breast cancer each year in the United States.	{ "pile_set_name": "NIH ExPorter" }
The overall objective of this study is to obtain a comprehensive analysis of the protein composition of the plasma membranes of the ocular lens and of related changes that may be associated with development. The specific objectives are to study the plasma membranes of the annular pad and fiber cells of the lens with respect to: a) protein composition, b) rate of membrane protein turnover, and c) related changes in a) and b) associated with lens development, including differentiation and ageing. The methods to be used include: polyacrylamide gel electrophoresis, immuno-electron microscopy, organ culture, radioisotope incorporation with liquid scintillation counting, immunology, and protein fractionation.	{ "pile_set_name": "NIH ExPorter" }
Na1BzoH (6-desoxy-6-benzoylhydrazido-N-allyl-14- hydroxydihydronormorphinone) is a novel opiate active at both mu and kappa receptors. In the presence of Mg++ ions, 3H-NalBzoH labels mu receptors pseudoirreversibly, with a remarkably prolonged rate of dissociation which is 145-fold slower than 3H-naloxone. The binding is not covalent since the GTP analog Gpp(NH)p rapidly dissociates the ligand, presumably by interrupting a stabilized receptor-G-protein complex. 3H-NalBzoH also labels a novel kappa receptor subtype present in high densities in calf, rat and mouse brains, kappa3. In vivo, NalBzoH is a potent, long-acting mu antagonist, blocking morphine analgesia for over 24 hr after a single dose. Low doses of NalBzoH also partially reverse the inhibition of GI transit produced by morphine, completely antagonize morphine lethality and precipitate withdrawal in morphine-dependent mice. Higher NalBzoH doses produce analgesia through a kappa receptor mechanism based upon its sensitivity towards a series of antagonists. NalBzoH has excellent oral activity, with a potency equivalent to the subcutaneous route. Thus, NalBzoH has several potential advantages over traditional opiates and could be useful as either a long-acting antagonist or a nonabusable analgesic. We propose to examine the features which distinguish NalBzoH from traditional opiates: 1) its prolonged duration of mu receptor antagonism, 2) its potent kappa analgesia and 3) its excellent oral potency. We will determine whether the prolonged mu receptor antagonism results from pharmacokinetic factors, such as a slow clearance, or whether it corresponds to its ability to label mu receptors pseudoirreversibily. We will determine half-lives of distribution and elimination, volumes of distribution and clearances. Efforts will be made to identify active major metabolites. Using 3H-NalBzoH, we also will investigate the formation of psuedoirreversible mu binding following NalBzoH administration in vivo. We will synthesize a series of analogs of NalBzoH to be examined in binding studies looking at their ability to label mu receptors pseudoirreversibly and their affinity at a variety of opiate receptor subtypes, especially kappa3. Selected derivatives will be synthesized in radiolabeled form and their binding directly examined. Additional derivatives will be examined in vivo. Finally, we will compare the metabolism and pharmacokinetics of NalBzoH following both oral and intravenous administration.	{ "pile_set_name": "NIH ExPorter" }
Humans are an intrinsically social species. Our advanced socio-emotional skills are thought to underpin our tremendous cultural advances as well as the evolution of language. Our superior social skills are underpinned by an increased proficiency in the identification of, and access to knowledge about other people. Social knowledge is supported by increased specialization in a number of cortical regions involved in face processing as well as higher-level regions involved in linking biographical knowledge as well as motivational value to specific faces. In this renewal of our prior R01, we propose to extend our work on the anterior temporal lobes (ATLs) to examine a more specialized circuit for person knowledge. This circuit involves a region in the ventral ATL (vATL) that when damaged, leads to person memory deficits. The vATL face patch appears to serve as an interface between face perception and person memory, linking perceptual representations of distinct identities with abstract person knowledge. However this region does not act alone. We will use multivariate fMRI as well as structural and functional connectivity analyses to examine the flexible interaction of this region with perceptual systems, episodic memory systems, and hedonic valuation systems. Success in this project will significantly advance our understanding of the basic neural mechanisms that contribute to high-level social processing and will have important clinical implications for disorders that have core social knowledge deficits such as autism spectrum disorder and schizophrenia.	{ "pile_set_name": "NIH ExPorter" }
The P-Glycoprotein multidrug transport protein is a membrane glycoprotein which is overexpressed in cell lines and tumors that are resistent to structurally unrelated cytotoxic agents. The exact mechanism of action is unknown; however, the overexpression of the protein results in reduced uptake of cytotoxic drugs. We have been investigating the interaction of forskolin, a naturally occurring diterpene, with the P-Glycoprotein. Forskolin photoaffinity agents have been synthesized which are capable of covalent labelling of the forskolin binding site. The location of the forskolin labelling sites on the P-Glycoprotein is being determined by digestion of labelled protein and immunoprecipitation with peptide-specific antibodies. Labelling sites have been identified on the C- and N-terminal halves of the protein. The labelling site on the N-terminal half of the protein has been associated with a 68 amino acid fragment that spans the fifth and sixth transmembrane regions of the P-Glycoprotein. The labelling patterns identified with forskolin photoaffinity labels are identical to those observed with photoaffinity labels derived from another drug, prazosin. The specific labelling site on the 68 amino acid fragment will be determined by amino acid sequencing of the isolated fragment.	{ "pile_set_name": "NIH ExPorter" }
Voltage-gated potassium (Kv) channels regulate cell membrane potential and control a variety of cellular processes. Kv1.3, a Shaker-related Kv channel is expressed in several tissues and believed to participate in cell volume regulation, apoptosis, T cell activation, and renal solute homeostasis. Channel activity is regulated, in a complex manner, by threonine and serine phosphorylation. Serotonin and insulin can both down-regulate the activity of the Kv1.3 channel. In the case of insulin, channel inhibition is observed in the olfactory bulb, and is mediated by phosphorylation of multiple threonine sites. To investigate the function of Kv1.3 in vivo, we carried out preliminary studies using Kv1.3 deficient mice (Kv1.3-/-) generated by gene targeting. Examination of these mice revealed that they weigh less and are more sensitive to the glucose-lowering action of insulin than control littermates. Furthermore, the phenotype observed in Kv1.3-/- mice was recapitulated by the pharmacological blockade of Kv1.3 in wild-type (wt) and diabetic mice, suggesting that Kv1.3's action on insulin sensitivity is independent of body weight. Moreover, inhibition of Kv1.3 channel activity in a skeletal muscle cell line (L6) significantly increased cell glucose uptake. Taken together, these data Kv1.3 strongly support the notion that Kv1.3 and its signaling pathway represent a novel component of the homeostatic mechanisms that regulate glucose metabolism. As such, we believe further investigation is warranted, from the standpoint of gaining a more detailed physiological understanding of the process and of uncovering potential targets for the development of drugs useful in the management of diabetes. While the effect of Kv1.3 on the regulation of body weight is of significant physiological and clinical interest, the current proposal is limited to studying its role in glucose homeostasis. The broad aims of the work are to elucidate the cellular and molecular mechanisms by which Kv1.3 modulates peripheral glucose metabolism and cause increased insulin sensitivity. In the context of our preliminary work, we propose to test if Kv1.3 modulate glucose uptake via an insulin-dependent pathway, and to examine the cellular mechanisms that mediate the action of Kv1.3 on glucose metabolism.	{ "pile_set_name": "NIH ExPorter" }
My current research agenda falls into two main areas. The first area is my focus on a population-based approach to examining the quality of care for women with breast cancer. The second area is the evaluation of online communication and service tools to improve the quality of cancer early detection initiatives in primary care settings. My research agenda over the next five years will extend my current work in several ways. First, I plan to expand my current SEER-based population research program to address the quality of care for women with breast cancer. This will include: 1) additional population-based studies of determinants and outcomes of other potentially marginalized patient populations; and 2) population-based intervention studies to improve communication, treatment processes and outcomes for women with breast cancer. Second, I will build and evaluate novel online communication and service tools for patients and providers to improve cancer screening processes and outcomes in primary care settings. The goals of my mentoring plan are 1) to increase the quantity and quality of NIH investigator-initiated proposals in oncology health services research from the University of Michigan; 2) to foster interest in oncology-related health services research by fellows and junior investigators through my leadership on clinical training programs at the University of Michigan; and 3) recruit new investigators to the medical school focused on health services research in oncology. In order to address the goals above I will lead the following initiatives: 1) to obtain intramural and extramural funding to recruit and support clinician investigators and non-clinician research scientists in oncology health services research; 2) to personally mentor these investigators towards a successful career; 3) to develop and direct educational programs to support this area of research; and 4) to coordinate efforts in this area across professional schools at Michigan. The structure, process and outcomes of my personal mentoring program are described in detail in the application. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]	{ "pile_set_name": "NIH ExPorter" }
The objectives of this project are to: (1) study structure and dynamics of membranes composed of lipids containing polyunsaturated fatty acids such as docosahexaenoic acid (DHA) 22:6n-3, (2) study lipid-protein interactions related to lipid polyunsaturation and alcoholism, and (3) investigate the interaction of alcohol with proteins and lipids in biological membranes. (1) The membranes of brain synaptosomes and retinal rod outer segments contain 30-50 mol% of the six-fold unsaturated docosahexaenoic acid (DHA) as lipid hydrocarbon chains. One possible role of DHA is to alter membrane mechanical properties important for activity of receptor proteins. Using a magic angle spinning NMR experiment which re-couples 13C-1H dipolar interactions, assigned DHA order parameters were obtained. A unique membrane probe - perdeuterated DHA - was synthesized and incorporated into the lipid matrix. Twelve distinct order parameters were measured. Furthermore, the dimensions of the DHA chain unit cell were determined by x-ray diffraction. Order parameters of all methylene segments between double bonds in the hydrocarbon chain, and the order of the majority of double bonds is very low. Only the two methylene segments near the carboxyl group of DHA have order parameters that are comparable to values of more saturated chains. The low order is a reflection of both a change in bond geometry and an increase in chain motions. Experimental results were combined with results of simulations. The analysis suggests that DHA chains in membranes can exchange between looped, tilted, and extended conformations in rapid succession, providing increased flexibility to receptor-rich neural membranes. We developed quantitative methods for interpretation of NMR NOESY cross-relaxation rates between lipid resonances. In addition to providing information on lipid structure, these rates are sensitive to the dynamics of membrane reorganization in the correlation time range form pico- to microseconds. The comparison of experimental rates and rates from molecular dynamics simulations suggests that distance variation between protons caused by lateral diffusion of lipid molecules is the primary mechanism of cross-relaxation in lipids. The analysis quantifies the high degree of molecular disorder in biological membranes, showing a finite probability of close approach between even the most distant segments of neighboring lipid molecules (e.g. the methyl groups in the choline headgroup and the terminal methyl groups of the fatty acid chains). Intermolecular cross-relaxation rates are an ideal tool to study lateral lipid organization in the liquid-crystalline phase of lipids. Inhomogeneous lipid distribution and preferences in the interaction of lipid species, as well as preferences in the location of substances that incorporate into membranes can be detected. We developed approaches to conduct experiments on membrane samples oriented at solid interfaces and in lipid mixtures that orient spontaneously in the strong magnetic field of NMR instruments. The analysis of NMR lineshapes revealed the variable degree of mosaic spread in bilayer orientation for the different membranes. (2) There is evidence that a high content of DHA in retinal membranes modulates physical properties of membranes, creating an environment that is optimal for function of rhodopsin, the primary visual receptor, and a member of the G-protein coupled receptor family. We investigated this hypothesis by solid-state NMR methods. Rhodopsin was reconstituted into fully hydrated, solid-supported oriented multi-bilayer samples. Using 2H-labeled lipids, we compared lipid order parameters in the absence, and in the presence of the protein. We obtained highly resolved spectra from deuterated acyl chains in membranes containing a reconstituted integral membrane protein under physiological conditions. Oriented samples also improve NMR sensitivity enabling work with milligram-size samples. We have studied the phase diagram of the polyunsaturated 18:0-22:6 PE and demonstrated that it forms inverse hexagonal phases at all temperatures above the gel-fluid transition. The results suggest the existence of polyunsaturated lipid-induced membrane curvature stress that is likely to modulate the degree of activation of membrane incorporated receptors like rhodopsin. (3) Ethanol can act at multiple sites, with variable emphasis on interaction via the lipid matrix or via direct interaction with the protein, depending on the specific protein system involved. We propose that the binding of ethanol molecules to the lipid matrix of biomembranes is an important event in the action of ethanol on biological matter. We studied the interaction of ethanol with saturated, mono-, and polyunsaturated membranes quantitatively by MAS NOESY NMR. The resolution of resonance lines allows detection of 13-16 proton signals from lipid, ethanol, and water. Results of NMR measurements were combined with atomic-level molecular dynamics simulations to provide a deeper interpretation of experimental results. The site of ethanol interaction with the lipids is the primary factor that determines NMR cross-relaxation rates. Differences in correlation times and motional amplitudes of lipid segments play a secondary role. In particular, magnetization transfer to the headgroup choline resonance was somewhat lower than expected, due to fast lipid dynamics. As observed previously for lipid-lipid cross-relaxation, the rates scale with translational diffusion rates of ethanol in the bilayer. Ethanol resides mostly in the water phase, but binds for brief periods of time, of the order of nanoseconds, to the polar groups of the lipid/water interface, primarily to lipid phosphate groups. This temporary interaction introduces anisotropy into the motion of ethanol, with ethanol methylene C-D order parameters of 0.06. Cross-relaxation is strongest between ethanol and lipid resonances from the lipid/water interface including the glycerol, upper hydrocarbon chain, and lipid headgroup regions. There is evidence that the ethanol molecule in membranes is oriented preferentially with its methyl group toward the hydrophobic bilayer core. Cross-relaxation between ethanol and lipid hydrocarbon chains methyl is a reflection of both lipid hydrocarbon chain upturns and brief excursions of ethanol molecules into the upper region of lipid hydrocarbon chains. Overall, the probability of ethanol penetration into the center of the hydrophobic core of membranes is extremely low.	{ "pile_set_name": "NIH ExPorter" }
This CDMAS project involves the cloning of the PTH receptor, mutation screening of the PTH receptor, and a genome-wide screen for a susceptibility gene for pseudohypoparathyroidism.	{ "pile_set_name": "NIH ExPorter" }
Our larger objective is to understand the forces that hold alpha-helices in that conformation and the forces that hold one helix to another in protein double helices. Our approach is: to remove or characterize the polydispersity of certain alpha-helix-forming synthetic polypeptides in order to render their physical properties, including the helix-coil transition, more interpretable; to explore conditions under which these synthetic polymers might form double alpha-helices; to investigate the helix-coil transition in the protein double helices; to seek conditions under which single, protein helices might form; and to produce and examine double helices, i.e., clam paramyosin - lobster paramyosin, paramyosin-tropomyosin, tropomyosin-synthetic polypeptide hybrids.	{ "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. We are investigating the structure of the U2-U6 snRNA complex from S. cerevisiae. This large RNA complex has been highly conserved throughout eukaryotic evolution, and forms the catalytic core of the spliceosome. The RNA structure will be investigated at a range of temperatures, pH and ionic conditions. NMR experiments that we will need to collect include: 1D (1H), 2D NOESY, 2D 1H-15N HSQC (normal and TROSY enhanced), 2D 1H-15N HNN COSY, 2D 1H-13C HSQC (normal and TROSY enhanced), 2D NOESY, 2D 1H,13C filter/edit NOESY. Residual dipolar couplings will be measured for samples in both isotropic and partially aligned states (via addition of Pf1 phage).	{ "pile_set_name": "NIH ExPorter" }
The Quantitative Morphology Core (QMC) provides technology supports to the members of the Conte Center. The QMC has developed tools to quantify morphologic changes in spines and synapses at the confocal , multiphoton, and electron microscopy levels. These methods are developed to provide to provide the necessary degree of resolution to quantify plasticity-related changes in molecular constituents of synapses, a critical step to link such modifications to information processing. The QMC has developed applications designed to increase the throughput and accuracy, and reduce bias of quantitative analysis of immunogold electron microscopy of synapses as well as new tools to image Lucifer Yellow-injected neurons at high resolution from automatized acquisition of entire cells using confocal and multiphoton data.. These tools involve deconvolution, smoothing, automatic tiling, and 3-dimensional rendition with high level of detail and capacity for morphometric analysis of dendrites at the spine level of resolution. We are also performing mathematical modeling of neuronal complexity and biophysical simulation of the properties of the reconstructed neurons. The QMC was also involved in the development of a new stereologic probe to assess the 3-dimensional spatial distribution of particles in a volume of interest. Recently, the QMC has begun producing high resolution magnetic resonance microscopy (MRM; 9.4 T) datasets of the human amygdala to provide users with a high level of detail digital atlas of the human amygdala to be used as a reference guide for fMRI and PET analyses of live subjects as well as anatomic reference for amygdala nuclear parcellation. These MRM data will also inform the users on the degree of anatomic variation that occurs in the human amygdala.	{ "pile_set_name": "NIH ExPorter" }

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